The interplay of nested biotic interactions and the abiotic environment regulates populations of a hypersymbiont.

The role of biotic interactions in shaping the distribution and abundance of species should be particularly pronounced in symbionts. Indeed, symbionts have a dual niche composed of traits of their individual hosts and the abiotic environment external to the host, and often combine active dispersal at finer scales with host-mediated dispersal at broader scales. The biotic complexity in the determinants of species distribution and abundance should be even more pronounced for hyper symbionts (symbionts of other symbionts). We use a chain of symbiosis to explore the relative influence of nested biotic interactions and the abiotic environment on occupancy and abundance of a hypersymbiont. Our empirical system is the epibiont ciliate Lagenophrys discoidea, which attaches to an ostracod that is itself ectosymbiotic on crayfish (the basal host). We applied multimodel selection and variance partitioning for GLMM to assess the relative importance of (a) traits of symbiotic hosts (ostracod sex and abundance), (b) traits of basal hosts (crayfish body weight, abundance and intermoult stage), (c) the abiotic environment (water chemistry and climate) and (d) geospatial autocorrelation patterns (capturing potential effects of crayfish dispersal among localities). Our models explained about half of the variation in prevalence and abundance of the hypersymbiont. Variation in prevalence was partly explained, in decreasing order of importance (18%-4%) by shared effects of symbiotic host traits and the abiotic environment, pure fixed effects of symbiotic hosts, abiotic environment and geospatial patterns (traits of basal hosts were not relevant). Hypersymbiont abundance was most strongly explained by random effects of host traits (mainly the symbiotic host), in addition to weaker fixed effects (mostly abiotic environment). Our results highlight the major role of the interplay between abundance of symbiotic hosts and water physico-chemistry in regulating populations of a hypersymbiotic ciliate, which is likely critical for dispersal dynamics, availability of attachment resources and suitability of on-host living conditions for the ciliate. We also found moderate signal of regulation by the basal host, for which we propose three mechanisms: (a) modulation of microhabitat suitability (crayfish-created water currents); (b) concentration of symbiotic hosts within crayfish; and (c) dispersal mediated by crayfish.

Phylogeny of the families Zoothamniidae and Epistylididae (Protozoa: Ciliophora: Peritrichia) based on analyses of three rRNA-coding regions.

Peritrichs are a major group of ciliates with worldwide distribution, and they play important roles in many habitats. Intrafamilial phylogeny of some peritrichs was investigated using information from three genes, which provided more robust interpretations than single-gene analyses. Sixty-seven new sequences including SSU rDNA, ITS1-5.8S-ITS2 and LSU rDNA were aligned with available sequences in GenBank to infer phylogenetic relationships within the families Zoothamniidae and Epistylididae. Results reveal the following relationships: (1) Epistylididae is polyphyletic, consisting of two clades that nest within the Zoothamniidae as part of the crown clade of peritrichs (order Vorticellida) and a third one that is part of the basal clade of peritrichs (order Opercularida); (2) Epistylis elongata falls within one of the clades of Zoothamnium rather than with congeners; (3) Zoothamnium is probably paraphyletic, consisting of three divergent clades, with the genera Myoschiston and Zoothamnopsis intermingled with species of Zoothamnium. The following evolutionary hypotheses can be inferred from these results: (1) the contractile stalk of Zoothamnium is plesiomorphic. (2) Myoschiston, Zoothamnopsis and clade II of Epistylididae are derived from the Zoothamnium morphotype by partial or incomplete development of the spasmoneme that forms the contractile center of the stalk around which the rigid cortex is secreted. (3) Clade I of the Epistylididae, which are primarily colonial forms that appear never to have evolved a spasmoneme of any sort, may represent the ancestral morphotype of peritrichs.

New insights into the nature of semi-soft elasticity and "mechanical-Fréedericksz transitions" in liquid crystal elastomers.

The mechanical properties of an all-acrylate liquid crystal elastomer (LCE) with a glass transition of 14 ± 1 °C are reported. The highly nonlinear load curve has a characteristic shape associated with semi-soft elasticity (SSE). Conversely, measurements of the director orientation throughout tensile loading instead indicate a "mechanical-Fréedericksz" transition (MFT). Values of the step length anisotropy, r, are independently calculated from the theories of SSE (r = 3.2 ± 0.4), MFT (9.3 < r < 30.0) and thermally-induced length change (r = 3.8 ± 0.5). From simultaneously recorded polarising microscopy textures, the consequences of the above discrepancies are considered. Further, a mechanically-induced negative order parameter is observed. Results show the tensile load curve shape cannot solely be used to determine the underlying physics. Consequently, the LCE properties cannot be fully described by theories of SSE or MFTs alone. This suggests that the theory of LCEs is not yet complete. The conclusions suggest that both the LC order parameter and r must be functions of the mechanical deformation.

