Characterization, cloning and immunogenicity of antigens released by lung-stage larvae of Schistosoma mansoni

Lung-stage schistosomula are the target of protective immunity in mice vaccinated with attenuated cercariae of Schistosoma mansoni. Therefore, proteins present at this developmental stage, and in particular those which are secreted, are a potential source of novel vaccine candidates. However, little information is available about such molecules. Here we describe the cDNA clones identified by screening expression libraries with serum raised against proteins released by lung-stage schistosomula. In total, 11 different cDNA species were identified, 6 of which have been described previously in S. mansoni; these included fructose 1,6-bisphosphate aldolase and Sm21.7 which together accounted for two-thirds of all positive clones. Of the 5 newly described schistosome genes, 1 cDNA had a high degree of homology to the s5a subunit of 26S proteasomes, most significant being with the human protein. The remaining 4 clones showed no significant homologies to any genes sequenced previously. Fructose 1,6-bisphosphate aldolase, Sm21.7, the proteasome homologue and 1 unknown clone (A26) have been expressed in a bacterial expression system and serum produced against each recombinant protein. Immunolocalization showed fructose 1,6-bisphosphate aldolase, Sm21.7 and the proteasome homologue to be most abundant in muscle cells whilst clone A26 was distributed throughout many tissues, but was most abundant in the tegument. Analysis of the cellular immune responses of vaccinated mice showed 3 of the 4 expressed clones to be highly immunogenic, inducing the secretion of large quantities of the Th1-type cytokine interferon gamma

Patterns of gene expression in schistosomes: localization by whole mount in situ hybridization

rom the identification of genes to the characterization of their functions and interactions. Developmental biologists have long used whole mount in situ hybridization (WISH) to determine gene expression patterns, as a vital tool for formulating and testing hypotheses about function. This paper describes the application of WISH to the study of gene expression in larval and adult schistosomes. Fixed worms were permeablized by proteinase K treatment for hybridization with digoxygenin-labelled RNA probes, with binding being detected by alkaline phosphatase-coupled anti-digoxygenin antibodies, and BM Purple substrate. Discrete staining patterns for the transcripts of the molecules Sm29, cathepsin L, antigen 10.3 and chorion were observed in the tegument cell bodies, gut epithelium, oesophageal gland and vitelline lobules, respectively, of adult worms. Transcripts of the molecules SGTP4, GP18-22 and cathepsin L were localized to tegument cell bodies and embryonic gut, respectively, of lung schistosomula. We also showed that Fast Red TR fluorescent substrate can refine the pattern of localization permitting use of confocal microscopy. We believe that method of WISH will find broad application, in synergy with other emerging post-genomic techniques, such as RNA interference, to studies focused at increasing our molecular understanding of schistosomes

A dc model for power switching transistors suitable for computer-aided design and analysis

A model for bipolar junction power switching transistors whose parameters can be readily obtained by the circuit design engineer, and which can be conveniently incorporated into standard computer-based circuit analysis programs is presented. This formulation results from measurements which may be made with standard laboratory equipment. Measurement procedures, as well as a comparison between actual and computed results, are presented

Using the local gyrokinetic code, GS2, to investigate global ITG modes in tokamaks. (I) s-α{\alpha} model with profile and flow shear effects

This paper combines results from a local gyrokinetic code with analytical theory to reconstruct the global eigenmode structure of the linearly unstable ion-temperature-gradient (ITG) mode with adiabatic electrons. The simulations presented here employ the s-${\alpha}$ tokamak equilibrium model. Local gyrokinetic calculations, using GS2 have been performed over a range of radial surfaces, x, and for ballooning phase angle, p, in the range -${\pi} {\leq} p {\leq\pi}$, to map out the complex local mode frequency, ${\Omega_0(x, p) = \omega_0(x, p) + i\gamma_0(x, p)}$. Assuming a quadratic radial profile for the drive, namely ${\eta_i = L_n/L_T}$, (holding constant all other equilibrium profiles such as safety factor, magnetic shear etc.), ${\Omega_0(x, p)}$ has a stationary point. The reconstructed global mode then sits on the outboard mid plane of the tokamak plasma, and is known as a conventional or isolated mode, with global growth rate, ${\gamma}$ ~ Max[${\gamma_0(x, p)}$], where ${\gamma_0(x, p)}$ is the local growth rate. Taking the radial variation in other equilibrium profiles (e.g safety factor q(x)) into account, removes the stationary point in ${\Omega_0(x, p)}$ and results in a mode that peaks slightly away from the outboard mid-plane with a reduced global growth rate. Finally, the influence of flow shear has also been investigated through a Doppler shift, ${\omega_0 \rightarrow \omega_0 + n\Omega^{\prime}x}$, where n is the toroidal mode number and ${\Omega^{\prime}}$ incorporates the effect of flow shear. The equilibrium profile variation introduces an asymmetry to the growth rate spectrum with respect to the sign of ${\Omega^{\prime}}$, consistent with recent global gyrokinetic calculations.Comment: 10 pages, 8 figures and 1 tabl

In vitro effect of canine hyperimmune sera on TNFa activity

[Introduction]: Septic shock in dogs is caused by cardiovascular and vasomotor failure associated with an uncontrolled intrinsic release of inflammatory mediators [1–5]. The syndrome is characterized by cardiovascular dysfunction, vascular permeability alterations, pulmonary oedema and tissue hypoxia resulting from microthrombi which may culminate in disseminated intravascular coagulation and catastrophic multiple organ failure [6,7]. Systemic bacterial infection, particularly by Gram-negative enterobacteria, haemorrhagic trauma, gastric dilation/volvulus and pancreatitis are the major underlying causes leading tosepsis [8,9]. Because of haemodynamic instability and associated hypovolemia, fluid replacement therapy is generally applied to restore effective circulating volume. The use of fresh frozen plasma has been recommended in cases of coagulopathies as it has been recognized to assist restoration of haemodynamic stability [1,5,10,11]. There is increasing evidence that the drivers of the haemodynamic instability are inflammatory mediators (particularly TNFa) activated primarily by bacterial endotoxin [3,4,12,13]

