PHYLOGENETIC ANALYSIS OF THE LARGE SUBUNIT RUBISCO GENE SUPPORTS THE EXCLUSION OF AVRAINVILLEA AND CLADOCEPHALUS FROM THE UDOTEACEAE (BRYOPSIDALES,CHLOROPHYTA)1

The placement of Avrainvillea and Cladocephalus in the family Udoteaceae (order Bryopsidales) has been questioned on the basis of nuclear, plastid, and other ultrastructural characteristics unique to these genera. Bayesian analysis of the chloroplast‐encoded LSU RUBISCO (rbcL) gene showed that the Udoteaceae is paraphyletic. Cladocephalus luteofuscus (P. Crouan et H. Crouan) Børgesen, Avrainvillea nigricans f. floridana D. Littler et Littler, and A. mazei G. Murray et Boodle form a clade with the freshwater alga Dichotomosiphon tuberosus (A. Braun ex Kütz.) A. Ernst that is basal to a clade that includes other members of the Udoteaceae, the Halimedaceae, and the Caulerpaceae. The noncalcified species Boodleopsis pusilla (Collins) W. R. Taylor, A. B. Joly et Bernat. groups with species of the calcified Udoteacean genera Penicillus, Rhipocephalus, Udotea, and Halimeda.     

Revision of Phymatolithon purpureum (Hapalidiales,Rhodophyta) based on ultrastructural and molecular data

ABSTRACT

To elucidate the taxonomy and phylogeny of Phymatolithon purpureum (P. Crouan & H. Crouan) Woelkerling & L.M. Irvine, we observed the type specimens and fresh samples using SEM (ultra-morphology) and analysed DNA sequences. Phymatolithon purpureum was originally described as Lithothamnion purpureum P. Crouan & H. Crouan from Mingant, Brittany, France. Our molecular phylogenetic analyses using psbA and COI–5P regions placed our collections from Ireland, UK and France in a clade with ‘P. borealis W.H. Adey, J.J. Hernandez-Kantun & P.W. Gabrielson (MH252286)’ from Iceland and ‘uncultured Corallinales (GQ917711 and GQ917512)’ from Roscoff, Brittany, France, near the type locality of P. purpureum. We show that P. purpureum is conspecific with P. borealis and P. polymorphum f. papillatum (Foslie) Foslie based on morphology and molecular data. Also, although P. purpureum has been often confused with P. laevigatum (Foslie) Foslie because of their similar morphology (e.g. smooth surface and sunken tetra/bisporangial conceptacles), our molecular phylogenetic analyses indicate that P. purpureum and P. laevigatum are sister taxa. Our sequences from lectotype material of P. laevigatum and syntype material of Lithothamnion emboloides Heydrich are identical.     

Rocky shore turfs dominated by Corallina (Corallinales,Rhodophyta) in northern Japan†

Three intertidal sites dominated by Corallina turfs were investigated in Hokkaido, Japan. The sites (A, B and C) differed in slope, wave exposure and length of time exposed to air during tidal cycles. Monthly samples were analyzed for frond morphology and other features. Site A, the most wave-exposed site, was dominated by Corallina sp. X, an unknown species, and sites B and C by Corallina pilulifera Postels et Ruprecht. At the different sites the populations differed in conceptacle abundance, coverage by epiphytic Titanoderma corallinae (P. Crouan et H. Crouan) Woelkerling, Chamberlain et Silva, amount of contained sediment, numbers of axes per quadrat, numbers of branch fusions, branch entanglement, frond dryweight, frond length, amount of adventitious branching, numbers of epiphytes (exclusive of T. corallinae), and numbers of animal species. Ninety-one animal species were recorded from the turfs. Corallina is affected morphologically by conditions inherent in its microhabitat, including desiccation, epiphyte loading and the abundance of herbivores.     

