Efficient separation and purification of allophycocyanin from <Emphasis Type=Italic>Spirulina</Emphasis> (<Emphasis Type=Italic>Arthrospira</Emphasis>) <Emphasis Type=Italic>platensis</Emphasis>

Allophycocyanin (APC) is a minor component of phycobiliproteins in cyanobacteria and red algae. This paper describes a simple and inexpensive extracting method for isolating APC from Spirulina (Arthrospira) platensis with high efficiency. The crude phycobiliprotein extract was pretreated by ammonium sulfate fractionation. Then, by adding hydroxylapatite into crude phycobiliprotein extract dissolved in 20 mM phosphate buffer (pH 7.0), APC was selectively adsorbed by hydroxylapatite but C-phycocyanin (C-PC) was not. The hydroxylapatite was collected and APC was extracted from the crude phycobiliprotein extract. Then, the enriched APC was washed off from the hydroxylapatite using 100 mM phosphate buffer (pH 7.0). In this simple extracting method it was easy to remove C-PC and isolate APC in large amounts. The absorbance ratio A ₆₅₀/A ₂₈₀ of extracted APC reached 2.0. The recovery yield was 70%, representing 4.61 mg · g⁻¹ wet weight. The extracted APC could be further purified by a simple anion-exchange chromatography with a pH gradient from 5.6 to 4.0. The absorbance ratio A ₆₅₀/A ₂₈₀ of the purified APC reached 5.0, and the overall recovery yield was 43%, representing 2.83 mg · g⁻¹ wet weight. Its purity was confirmed by native polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate-PAGE.     

Morphological and molecular characterization of two ITS groups of <Emphasis Type=Italic>Erysiphe</Emphasis> (Erysiphales) occurring on <Emphasis Type=Italic>Syringa</Emphasis> and <Emphasis Type=Italic>Ligustrum</Emphasis> (Oleaceae)

ITS sequences determined for 53 Erysiphe specimens on Syringa and Ligustrum collected in Europe, East Asia, and North and South America were divided into two ITS groups, S and K types. Phylogenetic analysis showed that these two ITS types do not share a common ancestor and form separate clades. The K type on Ligustrum was identified as Erysiphe ligustri based on the three-dimensional branching pattern of appendages. Morphological observations showed that there are some morphological differences—pigmentation of appendages and number of ascospores per ascus—between the S and K types on Syringa. Based on these morphological observations, the S and K types on Syringa were identified as E. syringae and E. syringae-japonicae, respectively. The recent abundant production of chasmothecia by lilac powdery mildew in Europe was caused by E. syringae-japonicae introduced from East Asia. DNA sequence analyses of the rDNA ITS region and the 28S rDNA, tub2, CYP51, and Chs1 genes did not support an interspecific hybrid origin for E. syringae-japonicae. Haplotype analysis suggested that E. syringae originated in North America and independently migrated to East Asia and Europe/South America.     

Genetic divergence in <Emphasis Type=Italic>Ditrema</Emphasis><Emphasis Type=Italic>jordani</Emphasis> (Perciformes: Embiotocidae) from the Pacific coast of southern Japan,as inferred from mitochondrial DNA sequences

Genetic divergence in Ditrema jordani was investigated from sequence variations on the mitochondrial cytochrome b gene. Clear genetic differences were found between specimens collected from the Mie and Shizuoka prefectures (westward of Izu Peninsula) and those from the Kanagawa and Chiba prefectures (eastward of Izu Peninsula). The uncorrected genetic distance between the two groups, which may represent separate taxa, was much greater (3.1–3.7%) than that between D. temminckii and D. viride (1.1–2.4%), and between two subspecies of D. temminckii (0.8–1.3%), suggesting that the Izu Peninsula acts as a stable geographic barrier to gene flow between the D. jordani groups. The clear genetic divergence between the two geographic populations of D. jordani may be partly related to direct development (viviparity) and low dispersal ability in the genus.     

Characteristics of <Emphasis Type=Italic>Phomopsis sclareae</Emphasis> obtained from sage (<Emphasis Type=Italic>Salvia officinalis</Emphasis>)

In the years 2004–2006 the species P. sclareae was isolated from sage stems showing the symptoms in the form of bark peeling off and breaking. On the basis of 5 isolates randomly chosen from the fungus population, morphology and the conditions of temperatures favourable for the most intensive growth and creation of the fungus infectious material were studied. The temperature of −6°C was viewed as unfavourable for the fungus growth, and that of 32°C was considered to prevent the formation of picnidia and conidia. The dynamic growth of the colonies and the formation of numerous picnidia and conidia were observed at the temperatures ranging from 20°C to 28°C.     

