Phylogenetic analysis of the outer-membrane-protein genes of Chlamydiae, and its implication for vaccine development

Examination of 18 complete and 6 partial sequences of the major outer- membrane protein from 24 chlamydiae isolates was used to reconstruct their evolutionary relationships. From this analysis, assuming that the clades with 100% bootstrap support are correct, come the following conclusions: (1) The tree of these sequences is not congruent with the phylogeny of the hosts, and thus host switching would seem to have occurred, thereby limiting the extent to which there has been coevolution of parasite and host. (2) The tree is also noncongruent with clustering by type of cell infected, thereby limiting the extent to which there has been coevolution of parasite and the cell type that it infects. (3) The tree is also noncongruent with clustering by the organ infected (eyes or genitalia), thereby limiting the extent to which there has been coevolution of parasite and the organ that it infects. (4) The tree is also noncongruent with genital strains arising from lymphogranuloma venereum strains. (5) The tree is also noncongruent with the geographic site at which the isolates were obtained, thereby limiting the extent of divergence explained by geographic separation. (6) There are estimated to be 185 amino acid positions that are invariable (as opposed to unvaried) in the major outer-membrane protein. There are 10 unvaried positions in the variable domains, of which 9 appear to be invariable, giving some reason to hope that development of a vaccine might be possible. (7) The rate of change of this protein is too small to see increased divergence over the time span of isolation of these genes, giving hope to any vaccine having longevity. Bootstrapping supports those portions of the tree on which the first five conclusions above depend. The picture that these results provide is more one of pathogen versatility than one of coevolutionary constraints. In addition, we examined 10 60-KDa, outer-membrane protein- 2 genes, all but one of which were from these same strains. The tree was not, among the trachomatis strains, congruent with the major-outer- membrane protein tree, suggesting that gene exchange could be occurring among strains. Moreover, there is an apparent slowdown in divergence in this gene, among the trachomatis strains.      

ANATOMY OF PSEUDOTRITONIA THIELE, 1912 AND NOTAEOLIDIA ELIOT, 1905 (GASTROPODA: OPISTHOBRANCHIA: NUDIBRANCHIA) FROM SIGNY ISLAND, ANTARCTICA

Several species of nudibranch molluscs are abundant amongstthe rich benthic hard substratum communities found close tothe British Antarctic Survey base at Signy Island, in the Atlanticsector of the Antarctic. Over the austral summer 1992/1993,two of the authors (LWB and DKAB) were able to collect someof the lesser known species of Nudibranchia at depths between3 and 40 metres, using SCUBA. New information on morphologyand anatomy of two of these species, Pseudotritonia gracilidensOdhner, 1944 (Charcotiidae, Arminoidea) and Notaeolidia gigasEliot, 1905(Notaeolidiidae, Aeolidoidea) are presented in thispaper. The validity of the species Notaeolidia gigas is strengthened. (Received 9 February 1994; accepted 18 September 1994)     

Negative feedback in the cold: ice retreat produces new carbon sinks in Antarctica

Feedbacks on climate change so far identified are predominantly positive, enhancing the rate of change. Loss of sea‐ice, increase in desert areas, water vapour increase, loss of tropical rain forest and the restriction of significant areas of marine productivity to higher latitude (thus smaller geographical zones) all lead to an enhancement of the rate of change. The other major feedback identified, changes in cloud radiation, will produce either a positive feedback, if high level clouds are produced, or a negative feedback if low level clouds are produced. Few significant negative feedbacks have been identified, let alone quantified. Here, we show that the loss of ice shelves and retreat of coastal glaciers around the Antarctic Peninsula in the last 50 years has exposed at least 2.4 × 10⁴ km² of new open water. We estimate that these new areas of open water have allowed new phytoplankton blooms containing a total standing stock of ～5.0 × 10⁵ tonnes of carbon to be produced. New marine zooplankton and seabed communities have also been produced, which we estimate contain ～4.1 × 10⁵ tonnes of carbon. This previously unquantified carbon sink acts as a negative feedback to climate change. New annual productivity, as opposed to standing stock, amounts to 3.5 × 10⁶ tonnes yr^?1 of carbon, of which 6.9 × 10⁵ tonnes yr^?1 deposits to the seabed. By comparison the total aboveground biomasses of lowland American tropical rainforest is 160–435 tonnes ha^?1. Around 50% of this is carbon. On this basis the carbon held in new biomass described here is roughly equivalent to 6000–17 000 ha of tropical rainforest. As ice loss increases in polar regions this feedback will become stronger, and eventually, over thousands to hundreds of thousands of years, over 50 Mtonnes of new carbon could be fixed annually in new coastal phytoplankton blooms and over 10 Mtonnes yr^?1 locked in biological standing stock around Antarctica.     