Beyond the "Code": A Guide to the Description and Documentation of Biodiversity in Ciliated Protists (Alveolata, Ciliophora).

Recent advances in molecular technology have revolutionized research on all aspects of the biology of organisms, including ciliates, and created unprecedented opportunities for pursuing a more integrative approach to investigations of biodiversity. However, this goal is complicated by large gaps and inconsistencies that still exist in the foundation of basic information about biodiversity of ciliates. The present paper reviews issues relating to the taxonomy of ciliates and presents specific recommendations for best practice in the observation and documentation of their biodiversity. This effort stems from a workshop that explored ways to implement six Grand Challenges proposed by the International Research Coordination Network for Biodiversity of Ciliates (IRCN-BC). As part of its commitment to strengthening the knowledge base that supports research on biodiversity of ciliates, the IRCN-BC proposes to populate The Ciliate Guide, an online database, with biodiversity-related data and metadata to create a resource that will facilitate accurate taxonomic identifications and promote sharing of data.

Electronic liquid crystal contact lenses for the correction of presbyopia.

Presbyopia, the age-related reduction in near vision acuity, is one of the leading issues facing the contact lens industry due to an increasingly ageing population and limitations associated with existing designs. A plastic-based liquid crystal contact lens is described which is designed to allow switchable vision correction. The device is characterized by low operating voltages (<5V(rms)) and has curvatures suitable for placement upon the cornea. Imaging and Point Spread Function analysis confirm that the lens provides an increase in optical power of + 2.00 ± 0.25 D when activated, ideal for presbyopia correction.

Ciliates and the rare biosphere: a review.

Here we provide a brief review of the rare biosphere from the perspective of ciliates and other microbial eukaryotes. We trace research on rarity from its lack of much in-depth focus in morphological and Sanger sequencing projects, to its central importance in analyses using high throughput sequencing strategies. The problem that the rare biosphere is potentially comprised of mostly errors is then discussed in the light of asking community-comparative, novel-diversity, and ecosystem-functioning questions.

An ITS-based phylogenetic framework for the genus Vorticella: finding the molecular and morphological gaps in a taxonomically difficult group.

Vorticella includes more than 100 currently recognized species and represents one of the most taxonomically challenging genera of ciliates. Molecular phylogenetic analysis of Vorticella has been performed so far with only sequences coding for small subunit ribosomal RNA (SSU rRNA); only a few of its species have been investigated using other genetic markers owing to a lack of similar sequences for comparison. Consequently, phylogenetic relationships within the genus remain unclear, and molecular discrimination between morphospecies is often difficult because most regions of the SSU rRNA gene are too highly conserved to be helpful. In this paper, we move molecular systematics for this group of ciliates to the infrageneric level by sequencing additional molecular markers-fast-evolving internal transcribed spacer (ITS) regions-in a broad sample of 66 individual samples of 28 morphospecies of Vorticella collected from Asia, North America and Europe. Our phylogenies all featured two strongly supported, highly divergent, paraphyletic clades (I, II) comprising the morphologically defined genus Vorticella. Three major lineages made up clade I, with a relatively well-resolved branching order in each one. The marked divergence of clade II from clade I confirms that the former should be recognized as a separate taxonomic unit as indicated by SSU rRNA phylogenies. We made the first attempt to elucidate relationships between species in clade II using both morphological and multi-gene approaches, and our data supported a close relationship between some morphospecies of Vorticella and Opisthonecta, indicating that relationships between species in the clade are far more complex than would be expected from their morphology. Different patterns of helix III of ITS2 secondary structure were clearly specific to clades and subclades of Vorticella and, therefore, may prove useful for resolving phylogenetic relationships in other groups of ciliates.

Optimization of refractive liquid crystal lenses using an efficient multigrid simulation.

