Microm In Length Research Articles

Controlled Introduction of Diameter Modulations in Arrayed Magnetic Iron Oxide Nanotubes

To date, no large-scale preparative method for arrays of nanotube enables the experimentalist to arbitrarily define changes in the tubes' diameter along their length. To this goal, we start with anodic alumina substrates displaying controlled modulations in pore diameter obtained by alternating "mild" and "hard" electrochemical etching conditions. We then utilize atomic layer deposition (ALD) to coat the internal pore walls with conformal layers of an oxide. Ferromagnetic Fe(3)O(4) tubes of 10 nm wall thickness and 10-30 microm in length are thus prepared, which replicate the modulated silhouette of the template. Their magnetic properties strongly depend on the presence of diameter modulations. Introducing one or several very short segments of large diameter (150 nm) into an otherwise thin tube (70 nm diameter) brings its initially large coercive field down to a value close to the case of a homogeneously thick tube. Theoretical modeling emphasizes the major influence of the magnetostatic interactions between neighboring tubes. They are enhanced locally at the sites of diameter modulations, which directly translates into a reduction in coercive field.

Exophiala xenobiotica infection in cultured striped jack, Pseudocaranx dentex (Bloch & Schneider), in Japan

This report describes Exophiala infection in cultured striped jack, Pseudocaranx dentex, in Japan in 2005. One hundred out of 35,000 fish died per day and mortalities continued for 1 month. Diseased fish showed swelling of the abdomen and kidney distension. Numerous septate hyphae, pale brown in colour, were seen in kidney in squash preparations. Histology revealed abundant fungal hyphae and conidia in gill, heart and kidney. Fungal hyphae were accompanied by cell necrosis and influx of inflammatory, mainly mononuclear cells. The fungus isolated from the diseased fish had septate hyphae, pale brown in colour and 1.8-3.0 microm in diameter. Conidiogenous cells were conspicuous annellides, short or cylindrical or fusiform in shape. Conidia were one-celled, ellipsoidal with smooth walls, accumulated in balls at the apices of annellides that tended to slide down, 1.5-2.0 microm in width and 3.0-5.0 microm in length. The fungus was classified into the genus Exophiala based on its morphology and as Exophiala xenobiotica based on the sequences of the ITS 1-5.8S-ITS 2 regions of rDNA. This is the first record of this fungus in a marine fish.

Micropatterning of single myotubes on a thermoresponsive culture surface using elastic stencil membranes for single-cell analysis

We have developed a micropatterning procedure for single myotubes and demonstrated recovery of patterned myotubes without the use of methods that might cause damage to the cells. Since skeletal muscle is a highly ordered tissue mainly composed of myotubes, analysis of single myotubes is one of the promising approaches for studying the various diseases related to skeletal muscle tissues. However, the analysis of single myotubes is quite complicated because of the difficulty in distinguishing individual myotubes differentiated on a normal cell culture surface. In the present study, thin polydimethylsiloxane (PDMS) membranes, which have rectangular holes (30, 50, 100, and 200 microm in width; 500, 750, and 1000 microm in length) through them, were fabricated by using a photolithography technique and used for single myotube micropatterning. A bovine serum albumin-coated (BSA-coated) stencil membrane was placed on a cell culture surface and C2C12 myoblasts were seeded on it. Since the cells could not attach to the surface of the stencil membrane, the cell proliferated and differentiated into myotubes in the hole areas specifically. By peeling off the membrane, a micropattern of myotubes was obtained. It was revealed that the optimum width of rectangular holes for a micropattern of single myotubes was between 30 to 50 microm. Furthermore, by placing a membrane on a thermoresponsive culture surface, recovery of the micropatterned myotubes was possible by lowering the temperature. This method involving the stencil membranes and a thermoresponsive culture surface is useful for analyzing subcellular or single myotubes.

