Phylogeny and systematics of demospongiae in light of new small-subunit ribosomal DNA (18S) sequences.

The most diverse and species-rich class of the phylum Porifera is Demospongiae. In recent years, the systematics of this clade, which contains more than 7000 species, has developed rapidly in light of new studies combining molecular and morphological observations. We add more than 500 new, nearly complete 18S sequences (an increase of more than 200%) in an attempt to further enhance understanding of the phylogeny of Demospongiae. Our study specifically targets representation of type species and genera that have never been sampled for any molecular data in an effort to accelerate progress in classifying this diverse lineage. Our analyses recover four highly supported subclasses of Demospongiae: Keratosa, Myxospongiae, Haploscleromorpha, and Heteroscleromorpha. Within Keratosa, neither Dendroceratida, nor its two families, Darwinellidae and Dictyodendrillidae, are monophyletic and Dictyoceratida is divided into two lineages, one predominantly composed of Dysideidae and the second containing the remaining families (Irciniidae, Spongiidae, Thorectidae, and Verticillitidae). Within Myxospongiae, we find Chondrosida to be paraphyletic with respect to the Verongida. We amend the latter to include species of the genus Chondrosia and erect a new order Chondrillida to contain remaining taxa from Chondrosida, which we now discard. Even with increased taxon sampling of Haploscleromorpha, our analyses are consistent with previous studies; however, Haliclona species are interspersed in even more clades. Haploscleromorpha contains five highly supported clades, each more diverse than previously recognized, and current families are mostly polyphyletic. In addition, we reassign Janulum spinispiculum to Haploscleromorpha and resurrect Reniera filholi as Janulum filholi comb. nov. Within the large clade Heteroscleromorpha, we confirmed 12 recently identified clades based on alternative data, as well as a sister-group relationship between the freshwater Spongillida and the family Vetulinidae. We transfer Stylissa flabelliformis to the genus Scopalina within the family Scopalinidae, which is of uncertain position. Our analyses uncover a large, strongly supported clade containing all heteroscleromorphs other than Spongillida, Vetulinidae, and Scopalinidae. Within this clade, there is a major division separating Axinellidae, Biemnida, Tetractinellida, Bubaridae, Stelligeridae, Raspailiidae, and some species of Petromica, Topsentia, and Axinyssa from Agelasida, Polymastiidae, Placospongiidae, Clionaidae, Spirastrellidae, Tethyidae, Poecilosclerida, Halichondriidae, Suberitidae, and Trachycladus. Among numerous results: (1) Spirophorina and its family Tetillidae are paraphyletic with respect to a strongly supported Astrophorina within Tetractinellida; (2) Agelasida is the earliest diverging lineage within the second clade listed above; and (3) Merlia and Desmacella appear to be the earliest diverging lineages of Poecilosclerida.

Relative increase in choline in the occipital cortex in chronic fatigue syndrome.

OBJECTIVE: To test the hypothesis that chronic fatigue syndrome (CFS) is associated with altered cerebral metabolites in the frontal and occipital cortices. METHOD: Cerebral proton magnetic resonance spectroscopy (1H MRS) was carried out in eight CFS patients and eight age- and sex-matched healthy control subjects. Spectra were obtained from 20 x 20 x 20 mm3 voxels in the dominant motor and occipital cortices using a point-resolved spectroscopy pulse sequence. RESULTS: The mean ratio of choline (Cho) to creatine (Cr) in the occipital cortex in CFS (0.97) was significantly higher than in the controls (0.76; P=0.008). No other metabolite ratios were significantly different between the two groups in either the frontal or occipital cortex. In addition, there was a loss of the normal spatial variation of Cho in CFS. CONCLUSION: Our results suggest that there may be an abnormality of phospholipid metabolism in the brain in CFS.

Recent introduction of the dominant tunicate, Pyura praeputialis (Urochordata, Pyuridae) to Antofagasta, Chile.

The large sessile tunicate Pyura stolonifera (Pleurogona: Stolibranchiata: Pyuridae), has been regarded as a complex taxon with disjointed distributions, including Australia (Pyura stolonifera praeputialis), South Africa (Pyura stolonifera stolonifera) and South America (Chile, Antofagasta: Pyura sp., the 'piure de Antofagasta'), and has been cited under at least five taxonomic combinations. The 'piure de Antofagasta' is a competitively dominant species in rocky intertidal habitats and shows a limited geographical range (60-70 km) exclusively inside the Bay of Antofagasta. Using cytochrome oxidase I (COI) mitochondrial sequence data from Pyura specimens of the three taxa we tested whether the Chilean taxon represents: (i) a Gondwana relict; (ii) a more recently divergent species; or (iii) a recently introduced species. The results suggest that the Chilean taxon is a recent introduction to Chile from Australian populations and that Pyura stolonifera praeputialis, from Australia, and the 'piure de Antofagasta' are geographical populations of a single species: Pyura praeputialis; whereas the South African taxon represents a second species: Pyura stolonifera.