A multigrid computational model has been developed to assess the performance of refractive liquid crystal lenses, which is up to 40 times faster than previous techniques. Using this model, the optimum geometries producing an ideal parabolic voltage distribution were deduced for refractive liquid crystal lenses with diameters from 1 to 9 mm. The ratio of insulation thickness to lens diameter was determined to be 1:2 for small diameter lenses, tending to 1:3 for larger lenses. The model is used to propose a new method of lens operation with lower operating voltages needed to induce specific optical powers. The operating voltages are calculated for the induction of optical powers between + 1.00 D and + 3.00 D in a 3 mm diameter lens, with the speed of the simulation facilitating the optimization of the refractive index profile. We demonstrate that the relationship between additional applied voltage and optical power is approximately linear for optical powers under + 3.00 D. The versatility of the computational simulation has also been demonstrated by modeling of in-plane electrode liquid crystal devices.

Molecular and morphological characterization of a poorly known marine ciliate, Myoschiston duplicatum precht 1935: implications for phylogenetic relationships between three morphologically similar genera -- Zoothamnium, Myoschiston, and Zoothamnopsis (Ciliophora, Peritrichia, Zoothamniidae).

We studied the morphology and molecular phylogeny of Myoschiston duplicatum, a peritrich ciliate that has been recorded as an epibiont of crustaceans, but which we also identified on marine algae from Korea. The important morphological characteristics revealed by silver staining of Myoschiston species have not been described because they are rarely collected. Using morphological methods, we redescribed the type species of the genus, Myoschiston duplicatum, and provided an improved diagnosis of Myoschiston. In addition, the coding regions for nuclear small subunit (SSU) rRNA and internal transcribed spacer 1-5.8S-internal transcribed spacer 2 sequences were sequenced. Phylogenetic analyses that included available SSU rDNA sequences of peritrichs from GenBank strongly supported a position of M. duplicatum within the family Zoothamniidae. In addition, phylogenetic analyses were performed with single datasets (ITS1-5.8S-ITS2) and combined datasets (SSU rDNA + ITS1-5.8S-ITS2) to explore further the phylogenetic relationship in the family Zoothamniidae between the three morphologically similar genera-Zoothamnium, Myoschiston, and Zoothamnopsis.

Evolution of variations in the common pattern of stomatogenesis in peritrich ciliates: evidence from a comparative Study including a new description of stomatogenesis in Pseudepistylis songi Peng et al., 2007.

The stomatogenesis of peritrich ciliates is an important developmental process but has been studied relatively little for such a large, diverse taxon. Complex oral structures and an inability of staining techniques to reveal them clearly have been the major factors hindering investigation of this process. In the present study, an improved method of staining with protargol was used to investigate the entire process of stomatogenesis in a large, colonial species of peritrich, Pseudepistylis songi, and to compare it to descriptions of stomatogenesis in several other species. We found that P. songi and other peritrichs have the same general type of ophryobuccokinetal stomatogenesis, with the parental oral complex being inherited by one daughter and the new oral complex by the other daughter; however, some differences between individual taxa appear to have arisen in the course of evolution. Reorganization of the entire germinal kinety (Gk) to form the germinal anlage may be plesiomorphic, with restriction of reorganization to its abstomal part being apomorphic. Development of the entire new haplokinety of one daughter cell (2Hk) from the germinal band or its homologue also appears to be plesiomorphic, with development of peristomial and infundibular parts of 2Hk from separate rudiments being apomorphic. Furthermore, development of the new Gk of the parental oral complex (1Gk) from the entire infundibular part of the parental haplokinety (1Hk) may be plesiomorphic, and development from just the abstomal part may be apomorphic. Finally, development of the Gk of the new oral complex (2Gk) from residual kinetosomes of the germinal band appears to be plesiomorphic.

Molecular phylogeny of the family Vorticellidae (Ciliophora, Peritrichia) using combined datasets with a special emphasis on the three morphologically similar genera Carchesium, Epicarchesium and Apocarchesium.

