The occipital lateral plate mesoderm is a novel source for vertebrate neck musculature.

In vertebrates, body musculature originates from somites, whereas head muscles originate from the cranial mesoderm. Neck muscles are located in the transition between these regions. We show that the chick occipital lateral plate mesoderm has myogenic capacity and gives rise to large muscles located in the neck and thorax. We present molecular and genetic evidence to show that these muscles not only have a unique origin, but additionally display a distinct temporal development, forming later than any other muscle group described to date. We further report that these muscles, found in the body of the animal, develop like head musculature rather than deploying the programme used by the trunk muscles. Using mouse genetics we reveal that these muscles are formed in trunk muscle mutants but are absent in head muscle mutants. In concordance with this conclusion, their connective tissue is neural crest in origin. Finally, we provide evidence that the mechanism by which these neck muscles develop is conserved in vertebrates.

Flammability and comfort properties of blended knit fabrics made from inherently fire-resistant fibers to use for fire fighters.

Achievement of better comfort properties in station suits of fire fighters without compromising flame retardancy is an utmost necessity. Inherently fire-resistant fibers play vital role in this scope. In this work twenty-three plain single jersey knitted fabric samples were developed by using five inherently fire-resistant fibers. The fibers used were meta-aramid Nomex, fire resistant (FR-Viscose, modacrylic (Protex), FR-polyester (Recron) and carbon fibers. All the fibers were blended in different blend ratios. Vertical flammability test was performed to investigate the flammability properties. Thermal characterization of the samples was done using thermogravimetric analysis (TGA). Thermophysiological and wearing comfort properties of all samples were evaluated. Scanning electron microscopy (SEM) was carried out for the evaluation of surface morphology of the fibers after charring. Results of vertical flammability test revealed that sample containing 100% Nomex fibers produced minimum char length. Nomex and carbon fiber blended fabric provided better moisture management along with better flame retardant behavior. Statistical tool named as Principal Component Analysis (PCA) was utilized for the optimization of all responses. All the samples were ranked as per principal component analysis. Sample containing 50/50 FR-polyester and FR-viscose fibers was found to be the top ranked, as this sample provided optimum flammability and comfort related performance.

Tribological Behaviour of Enamel Coatings Created by a Prototype Device for Local Repair of Inorganic Surfaces.

The ability of materials to withstand environmental influences is a frequent necessity in many industries. Special requirements are imposed by such industries where surfaces are affected by acidity during the processing or storage of products. In such cases, when the basic surface is exposed to chemical influences, it is possible to use enamel coatings, which, with their properties, guarantee the protection of the surface and achieve the required service life of the material. This article deals mainly with the interaction between the base material and the enamel and its resistance to wear between the original and the renovated surface caused by local heating. The article presents a methodical procedure for the preparation of test specimens with an enamel layer prepared by AWJ cutting, eliminating its damage. There are minimal differences in the microstructure between the original and the renovated surface due to the production technique. The renovated enamel surface had more bubbles of a larger size than the original surface. Good adhesion between the base metal material (substrate) and the ground coat was demonstrated. The tested surfaces demonstrated high resistance to intensive abrasion conditions with low linear wear increments.

Mechanical and Thermo-Mechanical Performance of Natural Fiber-Based Single-Ply and 2-Ply Woven Prepregs.

This paper presents a study conducted on prepregs manufactured by a novel method for the impregnation of a thermoplastic matrix. Different composite prepregs based on polypropylene and reinforced with natural fibers (e.g., basalt and jute fibers) were developed. The mechanical and dynamic mechanical properties were investigated. DMA tests were conducted at 1 Hz frequency and properties such as storage modulus and damping (tan δ) were evaluated. The overall mechanical properties of the basalt fiber composites were found to be superior to that of the jute fiber-based samples. Thermo-gravimetric analysis (TG/DTG) of the composite samples showed that the thermal degradation temperatures of the basalt-based composites shifted to higher temperature regions compared to the PP or jute fiber composites. The addition of basalt fiber considerably improved the thermal stability of the composite samples. Microscopic images of the tensile fractured composite samples illustrated better fiber-matrix interfacial interaction due to the novel technology of prepregs. Single-ply and 2-ply prepregs showed significantly superior mechanical, thermal, and thermo-dynamical performance compared to the control sample (pure PP). 2-Ply composites demonstrated higher modulus, tensile strength, and storage modulus due to the higher fiber volume fraction. Basalt-based samples showed a minimum weight loss of about 57% up to 700 °C in contrast to 96.05% weight loss in the jute-based samples and 98.4% in the case of pure PP. The heat resistance index (THRI) is more than twice for basalt compared to jute and PP. Furthermore, the superior thermal stability of basalt is reflected in its DSC curves, showing the highest endothermic peak. The technique of using the resin in the form of thermoplastic yarns offers cost effective and efficient alternatives for composite manufacturing.

