DNA Vaccine Encoding the Chimeric Form of Schistosoma mansoni Sm-TSP2 and Sm29 Confers Partial Protection against Challenge Infection

Schistosomiasis is an important parasitic disease worldwide that affects more than 207 million people in 76 countries and causes approximately 250,000 deaths per year. The best long-term strategy to control schistosomiasis is through immunization combined with drug treatment. Due to the ability of DNA vaccines to generate humoral and cellular immune responses, such vaccines are considered a promising approach against schistosomiasis. Sm29 and tetraspanin-2 (Sm-TSP2) are two proteins that are located in the S. mansoni tegument of adult worms and schistosomula and induce high levels of protection through recombinant protein immunization. In this study, we transfected BHK-21 cells with plasmids encoding Sm29, Sm-TSP2 or a chimera containing both genes. Using RT-PCR analysis and western blot, we confirmed that the DNA vaccine constructs were transcribed and translated, respectively, in BHK-21 cells. After immunization of mice, we evaluated the reduction in worm burden. We observed worm burden reductions of 17-22%, 22%, 31-32% and 24-32% in animals immunized with the pUMVC3/Sm29, pUMVC3/SmTSP-2, pUMVC3/Chimera and pUMVC3/Sm29 + pUMVC3/SmTSP-2 plasmids, respectively. We evaluated the humoral response elicited by DNA vaccines, and animals immunized with pUMVC3/Sm29 and pUMVC3/Sm29 + pUMVC3/SmTSP-2 showed higher titers of anti-Sm29 antibodies. The cytokine profile produced by the spleen cells of immunized mice was then evaluated. We observed higher production of Th1 cytokines, such as TNF-α and IFN-γ, in vaccinated mice and no significant production of IL-4 and IL-5. The DNA vaccines tested in this study showed the ability to generate a protective immune response against schistosomiasis, probably through the production of Th1 cytokines. However, future strategies aiming to optimize the protective response induced by a chimeric DNA construct need to be developed.     

Na+/H+ Exchanger Isoform 1-Induced Osteopontin Expression Facilitates Cardiomyocyte Hypertrophy

Enhanced expression and activity of the Na⁺/H⁺ exchanger isoform 1 (NHE1) has been implicated in cardiomyocyte hypertrophy in various experimental models. The upregulation of NHE1 was correlated with an increase in osteopontin (OPN) expression in models of cardiac hypertrophy (CH), and the mechanism for this remains to be delineated. To determine whether the expression of active NHE1-induces OPN and contributes to the hypertrophic response in vitro, cardiomyocytes were infected with the active form of the NHE1 adenovirus or transfected with OPN silencing RNA (siRNA-OPN) and characterized for cardiomyocyte hypertrophy. Expression of NHE1 in cardiomyocytes resulted in a significant increase in cardiomyocyte hypertrophy markers: cell surface area, protein content, ANP mRNA and expression of phosphorylated-GATA4. NHE1 activity was also significantly increased in cardiomyocytes expressing active NHE1. Interestingly, transfection of cardiomyocytes with siRNA-OPN significantly abolished the NHE1-induced cardiomyocyte hypertrophy. siRNA-OPN also significantly reduced the activity of NHE1 in cardiomyocytes expressing NHE1 (68.5±0.24%; P<0.05), confirming the role of OPN in the NHE1-induced hypertrophic response. The hypertrophic response facilitated by NHE1-induced OPN occurred independent of the extracellular-signal-regulated kinases and Akt, but required p90-ribosomal S6 kinase (RSK). The ability of OPN to facilitate the NHE1-induced hypertrophic response identifies OPN as a potential therapeutic target to reverse the hypertrophic effect induced by the expression of active NHE1.     

