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

The schistosome egg: development and secretions

We have investigated the development of the schistosome egg and its secretions in order to understand how it migrates through gut tissues and also initiates pathology in the liver. We show by electron microscopy that the subshell envelope is absent in the newly deposited egg, but appears very early and differentiates as development progresses. In the mature egg, this nucleated envelope contains extensive endoplasmic reticulum, suggestive of a protein synthetic capacity. Furthermore, Reynolds' layer only appears between the envelope and the egg-shell in the mature egg and may represent its accumulated secretions. We have biosynthetically labelled and collected the secretions (ESP) released by mature but not immature eggs during culture. Their fractionation by SDS–PAGE reveals a simple pattern of 6 bands, differing markedly in composition from soluble egg antigen preparations. Electrophoresis in casein substrate gels demonstrates the presence of 2 distinct proteases in the egg secretions. By immunocytochemistry, ESP localized predominantly to the envelope of the mature egg, suggesting that this layer rather than the miracidium is the source of egg secretions

Characterization, cloning and immunogenicity of antigens released by transforming cercariae of Schistosoma mansoni

A schistosome infection is initiated when the parasite penetrates the skin of a susceptible host. Relatively large quantities of protein are released by transforming cercariae compared to later larval stages. This represents the first parasite material to which the host's immune system is exposed, yet little is known about the proteins which are released during the first few hours post-transformation. We have shown that antiserum raised against such molecules was capable of imparting protection against a schistosome challenge infection upon passive transfer to naïve mice. By screening a cercarial cDNA library with this serum, 38 positive clones were identified. Sequence analysis showed these to represent 8 different molecules which included Schistosoma mansoni 21·7 kDa antigen, calcium-binding-protein and the vaccine candidate glutathione S-transferase (Sm28GST). In addition, 5 clones were isolated, 1 of which had significant homology to many cytochrome C proteins, another with leukocyte elastase inhibitors and 3 which represented novel molecules. Four clones were expressed in a prokaryotic high-level expression vector, sera produced against each purified recombinant protein and used subsequently to probe Western blots and parasite sections. The leukocyte elastase inhibitor homologue and 2 unknowns induced significant proliferation by lymph node cells recovered from mice vaccinated with irradiated cercariae. More strikingly, the 2 novel proteins stimulated very high levels of interferon [gamma] (IFN[gamma]) secretion both by lymph node cells and those recovered by broncho-alveolar lavage from the lungs of vaccinated mice. Such results will be discussed in the context of vaccine development

Potentiation of tumour growth by endotoxin in serum from syngeneic tumour-bearing mice.

The subcutaneous growth of 2 antigenically distinct syngeneic methylcholanthrene-induced murine fibrosarcomas, designated H1 and H7, were significantly augmented by the concomitant administration of E. coli endotoxin (LPS). Amounts as little as 0.2 micrograms i.p. potentiated tumour growth. The weakly antigenic tumour, H1, was more susceptible to provocation by LPS than the more strongly antigenic H7. Maximum provocation of H1 tumour growth occurred when LPS was injected 1 day before the administration of 5000 tumour cells. In contrast, significant anti-tumour resistance resulted if LPS was administered 6 days before the inoculation of tumour cells. Preliminary evidence indicates that low doses of LPS can facilitate the "sneaking through" phenomenon. Enhancement of tumour growth could not be demonstrated with sera or plasma from tumour-bearing mice, unless the samples were contaminated with endotoxin. The results illustrate the importance of excluding endotoxin from solutions used in studies of experimental tumours

Infrared spectroscopy of cataclysmic variables: III. Dwarf novae below the period gap and novalike variables

We present K-band spectra of the short-period dwarf novae YZ Cnc, LY Hya, BK Lyn, T Leo, SW UMa and WZ Sge, the novalike variables DW UMa, V1315 Aql, RW Tri, VY Scl, UU Aqr and GP Com, and a series of field dwarf stars with spectral types ranging from K2-M6. The spectra of the dwarf novae are dominated by emission lines of HI and HeI. The large velocity and equivalent widths of these lines, in conjunction with the fact that the lines are double-peaked in the highest inclination systems, indicate an accretion disc origin. In the case of YZ Cnc and T Leo, for which we obtained time-resolved data covering a complete orbital cycle, the emission lines show modulations in their equivalent widths which are most probably associated with the bright spot (the region where the gas stream collides with the accretion disc). There are no clear detections of the secondary star in any of the dwarf novae below the period gap, yielding upper limits of 10-30% for the contribution of the secondary star to the observed K-band flux. In conjunction with the K-band magnitudes of the dwarf novae, we use the derived secondary star contributions to calculate lower limits to the distances to these systems. The spectra of the novalike variables are dominated by broad, single-peaked emission lines of HI and HeI - even the eclipsing systems we observed do not show the double-peaked profiles predicted by standard accretion disc theory. With the exception of RW Tri, which exhibits NaI, CaI and 12CO absorption features consistent with a M0V secondary contributing 65% of the observed K-band flux, we find no evidence for the secondary star in any of the novalike variables. The implications of this result are discussed.Comment: 13 pages, 5 figures, to appear in MNRA

Early Surgery for Traumatic Spinal Cord Injury: Where Are We Now?

