Applying proteomics to tick vaccine development: where are we?

Introduction: Ticks are second to mosquitoes as a vector of human diseases and are the first vector of animal diseases with a great impact on livestock farming. Tick vaccines represent a sustainable and effective alternative to chemical acaricides for the control of tick infestations and transmitted pathogens. The application of proteomics to tick vaccine development is a fairly recent area, which has resulted in the characterization of some tick-host-pathogen interactions and the identification of candidate protective antigens.

Areas covered: In this article, we review the application and possibilities of various proteomic approaches for the discovery of tick and pathogen derived protective antigens, and the design of effective vaccines for the control of tick infestations and pathogen infection and transmission.

Expert commentary: In the near future, the application of reverse proteomics, immunoproteomics, structural proteomics, and interactomics among other proteomics approaches will likely contribute to improve vaccine design to control multiple tick species with the ultimate goal of controlling tick-borne diseases.     

Seed depletion and landscape structure affect aggregative response in two wintering passerine birds

Capsule: Seed abundance but also seed depletion during winter modifies habitat use and aggregation behaviour of Eurasian Sky Lark Alauda arvensis and Meadow Pipit Anthus pratensis.

Aims: To investigate the effect of seed abundance on the density of two passerine birds, the Eurasian Sky Larks and the Meadow Pipit, at an agricultural landscape scale during winter.

Methods: Bird counts as well as seed density measurements were carried out in winter to quantify the aggregative response of farmland birds to seed abundance from November to March.

Results: Seed resources varied by a factor of ten between crop types, but declined so sharply over the winter that seed abundance was low and similar among crop types by late winter. Sky Larks selected for higher seed density plots, but only at the end of winter when the resources were the lowest. Conversely, Meadow Pipits did not show any aggregative response to seed abundance.

Conclusion: These results show that an uptake of seed-rich habitats in agricultural landscape would be very beneficial for wintering granivorous birds, by fulfilling the late winter ‘hungry gap’. The inclusion of seed mixtures in habitats that attract high densities of birds and retain seeds until late winter is of primary interest.     

Serum tau fragments as predictors of death or poor neurological outcome after out-of-hospital cardiac arrest

Background: Anoxic brain injury is the primary cause of death after resuscitation from out-of-hospital cardiac arrest (OHCA) and prognostication is challenging. The aim of this study was to evaluate the potential of two fragments of tau as serum biomarkers for neurological outcome.

Methods: Single-center sub-study of 171 patients included in the Target Temperature Management (TTM) Trial randomly assigned to TTM at 33?°C or TTM at 36?°C for 24?h after OHCA. Fragments (tau-A and tau-C) of the neuronal protein tau were measured in serum 24, 48 and 72?h after OHCA. The primary endpoint was neurological outcome.

Results: Median (quartile 1 – quartile 3) tau-A (ng/ml) values were 58 (43–71) versus 51 (43–67), 72 (57–84) versus 71 (59–82) and 76 (61–92) versus 75 (64–89) for good versus unfavourable outcome at 24, 48 and 72?h, respectively (p_group = 0.95). Median tau C (ng/ml) values were 38 (29–50) versus 36 (29–49), 49 (38–58) versus 48 (33–59) and 48 (39–59) versus 48 (36–62) (p_group = 0.95). Tau-A and tau-C did not predict neurological outcome (area under the receiver-operating curve at 48?h; tau-A: 0.51 and tau-C: 0.51).

Conclusions: Serum levels of tau fragments were unable to predict neurological outcome after OHCA.     

Augmented simvastatin cytotoxicity using optimized lipid nanocapsules: a potential for breast cancer treatment

Context: We noticed paucity in exploiting solutol-based lipid nanocapsules in statins formulations though they carry all favorable properties that are needed for cancer passive targeting such as their small particle size, stealth properties, ability to highly accommodate lipophilic drugs, good internalization and P-gp pump inhibition.

Objective: The aim of this study was to design and optimize new simvastatin drug delivery systems; lipid nanocapsules intended for administration through the intravenous route as potential treatment for breast cancer.

