Phenotype selection reveals coevolution of muscle glycogen and protein and PTEN as a gate keeper for the accretion of muscle mass in adult female mice

We have investigated molecular mechanisms for muscle mass accretion in a non-inbred mouse model (DU6P mice) characterized by extreme muscle mass. This extreme muscle mass was developed during 138 generations of phenotype selection for high protein content. Due to the repeated trait selection a complex setting of different mechanisms was expected to be enriched during the selection experiment. In muscle from 29-week female DU6P mice we have identified robust increases of protein kinase B activation (AKT, Ser-473, up to 2-fold) if compared to 11- and 54-week DU6P mice or controls. While a number of accepted effectors of AKT activation, including IGF-I, IGF-II, insulin/IGF-receptor, myostatin or integrin-linked kinase (ILK), were not correlated with this increase, phosphatase and tensin homologue deleted on chromosome 10 (PTEN) was down-regulated in 29-week female DU6P mice. In addition, higher levels of PTEN phosphorylation were found identifying a second mechanism of PTEN inhibition. Inhibition of PTEN and activation of AKT correlated with specific activation of p70S6 kinase and ribosomal protein S6, reduced phosphorylation of eukaryotic initiation factor 2α (eIF2α) and higher rates of protein synthesis in 29-week female DU6P mice. On the other hand, AKT activation also translated into specific inactivation of glycogen synthase kinase 3? (GSK3?) and an increase of muscular glycogen. In muscles from 29-week female DU6P mice a significant increase of protein/DNA was identified, which was not due to a reduction of protein breakdown or to specific increases of translation initiation. Instead our data support the conclusion that a higher rate of protein translation is contributing to the higher muscle mass in mid-aged female DU6P mice. Our results further reveal coevolution of high protein and high glycogen content during the selection experiment and identify PTEN as gate keeper for muscle mass in mid-aged female DU6P mice.     

M13 bacteriophage display framework that allows sortase-mediated modification of surface-accessible phage proteins

We exploit bacterial sortases to attach a variety of moieties to the capsid proteins of M13 bacteriophage. We show that pIII, pIX, and pVIII can be functionalized with entities ranging from small molecules (e.g., fluorophores, biotin) to correctly folded proteins (e.g., GFP, antibodies, streptavidin) in a site-specific manner, and with yields that surpass those of any reported using phage display technology. A case in point is modification of pVIII. While a phage vector limits the size of the insert into pVIII to a few amino acids, a phagemid system limits the number of copies actually displayed at the surface of M13. Using sortase-based reactions, a 100-fold increase in the efficiency of display of GFP onto pVIII is achieved. Taking advantage of orthogonal sortases, we can simultaneously target two distinct capsid proteins in the same phage particle and maintain excellent specificity of labeling. As demonstrated in this work, this is a simple and effective method for creating a variety of structures, thus expanding the use of M13 for materials science applications and as a biological tool.     

Effect of capsaicin supplementation on repeated sprinting performance

Performance in many team sports is partially dependent on the ability to perform repeatedly at high intensity. Previous research demonstrates that capsaicin (CAP) has physiological and metabolic effects that could influence exercise performance and inflammation. The purpose of this study was to investigate the influence of CAP on performance of and the interleukin-6 (IL-6) response to repeated sprints. Nineteen healthy male experienced athletes, age 18-30 years, participated in a placebo (PCB)-controlled, crossover study. During 1 trial, they consumed 3 g·d(-1) cayenne (25.8 mg·d(-1) CAP) and the other a PCB for days. Directly after the supplementation period, they completed a repeated sprint test (RST) consisting of 15 30-m maximal effort sprints on 35-second intervals with sprint times measured via an electronic dual-beam timing system. Fasted blood draws for IL-6 were taken at baseline before supplementation, 45 minute pre-RST, and immediately post-RST. Rate of perceived exertion (RPE), muscle soreness (MS), and gastrointestinal distress (GD) for 5 symptom subscales were measured 1-minute pretest, during, posttest, and 1-minute posttest. The MS was additionally measured for 3-day posttest. Relative to the PCB, CAP significantly increased the sum of ratings of GD symptoms by 6.3-fold. There was no difference between treatments in fastest or mean sprint time, fatigue, IL-6 response, RPE, or MS. In summary, CAP did not influence repeated sprint performance or the IL-6 response but caused substantial GD. The CAP is not recommended for athletes involved in repeated sprinting.     