Cellular and molecular investigations into the development of the pectoral girdle.

The forelimbs of higher vertebrates are composed of two portions: the appendicular region (stylopod, zeugopod and autopod) and the less prominent proximal girdle elements (scapula and clavicle) that brace the limb to the main trunk axis. We show that the formation of the muscles of the proximal limb occurs through two distinct mechanisms. The more superficial girdle muscles (pectoral and latissimus dorsi) develop by the "In-Out" mechanism whereby migration of myogenic cells from the somites into the limb bud is followed by their extension from the proximal limb bud out onto the thorax. In contrast, the deeper girdle muscles (e.g. rhomboideus profundus and serratus anterior) are induced by the forelimb field which promotes myotomal extension directly from the somites. Tbx5 inactivation demonstrated its requirement for the development of all forelimb elements which include the skeletal elements, proximal and distal muscles as well as the sternum in mammals and the cleithrum of fish. Intriguingly, the formation of the diaphragm musculature is also dependent on the Tbx5 programme. These observations challenge our classical views of the boundary between limb and trunk tissues. We suggest that significant structures located in the body should be considered as components of the forelimb.

A hypoplastic model of skeletal muscle development displaying reduced foetal myoblast cell numbers, increased oxidative myofibres and improved specific tension capacity.

The major component of skeletal muscle is the myofibre. Genetic intervention inducing over-enlargement of myofibres beyond a certain threshold through acellular growth causes a reduction in the specific tension generating capacity of the muscle. However the physiological parameters of a genetic model that harbours reduced skeletal muscle mass have yet to be analysed. Genetic deletion of Meox2 in mice leads to reduced limb muscle size and causes some patterning defects. The loss of Meox2 is not embryonically lethal and a small percentage of animals survive to adulthood making it an excellent model with which to investigate how skeletal muscle responds to reductions in mass. In this study we have performed a detailed analysis of both late foetal and adult muscle development in the absence of Meox2. In the adult, we show that the loss of Meox2 results in smaller limb muscles that harbour reduced numbers of myofibres. However, these fibres are enlarged. These myofibres display a molecular and metabolic fibre type switch towards a more oxidative phenotype that is induced through abnormalities in foetal fibre formation. In spite of these changes, the muscle from Meox2 mutant mice is able to generate increased levels of specific tension compared to that of the wild type.

Influence of Alkali Treatment on the Microstructure and Mechanical Properties of Coir and Abaca Fibers.

Composite materials with natural fillers have been increasingly used as an alternative to synthetically produced materials. This trend is visible from a representation of polymeric composites with natural cellulose fibers in the automotive industry of the European Union. This trend is entirely logical, owing to a preference for renewable resources. The experimental program itself follows pronounced hypotheses and focuses on a description of the mechanical properties of untreated and alkali-treated natural vegetable fibers, coconut and abaca fibers. These fibers have great potential for use in composite materials. The results and discussion sections contribute to an introduction of an individual methodology for mechanical property assessment of cellulose fibers, and allows for a clear definition of an optimal process of alkalization dependent on the content of hemicellulose and lignin in vegetable fibers. The aim of this research was to investigate the influence of alkali treatment on the surface microstructure and tensile properties of coir and abaca fibers. These fibers were immersed into a 5% solution of NaOH at laboratory temperature for a time interval of 30 min, 1 h, 2 h, 3 h, 6 h, 12 h, 24 h, and 48 h, rinsed and dried. The fiber surface microstructures before and after the alkali treatment were evaluated by SEM (scanning electron microscopy). SEM analysis showed that the alkali treatment in the NaOH solution led to a gradual connective material removal from the fiber surface. The effect of the alkali is evident from the visible changes on the surface of the fibers.

Somitic origin of the medial border of the mammalian scapula and its homology to the avian scapula blade.

