RESURRECTION OF THE GENUS OSMUNDEA WITH AN EMENDATION OF THE GENERIC DELINEATION OF LAURENCIA (CERAMIALES, RHODOPHYTA)

Vegetative and reproductive development of some European and Californian species of Laurencia Lamouroux (Ceramiales, Rhodophyta), L. obtusa (Hudson) Lamouroux, L. spectabilis Postels et Ruprecht, L. crispa Hollenberg, L. osmunda (S.G. Gmelin) Maggs et Hommersand, L. pinnatifida (Hudson) Lamouroux and L. truncata Kutzing, is investigated on the basis of liquid-preserved and herbarium specimens. The latter five species share several features, but they differ distinctly from L. obtusa, the lectotype of the genus, in essential anatomical characters of vegetative and male reproductive structures and tetrasporangial development. In these five species each vegetative axial segment produces two rather than four pericentral cells, and spermatangial branches (filaments) are produced in apical pits of branchlets from apical and epidermal cells rather than from trichoblasts arising from axial cells. The spermatangial branches are usually branched alternately and usually terminate in a cluster of several large sterile vesicular cells, rather than being branched dichotomously and terminating in a single, or occasionally a row of two, large sterile vesicular cells as in L. obtusa. Apical spermatangial pits of fertile male branchlets (except for those in L. truncata) are pocket- (or urn)-shaped, with an ostiole-like upper opening, rather than cup- (or bowl)-shaped. In these five species tetrasporangia are produced laterally from random epidermal cells rather than abaxially from particular pericentral cells (the third and fourth ones) as in L. obtusa, and the two presporangial cover cells are aligned parallel rather than transverse to the stichidial axis in surface view. These important differences strongly suggest that L. spectabilis, L. crispa, L. osmunda, L. pinnatifida and L. truncata occupy a phylogenetically different position from L. obtusa, and lead to the conclusion that the genus Osmundea Stackhouse, which was based on 0. expansa Stackhouse, nom. illeg. (= Laurencia osmunda) and which has been a nomen rejiciendum as an earlier facultative synonym of Laurencia, should be resurrected. Emendations of the generic criteria of Laurencia and Osmundea are proposed here, and relevant nomenclatural changes for several Laurencia species are also included.

Langmuir probe measurements of plasma parameters in the late stages of a laser ablated plume

A simple Langmuir probe technique has been used to measure the electron density, electron temperature, and plasma potential in the late stages (>5 mu s) of a laser ablated plasma plume. In the plasma, formed following 248 nm laser irradiation of a copper target, in vacuum at a laser fluence of 2.5 J cm(-2), electron densities of similar to 10(18) m(-3) and temperatures of similar to 0.5 eV were measured. These values are comparable with those reported previously using Faraday cup detectors and optical emission spectroscopy, respectively. (C) 1997 American Institute of Physics.

What are RBC-transfusion-dependence and -independence?

The term RBC-transfusion-dependence is widely-used by hematologists to describe a condition of severe anemia typically arising when erythropoiesis is reduced such that a person continuously requires =1 RBC-transfusions over a specified interval. Defining a person as RBC-transfusion-dependent has important implications in diverse hematological disorders especially because it strongly-correlated with decreased survival. Conversely, becoming RBC-transfusion-independent or receiving fewer RBC-transfusions over a specified interval is defined as improvement or response in many disease- and/or therapy-setting. Whether this correlates with improved survival is controversial. We used a structured expert-panel consensus panel process to define RBC-transfusion-dependence and -independence or improvement. We suggest these definitions may prove useful to persons studying or treating these diseases.

Functional characterization of Gne (UDP-N-acetylglucosamine-4-epimerase), Wzz (chain length determinant), and Wzy (O-antigen polymerase) of Yersinia enterocolitica serotype O:8