Methods: Optimized nanocapsules were prepared by the phase-inversion method according to a D-optimal mixture design, characterized and assessed for their cytotoxicity.

Results: Three successful models for particle size, polydispersity index (PDI) and percentage of drug released after 48 h were generated. The prepared lipid nanocapsules acquired spherical and homogenous morphology, good stability and tolerance to sterilization. The obtained release profiles demonstrated desired sustained release pattern. Furthermore, testing selected formulations on human breast cancer adenocarcinoma cells showed augmented cytotoxicity of simvastatin reaching low IC50 values as 1.4?±?0.02 μg/ml compared to the pure drug.

Conclusion: The proposed lipid nanocapsules pose promising candidates as simvastatin carriers intended for the targeting of breast cancer.     

Association between polymorphisms in IL21 gene and risk for sepsis

Context: Sepsis is now the leading cause of death in the noncardiovascular intensive care unit (ICU).

Objective: To investigate whether polymorphisms in IL21 gene contribute to sepsis susceptibility.

Materials and methods: Three single-nucleotide polymorphisms of IL21 (rs907715, rs2055979, rs12508721) were genotyped by TaqMan assay in patients with sepsis and control subjects.

Results: Polymorphisms rs2055979 and rs12508721 in IL21 were more frequent in sepsis patients compared to general population. But allele frequency of rs907715 was not significantly different between sepsis patients and control subjects.

Conclusion: Polymorphisms in IL21 may be associated with sepsis risk.     

Impact of CT-apelin and NT-proBNP on identifying non-responders to cardiac resynchronization therapy

Context: Assessment of response to cardiac resynchronization therapy (CRT) is essential.

Objective: To assess the predictive value of CT-apelin together with NT-proBNP in patients undergoing CRT.

Methods: Serum CT-apelin and NT-proBNP were measured by ELISA before, and six months after CRT. Primary endpoint was non-response (<4% increase in LVEF) after six months.

Results: From 81 patients, 15 proved to be non-responders. Six-month CT-apelin was superior compared to NT-proBNP in identifying non-responders by multivariate ROC (CT-apelin: p?=?0.01, NT-proBNP: p?=?0.13) and by logistic regression (CT-apelin: p?=?0.01, NT-proBNP: p?=?0.41) analyses.

Conclusion: Six-month CT-apelin might be a valuable novel biomarker in identifying non-responders to CRT that was superior to NT-proBNP.     

The long and the short of it: long-styled florets are associated with higher outcrossing rate in Senecio vulgaris and result from delayed self-pollen germination

Background: It has been reported that some plants of the self-compatible species Senecio vulgaris produce capitula containing long-styled florets which fail to set seed when left to self-pollinate, although readily set seed when self-pollinated by hand.

Aims: To determine if production of long-styled florets is associated with higher outcrossing rate in S. vulgaris, and whether long-styles occur in non-pollinated florets, whereas short-styles are present in self-pollinated florets.

Methods: The frequency of long-styled florets was compared in the radiate and non-radiate variants of S. vulgaris, known to exhibit higher and lower outcrossing rates, respectively. In addition, style length was compared in emasculated florets that were either self-pollinated or left non-pollinated.

Results: Long-styled florets were more frequent in the higher outcrossing radiate variant. Following emasculation, long styles occurred in non-pollinated florets, while short styles were present in self-pollinated florets. The two variants did not differ in style length within the non-pollinated or within the self-pollinated floret categories.

Conclusions: A high frequency of long-styled florets is associated with higher outcrossing rate in S. vulgaris and results from delayed self-pollination and pollen germination on stigmas.     

Self-pollination,style length development and seed set in self-compatible Asteraceae: evidence from Senecio vulgaris L.

Background: Variation in style length has been reported in Senecio vulgaris and has been associated with outcrossing rate.

Aims: To determine if (i) long styles lack germinated pollen on stigmas left to self-pollinate, (ii) successful self-pollination causes styles to stop elongating and shrink in length and (iii) seed set increases with the amount of pollen deposited on stigmas.