Evaluating hypotheses of basal animal phylogeny using complete sequences of large and small subunit rRNA.

We studied the evolutionary relationships among basal metazoan lineages by using complete large subunit (LSU) and small subunit (SSU) ribosomal RNA sequences for 23 taxa. After identifying competing hypotheses, we performed maximum likelihood searches for trees conforming to each hypothesis. Kishino-Hasegawa tests were used to determine whether the data (LSU, SSU, and combined) reject any of the competing hypotheses. We also conducted unconstrained tree searches, compared the resulting topologies, and calculated bootstrap indices. Shimodaira-Hasegawa tests were applied to determine whether the data reject any of the topologies resulting from the constrained and unconstrained tree searches. LSU, SSU, and the combined data strongly contradict two assertions pertaining to sponge phylogeny. Hexactinellid sponges are not likely to be the basal lineage of a monophyletic Porifera or the sister group to all other animals. Instead, Hexactinellida and Demospongia form a well-supported clade of siliceous sponges, Silicea. It remains unclear, on the basis of these data alone, whether the calcarean sponges are more closely related to Silicea or to nonsponge animals. The SSU and combined data reject the hypothesis that Bilateria is more closely related to Ctenophora than it is to Cnidaria, whereas LSU data alone do not refute either hypothesis. LSU and SSU data agree in supporting the monophyly of Bilateria, Cnidaria, Ctenophora, and Metazoa. LSU sequence data reveal phylogenetic structure in a data set with limited taxon sampling. Continued accumulation of LSU sequences should increase our understanding of animal phylogeny.

Evolution of river dolphins.

The world's river dolphins (Inia, Pontoporia, Lipotes and Platanista) are among the least known and most endangered of all cetaceans. The four extant genera inhabit geographically disjunct river systems and exhibit highly modified morphologies, leading many cetologists to regard river dolphins as an unnatural group. Numerous arrangements have been proposed for their phylogenetic relationships to one another and to other odontocete cetaceans. These alternative views strongly affect the biogeographical and evolutionary implications raised by the important, although limited, fossil record of river dolphins. We present a hypothesis of river dolphin relationships based on phylogenetic analysis of three mitochondrial genes for 29 cetacean species, concluding that the four genera represent three separate, ancient branches in odontocete evolution. Our molecular phylogeny corresponds well with the first fossil appearances of the primary lineages of modern odontocetes. Integrating relevant events in Tertiary palaeoceanography, we develop a scenario for river dolphin evolution during the globally high sea levels of the Middle Miocene. We suggest that ancestors of the four extant river dolphin lineages colonized the shallow epicontintental seas that inundated the Amazon, Paraná, Yangtze and Indo-Gangetic river basins, subsequently remaining in these extensive waterways during their transition to freshwater with the Late Neogene trend of sea-level lowering.

Defining phyla: evolutionary pathways to metazoan body plans.

Phyla are defined by two sets of criteria, one morphological and the other historical. Molecular evidence permits the grouping of animals into clades and suggests that some groups widely recognized as phyla are paraphyletic, while some may be polyphyletic; the phyletic status of crown phyla is tabulated. Four recent evolutionary scenarios for the origins of metazoan phyla and of supraphyletic clades are assessed in the light of a molecular phylogeny: the trochaea hypothesis of Nielsen; the clonal hypothesis of Dewel; the set-aside cell hypothesis of Davidson et al.; and a benthic hypothesis suggested by the fossil record. It is concluded that a benthic radiation of animals could have supplied the ancestral lineages of all but a few phyla, is consistent with molecular evidence, accords well with fossil evidence, and accounts for some of the difficulties in phylogenetic analyses of phyla based on morphological criteria.

The significance of moulting in Ecdysozoan evolution.