Scanning Electron Microscopy of the Egg of <I>Ochlerotatus albifasciatus</I> (Diptera: Culicidae)

Ochlerotatus albifasciatus (Macquart) (Diptera: Culicidae) is a mosquito species of medical and veterinary importance, usually associated with temporary groundwater habitats. Eggs obtained from females collected in Córdoba city, in the center of Argentina, were studied using scanning electron microscopy. The eggs are elliptical in outline, measuring approximately 615.7 +/- 37.24 microm in length (1) and 180.7 +/- 22.87 microm in width (w), with an egg index (l/w ratio) of 2.9-3.7. The outer chorionic sculpture has racquet-shaped and wristwatch-shaped cells consisting of a small central tubercle surrounded by larger tubercles and followed by a row of smaller tubercles on one or both sides, respectively, although variations between regions of the egg were observed. The micropylar apparatus has a collar with evident molding and edges of determined length, albeit irregular, with defined margins for the transition area and a thickness of approximately 10.7 microm. The margins of the micropylar disc are raised and the disc measures approximately 21.1 microm in diameter. The micropyle is distinct.

Isolation and characterization of a new CO-utilizing strain, Thermoanaerobacter thermohydrosulfuricus subsp. carboxydovorans, isolated from a geothermal spring in Turkey

A novel anaerobic, thermophilic, Gram-positive, spore-forming, and sugar-fermenting bacterium (strain TLO) was isolated from a geothermal spring in Ayaş, Turkey. The cells were straight to curved rods, 0.4–0.6 μm in diameter and 3.5–10 μm in length. Spores were terminal and round. The temperature range for growth was 40–80°C, with an optimum at 70°C. The pH optimum was between 6.3 and 6.8. Strain TLO has the capability to ferment a wide variety of mono-, di-, and polysaccharides and proteinaceous substrates, producing mainly lactate, next to acetate, ethanol, alanine, H2, and CO2. Remarkably, the bacterium was able to grow in an atmosphere of up to 25% of CO as sole electron donor. CO oxidation was coupled to H2 and CO2 formation. The G + C content of the genomic DNA was 35.1 mol%. Based on 16S rRNA gene sequence analysis and the DNA–DNA hybridization data, this bacterium is most closely related to Thermoanaerobacter thermohydrosulfuricus and Thermoanaerobacter siderophilus (99% similarity for both). However, strain TLO differs from Thermoanaerobacter thermohydrosulfuricus in important aspects, such as CO-utilization and lipid composition. These differences led us to propose that strain TLO represents a subspecies of Thermoanaerobacter thermohydrosulfuricus, and we therefore name it Thermoanaerobacter thermohydrosulfuricus subsp. carboxydovorans.

Airborne Asbestos Concentrations Associated with Heavy Equipment Brake Removal

Asbestos-containing brake linings were used in heavy-duty construction equipment such as tractors, backhoes, and bulldozers prior to the 1980s. While several published studies have evaluated exposures to mechanics during brake repair work, most have focused on automobiles and light trucks, not on heavy agricultural or construction vehicles. The purpose of this study is to characterize the airborne concentration of asbestos to workers and bystanders from brake wear debris during brake removal from 12 loader/backhoes and tractors manufactured between 1960 and 1980. Asbestos content in brake lining (average 20% chrysotile by polarized light microscopy) and brake wear debris [average 0.49% chrysotile by transmission electron microscopy (TEM)] was also quantified. Breathing zone samples on the lapel of mechanics (n = 44) and area samples at bystander (n = 34), remote (n = 22), and ambient (n = 12) locations were collected during 12 brake changes and analyzed using phase contrast microscopy (PCM) [National Institute for Occupational Safety and Health (NIOSH) 7400] and TEM (NIOSH 7402). In addition, the fiber distribution by size and morphology was evaluated according to the International Organization for Standardization method for asbestos. Applying the ratio of asbestos fibers:total fibers (including non-asbestos) as determined by TEM to the PCM results, the average airborne chrysotile concentrations (PCM equivalent) were 0.024 f/cc for the mechanic and 0.009 f/cc for persons standing 1.2-3.1 m from the activity during the period of exposure ( approximately 0.5 to 1 h). Considering the time involved in the activity, and assuming three brake jobs per shift, these results would convert to an average 8-h time-weighted average of 0.009 f/cc for a mechanic and 0.006 f/cc for a bystander. The results indicate that (i) the airborne concentrations for worker and bystander samples were significantly less than the current occupational exposure limit of 0.1 f/cc; (ii) approximately 2% of respirable fibers were >20 microm in length; and (iii) approximately 95% of chrysotile in the brake linings degraded in the friction process. The industrial hygiene data presented here should be useful for conducting retrospective and current exposure assessments of individuals, as well as hazard assessments of work activities that involve repairing and replacing asbestos-containing brakes in heavy construction equipment.