The lipopolysaccharide (LPS) O-antigen of Yersinia enterocolitica serotype O:8 is formed by branched pentasaccharide repeat units that contain N-acetylgalactosamine (GalNAc), L-fucose (Fuc), D-galactose (Gal), D-mannose (Man), and 6-deoxy-D-gulose (6d-Gul). Its biosynthesis requires at least enzymes for the synthesis of each nucleoside diphosphate-activated sugar precursor; five glycosyltransferases, one for each sugar residue; a flippase (Wzx); and an O-antigen polymerase (Wzy). As this LPS shows a characteristic preferred O-antigen chain length, the presence of a chain length determinant protein (Wzz) is also expected. By targeted mutagenesis, we identify within the O-antigen gene cluster the genes encoding Wzy and Wzz. We also present genetic and biochemical evidence showing that the gene previously called galE encodes a UDP-N-acetylglucosamine-4-epimerase (EC 5.1.3.7) required for the biosynthesis of the first sugar of the O-unit. Accordingly, the gene was renamed gne. Gne also has some UDP-glucose-4-epimerase (EC 5.1.3.2) activity, as it restores the core production of an Escherichia coli K-12 galE mutant. The three-dimensional structure of Gne was modeled based on the crystal structure of E. coli GalE. Detailed structural comparison of the active sites of Gne and GalE revealed that additional space is required to accommodate the N-acetyl group in Gne and that this space is occupied by two Tyr residues in GalE whereas the corresponding residues present in Gne are Leu136 and Cys297. The Gne Leu136Tyr and Cys297Tyr variants completely lost the UDP-N-acetylglucosamine-4-epimerase activity while retaining the ability to complement the LPS phenotype of the E. coli galE mutant. Finally, we report that Yersinia Wzx has relaxed specificity for the translocated oligosaccharide, contrary to Wzy, which is strictly specific for the O-unit to be polymerized.

Effects of short term increase of intraocular pressure on optic disc cupping

Aims. To evaluate the effect of acute elevation of intraocular pressure (IOP) on optic disc cupping. Methods. 10 emmetropic and 10 myopic volunteers were included in this study. The cup area (CA) and cup volume (CV) of the optic disc were determined with the Heidelberg retina tomograph (HRT). After baseline determinations, a suction cup was used to increase the intraocular pressure (IOP) to 20-25 mmHg above the baseline and HRT images were obtained. Results. Baseline IOP was 13.5 (SD 1.3) mmHg and 12.6 (2.6) mmHg in the emmetropic and myopic groups, respectively. The IOP was elevated to 35.4 (3.3) mmHg and 34.4 (2.5) mmHg in the emmetropic and myopic groups, respectively. When compared with their baseline values, the cupping variables (CA and CV) were significantly increased (p <0.05) during the suction treatment in both emmetropic and myopic subjects. Conclusion. There was a significant enlargement in the optic disc cupping during the artificial increment of intraocular pressure in both emmetropic and myopic eyes. In non-glaucomatous eyes the optic nerve head has a partially dynamic topography dependent upon the level of IOP.

Quantitative estimation of retinal nerve fiber layer height in glaucoma and the relationship with optic nerve head topography and visual field

Purpose: The authors estimated the retinal nerve fiber layer height (RNFLH) measurements in patients with glaucoma compared with those in age-matched healthy subjects as obtained by the laser scanning tomography and assessed the relationship between RNFLH measurements and optic and visual field status. Methods: Parameters of optic nerve head topography and RNFLH were evaluated in 125 eyes of 21 healthy subjects and 104 patients with glaucoma using the Heidelberg Retina Tomograph ([HRT] Heidelberg Engineering GmbH, Heidelberg, Germany) for the entire disc area and for the superior 70°(50°temporal and 20°nasal to the vertical midline) and inferior 70°sectors of the optic disc. The mean deviation of the visual field, as determined by the Humphrey program 24-2 (Humphrey Instruments, Inc., San Leonardo, CA, U.S.A) was calculated in the entire field and in the superior and inferior Bjerrum area. Result: Retinal nerve fiber layer height parameters (mean RNFLH and RNFL cross-sectional area) were decreased significantly in patients with glaucoma compared with healthy individuals. Retinal nerve fiber layer height parameters was correlated strongly with rim volume, rim area, and cup/disc area ratio. Of the various topography measures, retinal nerve fiber layer (RNFL) parameters and cup/disc area ratio showed the strongest correlation with visual field mean deviation in patients with glaucoma. Conclusion: Retinal nerve fiber layer height measures were reduced substantially in patients with glaucoma compared with age-matched healthy subjects. Retinal nerve fiber layer height was correlated strongly with topographic optic disc parameters and visual field changes in patients with glaucoma.