Ecological contrasts across an Antarctic land–sea interface

Abstract We report the composition of terrestrial, intertidal and shallow sublittoral faunal communities at sites around Rothera Research Station, Adelaide Island, Antarctic Peninsula. We examined primary hypotheses that the marine environment will have considerably higher species richness, biomass and abundance than the terrestrial, and that both will be greater than that found in the intertidal. We also compared ages and sizes of individuals of selected marine taxa between intertidal and subtidal zones to test the hypothesis that animals in a more stressed environment (intertidal) would be smaller and shorter lived. Species richness of intertidal and subtidal communities was found to be similar, with considerable overlap in composition. However, terrestrial communities showed no overlap with the intertidal, differing from previous reports, particularly from further north on the Antarctic Peninsula and Scotia Arc. Faunal biomass was variable but highest in the sublittoral. While terrestrial communities were depauperate with low biomass they displayed the highest overall abundance, with a mean of over 3 × 10⁵ individuals per square metre. No significant differences in ages of intertidal and subtidal individuals of the same species were found, with bryozoan colonies of up to 4 years of age being present in the intertidal. In contrast with expectation and the limited existing literature we conclude that, while the Antarctic intertidal zone is clearly a suboptimal and highly stressful habitat, its faunal community can be well established and relatively diverse, and is not limited to short‐term opportunists or waifs and strays.     

NADPH-oxidase-driven oxygen radical production determines chondrocyte death and partly regulates metalloproteinase-mediated cartilage matrix degradation during interferon-γ-stimulated immune complex arthritis

In previous studies we have found that FcγRI determines chondrocyte death and matrix metalloproteinase (MMP)-mediated cartilage destruction during IFN-γ-regulated immune complex arthritis (ICA). Binding of immune complexes (ICs) to FcγRI leads to the prominent production of oxygen radicals. In the present study we investigated the contribution of NADPH-oxidase-driven oxygen radicals to cartilage destruction by using p47phox^-/- mice lacking a functional NADPH oxidase complex. Induction of a passive ICA in the knee joints of p47phox^-/- mice resulted in a significant elevation of joint inflammation at day 3 when compared with wild-type (WT) controls as studied by histology. However, when IFN-γ was overexpressed by injection of adenoviral IFN-γ in the knee joint before ICA induction, a similar influx of inflammatory cells was found at days 3 and 7, comprising mainly macrophages in both mouse strains. Proteoglycan depletion from the cartilage layers of the knee joints in both groups was similar at days 3 and 7. Aggrecan breakdown in cartilage caused by MMPs was further studied by immunolocalisation of MMP-mediated neoepitopes (VDIPEN). VDIPEN expression in the cartilage layers of arthritic knee joints was markedly lower (between 30 and 60%) in IFN-γ-stimulated arthritic p47phox^-/- mice at day 7 than in WT controls, despite significant upregulation of mRNA levels of various MMPs such as MMP-3, MMP-9, MMP-12 and MMP-13 in synovia and MMP-13 in cartilage layers as measured with quantitative RT-PCR. The latter observation suggests that oxygen radicals are involved in the activation of latent MMPs. Chondrocyte death, determined as the percentage of empty lacunae in articular cartilage, ranged between 20 and 60% at day 3 and between 30 and 80% at day 7 in WT mice, and was completely blocked in p47phox^-/- mice at both time points. FcγRI mRNA expression was significantly lower, and FcγRII and FcγRIII were higher, in p47phox^-/- mice than in controls. NADPH-oxidase-driven oxygen radical production determines chondrocyte death and aggravates MMP-mediated cartilage destruction during IFN-γ-stimulated IC-mediated arthritis. Upregulation of FcγRI by oxygen radicals may contribute to cartilage destruction.     