The scapula is the main skeletal element of the pectoral girdle allowing muscular fixation of the forelimb to the axial skeleton. The vertebrate limb skeleton has traditionally been considered to develop from the lateral plate mesoderm, whereas the musculature originates from the axial somites. However, in birds, the scapular blade has been shown to develop from the somites. We investigated whether a somitic contribution was also present in the mammalian scapula. Using genetic lineage-tracing techniques, we show that the medial border of the mammalian scapula develops from somitic cells. The medial scapula border serves as the attachment site of girdle muscles (serratus anterior, rhomboidei and levator scapulae). We show that the development of these muscles is independent of the mechanism that controls the formation of all other limb muscles. We suggest that these muscles be specifically referred to as medial girdle muscles. Our results establish the avian scapular blade and medial border of the mammalian scapula as homologous structures as they share the same developmental origin.

Neuromuscular junction morphology, fiber-type proportions, and satellite-cell proliferation rates are altered in MyoD(-/-) mice.

Gene compensation by members of the myogenic regulatory factor (MRF) family has been proposed to explain the apparent normal adult phenotype of MyoD(-/-) mice. Nerve and field stimulation were used to investigate contraction properties of muscle from MyoD(-/-) mice, and molecular approaches were used to investigate satellite-cell behavior. We demonstrate that MyoD deletion results in major alterations in the organization of the neuromuscular junction, which have a dramatic influence on the physiological contractile properties of skeletal muscle. Second, we show that the lineage progression of satellite cells (especially initial proliferation) in the absence of MyoD is abnormal and linked to perturbations in the nuclear localization of beta-catenin, a key readout of canonical Wnt signaling. These results show that MyoD has unique functions in both developing and adult skeletal muscle that are not carried out by other members of the MRF family.

The Influence of Modification with Natural Fillers on the Mechanical Properties of Epoxy Adhesive Compositions after Storage Time.

This article presents the initial test results examining basic technological factors, such as type of modifying agent and seasoning time, which influence properties of adhesive epoxide compositions. The aim of the study was to prepare adhesive compositions with 2% content of the selected natural fillers (montmorillonite NanoBent ZR-2, ground chalk (powder)-CaCO3, and activated carbon powder C) and to examine their strength properties. A polymeric matrix used to prepare an adhesive composition consisted of the epoxide resins used in industry: Epidian 5 and Epidian 53 cured by addition of an aminomethyl group, where curing occurred through the Mannich reaction. A composition of epoxide resins with a curing agent and without any modifying agents was used as reference. The examinations described in the present article aimed to show the significance of the impact of the fillers used on the strength properties of the examined compositions. A fracture surface of epoxide adhesive compositions modified with the selected fillers was tested by means of a scanning electron microscope.

Quasi-Static Tests of Hybrid Adhesive Bonds Based on Biological Reinforcement in the Form of Eggshell Microparticles.

The paper is focused on the research of the cyclic loading of hybrid adhesive bonds based on eggshell microparticles in polymer composite. The aim of the research was to characterize the behavior of hybrid adhesive bonds with composite adhesive layer in quasi-static tests. An epoxy resin was used as the matrix and microparticles of eggshells were used as the filler. The adhesive bonds were exposed to cyclic loading and their service life and mechanical properties were evaluated. Testing was performed by 1000 cycles at 5-30% (165-989 N) and 5-70% (165-2307 N) of the maximum load of the filler-free bond in the static test. The results of the research show the importance of cyclic loading on the service life and mechanical properties of adhesive bonds. Quasi-static tests demonstrated significant differences between measured intervals of cyclic loading. All adhesive bonds resisted 1000 cycles of the quasi-static test with an interval loading 5-30%. The number of completed quasi-static tests with the interval loading 5-70% was significantly lower. The filler positively influenced the service life of adhesive bonds at a higher amount of quasi-static tests, i.e., the safety of adhesive bonds increased. The filler had a positive effect on adhesive bonds ABF2, where the strength significantly increased up to 20.26% at the loading of 5-30% against adhesive bonds ABF0. A viscoelasticity characteristic (creep) of the adhesive layer occurred at higher values of loading, i.e., between loading 5-70%. The viscoelasticity behavior did not occur at lower values of loading, i.e., between loading 5-30%.

Material Utilization of Cotton Post-Harvest Line Residues in Polymeric Composites.