Synthesis, Purification and Opening of Short Cup-Stacked Carbon Nanotubes

A simple method is demonstrated to synthesize high-quality cup-stacked carbon nanotubes (CSCNTs) with short length. The as-synthesized CSCNTs are 0.2-3.2 microm in length, even shorter than the ball-milled long CSCNTs (approximately 7 microm). These CSCNTs have a diameter of 80-120 nm and a hollow channel of 60-100 nm along the nanotube axis. Moreover, a simple purification method is developed to remove iron particles and obtain short CSCNTs with two open ends. Such CSCNTs are very easily suspended in distilled water and tetrahydrofuran, which makes such material greatly promising as a candidate in many applications such as heterogeneous catalysis and photoelectrochemical cell.

The Effect of Microstructure of Octacalcium Phosphate on the Bone Regenerative Property

The present study was designed to investigate whether the microstructure of synthetic octacalcium phosphate (OCP) affects its intrinsic bone regenerative properties as a scaffold and its conversion process into hydroxyapatite (HA). Our previous studies indicated that an agregate of OCP crystals, consisting of randomly oriented plate-like crystals, are capable of enhancing both osteoblastic cell differentiation in vitro and bone regeneration. While the transformation of OCP into HA has been considered in relation to the stimulatory capacity of OCP in bone regeneration, little is known about the effect of the microstructure of OCP granules on these capabilities. Two types of OCP granules, with identical diameters (300-500 microm) but composed of crystals with distinct crystal dimensions (4.0 and 26.6 microm length), were prepared (hereafter referred to as fine OCP granules [F-OCP] and coarse OCP granules [C-OCP], respectively). The intergranule distances and the porosity, including the intergranule spaces, were 108.5 microm and 93.7% for F-OCP, and 67.5 microm and 95.7% for C-OCP, as estimated by mercury intrusion. The OCP granules were implanted in mouse critical-sized calvarial defects for up to 14 days. Histological examination demonstrated that osteoblastic cells aligned on the surface of F-OCP at day 7 and formed new bone around the granules up to day 14. On the other hand, cells around C-OCP were sparse at day 7, and resulted in only slight bone formation around the granules at day 14. X-ray diffraction showed that both OCP granules tended to be converted to an apatite structure with similar conversion velocity by the implantation. Adhesion of mouse bone marrow stromal ST-2 cells was markedly inhibited on C-OCP compared to F-OCP in vitro. These results suggested that the microstructure consisting of plate-like crystals of OCP controls cell adhesion on the crystal surfaces and their resultant bone regenerative properties as well as the physicochemical effect associated with the transitory nature of OCP previously reported.

Antibiofouling, Sustained Antibiotic Release by Si Nanowire Templates

Loading or filling nanostructures with antibiotics can be one of the relevant approaches for obtaining a controlled drug release rate. Vertically aligned silicon nanowire (SiNW) arrays with 10-40 nm diameter wires having 1-3 microm in length obtained by the electroless etching (EE) technique are used in this study as novel nanostructures for mediating drug delivery. Here we report controlled antibiotic activity and sustained bioavailability from SiNW arrays and also show microstructural manipulations for a tunable release rate. As well, we have demonstrated biodegradability of SiNWs in phosphate buffer saline (PBS) solution. Strikingly suppressed cell and protein adhesion was observed on our SiNW surface, which indicates a reduced probability for biofouling and drug release impediments. Such antibiotic release from the nanowire-structured surface can provide more reliable antibiotic protection at a targeted implantation or biosensor site.