Vitamin D and the Risk of Atrial Fibrillation - The Rotterdam Study

Atrial fibrillation (AF) is the most common chronic arrhythmia and it increases the risk of cardiovascular morbidity and mortality. Still there is not a complete understanding of its etiology and underlying pathways. Vitamin D might regulate renin-angiotensin-aldosterone system and might be involved in inflammation, both implicated in the pathophysiology of AF. The objective of this work was to investigate the association between vitamin D status with the risk of AF in the elderly. This study was conducted within the Rotterdam Study, a community-based cohort of middle-aged and elderly participants in Rotterdam, The Netherlands. We had 3,395 participants who were free of AF diagnosis at the start of our study and who had vitamin D data available. We analyzed the association between serum 25-hydroxivitamin D (25(OH)D) and incidence of AF using Cox regression models. Vitamin D deficiency was defined as serum 25(OH)D concentrations <50nmol/l, insufficiency between 50nmol/l and 75nmol/l, while serum 25(OH)D concentrations equal to and above 75nmol/l were considered as adequate. After mean follow-up of 12.0 years 263 (7.7%) participants were diagnosed with incident AF. Vitamin D status was not associated with AF in any of the 3 multivariate models tested (model adjusted for socio-demographic factors and life-style factors: HR per 10 unit increment in serum 25(OH)D 0.96, 95% CI: 0.91-1.02; HR for insufficiency: 0.82, 95%CI: 0.60-1.11,and HR for adequate status: 0.76, 95%CI: 0.52-1.12 compared to deficiency). This prospective cohort study does not support the hypothesis that vitamin D status is associated with AF.     

A sensory approach for multispecies anthropology

This special issue suggests that the need to examine the entangled lives of species, selves and other beings through a multisensory perspective is crucial and timely. Developing on a sensory analysis, one that emerges through what Anna Tsing refers to as the ‘arts of noticing’ (2015), this introductory paper explores how both nonhuman and human lives are intertwined, and how their close examination can guide anthropologists in their ability to capture the subtleties of more‐than‐human engagement, connection and relatedness. Through articles within this issue from Australian anthropology and beyond, we ask how becoming‐with more‐than‐humans helps us to construct a post‐humanist analysis in the combination of sensory anthropology and multispecies anthropology. Through a combination of these two fields, the paper suggests, anthropology can take up the opportunity to think about animals as subjects, through our ability to communicate beyond language and to engage in a more meaningful way through interspecies knowledge‐making.     

Light modulated cnidocyte discharge predates the origins of eyes in Cnidaria

Complex biological traits often originate by integrating previously separate parts, but the organismal functions of these precursors are challenging to infer. If we can understand the ancestral functions of these precursors, it could help explain how they persisted and how they facilitated the origins of complex traits. Animal eyes are some of the best studied complex traits, and they include many parts, such as opsin‐based photoreceptor cells, pigment cells, and lens cells. Eye evolution is understood through conceptual models that argue these parts gradually came together to support increasingly sophisticated visual functions. Despite the well‐accepted logic of these conceptual models, explicit comparative studies to identify organismal functions of eye precursors are lacking. Here, we investigate how precursors functioned before they became part of eyes in Cnidaria, a group formed by sea anemones, corals, and jellyfish. Specifically, we test whether ancestral photoreceptor cells regulated the discharge of cnidocytes, the expensive single‐use cells with various functions including prey capture, locomotion, and protection. Similar to a previous study of Hydra, we show an additional four distantly related cnidarian groups discharge significantly more cnidocytes when exposed to dim blue light compared with bright blue light. Our comparative analyses support the hypothesis that the cnidarian ancestor was capable of modulating cnidocyte discharge with light, which we speculate uses an opsin‐based phototransduction pathway homologous to that previously described in Hydra. Although eye precursors might have had other functions like regulating timing of spawning, our findings are consistent with the hypothesis that photoreceptor cells which mediate cnidocyte discharge predated eyes, perhaps facilitating the prolific origination of eyes in Cnidaria.     

Sortin2 enhances endocytic trafficking towards the vacuole in Saccharomyces cerevisiae

Background

A highly regulated trafficking of cargo vesicles in eukaryotes performs protein delivery to a variety of cellular compartments of endomembrane system. The two main routes, the secretory and the endocytic pathways have pivotal functions in uni- and multi-cellular organisms. Protein delivery and targeting includes cargo recognition, vesicle formation and fusion. Developing new tools to modulate protein trafficking allows better understanding the endomembrane system mechanisms and their regulation. The compound Sortin2 has been described as a protein trafficking modulator affecting targeting of the vacuolar protein carboxypeptidase Y (CPY), triggering its secretion in Saccharomyces cerevisiae.