This paper deals with a research focused on utilization of microparticle and short-fiber filler based on cotton post-harvest line residues in an area of polymeric composites. Two different fractions of the biological filler (FCR-reinforced cotton filler) of 20 and 100 µm and the filler with short fibers of a length of 700 µm were used in the research. The aim of the research was to evaluate mechanical characteristics of composites and adhesive bonds for the purpose of gaining new pieces of knowledge which will be applicable in the area of material engineering and assessing application possibilities of residues coming into being from agricultural products processing. Mechanical properties of the composite material produced by a vacuum infusion and tested at temperatures 20, 40, and 60 °C and adhesive bonds which were exposed to a low-cyclic loading, i.e., 1000 cycles at 30% to 70% from reference value of the maximum strength, were evaluated. Composite systems with the FCR adjusted in 5% water solution of NaOH showed higher strength values on average compared to untreated FCR. Unsuitable size of the FCR led to a deterioration of the strength. The filler in the form of 700 FCR microfibers showed itself in a positive way to composite materials, and the particle in the form of 20 FCR did the same to adhesive bonds. Results of adhesive bond cyclic tests at higher stress values (70%) demonstrated viscoelastic behavior of the adhesive layer.

The hypaxial origin of the epaxially located rhomboid muscles.

In vertebrates, skeletal muscles of the body are made up of epaxial and hypaxial muscles based on their innervation and relative position to the vertebral column. The epaxial muscles are innervated by the dorsal branches of the spinal nerves and comprise the intrinsic (deep) back muscles, while the hypaxial muscles are innervated by the ventral branches of the spinal nerves including the plexus and consist of a heterogeneous group of intercostal, abdominal, and limb as well as girdle muscles. The canonical view holds that the epaxial muscles are derived from the medial halves of the somites, whereas the hypaxial muscles are all derived from the lateral somitic halves. The rhomboid muscles are situated dorsal to the vertebral column and therefore in the domain typically occupied by epaxial muscles. However, they are innervated by a ventral branch of the brachial plexus called the N. dorsalis scapulae. Due to the apparent inappropriate position of the muscle in relation to its innervation we investigated its origin to help clarify this issue. To study the embryonic origin of the rhomboid muscles, we followed derivatives of the medial and lateral somite halves using quail-chick chimeras. Our results showed that the rhomboid muscles are made up of cells derived mainly from the lateral portion of the somite. Therefore the rhomboid muscles which lie within the epaxial domain of the body, originate from the hypaxial domain of the somites. However their connective tissue is derived from both medial and lateral somites.

Skeletal muscle translocation in vertebrates.

It is now over 30 years since Bodo Christ first demonstrated that the musculature of the limb originated from the somites and overturned the then prevailing view that limb muscle develops from a local source. Subsequently, using electron microscopy and histological procedures, Bodo Christ identified that cells of the somites undergo an epithelial to mesenchymal transition which enabled them to move from their paraxial point of origin to distal locations. These studies defined this translocation as one of the major mechanisms allowing myogenic cells to translocate around the body. The other means used to translocate muscle involves the movement of cells as a sheet. The deployment of one of these two mechanisms has been postulated to be involved in the formation of all the hypaxial musculature of the vertebrate body. In this paper we describe the formation of muscles both in the head and in the body, which use a translocatory mechanism during their development. We highlight recent data showing that muscle translocation is a far more complex process than first thought but which in itself can be used as a valuable tool to address questions regarding tissue patterning and development.

Expression and regulation of Nkd-1, an intracellular component of Wnt signalling pathway in the chick embryo.

The Wnt family of secreted signalling molecules control a wide range of developmental processes in all metazoans. The intracellular response to Wnt signalling depends on the choice of signalling cascade activated in the responding cell. Cells can activate either the canonical pathway that modulates gene expression to control cellular differentiation and proliferation, or the non-canonical pathway that controls cell polarity and movement. Recent work has identified the protein Naked Cuticle to act as an intracellular switch to promote the non-canonical pathway at the expense of the canonical pathway. We have cloned chick Naked Cuticle-1 (cNkd-1) and show that it is expressed in a dynamic manner during early embryogenesis. We show that it is expressed in the somites and in particular regions where cells are undergoing movement. Lastly, we show that the expression of cNkd-1 is regulated by Wnt expression originating from the neural tube. This study provides evidence that non-canonical Wnt signalling plays a part in somite development.