CYTOMORPHOLOGICAL ASPECTS OF WOUND HEALING IN SELECTED SIPHONOCLADALES (CHLOROPHYCEAE)1

Wound healing in most of the Siphonocladales investigated differs markedly from healing processes known in other coenocytic green algae. No wound plug is evident during healing in Ernodesmis verticillata (Kützing) Børgesen, Boergesenia forbesii (Harvey) Feldmann, Cladophoropsis membranacea (C. Agardh) Børgesen, Siphonocladus tropicus (Crouan) J. Agardh, Struvea elegans Børgesen, Struvea sp. and Valonia spp. The wound response in these genera involves substantial protoplasmic motility which includes retraction of the cell contents from the wound site. The protoplasm then either closes around the central vacuole (e.g. Ernodesmis) or breaks up into numerous spherical protoplasts (e.g. Valonia ventricosa J. Agardh). An intermediate pattern of healing is present in Cladophoropsis where the protoplasm usually separates into a small number of portions that individually heal in a manner similar to Ernodesmis. In all cases, the protoplasts resulting from wounding are viable. Struvea anastomosans (Harvey) Piccone, Chamaedoris sp. and Boodlea sp. are unusual in producing a distinct wound plug during healing, thereby resembling the mode of healing reported in various codialean and dasycladalean genera. It is hypothesized, that wound-induced protoplasmic motility in the Siphonocladales involves the same cellular mechanisms that segregate protoplasm during cell, division.     

TAXONOMY OF THE FLATTENED SOLIERIACEAE (RHODOPHYTA) IN BRAZIL: AGARDHIELLA AND MERISTIELLA1

Taxonomic studies of an extensive collection of flattened members of the Solieriaceae considered by many authors to be species of Agardhiella, Eucheuma, Meristotheca, and Meristiella lead us to conclude that they represent only two genera in Brazil: Agardhiella, including A. ramosissima (Harvey) Kylin and A. floridana (Kylin) Gabrielson ex Guimarães et Oliveira, and Meristiella, with only one species, M. gelidium (J. Agardh) Cheney et Gabrielson. Accordingly, we propose that 1) Meristotheca gigartinoides Joly et Ugadim be synonymized with Agardhiella floridana, and 2) Meristiella echinocarpa (Areschoug) Cheney et Gabrielson and M. schrammii (P. et H. Crouan) Cheney et Gabrielson be synonymized with Meristiella gelidium.     

SCHMITZIA EVANESCENS SP. NOV. (RHODOPHYTA,GIGARTINALES) A NEW SPECIES OF CALOSIPHONIACEAE FROM THE NORTHEASTERN NEW ZEALAND COAST1

A new red alga, Schmitzia evanescens sp. nov. (Gigartinales), is described from the northeastern coast of North I., New Zealand. Gametophytes are diminutive, delicate, mucilaginous spring annuals which grow subtidally on cobble substrata. The uniaxial construction and carposporophyte development place this new taxon in the Calosiphoniaceae, a family of only two genera, Calosiphonia H. M. Crouan et P. L. Crouan and Schmitzia P. C. Silva. This is the first report of this family in New Zealand. The new species has vegetative features of both these genera, but is assigned to Schmitzia on the basis of its unusual carposporophyte development in which the gonimoblast initial arises directly from the connecting filament at some distance from the undifferentiated intercalary auxiliary cell. Because recent investigators of other red algal genera with a similar carposporophyte development have questioned the validity of site of gonimoblast initiation as a generic criterion it is suggested that the separation of Schmitzia and Calosiphonia should be re-evaluated. These and several other genera in the Gymnophlaeaceae, Pseudoanemoniaceae, Dumontiaceae and Gloiosiphoniaceae, can be considered, on morphological and reproductive grounds, to be relatively simple and therefore perhaps primitive members of the Rhodophyta.     

