Hemagglutinin sequence conservation guided stem immunogen design from influenza A H3 subtype

Seasonal epidemics caused by influenza A (H1 and H3 subtypes) and B viruses are a major global health threat. The traditional, trivalent influenza vaccines have limited efficacy because of rapid antigenic evolution of the circulating viruses. This antigenic variability mediates viral escape from the host immune responses, necessitating annual vaccine updates. Influenza vaccines elicit a protective antibody response, primarily targeting the viral surface glycoprotein hemagglutinin (HA). However, the predominant humoral response is against the hypervariable head domain of HA, thereby restricting the breadth of protection. In contrast, the conserved, subdominant stem domain of HA is a potential ``universal'' vaccine candidate. We designed an HA stem-fragment immunogen from the 1968 pandemic H3N2 strain (A/Hong Kong/1/68) guided by a comprehensive H3 HA sequence conservation analysis. The biophysical properties of the designed immunogen were further improved by C-terminal fusion of a trimerization motif, ``isoleucine-zipper'', or ``foldon''. These immunogens elicited cross-reactive, antiviral antibodies and conferred partial protection against a lethal, homologous HK68 virus challenge in vivo. Furthermore, bacterial expression of these immunogens is economical and facilitates rapid scale-up.

Distribution, relative abundance, and conservation status of Asian elephants in Karnataka, southern India

Karnataka state in southern India supports a globally significant and the country's largest population of the Asian elephant Elephas maximus. A reliable map of Asian elephant distribution and measures of spatial variation in their abundance, both vital needs for conservation and management action, are unavailable not only in Karnataka, but across its global range. Here, we use various data gathered between 2000 and 2015 to map the distribution of elephants in Karnataka at the scale of the smallest forest management unit, the `beat', while also presenting data on elephant dung density for a subset of `elephant beats.' Elephants occurred in 972 out of 2855 forest beats of Karnataka. Sixty percent of these 972 beats and 55% of the forest habitat lay outside notified protected areas (PM), and included lands designated for agricultural production and human dwelling. While median elephant dung density inside protected areas was nearly thrice as much as outside, elephants routinely occurred in or used habitats outside PM where human density, land fraction under cultivation, and the interface between human-dominated areas and forests were greater. Based on our data, it is clear that India's framework for elephant conservation which legally protects the species wherever it occurs, but protects only some of its habitats while being appropriate in furthering their conservation within PM, seriously falters in situations where elephants reside in and/or seasonally use areas outside PAs. Attempts to further elephant conservation in production and dwelling areas have extracted high costs in human, elephant, material and monetary terms in Karnataka. In such settings, conservation planning exercises are necessary to determine where the needs of elephants or humans must take priority over the other, and to achieve that in a manner that is based not only on reliable scientific data but also on a process of public reasoning. (C) 2015 Elsevier Ltd. All rights reserved.

Weak conservation of structural features in the interfaces of homologous transient protein-protein complexes

Residue types at the interface of protein-protein complexes (PPCs) are known to be reasonably well conserved. However, we show, using a dataset of known 3-D structures of homologous transient PPCs, that the 3-D location of interfacial residues and their interaction patterns are only moderately and poorly conserved, respectively. Another surprising observation is that a residue at the interface that is conserved is not necessarily in the interface in the homolog. Such differences in homologous complexes are manifested by substitution of the residues that are spatially proximal to the conserved residue and structural differences at the interfaces as well as differences in spatial orientations of the interacting proteins. Conservation of interface location and the interaction pattern at the core of the interfaces is higher than at the periphery of the interface patch. Extents of variability of various structural features reported here for homologous transient PPCs are higher than the variation in homologous permanent homomers. Our findings suggest that straightforward extrapolation of interfacial nature and inter-residue interaction patterns from template to target could lead to serious errors in the modeled complex structure. Understanding the evolution of interfaces provides insights to improve comparative modeling of PPC structures.