Characterisation of Development and Electrophysiological Mechanisms Underlying Rhythmicity of the Avian Lymph Heart.

Despite significant advances in tissue engineering such as the use of scaffolds, bioreactors and pluripotent stem cells, effective cardiac tissue engineering for therapeutic purposes has remained a largely intractable challenge. For this area to capitalise on such advances, a novel approach may be to unravel the physiological mechanisms underlying the development of tissues that exhibit rhythmic contraction yet do not originate from the cardiac lineage. Considerable attention has been focused on the physiology of the avian lymph heart, a discrete organ with skeletal muscle origins yet which displays pacemaker properties normally only found in the heart. A functional lymph heart is essential for avian survival and growth in ovo. The histological nature of the lymph heart is similar to skeletal muscle although molecular and bioelectrical characterisation during development to assess mechanisms that contribute towards lymph heart contractile rhythmicity have not been undertaken. A better understanding of these processes may provide exploitable insights for therapeutic rhythmically contractile tissue engineering approaches in this area of significant unmet clinical need. Here, using molecular and electrophysiological approaches, we describe the molecular development of the lymph heart to understand how this skeletal muscle becomes fully functional during discrete in ovo stages of development. Our results show that the lymph heart does not follow the normal transitional programme of myogenesis as documented in most skeletal muscle, but instead develops through a concurrent programme of precursor expansion, commitment to myogenesis and functional differentiation which offers a mechanistic explanation for its rapid development. Extracellular electrophysiological field potential recordings revealed that the peak-to-peak amplitude of electrically evoked local field potentials elicited from isolated lymph heart were significantly reduced by treatment with carbachol; an effect that could be fully reversed by atropine. Moreover, nifedipine and cyclopiazonic acid both significantly reduced peak-to-peak local field potential amplitude. Optical recordings of lymph heart showed that the organ's rhythmicity can be blocked by the HCN channel blocker, ZD7288; an effect also associated with a significant reduction in peak-to-peak local field potential amplitude. Additionally, we also show that isoforms of HCN channels are expressed in avian lymph heart. These results demonstrate that cholinergic signalling and L-type Ca2+ channels are important in excitation and contraction coupling, while HCN channels contribute to maintenance of lymph heart rhythmicity.

Nonmyocytic androgen receptor regulates the sexually dimorphic development of the embryonic bulbocavernosus muscle.

The bulbocavernosus (BC) is a sexually dimorphic muscle observed only in males. Androgen receptor knockout mouse studies show the loss of BC formation. This suggests that androgen signaling plays a vital role in its development. Androgen has been known to induce muscle hypertrophy through satellite cell activation and myonuclei accretion during muscle regeneration and growth. Whether the same mechanism is present during embryonic development is not yet elucidated. To identify the mechanism of sexual dimorphism during BC development, the timing of morphological differences was first established. It was revealed that the BC was morphologically different between male and female mice at embryonic day (E) 16.5. Differences in the myogenic process were detected at E15.5. The male BC possesses a higher number of proliferating undifferentiated myoblasts. To identify the role of androgen signaling in this process, muscle-specific androgen receptor (AR) mutation was introduced, which resulted in no observable phenotypes. Hence, the expression of AR in the BC was examined and found that the AR did not colocalize with any muscle markers such as Myogenic differentiation 1, Myogenin, and paired box transcription factor 7. It was revealed that the mesenchyme surrounding the BC expressed AR and the BC started to express AR at E15.5. AR mutation on the nonmyocytic cells using spalt-like transcription factor 1 (Sall1) Cre driver mouse was performed, which resulted in defective BC formation. It was revealed that the number of proliferating undifferentiated myoblasts was reduced in the Sall1 Cre:AR(L-/Y) mutant embryos, and the adult mutants were devoid of BC. The transition of myoblasts from proliferation to differentiation is mediated by cyclin-dependent kinase inhibitors. An increased expression of p21 was observed in the BC myoblast of the Sall1 Cre:AR(L-/Y) mutant and wild-type female. Altogether this study suggests that the nonmyocytic AR may paracrinely regulate the proliferation of myoblast possibly through inhibiting p21 expression in myoblasts of the BC.

Limitations of the MRL mouse as a model for cardiac regeneration.