Multimodal communication in a noisy environment: a case study of the Bornean rock frog Staurois parvus

High background noise is an impediment to signal detection and perception. We report the use of multiple solutions to improve signal perception in the acoustic and visual modality by the Bornean rock frog, Staurois parvus. We discovered that vocal communication was not impaired by continuous abiotic background noise characterised by fast-flowing water. Males modified amplitude, pitch, repetition rate and duration of notes within their advertisement call. The difference in sound pressure between advertisement calls and background noise at the call dominant frequency of 5578 Hz was 8 dB, a difference sufficient for receiver detection. In addition, males used several visual signals to communicate with conspecifics with foot flagging and foot flashing being the most common and conspicuous visual displays, followed by arm waving, upright posture, crouching, and an open-mouth display. We used acoustic playback experiments to test the efficacy-based alerting signal hypothesis of multimodal communication. In support of the alerting hypothesis, we found that acoustic signals and foot flagging are functionally linked with advertisement calling preceding foot flagging. We conclude that S. parvus has solved the problem of continuous broadband low-frequency noise by both modifying its advertisement call in multiple ways and by using numerous visual signals. This is the first example of a frog using multiple acoustic and visual solutions to communicate in an environment characterised by continuous noise.     

TissuePatch™ as a novel synthetic sealant for repair of superficial lung defect: in vitro tests results

Controversies surrounding the efficacy of surgical sealants against alveolar air leaks (AAL) in lung surgery abound in the literature. We sought to test the sealing efficacy of a novel synthetic sealant, TissuePatch? in an in vitro lung model. The lower lobe of freshly excised swine lung (n?=?10) was intubated and ventilated. A superficial parenchymal defect (40?×?25 mm) was created, followed by AAL assessment. After sealant application, AAL was assessed again until burst failure occurred. The length of defect was recorded to evaluate the elasticity of the sealant. Superficial parenchymal defects resulted in AAL increasing disproportionally with ascending maximal inspiratory pressure (Pmax). Multiple linear regression analysis revealed strong correlation between AAL and Pmax, compliance, resistance. After sealant application, AAL was sealed in all ten tests at an inspired tidal volume (TVi) of 400 ml, in nine tests at TVi?=?500 ml, in seven at TVi?=?600 ml and in five at TVi?=?700 ml. The mean burst pressure was 42?±?9 mBar. Adhesive and cohesive sealant failures were found in six and three tests respectively. The length of defect before sealant failure was 8.9?±?4.9% larger than that at TVi?=?400 ml, demonstrating an adequate elasticity of this sealant film. TissuePatch? may be a reliable sealant for alternative or adjunctive treatment for repair of superficial parenchymal defects in lung surgery. The clinical benefits of this sealant should be confirmed by prospective, randomised controlled clinical trials. Die Wirksamkeit von chirurgischen Klebstoffen zur Prävention von alveolo-pleuralem Luftleck (APL) ist trotz zunehmenden klinischen Anwendungen in Lungenchirurgie immer noch kontrovers diskutiert. Wir evaluierten die Abdichtungswirksamkeit von einem neuartigen synthetischen Kleber, TissuePatch? mittels eines in vitro Lungenmodels. Der Unterlappen von frisch entnommenen Schweinlungen (n?=?10) wurde intubiert und beatmet. Eine pleurale Läsion (40?×?25 mm) wurde erstellt und APL mit steigendem inspiratorischem Tidalvolumen (TVi) untersucht. Nach Applikation von TissuePatch? wurde APL auf die gleiche Weise gemessen bis zur Auftritt von Kleberbruch. Zur Untersuchung der Elastizität des Klebers wurde die Länge der pleuralen Läsion gemessen. Pleurale Läsion führte bei aufsteigendem maximalem inspiratorischem Druck (Pmax) zu überproportionalem Anstieg von APL. Multiple lineare Regressionsanalyse ergab eine starke Korrelation zwischen APL und Pmax, Lungencompliance sowie Widerstand. Nach der Applikation von Klebstoff wurde APL bei TVi?=?400 ml in allen zehn Testen versiegelt, bei TVi?=?500 ml in neun Testen, bei TVi?=?600 ml in sieben und bei TVi?=?700 ml in fünf Testen. Der mittlere Pmax, der zu Kleberbruch führte, betrug 42?±?9 mBar. Bei den Versuchen wurden adhäsiver und kohäsiver Kleberbruch in jeweils sechs und drei Testen gefunden. Die Länge der pleuralen Läsion vor dem Kleberbruch war 8,9?±?4,9% größer als die bei TVi?=?400 ml. Unsere Versuche zeigten eine zuverlässige Versiegelung von TissuePatch? unter mechanischer Ventilation. Die klinische Nützlichkeit vom Kleber als unterstützende Maßnahme zur Prävention von alveolo-pleuralem Luftleck in Lungenchirurgie sollte durch prospektive, randomisierte kontrollierte klinische Studien bestätigt werden.     