Sulla presenza nell'Italia meridionale di alcune alghe marine bentoniche rare per il Mediterraneo

Abstract

On the occurrence in Southern Italy of some benthic marine algae rare to the Mediterranean Sea. - The Authors report the occurrence along the coast of Southern Italy of the following species that resulted rare to the Mediterranean Sea: Antithamnion piliferum Cormaci et Furnari; Bonnemaisonia asparagoides (Woodward) C. Agardh (tetrasporophyte); Corynophlaea flaccida Kuetzing; Fosliella farinosa (Lamouroux) Howe: of this very common species, the occurrence of thalli with propagules is reported; Hypnea cervicornis J. Agardh; Jania adherens Lamouroux, new to Italy; Mesogloia lanosa P.L. et H.M. Crouan, new to Italy; Pneophyllum confervicolum (Kuetzing) Chamberlain f. minutulum (Foslie) Chamberlain, new to the Mediterranean Sea; Polystrata fosliei (Weber van Bosse) Denizot, new to Italy; Rhodymenia delicatula Dangeard; Spermothamnion johannis G. Feldmann-Mazoyer: of this common species, the occurrence of bisporocysts in monoic gametophytes is reported; Vickersia baccata (J. Agardh) Karsakoff.     

MORPHOLOGY AND TAXONOMY OF MERISTIELLA GEN. NOV. (SOLIERIACEAE,RHODOPHYTA) 1

Eucheuma acanthocladum (Harvey) J. Agardh, E. gelidium (J Agardh) J. Agardh, E. echinocarpum Areschoug and E. schrammii(P. et H. Crouan) J. Agardh from the tropical and warm temperate waters of the western Atlantic Ocean and Caribbean Sea are transferred to a new genus, Meristiella. Meristiella exhibits the following Unique combination of characters among genera in the Solieriaceae: (1) rotated periaxial cells, (2) a loosely filamentous medulla. (3) an auxiliary cell complex, (4) Single and twin connecting filaments and (5) spinose cystocarps composed of a central, small-celled placentum, based on its reproductive features, Meristiella. is assigned to the tribe Agardhielleae. Culture experiments and herbarium studies provide evidence that E, gelidium and E. acantghocladum are conspecific. Lectotypes are designated for the included species.     

SYSTEMATICS OF THE GRACILARIACEAE (GRACILARIALES,RHODOPHYTA): A CRITICAL ASSESSMENT BASED ON RBCL SEQUENCE ANALYSES1

Generic concepts in the economically important agarophyte red algal family Gracilariaceae were evaluated based on maximum parsimony, Bayesian likelihood, and minimum evolution analyses of the chloroplast‐encoded rbc L gene from 67 specimens worldwide. The results confirm the monophyly of the family and identify three large clades, one of which corresponds to the ancestral antiboreal genera Curdiea and Melanthalia, one to Gracilariopsis, and one to Gracilaria sensu lato, which contains nine distinct independent evolutionary lineages, including Hydropuntia. The species currently attributed to Hydropuntia comprise a single well‐supported clade composed of two distinct lineages. The two most basal clades within Gracilaria sensu lato deserve generic rank: a new genus centered around G. chilensis Bird, McLachlan et Oliveira and G. aff. tenuistipitata Chang et Xia and a resurrected Hydropuntia encompassing primarily Indo‐Pacific (G. urvillei [Montagne] Abbott, G. edulis [S. Gmelin] P. Silva, G. eucheumatoides Harvey, G. preissiana [Sonder] Womersley, and G. rangiferina [Kützing] Piccone) and western Atlantic species (G. cornea J. Agardh, G. crassissima P. et H. Crouan in Mazé et Schramm, G. usneoides [C. Agardh] J. Agardh, G. caudata J. Agardh, and G. secunda P. et H. Crouan in Mazé et Schramm). Cystocarpic features within the Gracilaria sensu lato clades appear to be more phylogenetically informative than male characters. The textorii‐type spermatangial configuration is represented in two distinct clusters of Gracilaria. The rbc L genetic divergence among the Gracilariaceae genera ranged between 8.46% and 16.41%, providing at least 2.5 times more genetic variation than does the 18S nuclear rDNA. rbc L also resolves intrageneric relationships, especially within Gracilaria sensu lato. The current number of gracilariacean species is underestimated in the western Atlantic because of convergence in habit and apparent homoplasy in vegetative and reproductive anatomy.     