The importance of reproductive strategies in population genetic approaches to conservation: an example from the marine angiosperm genus Zostera

Knowledge of the levels of genetic diversity maintained in natural populations can play a central role in conservation programmes, particularly in threatened habitats or species. Fluctuations in population size can lead to loss of variation and, consequently, increase the risk of extinction. We have examined whether such a genetic bottleneck has occurred in populations of two species in the seagrass genus Zostera, which are believed to have been affected by an outbreak of wasting disease at the start of the last century. A test for heterozygote excess at five nuclear microsatellite loci did not suggest the occurrence of a genetic bottleneck, but analysis of seven chloroplast microsatellite loci and sequence data from two regions did suggest a bottleneck in the chloroplast genome. Extremely low levels of between-population diversity suggest that all subpopulations can be treated as a single management unit for each species. Comparable levels of nuclear genetic diversity were found in the three populations of the primarily sexual Zostera marina var. angustifolia studied but a wider range of within-population diversity was found in Zostera noltii, which displays both. sexual and vegetative reproductive strategies. This may be due to an increase in sexual recruitment due to localised fresh water inflow into the study site near to the most diverse population. Such populations should be prioritised as source material for any replanting or remediation due to natural or anthropogenic loss of Zostera beds in the area.

Covering over the cracks in conservation assessments at EU interfaces: A cross-jurisdictional ecoregion scale approach using the Eurasian otter (Lutra lutra)

Throughout the European Union, the EC Habitats Directive requires that member states undertake national surveillance of designated species. Despite biological connections between-populations across-borders, national assessments need not be co-ordinated in any way. We conducted a trans-boundary assessment of the status of Eurasian otters (Lutra lutra) aimed at providing consistency across a single biogeographical unit, i.e. the island of Ireland, comprising two states, i.e. the Republic of Ireland and the United Kingdom (Northern Ireland). Our aim was to ensure consistency with previous assessments conducted separately in each state, and permit each Government to fulfil their separate statutory reporting commitments. The species range increased by 23% from 1996–2006 and 2007–11. The population estimate of 9400 [95%CI 8700–12,200] breeding females during 2010/11 was not significantly different from 8300 [95%CI 7600–9800] breeding females established as a baseline during 1981–82. Modelling of species-habitat associations suggested that available habitat was not limiting and no putative pressures recorded at sites surveyed negatively affected species occurrence. Thus, under the statutory parameters for assessing a species’ conservation status, i.e. range, population, habitat and future prospects, the otter was judged to be in ‘Favourable’ status throughout Ireland and in both discrete political jurisdictions. Thus, we provide a trans-boundary test case for EU member states that share habitats and species across ecoregions, ensuring conservation assessment data are standardised, synchronised, spatially consistent and, therefore, biologically relevant without compromising legal and administrative autonomy within separate jurisdictions.

Caractérisation de la sous-unité bêta du translocon chez la levure Schizosaccharomyces pombe

La sécrétion des protéines est un processus essentiel à la vie. Chez les eucaryotes, les protéines sécrétées transitent dans le réticulum endoplasmique par le pore de translocation. Le translocon est composé de trois sous-unités fondamentales nommées Sec61α, β et γ chez les mammifères, ou Sec61p, Sbh1p et Sss1p chez les levures. Tandis que le rôle des sous-unités α et γ est bien connu, celui de la sous-unité β demeure énigmatique. Plusieurs phénotypes distincts sont associés à cette protéine dans différents organismes, mais le haut niveau de conservation de séquence suggère plutôt une fonction universelle conservée. Récemment, Feng et al. (2007) ont montré que le domaine transmembranaire (TMD) de Sbh1p était suffisant pour complémenter plusieurs phénotypes associés à la délétion du gène chez Saccharomyces cerevisiae, suggérant un rôle important de cette région. L’objectif de mon projet de recherche consiste à étudier la fonction biologique de la sous-unité β du translocon et de son TMD chez Schizosaccharomyces pombe. Dans cette levure, j’ai découvert que le gène sbh1+ n’était pas essentiel à la viabilité à 30oC, mais qu’il était requis pour la croissance à basse température. La délétion de sbh1+ entraîne une sensibilité aux stress de la paroi cellulaire et une diminution de la sécrétion des protéines à 23oC. La surexpression de Sbh1p diminue elle aussi la sécrétion des protéines et altère la morphologie cellulaire. Ces phénotypes sont distincts de ceux observés chez S. cerevisiae, où la délétion des deux paralogues de Sec61β entraîne une sensibilité à haute température plutôt qu’à basse température. Malgré cela, les homologues de Sec61β de S. pombe et de S. cerevisiae sont tout deux capables de complémenter la thermosensibilité respective de chaque levure. La complémentation est possible même avec l’homologue humain de Sec61β, indiquant la conservation d’une fonction de Sec61β de la levure à l’homme. Remarquablement, le TMD de Sec61β de S. pombe, de S. cerevisiae et de l’humain sont suffisants pour complémenter la délétion génomique autant chez la levure à fission que chez la levure à bourgeons. Globalement, ces observations indiquent que le TMD de Sec61β exerce une fonction cellulaire conservée à travers les espèces.