Little is known about the phylogeny of the family Vorticellidae at the generic level because few comprehensive analyses of molecular phylogenetic relationships between members of this group have, so far, been done. As a result, the phylogenetic positions of some genera that were based originally on morphological analyses remain controversial. In the present study, we performed phylogenetic analyses of vorticellids based on the sequence of the small-subunit (SSU) rRNA gene, including one species of the genus Apocarchesium, for which no sequence has previously been reported. Phylogenetic trees were reconstructed with SSU rRNA gene sequences by using four different methods (Bayesian analysis, maximum-likelihood, neighbour-joining and maximum-parsimony) and had a consistent branching pattern. Members of the genera Vorticella (except V. microstoma) and Carchesium formed a clearly defined, well supported clade that was divergent from the clade comprising members of the genera Pseudovorticella and Epicarchesium, suggesting that the differences in the silverline system (transverse vs reticulate) among vorticellids may be the result of genuine evolutionary divergence. Members of the newly established genus Apocarchesium clustered within the family Vorticellidae basal to the clade containing members of the genera Pseudovorticella and Epicarchesium and were distinct from members of the genus Carchesium, supporting the validity of Apocarchesium as a novel genus. Additional phylogenetic analyses of 21 strains representing seven genera from the families Vorticellidae and Zoothamniidae were performed with single datasets (ITS1-5.8S-ITS2, ITS2 alone) and combined datasets (SSU rRNA+ITS1-5.8S-ITS2, SSU rRNA+ITS2) to explore further the phylogenetic relationship between the three morphologically similar genera Carchesium, Epicarchesium and Apocarchesium, using characteristics not included in previous analyses. The phylogenetic trees reconstructed with combined datasets were more robust and therefore more reliable than those based on single datasets and supported the results of trees based on SSU rRNA sequences.

Analysis of the secondary structure of ITS transcripts in peritrich ciliates (Ciliophora, Oligohymenophorea): implications for structural evolution and phylogenetic reconstruction.

Despite extensive previous morphological work, little agreement has been reached about phylogenetic relationships among peritrich ciliates, making it difficult to study the evolution of the group in a phylogenetic framework. In this study, the nucleotide characteristics and secondary structures of internal transcribed spacers 1 and 2 (ITS1 and ITS2) of 26 peritrich ciliates in 12 genera were analyzed. Information from secondary structures of ITS1 and ITS2 then was used to perform the first systematic study of ITS regions in peritrich ciliates, including one species of Rhabdostyla for which no sequence has been reported previously. Lengths of ITS1 and ITS2 sequences varied relatively little among taxa studied, but their G+C content was highly variable. General secondary structure models of ITS1 and ITS2 were proposed for peritrich ciliates and their reliability was assessed by compensatory base changes. The secondary structure of ITS1 contains three major helices in peritrich ciliates and deviations from this basic structure were found in all taxa examined. The core structure of peritrich ITS2 includes four helices, with helix III as the longest and containing a motif 5'-MAC versus GUK-3' at its apex as well as a YU-UY mismatch in helix II. In addition, the structural motifs of both ITS secondary structures were identified. Phylogenetic analyses using ITS data were performed by means of Bayesian inference, maximum likelihood and neighbor joining methods. Trees had a consistent branching pattern that included the following features: (1) Rhabdostyla always clustered with members of the family Vorticellidae, instead of members of the family Epistylididae, in which it is now classified on the basis of morphology. (2) The systematically questionable genus Ophrydium closely associated with Carchesium, forming a clearly defined, monophyletic group within the Vorticellidae. This supported the hypothesis derived from previous study based on small subunit rRNA gene sequences that Ophrydium and its few relatives are morphologically anomalous vorticellids, not sufficiently distinct to be given familial status and should be placed within a more broadly defined family Vorticellidae. This study validated for the first time a secondary structure of ITS1 and ITS2 from peritrich ciliates and demonstrated its potential in helping to resolve deep phylogenetic relationships.

Description of Paratetrahymena parawassi n. sp. using morphological and molecular evidence and a phylogenetic analysis of Paratetrahymena and other taxonomically ambiguous genera in the order Loxocephalida (Ciliophora, Oligohymenophorea).