Results

A reverse chemical-genetics approach was used to identify key proteins for Sortin2 bioactivity. A genome-wide Sortin2 resistance screen revealed six yeast deletion mutants that do not secrete CPY when grown at Sortin2 condition where the parental strain does: met18, sla1, clc1, dfg10, dpl1 and yjl175w. Integrating mutant phenotype and gene ontology annotation of the corresponding genes and their interactome pointed towards a high representation of genes involved in the endocytic process. In wild type yeast endocytosis towards the vacuole was faster in presence of Sortin2, which further validates the data of the genome-wide screen. This effect of Sortin2 depends on structural features of the molecule, suggesting compound specificity. Sortin2 did not affect endocytic trafficking in Sortin2-resistant mutants, strongly suggesting that the Sortin2 effects on the secretory and endocytic pathways are linked.

Conclusions

Overall, the results reveal that Sortin2 enhances the endocytic transport pathway in Saccharomyces cerevisiae. This cellular effect is most likely at the level where secretory and endocytic pathways are merged. Them Sortin2 specificity over the endomembrane system places it as a powerful biological modulator for cell biology.

Electronic supplementary material

The online version of this article (doi:10.1186/s40659-015-0032-9) contains supplementary material, which is available to authorized users.     

How do disordered regions achieve comparable functions to structured domains?

The traditional structure to function paradigm conceives of a protein''s function as emerging from its structure. In recent years, it has been established that unstructured, intrinsically disordered regions (IDRs) in proteins are equally crucial elements for protein function, regulation and homeostasis. In this review, we provide a brief overview of how IDRs can perform similar functions to structured proteins, focusing especially on the formation of protein complexes and assemblies and the mediation of regulated conformational changes. In addition to highlighting instances of such functional equivalence, we explain how differences in the biological and physicochemical properties of IDRs allow them to expand the functional and regulatory repertoire of proteins. We also discuss studies that provide insights into how mutations within functional regions of IDRs can lead to human diseases.     

Effects of gradual-elastic compression stockings on running economy, kinematics, and performance in runners

We investigated the effect of gradual-elastic compression stockings (GCSs) on running economy (RE), kinematics, and performance in endurance runners. Sixteen endurance trained athletes (age: 34.73 ± 6.27 years; VO2max: 62.83 ± 9.03 ml·kg(-1)·min(-1); 38 minutes in 10 km; 1 hour 24 minutes in half marathon) performed in random order 4 bouts of 6 minutes at a recent half-marathon pace on a treadmill to evaluate RE with or without GCSs. Subsequently, 12 athletes were divided into 2 equal groups matched by their VO2max, and they performed a time limit test (T(lim)) on a treadmill at 105% of a recent 10-km pace with or without GCSs for evaluation of physiological responses and running kinematics. There were no significant differences in the RE test in all of the variables analyzed for the conditions, but a moderate reproducibility for some physiological responses was detected in the condition with GCSs. In the T(lim), the group that wore GCSs reached a lower % of maximum heart rate (HRmax) compared with the control group (96.00 ± 2.94 vs. 99.83 ± 0.40) (p = 0.01). Kinematics did not differ between conditions during the T(lim) (p > 0.05). There were improvement trends for time to fatigue (337 vs. 387 seconds; d = 0.32) and a lower VO2peak (≈53 vs. 62 ml·kg(-1)·min(-1); d = 1.19) that were detected with GCSs during the T(lim). These results indicate that GCSs reduce the % of HRmax reached during a test at competition pace. The lower reproducibility of the condition with GCSs perhaps suggests that athletes may possibly need an accommodation period for systematically experiencing the benefits of this garment, but this hypothesis should be further investigated.     

An assessment of mumps vaccine effectiveness by dose during an outbreak in Canada

Background

This investigation was done to assess vaccine effectiveness of one and two doses of the measles, mumps and rubella (MMR) vaccine during an outbreak of mumps in Ontario. The level of coverage required to reach herd immunity and interrupt community transmission of mumps was also estimated.