Studies on Energy Requirement and Conservation in Selected Fish Harvesting Systems

Present work deals with the studies on energy requirement and convervation in selected fish harvesting systems.Modem fishing is one of the most energy intensive methods of food production. Fossil fuels used for motorised and mechanised fishing are nonrenewable and limited. Most of the environmental problems that confront mankind today are connected to the use of energy in one way or another. Code of Conduct for Responsible Fisheries (FAO, 1995) highlights the need for efficient use of energy in the fisheries sector. Information on energy requirement in different fish harvesting systems, based on the principles of energy analysis, is entirely lacking in respect of Indian fisheries. Such an analysis will provide an unbiased decision making support for maximising the yield per unit of non-renewable energy use, from different fishery resource systems, by rational deployment of harvesting systems. In the present study, results of investigations conducted during 1997-2000 on energy requirement in selected fish harvesting systems and approaches to energy conservation in fishing, are presented along with a detailed description of the fish harvesting systems and their operation. The content of the thesis is organised into 8 Chapters.

Conservation ecology and breeding biology of the white-browed treecreeper, Climacteris affinis

The White-browed Treecreeper Climacteris affinis is one of many woodland-dependent birds that are at risk from the encroachment of human-dominated land-uses into natural landscapes. The White-browed Treecreeper inhabits semi-arid woodlands in north-west Victoria, Australia, a vegetation community that has undergone extreme modification in the last century due to the expansion of agriculture in the region. Extant woodlands represent only 10% of the original woodland cover in the region, and are highly fragmented and disturbed in many districts. Thus, the survival of the White-browed Treecreeper may depend on active management. However, current knowledge of the ecology and biology of this species is virtually non-existent, and inadequate for informed and effective conservation actions. The aim of this thesis is to redress this situation and provide the ecological basis for sound conservation management of the species. The thesis consists of two parts: an investigation of habitat use at three spatial scales and a study of the social organization, nesting requirements, breeding behaviour and reproductive success of a population of White-browed Treecreepers. Fifty-six patches of remnant woodland in north-west Victoria were surveyed to determine the factors affecting the occurrence of the White-browed Treecreeper at the regional scale. It was detected in 16 patches, and was largely confined to two core districts - Yarrara and, Wyperfeld (Pine Plains). The floristic composition of the dominant tree species was an important determinant of patch occupancy, with the results providing quantitative support for the previously suspected affinity for Belah Casuarina pauper and Slender Cypress-pine Callitris gracilis — Buloke Allocasuarina luehmannii woodlands. However, the absence of the White-browed Treecreeper from several districts was due to factors other than a lack of appropriate habitat. Demographic isolation - the distance from the focal patch to the nearest population of the White-browed Treecreeper - was the most important variable in explaining variation in patch occupancy. Patches isolated from other treecreeper populations by more than 8.3 km in landscapes of non-preferred native vegetation, and 3 km in agricultural landscapes, were unlikely to support the White-browed Treecreeper. The impact of habitat loss and fragmentation on the capacity of individuals to move through the landscape (i.e. functional connectivity) is considered in relation to disruption to dispersal and migration, and the potential collapse of local metapopulations. Habitat use was then examined in a network of patches and linear strips of Belah woodland embedded in a predominantly cultivated landscape. A minimum area of 18.5 ha of Belah woodland was identified as the most important criterion for patch occupancy at the local scale. This landscape appeared to be permeable to movement by the White-browed Treecreeper, facilitated by the extensive network of linear habitat, and clusters of small to medium fragments. The third scale of habitat use investigated the frequency of use of 1-ha plots within tracts of occupied woodland. It is important to discriminate between habitat traits that operate at the population level, and those that act as proximate cues for habitat selection by individuals. Woodlands that have high tree density, extensive cover of low-stature shrubs, abundant lichen, a complex vertical structure, and relatively low cover of grass and herbs are likely to support larger populations of the White-browed Treecreeper. However, individuals appeared to be using tree dominance (positive) and tall shrub cover (negative) as proximate environmental stimuli for habitat selectivity. A relatively high cover of ground lichen, which