Comparison between laser scanning tomography and computerised image analysis of the optic disc

Aims - To study the interchangeability of the measurements of the optic disc topography obtained by one computerised image analyser and one confocal laser tomographic scanner. Methods - One eye of 28 patients with glaucoma or glaucoma suspects was studied. All cases had simultaneous stereoscopic disc photographs taken with the fundus camera Topcon TRC-SS and optic disc examination with the Heidelberg retina tomograph (HRT) during the same visit. The optic disc photographs were digitised and analysed with the Topcon ImageNet (TI) system. Three variables of the optic disc topography provided by the TI and the HRT were compared - cup volume (CV), rim area (RA), and cup area to disc area ratio (CA/DA). Results - The mean values of CV and RA provided by the TI (0.52 (SD 0.32) mm and 1.58 (0.39) mm , respectively) were greater (p <0.01) than the mean values of CV and RA determined by the HRT (0.32 (0.25) mm , and 1.33 (0.47) mm , respectively). The mean value of CA/DA provided by the TI (0.42 (0.14)) and the HRT (0.42 (0.18)) was similar (p = 0.93). Correlation coefficients between measurements obtained by the two methods ranged from 0.53 to 0.73. Conclusion - There was a significant discrepancy in the measurements of rim area and cup volume of the optic disc obtained by a computerised image analyser and a laser scanning tomograph.

Three dimensional analysis of the lamina cribrosa in glaucoma

Background/aim: Structural changes in the lamina cribrosa have been implicated in the pathogenesis of glaucomatous optic atrophy. The aim of this study was to determine a measure the surface variability of the cup floor in normal subjects and patients with glaucoma. Methods: A sample of age matched normal subjects (NN), patients with low tension glaucoma (LTG), and primary open angle glaucoma (POAG) were included in the study. The glaucoma groups were matched for the severity of the visual field loss. Mean 10 degree topographic images of normal and glaucomatous eyes from the Heidelberg retina tomograph were imported into ERDAS image processing software where topographic analysis of the cup floor could be assessed. Each image was processed using customised spatial filters that calculated the surface depth variation in localised neighbourhood areas across each image. The local change in depth across the cup floor surface was determined and compared between the three clinical groups. Results: The depth variation in the cup floor was largest in normal subjects followed by LTG and POAG. Highly statistically significant differences in surface depth variability of the cup floor existed between normal and LTG (p=0.005), between normal and POAG (p

Reversal of optic disc cupping after glaucoma surgery analyzed with a scanning laser tomograph

Objective: To detect and quantitate changes in optic nerve morphology after glaucoma surgery using the Heidelberg Retina Tomograph (HRT, Heidelberg Instruments, Heidelberg, Germany). Design: Nonconsecutive observational case series. Participants and Intervention: The authors prospectively enrolled 21 adult patients undergoing incisional glaucoma surgery for progressive glaucoma damage. Quantitative analysis of the optic nerve head by scanning laser tomography and automated perimetry were performed before and after glaucoma surgery. Main Outcome Measures: Changes in optic nerve parameters were subjected to linear regression analysis with respect to percent of postoperative reduction of intraocular pressure (IOP), as well as with respect to age, refraction, preoperative cup:disc ratio, and change in visual field parameters. Results: Seventeen patients had pre- and postoperative images suitable for analysis. Mean IOP at the time of image acquisition before surgery was 30.5 ± 12 mmHg, and after surgery 11.8 ± 5.2 mmHg (mean follow-up, 26 ± 7 weeks). Eleven of 13 (85%) patients having IOP reduction of greater than 40% showed improvement in optic disc parameters. All four patients with less than 25% reduction in IOP showed worsening of most parameters. Changes in optic disc parameters were highly correlated with percent IOP reduction and with age. The parameters in which change most strongly correlated with percent change of IOP were cup area, rim area, cup:disc ratio, and mean cup depth (each, P <0.005). The age of the patient correlated highly with change in maximum cup depth (P <0.005). Refraction and clinically determined cup:disc ratio correlated poorly with changes in measured optic disc parameters. Clinical improvement in visual fields was correlated with the degree of improvement of cup:disc ratio (P = 0.025). Conclusion: Most patients showing a 40% lowering of IOP after glaucoma surgery show improved optic nerve morphology as measured by the HRT. The amount of improvement correlated highly with the percent reduction of IOP.