The marine scuticociliate Paratetrahymena parawassi n. sp. is described on the basis of morphology, especially infraciliature, and the sequence of its small subunit (SSU) rRNA gene to become the second known member of its genus. Paratetrahymena and other ciliates in the order Loxocephalida possess a mixture of morphological and morphogenetic features characteristic of the subclasses Hymenostomatia and Scuticociliatia. Accordingly, we used SSU rRNA sequences to analyze the phylogeny of Paratetrahymena and three other loxocephalid genera. Paratetrahymena and Cardiostomatella vermiformis formed a moderately well-supported clade that diverged at a deep level from all other scuticociliates, supporting separation of loxocephalids from other scuticociliates as a suprafamilial taxon. Sathrophilus holtae was a sister taxon to Paratetrahymena and Cardiostomatella in a poorly supported, unresolved relationship; nevertheless, association of all three genera into a single clade was supported by an approximately unbiased (AU) test. Any association of these genera singly or as a group with the Hymenostomatia was rejected decisively by AU tests and by a complete absence in the loxocephalids of the unique nucleotide identities that distinguish hymenostomes. Therefore, the morphological and morphogenetic similarities of loxocephalids to hymenostomes may be plesiomorphies, and the conflicting mix of scuticociliate and hymenostome characteristics seen in loxocephalids may result from differing rates of character evolution. Dexiotrichides pangi and Urocentrum, which is currently classified as a peniculid, formed a small clade that associated with hymenostomes and peritrichs. Monophyly of the Loxocephalida with Dexiotrichides and/or Urocentrum included was not rejected by AU; however, inclusion of Urocentrum in the Peniculia was rejected by AU tests. A hypothesis is offered to explain the lack of resolution of loxocephalid ciliates and Urocentrum in phylogenetic trees, namely that their phylogenetic positions are influenced by a combination of heterogeneous data and long-branch attraction caused by poor representation of taxa in analyses. The well-known genus Cyclidium, a member of the order Pleuronematida, was revealed to be polyphyletic as a byproduct of our analyses of loxocephalids. In particular, Cyclidium porcatum appears to fall outside the clade containing typical members of the subclass Scuticociliatia and thus invites investigation as a possible member of the order Loxocephalida.

Reconsideration of systematic relationships within the order Euplotida (Protista, Ciliophora) using new sequences of the gene coding for small-subunit rRNA and testing the use of combined data sets to construct phylogenies of the Diophrys-complex.

Comprehensive molecular analyses of phylogenetic relationships within euplotid ciliates are relatively rare, and the relationships among some families remain questionable. We performed phylogenetic analyses of the order Euplotida based on new sequences of the gene coding for small-subunit RNA (SSrRNA) from a variety of taxa across the entire order as well as sequences from some of these taxa of other genes (ITS1-5.8S-ITS2 region and histone H4) that have not been included in previous analyses. Phylogenetic trees based on SSrRNA gene sequences constructed with four different methods had a consistent branching pattern that included the following features: (1) the "typical" euplotids comprised a paraphyletic assemblage composed of two divergent clades (family Uronychiidae and families Euplotidae-Certesiidae-Aspidiscidae-Gastrocirrhidae), (2) in the family Uronychiidae, the genera Uronychia and Paradiophrys formed a clearly outlined, well-supported clade that seemed to be rather divergent from Diophrys and Diophryopsis, suggesting that the Diophrys-complex may have had a longer and more separate evolutionary history than previously supposed, (3) inclusion of 12 new SSrRNA sequences in analyses of Euplotidae revealed two new clades of species within the family and cast additional doubt on the present classification of genera within the family, and (4) the intraspecific divergence among five species of Aspidisca was far greater than those of closely related genera. The ITS1-5.8S-ITS2 coding regions and partial histone H4 genes of six morphospecies in the Diophrys-complex were sequenced along with their SSrRNA genes and used to compare phylogenies constructed from single data sets to those constructed from combined sets. Results indicated that combined analyses could be used to construct more reliable, less ambiguous phylogenies of complex groups like the order Euplotida, because they provide a greater amount and diversity of information.

Further consideration of the phylogeny of some "traditional" heterotrichs (Protista, Ciliophora) of uncertain affinities, based on new sequences of the small subunit rRNA gene.

The systematic relationships and taxonomic positions of the traditional heterotrich genera Condylostentor, Climacostomum, Fabrea, Folliculina, Peritromus, and Condylostoma, as well as the licnophorid genus Licnophora, were re-examined using new data from sequences of the gene coding for small subunit ribosomal RNA. Trees constructed using distance-matrix, Bayesian inference, and maximum-parsimony methods all showed the following relationships: (1) the "traditional" heterotrichs consist of several paraphyletic groups, including the current classes Heterotrichea, Armophorea and part of the Spirotrichea; (2) the class Heterotrichea was confirmed as a monophyletic assemblage based on our analyses of 31 taxa, and the genus Peritromus was demonstrated to be a peripheral group; (3) the genus Licnophora occupied an isolated branch on one side of the deepest divergence in the subphylum Intramacronucleata and was closely affiliated with spirotrichs, armophoreans, and clevelandellids; (4) Condylostentor, a recently defined genus with several truly unique morphological features, is more closely related to Condylostoma than to Stentor; (5) Folliculina, Eufolliculina, and Maristentor always clustered together with high bootstrap support; and (6) Climacostomum occupied a paraphyletic position distant from Fabrea, showing a close relationship with Condylostomatidae and Chattonidiidae despite of modest support.