probably reflects a ground layer with low disturbance and high structural complexity, was also a reliable indicator of habitat use. Predictive models were developed which could be used to plan vegetation management to enhance habitat for the White-browed Treecreeper. The results of the regional, landscape and patch-scale investigations emphasise that factors operating at multiple spatial scales influence the suitability of remnant vegetation as habitat for the White-browed Treecreeper. The White-browed Treecreeper is typical of many small Australian passerines in that it has high annual survival, small clutches, a long breeding season, multiple broods and relatively low reproductive rates. Reproductive effort is adjusted through the number of clutches laid rather than clutch size. They occupy relatively large, all-purpose territories throughout the year. However, unlike many group territorial birds, territory size was not related to the number of occupants. The White-browed Treecreeper nests in tree hollows. They select hollows with a southerly orientation where possible, and prefer hollows that were higher from the ground. At Yarrara, there was considerable spatial variation in hollow abundance that, in concert with territorial constraints, restricted the actual availability of hollows to less than the absolute abundance of hollows. Thus, the availability of suitable hollows may limit reproductive productivity in some territories, although the magnitude of this constraint on overall population growth is predicted to be small. However, lack of recruitment of hollow-bearing trees would increase the potential for hollow availability to limit population growth. This prospect is particularly relevant in grazed remnants and those outside the reserve system. Facultative cooperative breeding was confirmed, with groups formed through male philopatry. Consequently, natal dispersal is female-biased, although there was no skew in the sex ratio of the fledglings or the general adult population. Helpers were observed performing all activities associated with parenting except copulation and brooding. Cooperatively breeding groups enjoyed higher fledgling productivity than simple pairs, after statistically accounting for territory and parental quality. However, the difference reflected increased productivity in the 1999-breeding season only, when climatic conditions were more favourable than in 1998. Breeding commenced earlier in 1999, and all breeding units were more likely to attempt a second brood. However, only breeders with helpers were successful in fledging second brood young, and it was this difference that accounted for the overall discrepancy in productivity. The key mechanism for increased success in cooperative groups was a reduction hi the interval between first and second broods, facilitated by compensatory reductions in the level of care to the first brood. Thus, females with helpers probably achieved significant energetic savings during this period, which enabled them to re-lay sooner. Furthermore, they were able to recommence nesting when the fledglings from the first brood were younger because there were more adults to feed the dependent juveniles. The current utility, and possible evolutionary pathways, of cooperative breeding is examined from the perspective of both breeders and helpers. Breeders benefit through enhanced fledgling productivity in good breeding conditions and a reduction in the burden of parental care, which may impart significant energetic savings. Further, breeders may facilitate philopatry as a means for ensuring a minimum level of reproductive success. Helpers benefit through an increase in their inclusive fitness in the absence of opportunities for independent breeding (i.e. ecological constraints) and access to breeding vacancies in the natal or adjacent territories (i.e. benefits of philopatry). However, the majority of breeding unit-years comprised unassisted breeders, which suggests that pairs are selectively favoured under certain environmental or demographic conditions.

Vive la résistance: reviving resistance for 21st century conservation

Confronted with increasing anthropogenic change, conservation in the 21st century requires a sound understanding of how ecological systems change during disturbance. We highlight the benefits of recognizing two distinct components of change in an ecological unit (i.e., ecosystem, community, population): 'resistance', the ability to withstand disturbance; and 'resilience', the capacity to recover following disturbance. By adopting a 'resistance-resilience' framework, important insights for conservation can be gained into: (i) the key role of resistance in response to persistent disturbance, (ii) the intrinsic attributes of an ecological unit associated with resistance and resilience, (iii) the extrinsic environmental factors that influence resistance and resilience, (iv) mechanisms that confer resistance and resilience, (v) the post-disturbance status of an ecological unit, (vi) the nature of long-term ecological changes, and (vii) policy-relevant ways of communicating the ecological impacts of disturbance processes.