Thermus islandicus sp. nov., a mixotrophic sulfur-oxidizing bacterium isolated from the Torfajokull geothermal area

Strains PRI 2268 and PRI 3838(T) were isolated from two separate hot springs in the Torfajokull geothermal area of South Iceland. The cells were non-motile rods, approximately 0.3 microm in width and 1.5-2.5 microm in length. Electron microscopy revealed a Gram-negative cell-wall structure. The strains grew at 45-79 degrees C (optimum, 65 degrees C) and pH 5.5-10.5 (optimum, pH 6.0-7.0). 16S rRNA gene sequence analysis indicated that they formed a separate branch within the genus Thermus with 'Thermus kawarayensis' KW11 as their closest cultured relative (96.5 % similarity). The gene sequence similarities of both new isolates to Thermus aquaticus YT-1(T) and Thermus igniterrae RF-4(T) were 96.1 % and 95.5 %, respectively. DNA-DNA relatedness between strain PRI 3838(T) and 'T. kawarayensis' was 46.1 %. The DNA G+C content of strain PRI 3838(T) was 69.0 mol%. The predominant menaquinones, pigmentation, fatty acid profiles and phospholipid profiles of the novel strains were similar to those of other members of the genus Thermus. However, the new strains could be differentiated from the type strains of all other species of the genus Thermus by their lack of catalase activity and their utilization of only a few carbon sources. Furthermore, the novel strains exhibited mixotrophic growth with sulfur oxidation. On the basis of 16S rRNA gene sequence comparisons, DNA-DNA hybridization and physiological and biochemical characteristics, the new isolates represent a novel species. Since the species appears to be ubiquitous in Icelandic hot springs, the name Thermus islandicus sp. nov. is proposed. The type strain is PRI 3838(T) (=DSM 21543(T)=ATCC BAA-1677(T)).

Caldanaerovirga acetigignens gen. nov., sp. nov., an anaerobic xylanolytic, alkalithermophilic bacterium isolated from Trego Hot Spring, Nevada, USA

An anaerobic thermophilic bacterium, designated strain JW/SA-NV4(T), was isolated from a xylan-supplemented enrichment culture from Trego hot spring located within the Black Rock Desert (NV, USA). Cells were generally straight or slightly bent rod-shaped, 0.4-0.8 microm in width and 3-6 microm in length during exponential growth. Cells from stationary phase were variable in size and shape, showing curved or bent morphology. Motility was not seen and flagella were not observed in electron micrographs. Sporulation was not observed. Strain JW/SA-NV4(T) stained Gram-negative but is phylogenetically Gram-type positive. Growth occurred at pH(25 degrees C) 6.8-8.8, with optimum growth at pH 8.4; no growth occurred at pH 9.0 or above or at 6.5 or below. With glucose or xylose as the carbon source, strain JW/SA-NV4(T) grew at 44-74 degrees C; no growth occurred at 76 degrees C or above or at 42 degrees C or below. However, the optimum temperature was 62 and 66 degrees C when grown on glucose and xylose, respectively. The shortest doubling time observed with glucose was approximately 4 h, and with xylose approximately 3.4 h. Strain JW/SA-NV4(T) tolerated an atmosphere containing up to 0.1 % O(2); no growth occurred at a gas atmosphere of 0.2 % O(2). Chemo-organotrophic growth occurred with xylose, glucose, mannose, xylan, pyruvate, fructose, ribose, Casamino acids, mannitol, tryptone, peptone, cellobiose and yeast extract. When grown in mineral media containing 1 g yeast extract l(-1) as an electron donor, thiosulfate and sulfur were reduced to sulfide. The G+C content of the DNA was 38.6 mol% (HPLC). 16S rRNA gene sequence analysis placed strain JW/SA-NV4(T) within the order Thermoanaerobacterales and within the Thermoanaerobacterales Incertae Sedis Family III, specifically between taxa classified within the genera Thermosediminibacter and Thermovenabulum. The closest phylogenetic neighbours were Thermosediminibacter oceani JW/IW-1228P(T) (94.2 % 16S rRNA gene sequence similarity) and Thermosediminibacter litoriperuensis JW/YJL-1230-7/2(T) (94.0 %) [Lee, Y.-J., Wagner, I. D., Brice, M. E., Kevbrin, V. V., Mills, G. L., Romanek, C. S. & Wiegel, J. (2005). Extremophiles 9, 375-383]. Based on physiological and genotypic characteristics, strain JW/SA-NV4(T) (=DSM 18802(T)=ATCC BAA-1454(T)) is proposed to represent the type strain of a novel species in a novel genus, Caldanaerovirga acetigignens gen. nov., sp. nov.