New biostratigraphic, magnetostratigraphic and isotopic insights into the Middle Eocene Climatic Optimum in low latitudes

The Middle Eocene Climatic Optimum (MECO) was a warming event that interrupted the long-term Eocene cooling trend. While this event is well documented at high southern and mid-latitudes, it is poorly known from low latitudes and its timing and duration are not well constrained because of problems of hiati, microfossil preservation and weak magnetic polarity in key sedimentary sections. Here, we report the results of a study designed to improve the bio-, magneto- and chemostratigraphy of the MECO interval using high-resolution records from two low-latitude sections in the Atlantic Ocean, Ocean Drilling Program (ODP) Sites 1051 and 1260. We present the first detailed benthic foraminiferal stable isotope records of the MECO from the low latitudes as well as the biostratigraphic counts of Orbulinoides beckmanni and new magnetostratigraphic results. Our data demonstrate a ~ 750 kyr-long duration for the MECO characterized by increasing δ13C and decreasing δ18O, with minimum δ18O values lasting ~ 40 kyr at 40.1 Ma coincident with a short-lived negative δ13C excursion. Thereafter, δ18O and δ13C values recover rapidly. The shift to minimum δ18O values at 40.1 Ma is coincident with a marked increase in the abundance of the planktonic foraminifera O. beckmanni, consistent with its inferred warm-water preference. O. beckmanni is an important Eocene biostratigraphic marker, defining planktonic foraminiferal Zone E12 with its lowest and highest occurrences (LO and HOs). Our new records reveal that the LO of O. beckmanni is distinctly diachronous, appearing ~ 500 kyr earlier in the equatorial Atlantic than in the subtropics (40.5 versus 41.0 Ma). We also show that, at both sites, the HO of O. beckmanni at 39.5 Ma is younger than the published calibrations, increasing the duration of Zone E12 by at least 400 kyr. In accordance with the tropical origins of O. beckmanni, this range expansion to higher latitudes may have occurred in response to sea surface warming during the MECO and subsequently disappeared with cooling of surface waters

The effect of equine hyperimmune sera on TNF alpha activity in a L929 cell bioassay

In this pilot study we examined the effect of sera from a proprietary hyperimmune plasma product (Equiplas) on the activity of TNF: in an in vitro L929 cell bioassay. In brief, we report observations from 2 accessions of sera. Accession 1 describes the antiTNF: activity of 3 hyperimmune sera and an untreated serum sample that were provided blind to the study. Accession 2 reports a comparison of antiTNFalpha activity found in 3 paired hyperimmune sera collected following a multiple endotoxin vaccination regimen

Radio Observations of the Black Hole Candidate GX 339-4

The black hole candidate GX 339-4 was first detected as a variable radio source by Sood & Campbell-Wilson in May 1994 with the Molonglo Observatory Synthesis Telescope (MOST). Since then, several observations have been obtained with the Australian Telescope Compact Array (ATCA) in order to study the radio behavior of this source in relation to its soft and hard X-ray activity. We present new results of high resolution radio observations performed with the ATCA in order to study the jet-like feature observed in GX 339-4 by Fender et al (1997). From the ATCA lightcurve at 8640 MHz, we find evidence of quenched radio emission from GX 339-4.Comment: To appear in Proc. 4th Compton Symposium, AIP pres

Photometric Solutions for Detached Eclipsing Binaries: selection of ideal distance indicators in the SMC

Detached eclipsing binary stars provide a robust one-step distance determination to nearby galaxies. As a by-product of Galactic microlensing searches, catalogs of thousands of variable stars including eclipsing binaries have been produced by the OGLE, MACHO and EROS collaborations. We present photometric solutions for detached eclipsing binaries in the Small Magellanic Cloud (SMC) discovered by the OGLE collaboration. The solutions were obtained with an automated version of the Wilson-Devinney program. By fitting mock catalogs of eclipsing binaries we find that the normalized stellar radii (particularly their sum) and the surface brightness ratio are accurately described by the fitted parameters and estimated standard errors, despite various systematic uncertainties. In many cases these parameters are well constrained. In addition we find that systems exhibiting complete eclipses can be reliably identified where the fractional standard errors in the radii are small. We present two quantitatively selected sub-samples of eclipsing binaries that will be excellent distance indicators. These can be used both for computation of the distance to the SMC and to probe its structure. One particularly interesting binary has a very well determined solution, exhibits complete eclipses, and is comprised of well detached G-type, class $II$ giants.Comment: 29 pages, 12 figures. To be published in Ap

Dispersion strengthening in vanadium microalloyed steels processed by simulated thin slab casting and direct charging. Part 2 - chemical characterisation of dispersion strengthening precipitates

The composition of the sub-15 nm particles in six related vanadium high strength low alloy steels, made by simulated thin slab direct charged casting, has been determined using electron energy loss spectroscopy (EELS). Such particles are considered to be responsible for dispersion hardening. For the first time, particles down to 4 nm in size have had their composition fully determined. In all the steels, the particles were nitrogen and vanadium rich and possibly slightly sub-stoichiometric carbonitrides. Equilibrium thermodynamics predicted much higher carbon to metal atomic ratios than observed in all cases so that kinetics and mechanical deformation clearly control the precipitation process. Thus it is important to formulate the steel with this in mind