Genes of the twist family encode bHLH transcription factors known to be involved in the regulation and differentiation of early mesoderm. Here, we report our characterization of Hau-twist, a twist homolog from the leech Helobdella austinensis, a tractable lophotrochozoan representative. Hau-twist was expressed in segmental founder cells of the mesodermal lineage, in subsets of cells within the mesodermal lineage of the germinal plate, in circumferential muscle fibers of a provisional integument during segmentation and organogenesis stages and on the ventral side of the developing proboscis. Thus, consistent with other systems, our results suggest that twist gene of the leech Helobdella might function in mesoderm differentiation.

snail gene family members are zinc-finger transcription factors with key roles in morphogenesis. Involvement of snail family genes in mesoderm formation has been observed in insects and mammals. The snail genes are also involved in cell motility, neural differentiation, cell fate, survival decision, and left-right identity. The functions of snail genes have been studied primarily among ecdysozoans and deuterostomes, with relatively little work carried out in lophotrochozoans. In this study, we isolated two snail homologs (Hau-snail1 and Hau-snail2) from the leech Helobdella austinensis. We characterized the temporal and spatial expression patterns of these two genes by semi-quantitative RT-PCR and in situ hybridization. The expression of Hau-snail1 and Hau-snail2 correlates with ventral nerve cord (VNC) development, segmental mesoderm, and with a ring of cells that comes to lie at the base of the leech proboscis, respectively, showing similarity to the divergent expression of duplicated snail genes in polychaetes. Our results do not support the function of lophotrochozoan snail genes in mesoderm specification.

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The earthworm Perionyx excavatus is a species highly capable of bidirectional regeneration. Pair-rule genes are thought to have an ancestral function in arthropod segmentation. However, orthologs in annelids (i.e. Capitella teleta) do not exhibit segmental expression in the ectoderm or mesoderm. Their role in regeneration is currently unclear. Here, we report the expression profile of primary pair-rule genes (Pex-EvxA, Pex-EvxB, Pex-RuntA, Pex-RuntB, Pex-Hes1A, Pex-Hes1B, Pex-Hes4A, and Pex-Hey) found in P. excavatus using semi-quantitative reverse transcription-polymerase chain reaction. Our results indicated these genes showed variable expression during bidirectional regeneration. Six of these genes might play diverse and potentially critical roles in head and/or tail regeneration.

The finite element method (FEM) is advantageous because it can save time and cost by reducing the number of samples and experiments in the effort to identify design factors. In computational problem-solving it is necessary that the exact material properties are input for achieving a reliable analysis. However, in the case of fiber boards, it is difficult to measure their cross-directional material properties because of their small thickness. In previous research studies, the Poisson's ratio was measured by analyzing ultrasonic wave velocities. Recently, the Poisson's ratio was measured using a high-speed digital camera. In this study, we measured the transverse strain of a fiber board and calculated its Poisson's ratio using a high-speed digital camera in order to apply these estimates to a FEM analysis of a fiber board, a corrugated board, and a corrugated box. Methods: Three different fiber board samples were used in a uniaxial tensile test. The longitudinal strain was measured using the Universal Testing Machine. The transverse strain was measured using an image processing method. To calculate the transverse strain, we acquired images of the fiber board before the test onset and before the fracture occurred. Acquired images were processed using the image processing program MATLAB. After the images were converted from color to binary, we calculated the width of the fiber board. Results: The calculated Poisson's ratio ranged between 0.2968-0.4425 (Machine direction, MD) and 0.1619-0.1751 (Cross machine direction, CD). Conclusions: This study demonstrates that measurement of the transverse properties of a fiber board is possible using image processing methods. Correspondingly, these processing methods could be used to measure material properties that are difficult to measure using conventional measuring methodologies that employ strain gauge extensometers.