Sexuelle Fortpflanzung bei der braunen KrustenalgePseudolithoderma extensum

Reproduction was studied inPseudolithoderma extensum (Crouan et Crouan) Lund (Lithodermataceae, Phaeophyceae) from Helgoland. Both gametophytes and sporophytes are crustose. Fertile gametophytes were found in nature in January and February. They released motile isogametes from plurilocular gametangia, each gamete leaving its locule through a separate opening. Plasmogamy was observed directly under the microscope. In laboratory culture, zygotes developed to crustose thalli, whereas unfused gametes died soon after germination. Sporophytes were found in nature with fertile unilocular sporangia in March. In culture, zoospores released from these sporangia also developed to crusts. Kuckuck’s observation at the beginning of the century (1912a) thatP. extensum exhibits an alternation of isomorphic generations and isogamy is thus confirmed.

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Anschrift bis Ende 1989: Instituto de Botánica, Universidad Austral de Chile, Cas. 567, Valdivia, Chile     

Contribution to the seaweed flora of Namibia: new records of Chlorophyta and Phaeophyceae

Six species of benthic marine Chlorophyta and Phaeophyceae are recorded for the first time from the coast of Namibia: Ectocarpus fasciculatus Harvey, Feldmannia irregularis (Kützing) G. Hamel, Bachelotia antillarum (Grunow) Gerloff, Straguloria clavata (Harvey in Hooker) Hamel, Entocladia leptochaete (Huber) Burrows and Ulvella lens P. Crouan & H. Crouan. Data concerning local and world distribution of these species as well as some taxonomical remarks are provided.     

Studies in the Liagoraceae (Nemaliales, Rhodophyta) I. The genus Trichogloea

The calcified, though strongly mucosoid gametophytes of Trichogloea (Nemaliales, Liagoraceae) usually appear seasonally for a short period in tropical and subtropical regions. Vegetative and reproductive characteristics of the presently recognized species, Trichogloea requienii (Mont.) Kützing (the generitype), Trichogloea lubrica J. Agardh, and Trichogloea herveyi W. R. Taylor, are described and compared. In addition, the identities of two previously synonymized species, Trichogloea jadinii Børgesen and Trichogloea javensis B0rgesen are clarified. Species of the genus display considerable variation in habit and branching patterns, necessitating an assessment of reproductive structures for accurate identification. Reliable features of Trichogloea include the structure of the medulla and the cortical (assimilatory) filaments, location and structure of carpogonial branches and spermatangia, and the nature of sterile filaments in the vicinity of the cystocarp. Secondary features include both external and internal branching pattern, and the proportion of calcium carbonate to lubricous material, the latter having to be assessed from fresh collections. The present study provides a comparative morphological account of the species of Trichogloea. As a result, T. requienii, T. lubrica, T. herveyi are confirmed as independent species; T. jadinii is removed from synonymy with T. lubrica and placed in that of T. requienii, and T. javensis is confirmed as a synonym of Izziella orientalis (J. Agardh) Huisman et Schils.     

The occurrence of shell-boring Phaeophila species in Britain

Phaeophila tenuis (Kylin) R. Nielsen (= Entocladia tenuis Kylin) is reported for the first time from British shores. The existence in Britain of a form intermediate in morphology between P. tenuis and Phaeophila engleri Reinke is also noted. The relationships of this form and P. tenuis to each other and to Phaeophila dendroides (Crouan frat.) Batt. are discussed with reference to the effect of host-shell species and habitat on morphology.     