Anodised Titania Nanotubes Prepared in a Glycerol/NaF Electrolyte

This paper discusses the preparation of titania nanotubes by anodisation of Ti in a glycerol-based electrolyte containing 0.5% wt of sodium fluoride (NaF). The influence of anodisation voltage and anodisation time on nanotube wall thickness, diameter and length has been investigated. The results indicate that nanotubes can be formed within a voltage range 10-40 V and that the tubular structure is lost when using a higher voltage. The diameter of the nanotubes is voltage dependent, with the widest tubes being obtained at the highest possible applied voltage of 40 V. An initial voltage ramp which increases at 100 mV/s to the anodisation voltage, rather than an instantaneous step, was observed to stabilise the metal-oxide interface. This enabled the growth of anodic films up to 5.5 microm in length by anodising for approximately 48 h. In the absence of a voltage ramp the films tended to collapse and become detached from the titanium electrode after 15-20 h. Electron microscopy observation suggests that the nanotubes in glycerol develop in a similar way to those produced in water-based media. The nanotubes formed using glycerol also exhibit ripples along the tube wall, although, growing at a slower rate, they are generally smoother than those formed in water.

Two New Dicyemids from Sepia longipes (MollUsca: Cephalopoda: Decapoda)

Two new species of dicyemid mesozoan are described from Sepia longipes, Sasaki, 1913, collected from Tosa Bay in Japan. Dicyema oxycephalum n. sp. is a medium-sized species that is about 1,800 microm in length. It lives in folds of the renal appendages. The vermiform stages are characterized as having 28-34 peripheral cells, a conical calotte, and an axial cell that extends to the base of the propolar cells. Infusoriform embryos consist of 39 cells; 2 nuclei are present in each urn cell and the refringent bodies are solid. Pseudicyema cappacephalum n. sp. is also a medium-size species; it is about 1,000 microm in length. It too lives in folds of the renal appendages. The vermiform stages are characterized as having 32-34 peripheral cells, a cap-shaped calotte, and an axial cell that extends to the base of propolar cells. Infusoriform embryos consist of 39 cells; 2 nuclei are present in each urn cell and the refringent bodies are solid. This is the first description of dicyemids from S. longipes.

Paramarteilia canceri sp. n. (Cercozoa) in the European edible crab (Cancer pagurus) with a proposal for the revision of the order Paramyxida Chatton, 1911

A new species, Paramarteilia canceri sp. n., is described using light and electron microscopy from the edible crab Cancer pagurus L. captured from the English Channel. No external symptoms were noted, although infected animals were typically lethargic and unresponsive to external stimuli. Organs of infected animals were shrunken and collapsed compared with apparently healthy individuals. Although the infection was systemic, marked host responses were only noted in the hepatopancreas where the parasite induced a pronounced haemocytic infiltration. Prevalence of infection throughout the study was 1.1%, with a maximum monthly prevalence of 3%. The intracellular parasite was typically 15 microm in length and composed of a primary cell containing up to three secondary cells derived by internal cleavage. Each secondary cell contains two bicellular spores. The parasite is readily differentiated from the other described paramyxean species by a combination of the number of secondary and tertiary cells. In light of this new discovery, a revision of the order Paramyxida Chatton, 1911 is proposed based upon comparison to the original descriptions of this parasite group in various species of invertebrate hosts. The proposed classification is based on the number of cells within the spores (tertiary cells), so that only three genera remain within the order, namely Marteilia Grizel, Comps, Bonami, Cousserans, Duthoit et Le Pennec, 1974, Paramarteilia Ginsburger-Vogel et Desportes, 1979 and Paramyxa Chatton, 1911. Subsequent discrimination of species is based on a combination of the number of secondary cells within the primary cell and the number of tertiary cells within secondary cells. It is proposed that the genus Marteilioides Comps, Park et Desportes, 1986 is suppressed and the type species of the genus, M. chungmuensis Comps, Park et Desportes, 1986, is transferred to Marteilia and that the other representative of the genus, M. branchialis Anderson et Lester, 1992, is transferred to Paramarteilia. Further, Paramyxoides Larsson et Køie, 2005 is considered as a junior synonym of Paramyxa and its type and only species, Paramyxoides nephtys Larsson et Køie, 2005, is transferred to Paramyxa.