Copper(II) coordination polymers derived from triethanolamine and pyromellitic acid for bioinspired mild peroxidative oxidation of cyclohexane

The new inorganic 1D coordination polymer [Cu₂(H₃tea)₂(μ₄-pma)]_n has been prepared, via self-assembly in aqueous medium, from copper(II) nitrate, triethanolamine (H₃tea), pyromellitic acid (H₄pma) and lithium hydroxide, and characterized by IR spectroscopy, elemental and single-crystal X-ray diffraction analyses. This compound and the related 2D polymer [Cu₂(μ-H₂tea)₂{μ₃-Na₂(H₂O)₄}(μ₆-pma)]_n · 10nH₂O are shown to mimic the alkane partial oxidation activity of the multicopper particulate methane monooxygenase, acting as catalysts precursors for the peroxidative oxidation of cyclohexane into cyclohexanol and cyclohexanone, by hydrogen peroxide (as green oxidant) and at room temperature in acidic MeCN/H₂O medium. An overall yield (based on cyclohexane) of 29% has been achieved.     

Modelling the hydro-geochemistry of acid-sensitive catchments in Finland under atmospheric deposition and biomass harvesting scenarios

The dynamic hydro-geochemical Model of Acidification of Groundwater in Catchments (MAGIC) was used to predict the response of 163 Finnish lake catchments to historic and future atmospheric deposition (1880–2100) and future tree harvesting practices. Deposition was assumed to follow current legislated European emission reduction policies (CLe) and a scenario based on maximum (technically) feasible reductions (MFR). Future harvesting was assumed to shift from stem-only harvesting (SOH) to whole-tree harvesting (WTH) owing to the potential increased utilisation of biofuels. Despite the modest changes in atmospheric deposition under CLe (compared to current day), these reductions are predicted to halt the decline in soil base saturation; however, further reductions are required to improve soil and lake water chemistry. The MFR scenario predicted a significant long-term improvement in soil base saturation leading to continued long-term recovery in surface waters (all lakes with ANC > 0 by 2100). However under the WTH scenario, significant long-term impacts (re-acidification) were predicted for soil and surface water chemistry (14 lakes with ANC < 0 by 2100). To some extent the long-term negative impacts were reduced under MFR, indicating that increased utilisation of biofuels will necessitate ‘trading emissions for timber’, or soil amendment, to maintain ecosystem quality and sustainable forest growth. The current practice of SOH is close to the sustainable maximum harvesting under current (legislated) atmospheric deposition in Finland.