Non-Orthogonal Refractive Lenses for Non-Orthogonal Astigmatic Eyes.

Purpose: To present a novel design method for non-orthogonal lenses to reduce the problem of residual astigmatism in non-orthogonal, astigmatic eyes Methods: A method to create spectacle trial lenses with non-orthogonal power axes was developed based on a novel optimised light ray-tracing algorithm rather than conventional lens design methods which could not fully eliminate spherical aberration. Using this method, three sets of refraction trial lenses were made with the angles between power axes of each set controlled at 80°, 70° and 60°, respectively. Within each set, the cylindrical power varied from -1.00 D to -6.00 D in 1.00 D steps in addition to a -0.50 D lens. Computer-based numerical simulation of the lenses optical performance was carried out to apply orthogonal and non-orthogonal lenses on simulated astigmatic eyes. Subsequently, three clinical trial cases were investigated. Results: Computer-simulated optical performance of non-orthogonal lenses showed the ability to achieve high performance in correcting non-orthogonal astigmatism. Subsequently, three patients with irregular astigmatism were refracted with the non-orthogonal lens sets, and clinically observed improvement at least two lines in the LogMAR chart was achieved in all three cases, compared with correction with orthogonal lenses, along with subjective improvement in image quality. Conclusions: Non-orthogonal astigmatism, which is commonly ignored by current eye prescription systems, is taken into account in this study in the design of spectacle and soft contact lenses. The new approach considers the possible non-orthogonal positions of the eye's two optical power meridians and appears to be better able to correct the vision of irregular astigmatic eyes and significantly reduce residual astigmatism.

Artefact-free topography based scleral-asymmetry.

PURPOSE: To present a three-dimensional non-parametric method for detecting scleral asymmetry using corneoscleral topography data that are free of edge-effect artefacts. METHODS: The study included 88 participants aged 23 to 65 years (37.7±9.7), 47 women and 41 men. The eye topography data were exported from the Eye Surface Profiler software in MATLAB binary data container format then processed by custom built MATLAB codes entirely independent from the profiler software. Scleral asymmetry was determined initially from the unprocessed topography before being determined again after removing the edge-effect noise. Topography data were levelled around the limbus, then edge-effect was eliminated using a robust statistical moving median technique. In addition to comparing raw elevation data, scleral elevation was also compared through fitting a sphere to every single scleral surface and determining the relative elevation from the best-fit sphere reference surface. RESULTS: When considering the averaged raw topography elevation data in the scleral section of the eye at radius 8 mm, the average raw elevations of the right eyes' sclera were -1.5±1.77, -1.87±2.12, -1.36±1.82 and -1.57±1.87 mm. In the left eyes at the same radius the average raw elevations were -1.62±1.78, -1.82±2.07, -1.28±1.76 and -1.68±1.93 mm. While, when considering the average raw elevation of the sclera after removing the edge effect, the average raw elevations of the right eyes were -3.71±0.25, -4.06±0.23, -3.95±0.19 and -3.95±0.23 mm. In the left eyes at the same radius the average raw elevations were -3.71±0.19, -3.97±0.22, -3.96±0.19 and -3.96±0.18 mm in the nasal, temporal, superior and inferior sides respectively. Maximum raw elevation asymmetry in the averaged scleral raw elevation was 1.6647±0.9015 mm in right eyes and 1.0358±0.6842 mm in left eyes, both detected at -38° to the nasal side. Best-fit sphere-based relative elevation showed that sclera is more elevated in three main meridians at angles -40°, 76°, and 170° in right eyes and -40°, 76°, and 170° in left eyes, all measured from the nasal meridian. Maximum recorded relative elevation asymmetries were 0.0844±0.0355 mm and 0.068±0.0607 mm at angular positions 76° and 63.5° for right and left eyes in turn. CONCLUSIONS: It is not possible to use corneoscleral topography data to predict the scleral shape without considering a method of removing the edge-effect from the topography data. The nasal side of the sclera is higher than the temporal side, therefore, rotationally symmetric scleral contact lenses are more likely to be translated towards the temporal side. The scleral shape is best described by levelled raw elevation rather than relative elevation.