<Emphasis Type=Italic>Barleria compacta</Emphasis>: a new species in <Emphasis Type=Italic>Barleria</Emphasis> sect. <Emphasis Type=Italic>Prionitis</Emphasis> (Acanthaceae) from Somalia

Following a re-examination of the material treated under Barleria brevispina (Fiori) Hedrén in the recent Flora of Somalia account of the Acanthaceae, it is concluded that two distinct species are involved and Barleria compacta Malombe & I. Darbysh. is described here from north-eastern Somalia. Its affinities and conservation status are discussed.     

Attenuation of fibrosis <Emphasis Type=Italic>in vitro</Emphasis> and <Emphasis Type=Italic>in vivo</Emphasis> with <Emphasis Type=Italic>SPARC</Emphasis> siRNA

Introduction  

SPARC is a matricellular protein, which, along with other extracellular matrix components including collagens, is commonly over-expressed in fibrotic diseases. The purpose of this study was to examine whether inhibition of SPARC can regulate collagen expression in vitro and in vivo, and subsequently attenuate fibrotic stimulation by bleomycin in mouse skin and lungs.     

Novel molecular markers of <Emphasis Type=Italic>Chlamydia pecorum</Emphasis> genetic diversity in the koala (<Emphasis Type=Italic>Phascolarctos cinereus</Emphasis>)

Background  

Chlamydia pecorum is an obligate intracellular bacterium and the causative agent of reproductive and ocular disease in several animal hosts including koalas, sheep, cattle and goats. C. pecorum strains detected in koalas are genetically diverse, raising interesting questions about the origin and transmission of this species within koala hosts. While the ompA gene remains the most widely-used target in C. pecorum typing studies, it is generally recognised that surface protein encoding genes are not suited for phylogenetic analysis and it is becoming increasingly apparent that the ompA gene locus is not congruent with the phylogeny of the C. pecorum genome. Using the recently sequenced C. pecorum genome sequence (E58), we analysed 10 genes, including ompA, to evaluate the use of ompA as a molecular marker in the study of koala C. pecorum genetic diversity.     

Genetic diversity and domestication history of African rice (<Emphasis Type=Italic>Oryza glaberrima</Emphasis>) as inferred from multiple gene sequences

Nucleotide variation in 14 unlinked nuclear genes was investigated in species-wide samples of African rice (Oryza glaberrima) and its wild progenitor (O. barthii). Average estimates of nucleotide diversity were extremely low in both species (θ _sil = 0.0007 for O. glaberrima; θ _sil = 0.0024 for O. barthii). About 70% less diversity was found in O. glaberrima than in its progenitor O. barthii. Coalescent simulation indicated that such dramatic reduction of nucleotide diversity in African rice could be explained mainly by a severe bottleneck during its domestication. The progenitor of African rice maintained also low genetic diversity, which may be attributed to small effective population size in O. barthii. Self-pollinating would be another factor leading to the unusually low diversity in both species. Genealogical analyses showed that all O. glaberrima accessions formed a strongly supported cluster with seven O. barthii individuals that were sampled exclusively from the proposed domestication centers of African rice. Population structure and principal component analyses found that the O. glaberrima group was homogeneous with no obvious genetic subdivision, in contrast to the heterogeneous O. barthii cluster. These findings support a single domestication origin of African rice in areas of the Upper Niger and Sahelian Rivers.     

Mining metadata from unidentified ITS sequences in GenBank: A case study in <Emphasis Type=Italic>Inocybe</Emphasis> (<Emphasis Type=Italic>Basidiomycota</Emphasis>)

Background  

The lack of reference sequences from well-identified mycorrhizal fungi often poses a challenge to the inference of taxonomic affiliation of sequences from environmental samples, and many environmental sequences are thus left unidentified. Such unidentified sequences belonging to the widely distributed ectomycorrhizal fungal genus Inocybe (Basidiomycota) were retrieved from GenBank and divided into species that were identified in a phylogenetic context using a reference dataset from an ongoing study of the genus. The sequence metadata of the unidentified Inocybe sequences stored in GenBank, as well as data from the corresponding original papers, were compiled and used to explore the ecology and distribution of the genus. In addition, the relative occurrence of Inocybe was contrasted to that of other mycorrhizal genera.     