Methods: Determined germinated self-pollen on stigmas of long and short styles after auto-self-pollination; scored style length over 48 h in self-pollinated and non-pollinated florets; recorded seed set after placing different amounts of pollen on stigmas.

Results: Most long-styled florets had zero or low amounts of germinated pollen on stigmas in contrast to most short-styled florets. Styles initially elongated to the same length in self-pollinated and non-pollinated florets, then shrank in length in self-pollinated florets while continuing to elongate in non-pollinated florets. Seed set increased with number of pollen grains deposited on stigmas.

Conclusions: Successful self-pollen deposition and/or germination on stigmas of S. vulgaris are indicated by presence of short styles, whereas the opposite is indicated by presence of long styles in florets left to self-pollinate. Self-pollination causes styles to shrink after initially elongating. Seed set is dependent on the amount of pollen deposited on stigmas.     

Mutation in the beta3 subunit of Guanine nucleotide-binding protein (GNB3) gene is not associated with Type II diabetes mellitus risk: a case–control study of a North Indian population

Context: Genetics play a major role in development and pathophysiology of Type 2 diabetes mellitus (T2DM).

Objective: To asses the association of Guanine nucleotide-binding protein (GNB3) (C825T) gene's polymorphism with T2DM.

Materials and methods: A case–control study including 400 North Indians was performed using Polymerase Chain Reaction–Restriction Fragment Length Polymorphism (PCR-RFLP) approach to analyze genetic polymorphism.

Results: No significant difference was observed in genotype and allele frequencies of GNB3 gene on comparing cases with controls.

Discussion: Our study is in agreement with studies on Polish, Japanese, Hispanic-American and Danish populations who observed no significant association between GNB3 (C825T) polymorphism and T2DM.

Conclusion: GNB3 (C825T) polymorphism is not associated with T2DM.     

The diversity and community dynamics of hazelwood lichens and bryophytes along a major gradient of human impact

Background: Oceanic hazelwoods in western Scotland are hypothesised to be unmanaged post-glacial relicts, representing an unusual type of old-growth forest habitat in Europe. They are characterised by an exceptionally high epiphytic diversity, including their status as ‘hotspots’ for indicators of woodland ecological continuity.

Aims: The aim of this study was to evaluate the effects of climate, pollution and management on the occurrence of epiphytic lichens and bryophytes on Corylus avellana.

Methods: Thirteen hazelwood study sites were systematically sampled along a climate, pollution and management gradient in Britain. Epiphyte composition and richness were examined in a strict hierarchical framework, and compared against site-, stool- and stem-scale environmental predictors.

Results: The study showed that along the gradient from ‘clean-air’ relict sites to ‘polluted’ coppiced sites: (i) epiphytic local stem-scale diversity declined, (ii) there was a loss of late-successional species including foliose cyano- and tripartite lichens and bryophytes and (iii) stem sizes were reduced, providing a further limit to the accumulation of species richness within a site.

Conclusions: Relict hazelwoods in western Scotland are confirmed as an example of the most intact epiphyte communities. In particular, we show that the transition to coppicing can be clearly linked to ecological processes causing species loss.     

Plant sex and phenological stage affect interactions with rhizosphere nematode communities

Background: In dioecious plants, females and males associate differently with mycorrhizal fungi, but interactions with other rhizosphere organisms are less well studied.

Aims: We investigated the effect of plant sex on rhizosphere nematode communities associated with Corema album, a dioecious shrub occurring in coastal habitats.

Methods: Rhizosphere samples were collected from males and females in three populations (150 plants), during fruiting and flowering. Nematode communities were characterised and compared between plant sexes through statistical analyses of the abundance of trophic groups, plant parasitic nematode (PPN) genera and ecological indices.

Results: Free-living nematodes showed no statistically significant differences owing to plant sex. Conversely, PPN community composition was significantly different between plant sexes during fruiting but not flowering, suggesting that physiological requirements over the annual phenological cycle of the plant influence ecological interactions with the rhizosphere.