DNA alignment and Bayesian trees of partial sequences of COI, COIII and the haemocyanin functional unit f-g of 28 octopod species

Background: Octopods have successfully colonised the world's oceans from the tropics to the poles. Yet, successful persistence in these habitats has required adaptations of their advanced physiological apparatus to compensate impaired oxygen supply. Their oxygen transporter haemocyanin plays a major role in cold tolerance and accordingly has undergone functional modifications to sustain oxygen release at sub-zero temperatures. However, it remains unknown how molecular properties evolved to explain the observed functional adaptations. We thus aimed to assess whether natural selection affected molecular and structural properties of haemocyanin that explains temperature adaptation in octopods. Results: Analysis of 239 partial sequences of the haemocyanin functional units (FU) f and g of 28 octopod species of polar, temperate, subtropical and tropical origin revealed natural selection was acting primarily on charge properties of surface residues. Polar octopods contained haemocyanins with higher net surface charge due to decreased glutamic acid content and higher numbers of basic amino acids. Within the analysed partial sequences, positive selection was present at site 2545, positioned between the active copper binding centre and the FU g surface. At this site, methionine was the dominant amino acid in polar octopods and leucine was dominant in tropical octopods. Sites directly involved in oxygen binding or quaternary interactions were highly conserved within the analysed sequence. Conclusions: This study has provided the first insight into molecular and structural mechanisms that have enabled octopods to sustain oxygen supply from polar to tropical conditions. Our findings imply modulation of oxygen binding via charge-charge interaction at the protein surface, which stabilize quaternary interactions among functional units to reduce detrimental effects of high pH on venous oxygen release. Of the observed partial haemocyanin sequence, residue 2545 formed a close link between the FU g surface and the active centre, suggesting a role as allosteric binding site. The prevalence of methionine at this site in polar octopods, implies regulation of oxygen affinity via increased sensitivity to allosteric metal binding. High sequence conservation of sites directly involved in oxygen binding indicates that functional modifications of octopod haemocyanin rather occur via more subtle mechanisms, as observed in this study.

Energy conservation through improved solid waste management /

"Solid waste management series (SW-125)"

Watershed Protection and Flood Prevention Training Conference, [held at] Soil Conservation Service Training Center, Fort Robinson, Nebraska, January 21-25, 1957.

Mode of access: Internet.

Energy conservation technical assistance update.

The Energy Conservation Act (20 ILCS 1115) was signed into law in September 1979. The law requires that the Department provide technical assistance in the development of thermal efficiency standards and lighting efficiency standards to units of local government, upon request by such unit. The law also states that the Department shall provide technical assistance in the development of a program for energy efficiency in procurement to units of local government, upon request by such unit. The Technical Assistance Programs provided under the law are supported by funds provided to the State pursuant to federal acts that provide funds for energy conservation efforts through the use of building codes.

Targhee National Forest ecological unit inventory /

Includes index to map sheets.

The crystal structure of a bacterial Class II ketol-acid reductolsomerase: Domain conservation and evolution

Ketol-acid reductoisomerase (KARI; EC 1.1.1.86) catalyzes two steps in the biosynthesis of branched-chain amino acids. Amino acid sequence comparisons across species reveal that there are two types of this enzyme: a short form (Class 1) found in fungi and most bacteria, and a long form (Class 11) typical of plants. Crystal structures of each have been reported previously. However, some bacteria such as Escherichia coli possess a long form, where the amino acid sequence differs appreciably from that found in plants. Here, we report the crystal structure of the E. coli enzyme at 2.6 A resolution, the first three-dimensional structure of any bacterial Class 11 KARI. The enzyme consists of two domains, one with mixed alpha/beta structure, which is similar to that found in other pyridine nucleotide-dependent dehydrogenases. The second domain is mainly alpha-helical and shows strong evidence of internal duplication. Comparison of the active sites between KARI of E. coli, Pseudomonas aeruginosa, and spinach shows that most residues occupy conserved positions in the active site. E. coli KARI was crystallized as a tetramer, the likely biologically active unit. This contrasts with P. aeruginosa KARI, which forms a dodecamer, and spinach KARI, a dimer. In the E. coli KARI tetramer, a novel subunit-to-subunit interacting surface is formed by a symmetrical pair of bulbous protrusions.