Development of the human heart

In 2014, an extensive review discussing the major steps of cardiac development focusing on growth, formation of primary and chamber myocardium and the development of the cardiac electrical system, was published. Molecular genetic lineage analyses have since furthered our insight in the developmental origin of the various component parts of the heart, which currently can be unambiguously identified by their unique molecular phenotype. Moreover, genetic, molecular and cell biological analyses have driven insights into the mechanisms underlying the development of the different cardiac components. Here, we build on our previous review and provide an insight into the molecular mechanistic revelations that have forwarded the field of cardiac development. Despite the enormous advances in our knowledge over the last decade, the development of congenital cardiac malformations remains poorly understood. The challenge for the next decade will be to evaluate the different developmental processes using newly developed molecular genetic techniques to further unveil the gene regulatory networks operational during normal and abnormal cardiac development.     

Gular pouch diversity in the Chamaeleonidae

Numerous chameleon species possess an out-pocketing of the trachea known as the gular pouch. After surveying more than 250 specimens, representing nine genera and 44 species, we describe two different morphs of the gular pouch. Species of the genera Bradypodion and Chamaeleo, as well as Trioceros goetzei, all possess a single gular pouch (morph one) formed from ventral expansion of soft tissue where the larynx and trachea meet. Furcifer oustaleti and Furcifer verrucosus possess from one to four gular pouches (morph two) formed by the expansion of soft tissue between sequential hyaline cartilage rings of the trachea. In Trioceros melleri, examples of both morphs of the gular pouch were observed. Morphometric data are presented for 100 animals representing eight species previously known to possess a gular pouch and two additional species, Bradypodion thamnobates and Bradypodion transvaalense. In the species with the absolutely and relatively largest gular pouch, Chamaeleo calyptratus, a significant difference was found between sexes in its width and volume, but not its length. In C. calyptratus, we show that an inflated gular pouch is in contact with numerous hyoid muscles and the tongue. Coupled with the knowledge that C. calyptratus generates vibrations from the throat region, we posit that the tongue (M. accelerator linguae and M. hyoglossus) and supporting hyoid muscles (i.e., Mm. sternohyoideus profundus et superficialis and Mm. mandibulohyoideus) are involved in the production of vibrations to produce biotremors that are amplified by the inflated gular pouch and used in substrate-borne communication.     

Improved Sensitivity for Molecular Detection of Bacterial and Candida Infections in Blood

The rapid identification of bacteria and fungi directly from the blood of patients with suspected bloodstream infections aids in diagnosis and guides treatment decisions. The development of an automated, rapid, and sensitive molecular technology capable of detecting the diverse agents of such infections at low titers has been challenging, due in part to the high background of genomic DNA in blood. PCR followed by electrospray ionization mass spectrometry (PCR/ESI-MS) allows for the rapid and accurate identification of microorganisms but with a sensitivity of about 50% compared to that of culture when using 1-ml whole-blood specimens. Here, we describe a new integrated specimen preparation technology that substantially improves the sensitivity of PCR/ESI-MS analysis. An efficient lysis method and automated DNA purification system were designed for processing 5 ml of whole blood. In addition, PCR amplification formulations were optimized to tolerate high levels of human DNA. An analysis of 331 specimens collected from patients with suspected bloodstream infections resulted in 35 PCR/ESI-MS-positive specimens (10.6%) compared to 18 positive by culture (5.4%). PCR/ESI-MS was 83% sensitive and 94% specific compared to culture. Replicate PCR/ESI-MS testing from a second aliquot of the PCR/ESI-MS-positive/culture-negative specimens corroborated the initial findings in most cases, resulting in increased sensitivity (91%) and specificity (99%) when confirmed detections were considered true positives. The integrated solution described here has the potential to provide rapid detection and identification of organisms responsible for bloodstream infections.     

Myocardial contraction and hyaluronic acid mechanotransduction in epithelial-to-mesenchymal transformation of endocardial cells

Epithelial-to-mesenchymal transition (EMT) of endocardial cells is a critical initial step in the formation of heart valves. The collagen gel in vitro model has provided significant information on the role of growth factors regulating EMT but has not permitted investigation of mechanical factors. Therefore we sought to develop a system to probe the effects of mechanical inputs on endocardial EMT by incorporating hyaluronic acid (HA), the primary component of endocardial cushions in developing heart valves, into the gel assay. This was achieved using a combination collagen and crosslinkable methacrylated HA hydrogel (Coll-MeHA). Avian atrioventricular canal explants on Coll-MeHA gels showed increased numbers of transformed cells. Analysis of the mechanical properties of Coll-MeHA gels shows that stiffness does not directly affect EMT. Hydrogel deformation from the beating myocardium of explants directly led to higher levels of regional gel deformation and larger average strain magnitudes associated with invaded cells on Coll-MeHA gels. Inhibition of this contraction reduced EMT on all gel types, although to a lesser extent on Coll-MeHA gels. Using the system we have developed, which permits the manipulation of mechanical factors, we have demonstrated that active mechanical forces play a role in the regulation of endocardial EMT.