In silico prediction of the effects of mutations in the human UDP-galactose 4'-epimerase gene:Towards a predictive framework for type III galactosemia

The enzyme UDP-galactose 4'-epimerase (GALE) catalyses the reversible epimerisation of both UDP-galactose and UDP-N-acetyl-galactosamine. Deficiency of the human enzyme (hGALE) is associated with type III galactosemia. The majority of known mutations in hGALE are missense and private thus making clinical guidance difficult. In this study a bioinformatics approach was employed to analyse the structural effects due to each mutation using both the UDP-glucose and UDP-N-acetylglucosamine bound structures of the wild-type protein. Changes to the enzyme's overall stability, substrate/cofactor binding and propensity to aggregate were also predicted. These predictions were found to be in good agreement with previous in vitro and in vivo studies when data was available and allowed for the differentiation of those mutants that severely impair the enzyme's activity against UDP-galactose. Next this combination of techniques were applied to another twenty-six reported variants from the NCBI dbSNP database that have yet to be studied to predict their effects. This identified p.I14T, p.R184H and p.G302R as likely severely impairing mutations. Although severely impaired mutants were predicted to decrease the protein's stability, overall predicted stability changes only weakly correlated with residual activity against UDP-galactose. This suggests other protein functions such as changes in cofactor and substrate binding may also contribute to the mechanism of impairment. Finally this investigation shows that this combination of different in silico approaches is useful in predicting the effects of mutations and that it could be the basis of an initial prediction of likely clinical severity when new hGALE mutants are discovered.

Comparison of dynamics of wildtype and V94M human UDP-galactose 4-epimerase-A computational perspective on severe epimerase-deficiency galactosemia

UDP-galactose 4'-epimerase (GALE) catalyzes the interconversion of UDP-galactose and UDP-glucose, an important step in galactose catabolism. Type III galactosemia, an inherited metabolic disease, is associated with mutations in human GALE. The V94M mutation has been associated with a very severe form of type III galactosemia. While a variety of structural and biochemical studies have been reported that elucidate differences between the wildtype and this mutant form of human GALE, little is known about the dynamics of the protein and how mutations influence structure and function. We performed molecular dynamics simulations on the wildtype and V94M enzyme in different states of substrate and cofactor binding. In the mutant, the average distance between the substrate and both a key catalytic residue (Tyr157) and the enzyme-bound NAD(+) cofactor and the active site dynamics are altered making substrate binding slightly less stable. However, overall stability or dynamics of the protein is not altered. This is consistent with experimental findings that the impact is largely on the turnover number (kcat), with less substantial effects on Km. Active site fluctuations were found to be correlated in enzyme with substrate bound to just one of the subunits in the homodimer suggesting inter-subunit communication. Greater active site loop mobility in human GALE compared to the equivalent loop in Escherichia coli GALE explains why the former can catalyze the interconversion of UDP-N-acetylgalactosamine and UDP-N-acetylglucosamine while the bacterial enzyme cannot. This work illuminates molecular mechanisms of disease and may inform the design of small molecule therapies for type III galactosemia.

A novel device to measure acetabular inclination with patients in lateral decubitus

The ability to measure acetabular cup orientation accurately during total hip arthroplasty represents a significant challenge. The aim of this research was to develop and evaluate a novel low cost mechanical device for measuring operative acetabular inclination. Cup implantation was simulated in two trials using the novel device: firstly involving surgeons and engineers orientating acetabular cups with sawbone pelves at a range of inclination angles (20°-55° in 5° increments); secondly in a simulated intra-operative scenario with surgeons. Target angles were compared with achieved angles and deviations from desired angles were recorded. In addition, all participants orientated cups under the same conditions using two other techniques: freehand and with a propriatory Mechanical Alignment Guide. In the first trial, the mean errors (deviations) using freehand technique, the mechanical alignment guide and the new device were 5.2° +/- 4.3° (range 0.1-22.0), 3.6° +/- 3.9° (range 0.1°-33.6°) and 0.5° +/- 0.4° (range 0.0-1.9) respectively. In the second trial, the mean error for freehand technique, mechanical alignment guide and the new device were 6.2° +/- 4.2° (range 0.2-18.2), 3.8° +/- 3.3° (range 0.0-19.1) and 0.6° +/- 0.5° (range 0.0-1.8) respectively. The new device has the potential to allow the surgeon to choose and record operative inclination accurately during total hip arthroplasty in the lateral decubitus position.