Dynamics of proteins: Light scattering study of dilute and dense colloidal suspensions of eye lens homogenates

We report a dynamic light scattering study on protein suspensions of bovine lens homogenates at conditions (pH and ionic strength) similar to the physiological ones. Light scattering data were collected at two temperatures, 20 oC and 37 oC, over a wide range of concentrations from the very dilute limit up to the dense regime approaching to the physiological lens concentration. A comparison with experimental data from intact bovine lenses was advanced revealing differences between dispersions and lenses at similar concentrations. In the dilute regime two scattering entities were detected and identified with the long-time, self-diffusion modes of alpha-crystallins and their aggregates, which naturally exist in lens nucleus. Self-diffusion coefficients are temperature insensitive, whereas the collective diffusion coefficient depends strongly on temperature revealing a reduction of the net repulsive interparticle forces with lowering temperature. While there are no rigorous theoretical approaches on particle diffusion properties for multi-component, non-ideal hard-sphere, polydispersed systems, as the suspensions studied here, a discussion of the volume fraction dependence of the long-time, self-diffusion coefficient in the context of existing theoretical approaches was undertaken. This study is purported to provide some insight into the complex light scattering pattern of intact lenses and the interactions between the constituent proteins that are responsible for lens transparency. This would lead to understand basic mechanisms of specific protein interactions that lead to lens opacification (cataract) under pathological conditions.Comment: To appear in J. Chem. Phy

Prevalence of disordered eating in elite female athletes in team sports in Greece

The purpose of the study was to assess a) the prevalence of disordered eating (DE) in elite female team sports players compared to non-athletes and b) to compare DE prevalence in elite female players in basketball, volleyball and water polo. One hundred and seventy-five females were recruited (age 23.10 ± 5.4, BMI 21.85 ± 2.3 kg/m2), 53 were elite basketball players, 42 were elite volleyball players, 34 were elite water polo players and 46 were non-athletes. Participants completed the Eating Disorders Questionnaire (EDE-Q) and a physical activity questionnaire. The EDE-Q incorporates 36 statements which relate to the occurrence and frequency of key behaviours of eating disorders, under the following four subscales: Restraint, eating concern, shape concern and weight concern and a global score of disordered eating. No differences were found in the EDE-Q subscale score and global score between athletes and non-athletes. Only 6.2% of the total number of participants exhibited DE using the global score >2.3. Water polo players had significantly higher scores in the ‘eating concern’ subscale and in the frequency of key behavioural features of DE such as binge eating episodes and objective and subjective bulimic episodes, compared to volleyball and basketball players. In conclusion, team sport elite female players do not exhibit greater prevalence of DE compared to non-athletes. Water polo, a sport that emphasizes leanness and control of body weight for international distinctions, is associated with a higher tendency to exhibit DE, when compared to other team sports

Predicting the targeting of tail-anchored proteins to subcellular compartments in mammalian cells

This is the author accepted manuscript. The final version is available from Company of Biologists via the DOI in this record.Tail-anchored (TA) proteins contain a single transmembrane domain (TMD) at the Cterminus, anchoring them to organelle membranes where they mediate a variety of critical cellular processes. Mutations in individual TA proteins cause a number of severe inherited disorders. However, the molecular mechanisms and signals facilitating proper TA protein targeting are not fully understood, in particular in mammals. Here, we identify additional TA proteins at peroxisomes or shared by multiple organelles in mammals and reveal that a combination of TMD hydrophobicity and tail charge determines targeting to distinct organelles. Specifically, an increase in tail charge can override a hydrophobic TMD signal and re-direct a protein from the ER to peroxisomes or mitochondria and vice versa. We demonstrate that subtle alterations in those physicochemical parameters can shift TA protein targeting between organelles, explaining why peroxisomes and mitochondria share many TA proteins. Our analyses enabled us to allocate characteristic physicochemical parameters to different organelle groups. This classification allows for the first time, successful prediction of the location of uncharacterized TA proteins.We thank colleagues who provided materials (see Tables S1-S4) and acknowledge support from A. C. Magalhães, M. Almeida, D. Tuerker, S. Kuehl and C. Davies. This work was supported by the Biotechnology and Biological Sciences Research Council (BB/K006231/1 to M.S.), a Wellcome Trust Institutional Strategic Support Award (WT097835MF, WT105618MA to M.S.), the Portuguese Foundation for Science and Technology and FEDER/COMPETE (PTDC/BIA-BCM/118605/2010 to M.S.; SFRH/BD/37647/2007 to N.B.; SFRH/BPD/77619/2011 and UID/BIM/04501/2013 to D.R.). M.W., E.A.G., and M.S. are supported by Marie Curie Initial Training Network (ITN) action PerFuMe (316723)