Phylogeny of six genera of the subclass Haptoria (Ciliophora, Litostomatea) inferred from sequences of the gene coding for small subunit ribosomal RNA.

The small subunit ribosomal RNA genes of nine species belonging to six genera of litostome ciliates, namely Amphileptus aeschtae, Chaenea teres, Chaenea vorax, Lacrymaria marina, Litonotus paracygnus, Loxophyllum sp.-GD-070419, Loxophyllum jini, Loxophyllum rostratum, and Phialina salinarum, were sequenced for the first time. Phylogenetic trees were constructed using different methods to assess the inter- and intra-generic relationships of haptorians, of which Chaenea, Lacrymaria, Litonotus, and Phialina were analyzed for the first time based on molecular data. Monophyly of the order Pleurostomatida was strongly confirmed, and the two existing families of pleurostomatids, created on the basis of morphology, were confirmed by molecular evidence. Within the Pleurostomatida, Siroloxophyllum utriculariae occupied a well-supported position basal to the Loxophyllum clade, supporting the separation of these genera from one another. Both the subclass Haptoria and the order Haptorida were partially unresolved, possibly paraphyletic assemblages of taxa in all analyses, creating doubts about the traditional placement of some haptorid taxa. The existing sequence of L. rostratum in GenBank (DQ411864) was conspicuously different from that of the isolate from Qingdao, China sequenced in the present work, indicating that they are different species. The isolate from Qingdao was verified as L. rostratum by morphological analysis, and the published morphology of existing GenBank record of L. rostratum is different from it. Based on both morphological and molecular evidence, the latter may be congeneric with an undescribed species of Loxophyllum from Guangdong Province, China.

Morphology, morphogenesis, and phylogeny of an Anteholosticha intermedia (Ciliophora, Urostylida) population from the United States.

A distinct population of Anteholosticha intermedia was isolated from soil in the Great Smoky Mountains of North Carolina, USA, and its morphology, morphogenesis and molecular phylogeny investigated by microscopic observations of live and protargol-prepared specimens and analyses of the sequence of small subunit (SSU) rDNA. Our population closely resembles the populations from Austria and Korea. Members of the genus Anteholosticha have been regarded as ontogenetically diverse, which is confirmed by the present work. The most noteworthy ontogenetic feature of the American population of A. intermedia is that the oral primordium in the proter appears apokinetally at the posterior end of the undulating membranes anlage at the beginning of division and then dedifferentiates midway through morphogenesis. Molecular phylogenetic analyses demonstrate, with high support, that the American population of A. intermedia is clearly distinct from congeners and branches as part of a sister lineage to the Bakuella-Urostyla clade that belongs to the major clade comprising the order Urostylida.

Non-Orthogonal Corneal Astigmatism among Normal and Keratoconic Brazilian and Chinese populations.

PURPOSE: To investigate the prevalence of non-orthogonal astigmatism among normal and keratoconic Brazilian and Chinese populations. METHODS: Topography data were obtained using the Pentacam High Resolution (HR) system ® from 458 Brazilian (aged 35.6 ± 15.8 years) and 505 Chinese (aged 31.6 ± 10.8 years) eyes with no history of keratoconus or refractive surgery, and 314 Brazilian (aged 24.2 ± 5.7 years) and 74 Chinese (aged 22.0 ± 5.5 years) keratoconic eyes. Orthogonal values of optical flat and steep powers were determined by finding the angular positions of two perpendicular meridians that gave the maximum difference in power. Additionally, the angular positions of the meridians with the minimum and maximum optical powers were located while being unrestricted by the usual orthogonality assumption. Eyes were determined to have non-orthogonal astigmatism if the angle between the two meridians with maximum and minimum optical power deviated by more than 5° from 90°. RESULTS: Evidence of non-orthogonal astigmatism was found in 39% of the Brazilian keratoconic eyes, 26% of the Chinese keratoconic eyes, 29% of the Brazilian normal eyes and 20% of the Chinese normal eyes. CONCLUSIONS: The large percentage of participants with non-orthogonal astigmatism in both normal and keratoconic eyes illustrates the need for the common orthogonality assumption to be reviewed when correcting for astigmatism. The prevalence of non-orthogonality should be considered by expanding the prescription system to consider the two power meridians and their independent positions.