Effect of H4R antagonist N-(2-aminoethyl)-5-chloro-1H-indol-2-carboxamides and 5-chloro-2-(piperazin-1-ylmethyl)-1H-benzimidazole on histamine and 4-methylhistamine-induced mast cell response

Context: The histamine plays a decisive role in acute and chronic inflammatory responses and is regulated through its four types of distinct receptors designated from H1 to H4. Recently histamine 4 receptor (H4R) antagonists have been reported to possess various pharmacological effects against various allergic diseases.

Objective: To investigate the inhibitory effect of N-(2-aminoethyl)-5-chloro-1H-indol-2-carboxamide (Compound A) and 5-chloro-2-(piperazin-1-ylmethyl)-1H-benzimidazole (Compound L) on H4R-mediated calcium mobilization, cytokine IL-13 production, ERK1/2, Akt and NF-κB activation in human mastocytoma cells-1 (HMC-1).

Materials and methods: Compounds A and L were synthesized chemically and their inhibitory effect on intracellular calcium release was analyzed by Fluo-4 calcium assay, cytokine measurement through ELISA and activation of signaling molecules by western blot.

Results: Pre-treatment with compounds A and L significantly reduced the H4R-mediated intracellular calcium release. Histamine and 4-methylhistamine (4-MH) induced Th2 cytokine IL-13 production in HMC-1 cells, was inhibited by compound A (77.61%, 74.25% at 1?μM concentration) and compound L (79.63%, 81.70% at 1?μM concentration). Furthermore, histamine induced the phosphorylation of ERK1/2, Akt and NF-κB was suppressed by compounds A and L at varying levels, ERK1/2 (88%, 86%), Akt (88%, 89%) and NF-κB (89%, 87%) in HMC-1 cells.

Discussion and conclusions: Taken together these data demonstrate that compound A and compound L may block H4R-mediated downstream signaling events.     

LIGHT AND ELECTRON MICROSCOPIC OBSERVATIONS OF PREFERENTIAL DESTRUCTION OF CHLOROPLAST AND MITOCHONDRIAL DNA AT EARLY MALE GAMETOGENESIS OF THE ANISOGAMOUS GREEN ALGA DERBESIA TENUISSIMA (CHLOROPHYTA)1

Gametogenesis in male and female gametophytes was studied by light microscopy and EM in the dioecious multinucleate green alga Derbesia tenuissima (Moris & De Notaris) P. Crouan & H. Crouan, where male and female gametes differ in size. Gametogenesis was divided into five stages: 32 h (stage 1), 24 h (stage 2), 16 h (stage 3), 8 h (stage 4), and 0.5 h (stage 5) before gamete release. At stage 1, the first sign of gametogenesis observed was the aggregation of gametophyte protoplasm to form putative gametangia. At stage 2, gametangia were separated from the vegetative protoplasm of gametophytes. Morphological changes of nuclei and organelles occurred at this early stage of male gametogenesis, and organelle DNA degenerated. At stage 3, male organelle DNA had completely degenerated, whereas in female gametangia, organelle DNA continued to exist in both chloroplasts and mitochondria. Gametogenesis was almost completed at stage 4 and fully at stage 5. Small male gametes had a DNA‐containing nucleus and a large mitochondrion and one or several degenerated chloroplasts. The mitochondria and plastids were devoid of DNA. The large female gametes had a nucleus and multiple organelles, all of which contained their own DNA. Thus, degeneration of chloroplast DNA along with morphological changes of organelles occurred at male gametogenesis in anisogamous green algae (Bryopsis and D. tenuissima), in contrast with previous studies in isogamous green algae (Chlamydomonas, Acetabularia caliculus, and Dictyosphaeria cavernosa) in which degeneration of chloroplast DNA occurred after zygote formation.     