Excimer Laser Induced Nanostructuring of Silicon Surfaces

The effect of KrF excimer laser energy density (below and above the ablation threshold), number of shots and angle of laser incidence on the morphological reconstruction, structure and specular reflectance of Si[311] surfaces is reported. At low energy densities (0.1 to 0.3 J/cm2) laser irradiation results in a variety of nanostructures, depending on laser energy density and number of shots, such as nanopores (40-60 nm dia) and nanoparticles (40-80 nm dia). At energies greater than the laser ablation threshold (2 to 5 J/cm2) the formation of nanowires (200 nm dia, 6-8 microm length), and closely spaced silicon nanograins (100-150 nm dia) is observed. Experiments to study the effect of laser irradiation in the proximity of a fixed shape such as a linear step edge in the form of a stainless steel blade and a cylindrical cross-section Cu wire were also carried out. In both cases, linearly organized nanoparticles (150-200 nm diameter) and nanowires (60-80 nm diameter) formed close to the edge. There is a systematic degradation of long-range order with the number of shots and laser energy density as evidenced from X-ray diffraction studies. At an energy density of 2 J/cm2, and 100 shots the [311] oriented silicon surface made a transition to a randomly oriented nanocrystalline state.

In Vivo Confocal Microscopy and Anterior Segment Optical Coherence Tomography Analysis of the Cornea in Nephropathic Cystinosis

To analyze the corneas of patients with nephropathic cystinosis using in vivo confocal microscopy (IVCM) and anterior segment optical coherence tomography (AS-OCT). Prospective case series. Sixteen eyes of 8 patients with nephropathic cystinosis aged 8 to 21 years. The ophthalmologic evaluation included best-corrected visual acuity, evaluation of photophobia (0-4), slit-lamp biomicroscopy analysis, intraocular pressure measurement, evaluation of crystal density using a slit-lamp-based scoring of the cornea, as well as AS-OCT and IVCM analysis. The depth of crystal deposition (DCD) in the central cornea and the central cornea thickness (CCT) were assessed using AS-OCT and IVCM. The IVCM images were evaluated for crystal density in each corneal layer and an IVCM score was calculated for each eye. All eyes had corneal crystal deposits, with a mean slit-lamp photography score of 2.90+/-0.13 (range, 2.75-3.00). Using AS-OCT, corneal crystals were observed in all eyes. These crystals appeared as hyperreflective punctuate deposits, predominantly observed within the anterior stroma. Measured with AS-OCT, the mean depth of DCD in the central cornea was 291.40+/-81.42 microm (range, 200-531 microm); the mean CCT was 543.47+/-29.62 microm. Using IVCM, the crystals appeared as spindle, needle-shaped, and fusiform hyperreflective bodies measuring from 1 to 175 microm in length and 1 to 30 microm in thickness. Except for the endothelium, randomly oriented crystals were observed in all corneal layers, with the greatest density observed within the anterior stroma. Measured with IVCM, the mean DCD was 426.07+/-88.19 microm (range, 284-531 microm); the mean CCT was 531.87+/-34.77 microm. There was no significant difference between the CCT measurements obtained with IVCM and with AS-OCT (mean difference, 11.31 microm; P = 0.07). Nevertheless, the DCD was significantly higher with IVCM than with AS-OCT (mean difference, 126.25 microm; P<0.0001). In patients with nephropathic cystinosis, IVCM could precisely quantify the density of crystals within the central cornea. Anterior segment OCT and IVCM should be used in further studies evaluating crystal deposition in patients with nephropathic cystinosis. The authors have no proprietary or commercial interest in any materials discussed in this article.

Core-shell InGaAs/GaAs quantum well nanoneedles grown on silicon with silicon-transparent emission

In(x)Ga(1-x)As wurtzite nanoneedles are grown without catalysts on silicon substrates with x ranging from zero to 0.15 using low-temperature metalorganic chemical vapor deposition. The nanoneedles assume a 6 degrees - 9 degrees tapered shape, have sharp 2-5 nm tips, are 4 microm in length and 600 nm wide at the base. The micro-photoluminescence peaks exhibit redshifts corresponding to their increased indium incorporation. Core-shell InGaAs/GaAs layered quantum well structures are grown which exhibit quantum confinement of carriers, and emission below the silicon bandgap.