New species of <Emphasis Type=Italic>Dypsis</Emphasis> and <Emphasis Type=Italic>Ravenea</Emphasis> (Arecaceae) from Madagascar

Fourteen new species of palms (Arecaceae) from Madagascar are described and named, based on material collected over the last 15 years. Twelve species belong to the genus Dypsis, namely D. andilamenensis Rakotoarin. & J. Dransf., D. anjae Rakotoarin. & J. Dransf., D. betsimisarakae Rakotoarin. & J. Dransf., D. culminis Rakotoarin. & J. Dransf., D. dracaenoides Rakotoarin. & J. Dransf., D. gautieri Rakotoarin. & J. Dransf., D. gronophyllum Rakotoarin. & J. Dransf., D. jeremiei Rakotoarin. & J. Dransf., D. metallica Rakotoarin. & J. Dransf., D. reflexa Rakotoarin. & J. Dransf., D. sancta Rakotoarin. & J. Dransf. and D. vonitrandambo Rakotoarin. & J. Dransf. and two species belong to the genus Ravenea: R. beentjei Rakotoarin. & J. Dransf. and R. hypoleuca Rakotoarin. & J. Dransf. Despite the fact that most of these species have been recorded from protected areas that are difficult to access in the eastern region of Madagasacar, they are all threatened. Based on IUCN categories and criteria, seven are Critically Endangered and seven are Vulnerable.     

Presence of <Emphasis Type=Italic>C. albidus</Emphasis>, <Emphasis Type=Italic>C. laurentii</Emphasis> and <Emphasis Type=Italic>C. uniguttulatus</Emphasis> in Crop and Droppings of Pigeon Lofts (<Emphasis Type=Italic>Columba livia</Emphasis>)

Columba livia is an important reservoir and carrier of Cryptococcus neoformans, Cryptococcus uniguttulatus, Cryptococcus laurentii and Cryptococcus albidus. Upper digestive tract of this species is also known as a habitat for Cryptococcus neoformans. Given the increasing clinical interest of this microorganism, 331 swabs from crop and 174 dropping samples from pigeon lofts in Grand Canary Island have been studied. The obtained results show an extensive presence samples 81 positive (24.47%) of Cryptococcus spp. in analysed crops: 32 (9.66%) for C. neoformans, 24 (7.2%) for C. uniguttulatus, 23 (6.9%) for C. albidus and 2 (0.6%) for C. laurentii. In the same way, Cryptococcus spp was also isolated in 82 (47.13%), dropping samples: C. neoformans in 59 (33.9%), C. uniguttulatus, in 9 (5.17%), C. laurentii in 8 (4.59%) and C. albidus in 6 (3.44%) of the investigated samples, respectively. The cryptococcosis produced by species of cryptococci other than C. neoformans has become more important during the last decade, supporting the study on the role of pigeon in the epidemiology of this disease.     

Life cycle of <Emphasis Type=Italic>Aylax hypecoi</Emphasis> (<Emphasis Type=Italic>Insecta: Hymenoptera: Cynipidae</Emphasis>), a gall inducer on <Emphasis Type=Italic>Hypecoum</Emphasis> ssp. (<Emphasis Type=Italic>Papaveraceae</Emphasis>)

The life cycle and developmental stages of Aylax hypecoi (Trotter, 1913, Hymenoptera: Cynipidae: Aylacini) were studied in detail. Aylax hypecoi is known to induce galls in fruits of two Hypecoum species — H. imberbe and H. geslini (Papaveraceae) and the larva develops in host plant fruits. The morphology and development of egg, larva and pupa were investigated, which has previously not been done. The shape and size of terminal-instar larvae and associated galls are sex-specific. Overwintering stage, adult emergence and flying periods, and egg productivity were studied also.     

Probiotic <Emphasis Type=Italic>Lactobacillus</Emphasis> strains: <Emphasis Type=Italic>in vitro</Emphasis> and <Emphasis Type=Italic>in vivo</Emphasis> studies

Three Lactobacillus strains (LOCK 0900, LOCK 0908, LOCK 0919) out of twenty-four isolates were selected according to their antagonistic activity against pathogenic bacteria, resistance to low pH and milieu of bile salts. Intragastric administration of a mixture of these strains to Balb/c mice affected cytokine T_H1-T_H2 balance toward nonallergic T_H1 response. Spleen cells, isolated from lactobacilli-treated mice and re-stimulated in vitro with the mixture of heat-inactivated tested strains, produced significantly higher amounts of anti-allergic tumor necrosis factor- and interferon-γ than control animals whereas the level of pro-allergic interleukin-5 was significantly lower. Lactobacillus cells did not translocate through the intestinal barrier into blood, liver and spleen; a few Lactobacillus cells found in mesenteric lymph nodes could create antigenic reservoir activating the immune system. The mixture of Lactobacillus LOCK 0900, LOCK 0908 and LOCK 0919 strains represents a probiotic bacterial preparation with possible use in prophylaxis and/or therapy of allergic diseases.     

A new name in <Emphasis Type=Italic>Pavetta</Emphasis> L. (<Emphasis Type=Italic>Rubiaceae</Emphasis>)

Summary  The name Pavetta modesta (Hiern) S. E. Dawson is a later homonym of P. modesta Bremek. Pavetta crystalensis is proposed as a new name.