HGF Mediates the Anti-inflammatory Effects of PRP on Injured Tendons

Platelet-rich plasma (PRP) containing hepatocyte growth factor (HGF) and other growth factors are widely used in orthopaedic/sports medicine to repair injured tendons. While PRP treatment is reported to decrease pain in patients with tendon injury, the mechanism of this effect is not clear. Tendon pain is often associated with tendon inflammation, and HGF is known to protect tissues from inflammatory damages. Therefore, we hypothesized that HGF in PRP causes the anti-inflammatory effects. To test this hypothesis, we performed in vitro experiments on rabbit tendon cells and in vivo experiments on a mouse Achilles tendon injury model. We found that addition of PRP or HGF decreased gene expression of COX-1, COX-2, and mPGES-1, induced by the treatment of tendon cells in vitro with IL-1β. Further, the treatment of tendon cell cultures with HGF antibodies reduced the suppressive effects of PRP or HGF on IL-1β-induced COX-1, COX-2, and mPGES-1 gene expressions. Treatment with PRP or HGF almost completely blocked the cellular production of PGE2 and the expression of COX proteins. Finally, injection of PRP or HGF into wounded mouse Achilles tendons in vivo decreased PGE2 production in the tendinous tissues. Injection of platelet-poor plasma (PPP) however, did not reduce PGE2 levels in the wounded tendons, but the injection of HGF antibody inhibited the effects of PRP and HGF. Further, injection of PRP or HGF also decreased COX-1 and COX-2 proteins. These results indicate that PRP exerts anti-inflammatory effects on injured tendons through HGF. This study provides basic scientific evidence to support the use of PRP to treat injured tendons because PRP can reduce inflammation and thereby reduce the associated pain caused by high levels of PGE2. © 2013 Zhang et al

Identification of cancer predisposition variants in apparently healthy individuals using a next-generation sequencing-based family genomics approach

Cancer, like many common disorders, has a complex etiology, often with a strong genetic component and with multiple environmental factors contributing to susceptibility. A considerable number of genomic variants have been previously reported to be causative of, or associated with, an increased risk for various types of cancer. Here, we adopted a next-generation sequencing approach in 11 members of two families of Greek descent to identify all genomic variants with the potential to predispose family members to cancer. Cross-comparison with data from the Human Gene Mutation Database identified a total of 571 variants, from which 47 % were disease-associated polymorphisms, 26 % disease-associated polymorphisms with additional supporting functional evidence, 19 % functional polymorphisms with in vitro/laboratory or in vivo supporting evidence but no known disease association, 4 % putative disease-causing mutations but with some residual doubt as to their pathological significance, and 3 % disease-causing mutations. Subsequent analysis, focused on the latter variant class most likely to be involved in cancer predisposition, revealed two variants of prime interest, namely MSH2 c.2732T>A (p.L911R) and BRCA1 c.2955delC, the first of which is novel. KMT2D c.13895delC and c.1940C>A variants are additionally reported as incidental findings. The next-generation sequencing-based family genomics approach described herein has the potential to be applied to other types of complex genetic disorder in order to identify variants of potential pathological significance

Constraint-Flow Nets: A Model for Building Constraints from Resource Dependencies

The major research in the resource management literature focuses primarily on two complementary sub-problems: (1) specification languages for formulating resource requests and (2) constraint problems modelling allocation and scheduling. Both directions assume the knowledge of the underlying platform architecture and the dependencies it induces on the usage of the various resources. In this paper, we bridge this gap by introducing constraint-flow nets (cfNets). A cfNet is defined by a set of resources and dependencies between them, each dependency having an associated constraint schema. The model is inspired by Petri nets, with resources corresponding to places and dependencies—to transitions. Given an architecture of dependent resources, an initial resource request is propagated through the dependencies. The generated constraints are then conjuncted into the global allocation constraint. We study the notion of conflicts in cfNets and prove that for conflict-free cfNets the global allocation constraint can be constructed unambiguously. Furthermore, we provide an SMT-based algorithm for conflict detection and discuss the use of priorities to dynamically resolve conflicts at run-time. Finally, we illustrate the use of cfNets on a case study inspired by the Kalray MPPA architecture