Genetic characterization of Hawaiian isolates of <Emphasis Type="Italic">Plasmodium relictum</Emphasis>reveals mixed-genotype infections

Background

The relatively recent introduction of a highly efficient mosquito vector and an avian pathogen (Plasmodium relictum) to an isolated island ecosystem with naïve, highly susceptible avian hosts provides a unique opportunity to investigate evolution of virulence in a natural system. Mixed infections can significantly contribute to the uncertainty in host-pathogen dynamics with direct impacts on virulence. Toward further understanding of how host-parasite and parasite-parasite relationships may impact virulence, this study characterizes within-host diversity of malaria parasite populations based on genetic analysis of the trap (thrombospondin-related anonymous protein) gene in isolates originating from Hawaii, Maui and Kauai Islands.

Methods

A total of 397 clones were produced by nested PCR amplification and cloning of a 1664 bp fragment of the trap gene from two malarial isolates, K1 (Kauai) and KV115 (Hawaii) that have been used for experimental studies, and from additional isolates from wild birds on Kauai, Maui and Hawaii Islands. Diversity of clones was evaluated initially by RFLP-based screening, followed by complete sequencing of 33 selected clones.

Results

RFLP analysis of trap revealed a minimum of 28 distinct RFLP haplotypes among the 397 clones from 18 birds. Multiple trap haplotypes were detected in every bird evaluated, with an average of 5.9 haplotypes per bird. Overall diversity did not differ between the experimental isolates, however, a greater number of unique haplotypes were detected in K1 than in KV115. We detected high levels of clonal diversity with clear delineation between isolates K1 and KV115 in a haplotype network. The patterns of within-host haplotype clustering are consistent with the possibility of a clonal genetic structure and rapid within-host mutation after infection.

Conclusion

Avian malaria (P. relictum) and Avipoxvirus are the significant infectious diseases currently affecting the native Hawaiian avifauna. This study shows that clonal diversity of Hawaiian isolates of P. relictum is much higher than previously recognized. Mixed infections can significantly contribute to the uncertainty in host-pathogen dynamics with direct implications for host demographics, disease management strategies, and evolution of virulence. The results of this study indicate a widespread presence of multiple-genotype malaria infections with high clonal diversity in native birds of Hawaii, which when coupled with concurrent infection with Avipoxvirus, may significantly influence evolution of virulence.

Reviewers

This article was reviewed by Joseph Schall (nominated by Laura Landweber), Daniel Jeffares (nominated by Anthony Poole) and Susan Perkins (nominated by Eugene Koonin).

     

Thymol and Carvacrol Prevent Cisplatin‐Induced Nephrotoxicity by Abrogation of Oxidative Stress,Inflammation, and Apoptosis in Rats

The aim of the present study is to assess the possible protective effects of thymol and carvacrol against cisplatin (CP)‐induced nephrotoxicity. A single dose of CP {6 mg/kg, intraperitoneally (i.p.)} injected to male rats revealed significant increases in serum urea, creatinine, and tumor necrosis factor alpha levels. It also increased kidney contents of malondialdehyde and caspase‐3 activity with significant reduction in serum albumin, kidney content of reduced glutathione as well as catalase, and superoxide dismutase activity as compared to that of the control group. In contrast, administration of thymol {20 mg/kg, orally (p.o.)} and/or carvacrol (15 mg/kg, p.o.) for 14 days before CP injection and for 7 days after CP administration restored the kidney function and examined oxidative stress parameters. In conclusion, thymol was more effective nephroprotective than carvacrol. Moreover, a combination of thymol and carvacrol had a synergistic nephroprotective effect that might be attributed to antioxidant, anti‐inflammatory, and antiapoptotic activities.