Synthesis and evaluation of biological and antitumor activities of 5,7-dimethyl- oxazolo[5,4-d]pyrimidine-4,6(5H,7H)-dione derivatives as novel inhibitors of FGFR1

Abstract

A series of 5,7-dimethyl-oxazolo[5,4-d]pyrimidine-4,6(5H,7H)-dione derivatives, N5a–5l, was designed, synthesized and evaluated for their FGFR1-inhibition ability as well as cytotoxicity against three cancer cell lines (H460, B16F10 and A549) in vitro. Several compounds displayed good-to-excellent potency against these cancer cell lines compared to SU5402. Structure–activity relationship analyses indicated that compounds with a rigid structure and more heteroatoms at the side chain of the parent ring were more effective than those without these substitutions. The compound N5g (37.4% FGFR1 inhibition at 1.0?μM) was identified to have the most potent antitumor activities, with IC₅₀ values of 5.472, 4.260 and 5.837?μM against H460, B16F10 and A549 cell lines, respectively. Together, our results suggest that 5,7-dimethyl-oxazolo[5,4-d]pyrimidine-4,6(5H,7H)-dione derivatives may serve as potential agents for the treatment of FGFR1-mediated cancers.     

Annual growth rate of the calcareous red alga Lithothamnion corallioides (Corallinales,Rhodophyta) in the Bay of Brest,France

Lithothamnion corallioides Crouan et Crouan (Rhodophyta, Corallinales) is the main constituent of the maerl beds of the Bay of Brest (Atlantic coast of western Brittany). Its growth rate was measured monthly in situ during one year. Growth rates were obtained by an adaptation of the buoyant weight technique. The highest daily growth rate was observed in July and reached 0.26 % d^–1 (S.D. = 0.06), when expressed as the increase of calcium carbonate weight. The average daily growth rate was 0.12% d^–1 (S.D. = 0.04) for a period of 275 days (summer and autumn 1988, winter 1989). Using this preliminary data, the calcium carbonate accretion rate can be estimated provisionally: 876 g m^–2 year^–1, a rate much lower than that of tropical reef coralline algae, but higher than that of Lithophyllum incrustans, the well known temperate European reef-builder.     

Karyological studies of three species of Cladophora (Cladophorales,Chlorophyta) from Bermuda

Abstract

Chromosome numbers for three species of Cladophora from Bermuda are presented. C. laetevirens (Dillw.) Kuetz., C. prolifera (Roth) Kuetz., and C. conferta Crouan frat. ex Schramm & Maze were found to have similar karyotypes with 1 N = 2 X = 12 chromosomes belonging to size class III (0.8–2.2 μm). Correlations between each species' karyotype and its morphology and phytogeographic affinity are discussed. Estimates of the basic genome (1 X) for these and other Cladophora species indicate that nuclear DNA content, which shows a threefold variation in the genus, occurs in discontinuous increments. These findings are discussed in relation to reports of large scale, discontinuous DNA variation in vascular plant genera.     

Synthesis and antitumor activity of novel 2, 3-didithiocarbamate substituted naphthoquinones as inhibitors of pyruvate kinase M2 isoform

The M2 isoform of pyruvate kinase (PKM2) is a potential antitumor therapeutic target. In this study, we designed and synthesised a series of 2, 3-didithiocarbamate substituted naphthoquinones as PKM2 inhibitors based on the lead compound 3k that we previously reported. Among them, compound 3f (IC₅₀?=?1.05?±?0.17 µM) and 3h (IC₅₀?=?0.96?±?0.18 µM) exhibited potent inhibition of PKM2, and their inhibitory activities are superior to compound 3k (IC₅₀?=?2.95?±?0.53 µM) and the known PKM2 inhibitor shikonin (IC₅₀?=?8.82?±?2.62 µM). In addition, we evaluated in vitro antiproliferative effects of target compounds using MTS assay. Most target compounds exhibited dose-dependent cytotoxicity with IC₅₀ values in nanomolar concentrations against HCT116, MCF7, Hela, H1299 and B16 cells. These small molecule PKM2 inhibitors not only provide candidate compounds for cancer therapy, but also offer a tool to probe the biological effects of PKM2 inhibition on cancer cells.