Of the 13 PPN genera identified, the ectoparasitic Criconema and Hemicriconemellawere more abundant in the rhizosphere of males during fruiting, whereas the endoparasitic Meloidogyne associated more frequently with females, suggesting that plant host suitability is related to PPN feeding strategy.

Conclusions: It appears that interactions of individuals of different sexes of C. album with the rhizosphere nematode community vary with phenological stage, especially for PPN.     

Nanostructured lipid carriers as semisolid topical delivery formulations for diflucortolone valerate

Context: Topical treatment of skin disease needs to be strategic to ensure high drug concentration in the skin with minimum systemic absorption.

Objective: The aim of this study was to produce semisolid nanostructured lipid carrier (NLC) formulations, for topical delivery of the corticosteroid drug, diflucortolone valerate (DFV), with minimum systemic absorption.

Method: NLC formulations were developed using a high shear homogenization combined with sonication, using Precirol® ATO5 or Tristearin® as the solid lipid, Capryol? or isopropyl myristate as the liquid lipid and Poloxamer® 407 as surfactant. The present study addresses the influence of different formulations composition as solid lipid, liquid lipid types and concentrations on the physicochemical properties and drug release profile from NLCs.

Results and discussion: DFV-loaded NLC formulations possessed average particle size ranging from 160.40?nm to 743.7?nm with narrow polydispersity index. The encapsulation efficiency was improved by adding the lipid-based surfactants (Labrasol® and Labrafil® M1944CS) to reach 68%. The drug release from the investigated NLC formulations showed a prolonged release up to 12?h. The dermatopharmacokinetic study revealed an improvement in drug deposition in the skin with the optimized DFV-loaded NLC formulation, in contrast to a commercial formulation.

Conclusion: NLC provides a promising nanocarrier system that work as reservoir for targeting topical delivery of DFV.     

H19 non-coding RNA in urine cells detects urothelial carcinoma: a pilot study

Context: Urothelial carcinoma (UC) is common and highly recurrent. Diagnosis and follow-up involve invasive cystoscopies.

Objective: To evaluate H19 RNA in urine cells as diagnostic tool for UC.

Materials and methods: RT-PCR analysis of urine samples from healthy volunteers and UC patients.

Results: H19 RNA was unequivocally detected in the urine of 90.5% of patients and 25.9% of controls. H19 copies were three orders of magnitude higher in patients. Receiver operating characteristic analysis showed an area under the curve of 0.933.

Conclusions: This pilot study shows that urinary cell H19 is a highly sensitive test for UC and pending verification could transform patient management.     

Habitat associations of young Black Grouse Tetrao tetrix broods

Capsule: In the Scottish Highlands, Black Grouse Tetrao tetrix females with young broods selected areas rich in wet flushes, grasses and Sphagnum mosses.

Aims: To identify associations between young Black Grouse broods and habitat characteristics.

Methods: We located brood-rearing Black Grouse females in four areas of moorland-forest mosaic in the Scottish Highlands. Each brood was paired with a random reference location 100?m away. We recorded cover of different habitats and fine-scale vegetation and structure within squares of 1 and 0.25?ha, respectively. Associations between brood occurrence and habitat and vegetation variables were examined at the two scales.

Results: Black Grouse broods were positively associated with wet flushes (1?ha), and with Heather Calluna vulgaris, sedges Carex spp., grasses, Sphagnum mosses and taller vegetation with intermediate levels of height variability (0.25?ha).

Conclusions: This study highlights the importance of wet habitats for young Black Grouse chicks. In addition, relatively tall field-layer vegetation comprising Heather, sedges, grasses over a layer of Sphagnum mosses attracted brood-rearing Black Grouse females. Many management practices associated with peatland restoration, including drain-blocking, Sphagnum re-establishment and the avoidance of burning on deep peat soils, are therefore consistent with favourable habitat management for Black Grouse.     