Three major bilaterian clades first appear in the Early Cambrian fossil record: Deuterostomia, Lophotrochozoa, and Ecdysozoa. The taxa placed in Ecdysozoa are characterized by a moulting habit, unknown in the other major clades. The origin and consequences of moulting are of fundamental importance to the history of the ecdysozoan clade, chiefly because moulting precludes motile ectodermal cilia. Moulting may have originated as an adaptation to permit the enlargement, during growth, of secreted cuticular spines, flanges, and other structures used as ancillary locomotory devices. A combination of phylogenetic and fossil evidence suggests that the early members of these clades were small vermiform paracoelomates that likely lacked indirect-developing planktotrophic larvae. Thus, the evolution of planktotrophic larvae may have been independently achieved at least three times within Bilateria. The nonmoulting clades evolved larvae that swim and feed via ciliated tufts and bands, presumably intercalating these forms within their early developmental systems. Within Ecdysozoa, feeding larvae lacked ciliary feeding tracts and evolved by modification of early instars, employing limbs or setae to generate feeding currents. The setting aside during larval life of cells that give rise to adult features is probably an adaptation associated with metamorphosis.

Evaluating multiple alternative hypotheses for the origin of Bilateria: an analysis of 18S rRNA molecular evidence.

Six alternative hypotheses for the phylogenetic origin of Bilateria are evaluated by using complete 18S rRNA gene sequences for 52 taxa. These data suggest that there is little support for three of these hypotheses. Bilateria is not likely to be the sister group of Radiata or Ctenophora, nor is it likely that Bilateria gave rise to Cnidaria or Ctenophora. Instead, these data reveal a close relationship between bilaterians, placozoans, and cnidarians. From this, several inferences can be drawn. Morphological features that previously have been identified as synapomorphies of Bilateria and Ctenophora, e.g., mesoderm, more likely evolved independently in each clade. The endomesodermal muscles of bilaterians may be homologous to the endodermal muscles of cnidarians, implying that the original bilaterian mesodermal muscles were myoepithelial. Placozoans should have a gastrulation stage during development. Of the three hypotheses that cannot be falsified with the 18S rRNA data, one is most strongly supported. This hypothesis states that Bilateria and Placozoa share a more recent common ancestor than either does to Cnidaria. If true, the simplicity of placozoan body architecture is secondarily derived from a more complex ancestor. This simplification may have occurred in association with a planula-type larva becoming reproductive before metamorphosis. If this simplification took place during the common history that placozoans share with bilaterians, then placozoan genes that contain a homeobox, such as Trox2, should be explored, for they may include the gene or genes most closely related to Hox genes of bilaterians.

The use of aliases by psychiatric patients.

We investigated the incidence and clinical characteristics associated with the use of aliases within a population attending a psychiatric unit over a 12-year period. The use of aliases was uncommonly recognised (0.7%). Alias users tended to be young. Many were non-nationals (43%). Identification usually occurred early in the admission and was often by self-admission (37%). A broad range of underlying diagnoses were noted with schizophrenia (31%) and factitious disorder (22%) most common. A previous psychiatric (85%) or forensic history (30%) was frequently noted. Recurrence was noted in one third of cases and was significantly associated with a diagnosis of factitious disorder.

Induction of microcurrents in critically ill patients in magnetic resonance systems.

OBJECTIVE: To determine whether electric current can be induced in intracardiac catheters, thermistor wires and pacing electrodes in patients during magnetic resonance imaging (MRI). DESIGN: Prospective laboratory study. SETTING: Postgraduate medical school hospital. SUBJECTS: A sheep heart model. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Voltage generated by saline 0.9% flowing through a magnetic field and distribution of current from a catheter tip within a sheep heart model were measured in a 0.15 Tesla MRI system. Resistance of loops formed by pacing wires, a pacing electrode, and a thermistor wire were measured in saline 0.9%. Effects of rapidly changing magnetic fields and the movement of the beating heart on epicardial pacing wires were calculated theoretically. A flow of 200 mL/min of saline 0.9% induced a current of 0.1 microampere (microA) (at 0.15 Tesla). From magnetic resonance images we derived a current density of approximately 0.004 microA/mm2 (at 0.15 Tesla). Internal resistance of pacing catheters and thermistor wires was > 1 megaohm (M omega). The maximum currents calculated (for a higher field strength of 1.5 Tesla) in a circuit formed by epicardial pacing wires were 80 microA, induced by the beating heart moving the wires through the magnetic field and 46 microA, induced by the rapidly changing magnetic fields. CONCLUSIONS: Current generated by flow of conducting fluid should be safe. Pacing catheters and thermistor wires should be safe if well insulated and disconnected from external electric connections. However, current induced in epicardial pacing wires may be a hazard, and precautions should be taken. External wire tips must be separated, insulated, and coiled to lie along the axis of the magnetic field. Electrocardiography is required, and defibrillation equipment should be available.