Study Design: Narrative review. Objective: There is a strong biological rationale to perform early decompression after traumatic spinal cord injury (SCI). With an enlarging clinical evidence base, most spine surgeons internationally now favor early decompression for the majority of SCI patients; however, a number of pertinent questions remain surrounding this therapy. Methods: A narrative review evaluating the status of early surgery for SCI. In particular, we addressed the following questions: (1) Which patients stand to benefit most from early surgery? 2) What is the most appropriate time threshold defining early surgery? Results: Although heterogeneity exists, the evidence generally seems to support early surgery. While the best evidence exists for cervical SCI, there is insufficient data to support a differential effect for early surgery depending on neurological level or injury severity. When comparing thresholds to define early versus late surgery-including a later threshold (48-72 hours), an earlier threshold (24 hours), and an ultra-early threshold (8-12 hours)-the 2 earlier time points seem to be associated with the greatest potential for improved outcomes. However, existing prehospital and hospital logistics pose barriers to early surgery in a significant proportion of patients. An overview of recommendations from the recent AOSpine guidelines is provided. Conclusion: In spite of increasing acceptance of early surgery post SCI, further research is needed to (1) identify subgroups of patients who stand to derive particular benefit-in particular to develop more evidence-based approaches for central cord syndrome and (2) investigate the efficacy and feasibility of ultra-early surgery targeting more aggressive timelines

Prevalence and associated harm of engagement in self-Asphyxial behaviours ('choking game') in young people:A systematic review

OBJECTIVE: To assess the prevalence of engagement in self-asphyxial (risk-taking) behaviour (SAB) (‘choking game’) and associated morbidity and mortality in children and young people up to age 20. DESIGN: Systematic literature review. SEARCH STRATEGY: Electronic database search of MEDLINE, Embase, PsycINFO, CINAHL, PubMed, Web of Science Core Collection, BIOSIS citation index and the Cochrane register with no language or date limits applied. References of key papers were reviewed, and experts were contacted to identify additional relevant papers. ELIGIBILITY CRITERIA: Systematic reviews, cross-sectional, cohort and case–control studies, and case reports examining SAB with regard to individuals aged 0–20 years, without explicitly stated autoerotic, suicidal or self-harm intentions were included. RESULTS: Thirty-six relevant studies were identified, and SAB was reported in 10 countries. In North America, France and Colombia, awareness of SAB ranged from 36% to 91% across studies/settings, and the median lifetime prevalence of engagement in SAB was 7.4%. Six studies identified the potential for SAB to be associated with engagement in other risk behaviours. Ninety-nine fatal cases were reported. Of the 24 cases described in detail, most occurred when individuals engaged in SAB alone and used a ligature. CONCLUSIONS: The current evidence on SAB among young people is limited, and stems predominantly from North America and France. Awareness of SAB among young people is high, and engagement varies by setting. Further research is needed to understand the level of risk and harm associated with SAB, and to determine the appropriate public health response

Indirect costs associated with surgery for low back pain-a secondary analysis of clinical trial data.

This study examines the indirect costs associated with surgery for axial low back pain using data obtained from a prospective multicenter clinical trial that compared Charité artificial disc replacement with anterior lumbar interbody fusion using iliac crest bone graft. While 75% of study subjects reported full- or part-time employment prior to surgery, this percentage dropped to 45% at 6 weeks postoperatively. Return to preoperative employment levels occurred at approximately 6 months postoperatively. Two years after surgery, employment levels were 16% higher than preoperative levels. Lost productivity related to absenteeism resulted in lost wages averaging $2884 per patient during the first postoperative year. Although short-term indirect costs of surgery are substantial from a societal perspective, the higher employment rate at 2 years suggests a long-term economic benefit. The findings demonstrate the significant, though not surprising, impact of spinal disability on productivity, and the importance of including measurement of lost productivity and return to work in the economic evaluation of related interventions

Guidelines and the adoption of ‘lipid rescue’ therapy for local anaesthetic toxicity

Accepted versio