Polymorphisms and expression of IL-32: impact on genetic susceptibility and clinical outcome of lung cancer

Context: Polymorphisms of IL-32 related closely to tumoregenesis.

Materials and methods: Two IL-32 polymorphisms (rs12934561 and rs28372698) and mRNA expression were conducted by SNP genotype assay and real-time PCR in 423 lung cancer patients and 437 controls.

Results: T allele of rs28372698 associated significantly with poor prognosis in moderate and well-differentiated lung cancer patients. TT genotype of rs12934561 related closely to poor survival status in squamous carcinoma. IL-32 mRNA expression decreased in lung cancer.

Discussion and conclusion: Our study indicates the importance of IL-32 polymorphism and mRNA expression in susceptibility and influence of survival status in lung cancer.     

Morphology,geographical variation and the subspecies of Marsh Tit Poecile palustris in Britain and central Europe

Capsule All British Marsh Tits belong to subspecies Poecile palustris dresseri, being smaller than nominate P. p. palustris of central Europe.

Aims Determining the subspecies of Marsh Tit in Britain to test whether ssp. P. p. palustris occurs in northern England and Scotland, by assessing regional variation in size compared with central European birds.

Methods 1147 wing length and 250 tail length measurements from 953 Marsh Tits were compared between eight British locations to test for regional variation. Biometrics were compared between birds from Britain and six locations within the continental European range of ssp. palustris.

Results There was no regional variation in wing or tail lengths among British Marsh Tits, indicating that all resident birds belong to ssp. dresseri. There was no evidence supporting the existence of ssp. palustris in northern England. British birds were significantly smaller than those from continental Europe, with proportionately shorter tails, consistent across all age and sex classes.

Conclusion All British Marsh Tits should be considered as ssp. dresseri, with ssp. palustris being limited to continental Europe. With no evidence of regional variation in size within Britain, reliable sexing methods based on biometrics could be applied in demographic studies throughout the country.     

Edge effects on epiphytic lichen diversity in the forest-steppe of the Kazakh Altai

Background: Forests in forest-steppe ecotones are usually highly fragmented and much of the forested area is exposed to climate and land-use-related edge effects.

Aim: To test the hypothesis that the epiphytic lichen diversity at the forest edges was reduced compared with that in the forest interior, and to analyse lichen diversity in comparison with the more highly elevated and more continental Mongolian Altai.

Methods: Six plots each in the interior and the edge with a total of 240 Larix sibirica trees were studied in the Katon-Karagai National Park, East Kazakhstan.

Results: Species richness and evenness at the tree level were higher in the interior than at the edge. The epiphytic lichen diversity in the forest interior was similar in the Kazakh and Mongolian Altai, whereas that at the forest edge was lower in the Mongolian Altai.

Conclusions: Strong degradation of the forest edges in the Kazakh Altai is the probable cause of the reduced epiphytic lichen diversity compared with the interior. The similar species richness in the forest interiors of the Kazakh and Mongolian Altai suggests that the differences at the forest edge are probably, at least partly, due to different land-use regimes and not to differences in macroclimate.     

Evaluation of genetic diversity and population structure of Macqueen’s Bustard Chlamydotis macqueenii in Iran

Capsule: There is low genetic diversity in the Macqueen’s Bustard Chlamydotis macqueenii in Iran.

Aims: To investigate the genetic diversity and population structure of Macqueen’s Bustard in Iran, using two mitochondrial DNA loci.

Methods: Molecular diversity of the mitochondrial cytochrome oxidase c subunit I (COI) gene and part of the mitochondrial control region D-loop (in total 1183 base pairs) were analysed from 26 individual Macqueen’s Bustards from three regions of Iran.

Results: There was little variation in nucleotides and haplotypes in the populations for genes of both CR and COI. The population had free breeding and gene flow between the three study regions in Iran: Petregan, Ferdows and Yazd.

Conclusion: The use of molecular and genetic studies is essential to strengthen the protection of genetic diversity of the Macqueen’s Bustard.