Imaging of human brain activity at 0.15 T using fluid attenuated inversion recovery (FLAIR) pulse sequences.

A 3-4% change in signal intensity correlated with visual stimulation was observed in the occipital lobes of three normal volunteers examined with MRI at 0.15 T using fluid attenuated inversion recovery pulse sequences. Similar results were observed at 1.0 T. A double difference technique in which difference images are themselves opposed provided an increase in sensitivity.

Phosphorus-31 magnetic resonance spectroscopy of the human liver using chemical shift imaging techniques.

Phosphorus-31 magnetic resonance spectroscopy of the human liver was undertaken in 28 healthy adult individuals and in 49 patients with liver disease of varying aetiology. Data localised to the liver were obtained using chemical shift imaging techniques. The mean (+/- 1 S.D.) of the peak area ratio phosphomonoesters (PME)/phosphodiesters (PDE) in healthy adult individuals, from spectra obtained with pulse angle 45 degrees and repetition time 1 s, was 0.24 +/- 0.07. The intra-examination variability of this ratio was 20%, the intra-subject variability 27% and the inter-subject variability 32%. An increase in the PME/PDE was observed in the 31P hepatic MR spectrum from primary or secondary tumours in all 17 patients studied, which invariably represented an increase in PME/ATP and, in some cases, a reduction in PDE/ATP. The spectra did not show aetiological characteristics. A non-specific elevation in PME/PDE was also observed in the 31P hepatic MR spectra of 10 (40%) of 25 patients studied who had diffuse liver diseases, such as cirrhosis and infiltrating malignancies. The spectral pattern did not distinguish between diseases of varying aetiologies, but there was a linear correlation between increasing PME/PDE and a reduction in plasma albumin concentrations (p = 0.03). In three patients with hepatic malignancy and abnormal hepatic 31P-MRS, marked spectral changes were observed after successful treatment to debulk the tumour. Only minor changes were observed in the abnormal spectrum of a fourth patient in whom treatment was unsuccessful. Hepatic 31P-MR spectroscopy may prove useful for monitoring disease processes and treatment effects in well characterised patient populations.

MR imaging of anisotropically restricted diffusion of water in the nervous system: technical, anatomic, and pathologic considerations.

The use of MR imaging to image anisotropically restricted diffusion (ARD) of water in the nervous system is described. The theoretical basis for the use of the pulsed gradient spin echo sequences is outlined, including an estimate of the range of cell dimensions that can be studied with this technique. The importance of restricted diffusion across myelinated white matter fibre tracts is emphasised and the capacity of MR imaging to demonstrate fibre pathways as a function of their direction is illustrated. Technical developments that have been implemented include 256 x 256 spatial resolution, a wider range of diffusion times Td, and an increased range of diffusion sensitivity parameters b. Effects of these are illustrated together with the use of gradient moment nulling methods, oblique sensitisation, and a smaller set of gradient coils that enable shorter values of echo time to be used with the same value of b. The anatomical basis for ARD imaging is analysed, and association, commissural, and projection fibre tracts are demonstrated in different planes. The published literature on variations of the apparent diffusion coefficient from normal is reviewed and examples where diffusion weighted images revealed information that was not necessarily apparent with conventional sequences are illustrated. These include cases of multiple sclerosis, chronic head injury, progressive multifocal leucoencephalopathy, cerebrovascular disease, astrocytoma, and probable metastases to the brain. Imaging of ARD affords a fascinating conjunction between the microscopic movement of water, the properties of myelinated white matter fibres, gross anatomy of the brain, and changes of the diffusion of water in disease.

Necrobiotic granulomas localised to the penis: a possible variant of subcutaneous granuloma annulare.

Three young men developed multiple firm asymptomatic nodules confined to the shaft of the penis. Skin biopsies from all 3 cases showed prominent necrobiotic granulomas indistinguishable from granuloma annulare. One of the patients had repeated recurrent nodules confined to the penis which were removed surgically over a period of 20 months. This unusual, but benign, presentation of granuloma annulare has to be distinguished from other granulomatous and malignant processes which may present as penile nodules.

Assessment of phase and amplitude effects due to susceptibility variations in MR imaging of the brain.

Susceptibility effects due to the presence of hemorrhage have been observed in a substantial number of patients and are a potential source of artifact in in vivo spectroscopy. The studies so far have concentrated on observations of phase differences, but amplitude variations also exist. This article reviews the degree to which the two are independent and suggests that at least three different pulse sequences are necessary to characterize effects due to local field inhomogeneities arising in clinical studies.