Widespread occurrence of an intranuclear bacterial parasite in vent and seep bathymodiolin mussels

Many parasitic bacteria live in the cytoplasm of multicellular animals, but only a few are known to regularly invade their nuclei. In this study, we describe the novel bacterial parasite "Candidatus Endonucleobacter bathymodioli" that invades the nuclei of deep-sea bathymodiolin mussels from hydrothermal vents and cold seeps. Bathymodiolin mussels are well known for their symbiotic associations with sulfur- and methane-oxidizing bacteria. In contrast, the parasitic bacteria of vent and seep animals have received little attention despite their potential importance for deep-sea ecosystems. We first discovered the intranuclear parasite "Ca. E. bathymodioli" in Bathymodiolus puteoserpentis from the Logatchev hydrothermal vent field on the Mid-Atlantic Ridge. Using primers and probes specific to "Ca. E. bathymodioli" we found this intranuclear parasite in at least six other bathymodiolin species from vents and seeps around the world. Fluorescence in situ hybridization and transmission electron microscopy analyses of the developmental cycle of "Ca. E. bathymodioli" showed that the infection of a nucleus begins with a single rod-shaped bacterium which grows to an unseptated filament of up to 20 microm length and then divides repeatedly until the nucleus is filled with up to 80,000 bacteria. The greatly swollen nucleus destroys its host cell and the bacteria are released after the nuclear membrane bursts. Intriguingly, the only nuclei that were never infected by "Ca. E. bathymodioli" were those of the gill bacteriocytes. These cells contain the symbiotic sulfur- and methane-oxidizing bacteria, suggesting that the mussel symbionts can protect their host nuclei against the parasite. Phylogenetic analyses showed that the "Ca. E. bathymodioli" belongs to a monophyletic clade of Gammaproteobacteria associated with marine metazoans as diverse as sponges, corals, bivalves, gastropods, echinoderms, ascidians and fish. We hypothesize that many of the sequences from this clade originated from intranuclear bacteria, and that these are widespread in marine invertebrates.

Retinal Vessels in Patients with Multiple Sclerosis: Baseline Diameter and Response to Flicker Light Stimulation

Transparency of ocular media enables the precise quantitative analysis of vessels of retina, a neuronal tissue which can be affected by multiple sclerosis (MS). Eyes with no history of optic neuritis (non-ON eyes) of 21 patients with MS were examined with Retinal Vessel Analyzer. Segments of vessels of 500 microm length were measured proximal and distal from the optic disc and compared to those of 21 age- and gender-matched controls. Baseline diameters and peak response to flicker light stimulation of retinal vessels were analyzed. MS eyes had thinner arterioles (p = 0.02) and thicker venules (p = 0.008) than controls: arterioles 111 +/- 14 microm (proximal), 99 +/- 11 microm (distal) in MS eyes and 121 +/- 15 and 107 +/- 9 in controls, respectively. Values for venules were 157 +/- 18 and 136 +/- 20 (MS); 147 +/- 15 and 119 +/- 20 (controls). Peak response was higher in MS eyes than in controls for arterioles (p = 0.007), but comparable for venules (p = 0.35). Narrower arterioles and wider venules might be a consequence of subclinical swelling of optic nerve axons in eyes with negative history of ON in MS patients.

Haplosporidian parasite in Diporeia spp. amphipods from the Great Lakes region, USA

Light microscope observations of a haplosporidian in Diporeia spp. amphipods from Lakes Michigan and Huron, USA, found that the parasite spore is operculate and measures 8.1 microm in length and 6.1 microm in width. Round to amorphous sporocysts averaging 23.6 microm were present throughout hemal sinuses of infected amphipods. The number of developing spores within a sporocyst was highly variable and spores were observed in various stages of development. Electron microscopy revealed a lid over the operculum of developing spores and documented spores with fairly large and extensive ornamentation. This is the first haplosporidian reported in Diporeia spp. amphipods and only the second haplosporidian from freshwater species.