OBJECTIVE: Myocardial repair following injury in mammals is restricted such that damaged areas are replaced by scar tissue, impairing cardiac function. MRL mice exhibit exceptional regenerative healing in an ear punch wound model. Some myocardial repair with restoration of heart function has also been reported following cryoinjury. Increased cardiomyocyte proliferation and a foetal liver stem cell population were implicated. We investigated molecular mechanisms facilitating myocardial repair in MRL mice to identify potential therapeutic targets in non-regenerative species. METHODS: Expressions of specific cell-cycle regulators that might account for regeneration (CDKs 1, 2, 4 and 6; cyclins A, E, D1 and B1; p21, p27 and E2F5) were compared by immunoblotting in MRL and control C57BL/6 ventricles during development. Flow cytometry was used to investigate stem cell populations in livers from foetal mice, and infarct sizes were compared in coronary artery-ligated and sham-treated MRL and C57BL/6 adult mice. KEY FINDINGS: No differences in the expressions of cell cycle regulators were observed between the two strains. Expressions of CD34+Sca1+ckit-, CD34+Sca1+ckit+ and CD34+Sca1-ckit+ increased in livers from C57BL/6 vs MRL mice. No differences were observed in infarct sizes, levels of fibrosis, Ki67 staining or cardiac function between MRL and C57BL/6 mice. CONCLUSIONS: No intrinsic differences were observed in cell cycle control molecules or stem cell populations between MRL and control C57BL mouse hearts. Pathophysiologically relevant ischaemic injury is not repaired more efficiently in MRL myocardium, questioning the use of the MRL mouse as a reliable model for cardiac regeneration in response to pathophysiologically relevant forms of injury.

Canonical Wnt signalling induces satellite-cell proliferation during adult skeletal muscle regeneration.

Satellite cells represent the stem cell population of adult skeletal muscle. The molecular mechanisms that control the proliferation of satellite cells are not well understood. In this study, we show that in response to injury, myofibres activate Wnt ligand transcription and activate a reporter cell line that is sensitive to the canonical Wnt-signalling pathway. Activated satellite cells on isolated cultured myofibres show robust expression of activated-beta-catenin (Act-beta-Cat), a key downstream transcriptional coactivator of canonical Wnt signalling. We provide evidence that the Wnt family of secreted glycoproteins act on satellite cells in a ligand-specific manner. Overexpression of Wnt1, Wnt3a or Wnt5a protein causes a dramatic increase in satellite-cell proliferation. By contrast, exposure of satellite cells to Wnt4 or Wnt6 diminishes this process. Moreover, we show that the prolonged satellite-cell quiescence induced by inhibitory Wnt is reversible and exposing inhibited satellite cells to stimulatory Wnt signalling restores their proliferation rate. Stimulatory Wnt proteins induce premature satellite cell BrdU incorporation as well as nuclear translocation of Act-beta-Cat. Finally, we provide evidence that the Act-beta-Cat translocation observed in single fibres during in vitro culture also occurs in cases of acute and chronic skeletal muscle regeneration in rodents and humans. We propose that Wnt proteins may be key factors that regulate the rate of satellite-cell proliferation on adult muscle fibres during the wound-healing response.

Lymph heart in chick--somitic origin, development and embryonic oedema.

The lymph heart is a sac-like structure on either side of avian tail. In some adult birds, it empties the lymph from the copulatory organ; however, during embryonic development, it is thought to circulate extra-embryonic lymph. Very little is known about the origin, innervation and the cellular changes it undergoes during development. Using immunohistochemistry and gene expression profiling we show that the musculature of the lymph heart is initially composed solely of striated skeletal muscle but later develops an additional layer composed of smooth myofibroblasts. Chick-quail fate-mapping demonstrates that the lymph heart originates from the hypaxial compartments of somites 34-41. The embryonic lymph heart is transiently innervated by somatic motoneurons with no autonomic input. In comparison to body muscles, the lymph heart has different sensitivity to neuromuscular junction blockers (sensitive only to decamethonium). Furthermore, its abundant bungarotoxin-positive acetylcholinesterase receptors are unique as they completely lack specific acetylcholinesterase activity. Several lines of evidence suggest that the lymph heart may possess an intrinsic pacing mechanism. Finally, we assessed the function of the lymph heart during embryogenesis and demonstrate that it is responsible for preventing embryonic oedema in birds, a role previously thought to be played by body skeletal muscle contractions.