Methods

Information on confirmed cases of mumps was retrieved from Ontario’s integrated Public Health Information System. Cases that occurred between Sept. 1, 2009, and June 10, 2010, were included. Selected health units supplied coverage data from the Ontario Immunization Record Information System. Vaccine effectiveness by dose was calculated using the screening method. The basic reproductive number (R₀) represents the average number of new infections per case in a fully susceptile population, and R₀ values of between 4 and 10 were considered for varying levels of vaccine effectiveness.

Results

A total of 134 confirmed cases of mumps were identified. Information on receipt of MMR vaccine was available for 114 (85.1%) cases, of whom 63 (55.3%) reported having received only one dose of vaccine; 32 (28.1%) reported having received two doses. Vaccine effectiveness of one dose of the MMR vaccine ranged from 49.2% to 81.6%, whereas vaccine effectiveness of two doses ranged from 66.3% to 88.0%. If we assume vaccine effectiveness of 85% for two doses of the vaccine, vaccine coverage of 88.2% and 98.0% would be needed to interrupt community transmission of mumps if the corresponding reproductive values were four and six.

Interpretation

Our estimates of vaccine effectiveness of one and two doses of mumps-containing vaccine were consistent with the estimates that have been reported in other outbreaks. Outbreaks occurring in Ontario and elsewhere serve as a warning against complacency over vaccination programs.Between September 2009 and June 2010, there was an outbreak of mumps in Ontario, Canada. Outbreaks of mumps were also taking place in the United States (New York and New Jersey) and in Israel during the same period.^1,2 These outbreaks shared several features, including the age distribution of the cases, the predominance of male cases and the occurrence of the disease among people who had received vaccinations against mumps.^1,2 This latter issue can lead to questions from the public and health care providers regarding the effectiveness of the vaccine. Rapid assessment of vaccine effectiveness is, therefore, an important component of outbreak management for vaccine-preventable diseases, and knowledge of the history of the vaccination program is necessary for interpreting the results.A live attenuated mumps vaccine was licensed in Canada in 1969 and introduced in Ontario shortly thereafter. In 1975, a single dose of the combined vaccine for measles, mumps and rubella (MMR) was implemented. A single-dose program continued until 1996, when a second dose of the MMR vaccine was introduced as part of a plan to eradicate measles. The first dose of the MMR vaccine is routinely given at 12 months of age; until 2007, the second dose was recommended for children between four and six years of age. The recommended age for the second dose is now 18 months.Two MMR vaccines are available for use in Ontario. A single dose of monovalent measles vaccine was offered to all students aged 4–18 years in 1996 as part of a measles catch-up campaign. Because mumps-containing vaccine was not used, a cohort of individuals born before 1992 who had only received one dose of mumps-containing vaccine were therefore potentially susceptible to the virus.Epidemiologic data show that the mumps virus circulated relatively widely throughout Ontario until approximately 1980 (M.A. Simpson, Ontario Ministry of Health and Long-Term Care, Toronto, Ont.: personal communication, 2011 Feb. 1), which would have resulted in the natural boosting of the population’s immunity to the disease. The combination of changes in the policy governing vaccination against mumps and the circulation of the disease thus resulted in a susceptible cohort of individuals born between approximately 1980 and 1992 (i.e., people currently between 19 and 31 years of age).A person’s susceptibility to mumps is also influenced by the effectiveness of the vaccine he or she received. Clinical trials have reported vaccine efficacy of approximately 95% after one dose of mumps vaccine under controlled conditions, although estimates of vaccine effectiveness that were conducted in the field (as opposed to clinical trials) have been much lower (i.e., 62%–85%).³ There is less information available about vaccine effectiveness after two doses, but estimates have ranged from 76% to 95%,^3–5 with accumulating evidence of waning immunity.^2–6The objectives of this investigation were to assess the vaccine effectiveness of MMR vaccine by dose and by birth cohort during the outbreak, and to estimate the level of vaccine coverage required to reach herd immunity and interrupt community transmission of mumps.