Reconstruction of the muscle system in Antygomonas sp. (Kinorhyncha,Cyclorhagida) by means of phalloidin labeling and cLSM

In the present investigation the entire muscle system of the cyclorhagid kinorhynch Antygomonas sp. was three-dimensionally reconstructed from whole mounts by means of FITC-phalloidin labeling and confocal scanning microscopy. With this technique, which proved to be especially useful for microscopically small species, we wanted to reinvestigate and supplement the findings obtained by histological and electron microscopical methods. The organization of the major muscle systems can be summarized as follows: 1) All muscle fibers, apart from the intestinal ones, the spine, and the mouth cone muscles, show a cross-striated pattern; 2) Dorsal longitudinal muscle fibers as well as segmentally arranged dorsoventral fibers occur from segment III to XIII; 3) Diagonal muscle fibers are located laterally in segments III to X; 4) Two rings of circular fibers are present in segment II, forming the closing apparatus in Cyclorhagida. Further circular muscles are present in segment I, forming the mouth cone and the eversible introvert, and in the pharyngeal bulb.     

Muscular anatomy of the millipede Phyllogonostreptus nigrolabiatus (Diplopoda: Spirostreptida) and its bearing on the millipede "thorax"

The muscular anatomy of the millipede Phyllogonostreptus nigrolabiatus (Newport, 1844) (Diplopoda; Spirostreptida; Harpagophoridae) is comprehensively surveyed. The musculature of the first three postcollum pleurotergites, the "thorax," and their associated appendages was found to be more complex than that of the postthoracic rings. It is hypothesized that the musculature of the postthoracic segments is derived relative to that of the thoracic segments, which retain primitively free sternites and are not diplosegments. This hypothesis is discussed relative to previous hypotheses positing that the anteriormost three leg-bearing rings in millipedes are diplosegments. The musculature of spirostreptid gonopods is described in detail for the first time. Comparison of the cephalic musculature is made with previously described musculature in Julida showing that, while many aspects of the musculature are conserved, there exist interordinal differences, documenting the potential utility of comparative anatomical studies for resolving millipede phylogeny.     

Increased glycated calmodulin in the submandibular salivary glands of streptozotocin‐induced diabetic rats

Non‐enzymatic glycosylation, a post translational protein modification may be implicated in the diabetes complications. Calmodulin is an important calcium binding protein that complexed with Ca²⁺ may be implicated in salivary gland secretory process. Glycated calmodulin has shown to be less effective in binding calcium. The aim of this study was to determine whether the concentration of glycated‐calmodulin may be elevated in the submandibular salivary glands of streptozotocin‐induced diabetic rats. Diabetes was induced by an intraperitoneal injection of spreptozotocin, and hyperglycemia was confirmed 72 h after injection using a glucosimeter. Thirty days after the induction of diabetes, submandibular salivary glands were used for the analysis of glycated and non‐glycated calmodulin, using a glycogel B columns for separation. Glycated and non‐glycated calmodulin were assayed by an enzymatic method and by ELISA. The overall concentration of CaM (non‐glycated + glycated) in induced diabetic rats was significantly lower than in controls (p < 0.05). The concentration of non‐glycated CaM in controls was significantly higher than in experimental group (p < 0.05), while the concentration of glycated calmodulin between these groups was statistically similar (p > 0.05). Copyright © 2009 John Wiley & Sons, Ltd.     

Development of the swimbladder and its innervation in the zebrafish, Danio rerio

Many teleosts including zebrafish, Danio rerio, actively regulate buoyancy with a gas-filled swimbladder, the volume of which is controlled by autonomic reflexes acting on vascular, muscular, and secretory effectors. In this study, we investigated the morphological development of the zebrafish swimbladder together with its effectors and innervation. The swimbladder first formed as a single chamber, which inflated at 1-3 days posthatching (dph), 3.5-4 mm body length. Lateral nerves were already present as demonstrated by the antibody zn-12, and blood vessels had formed in parallel on the cranial aspect to supply blood to anastomotic capillary loops as demonstrated by Tie-2 antibody staining. Neuropeptide Y-(NPY-) like immunoreactive (LIR) fibers appeared early in the single-chambered stage, and vasoactive intestinal polypeptide (VIP)-LIR fibers and cell bodies developed by 10 dph (5 mm). By 18 dph (6 mm), the anterior chamber formed by evagination from the cranial end of the original chamber; both chambers then enlarged with the ductus communicans forming a constriction between them. The parallel blood vessels developed into an arteriovenous rete on the cranial aspect of the posterior chamber and this region was innervated by zn-12-reactive fibers. Tyrosine hydroxylase- (TH-), NPY-, and VIP-LIR fibers also innervated this area and the lateral posterior chamber. Innervation of the early anterior chamber was also demonstrated by VIP-LIR fibers. By 25-30 dph (8-9 mm), a band of smooth muscle formed in the lateral wall of the posterior chamber. Although gas in the swimbladder increased buoyancy of young larvae just after first inflation, our results suggest that active control of the swimbladder may not occur until after the formation of the two chambers and subsequent development and maturation of vasculature, musculature and innervation of these structures at about 28-30 dph.     

Dysregulation of the Bmi‐1/p16Ink4a pathway provokes an aging‐associated decline of submandibular gland function

Bmi‐1 prevents stem cell aging, at least partly, by blocking expression of the cyclin‐dependent kinase inhibitor p16^Ink4a. Therefore, dysregulation of the Bmi‐1/p16^Ink4a pathway is considered key to the loss of tissue homeostasis and development of associated degenerative diseases during aging. However, because Bmi‐1 knockout (KO) mice die within 20 weeks after birth, it is difficult to determine exactly where and when dysregulation of the Bmi‐1/p16^Ink4a pathway occurs during aging in vivo. Using real‐time in vivo imaging of p16^Ink4a expression in Bmi‐1‐KO mice, we uncovered a novel function of the Bmi‐1/p16^Ink4a pathway in controlling homeostasis of the submandibular glands (SMGs), which secrete saliva into the oral cavity. This pathway is dysregulated during aging in vivo, leading to induction of p16^Ink4a expression and subsequent declined SMG function. These findings will advance our understanding of the molecular mechanisms underlying the aging‐related decline of SMG function and associated salivary gland hypofunction, which is particularly problematic among the elderly.     

Differential activation of parts of the serratus anterior muscle during push-up variations on stable and unstable bases of support

No studies have examined the effects of an unstable surface on push-up and push-up plus exercises in terms of the two parts of the serratus anterior muscle. We hypothesized that the lower part of the serratus anterior would have greater activity with an unstable surface, which requires stabilizing the scapular position. The present study was performed to investigate the intramuscular differences between parts of the serratus anterior muscle during push-up and push-up plus exercises. Twelve healthy subjects were included in the study. The upper and lower parts of the serratus anterior and upper and lower parts of the trapezius were investigated by surface EMG during four types of exercise. Repeated one-way ANOVA was used for statistical analyses. Maintaining the push-up plus phase caused significant increases in EMG activity of the upper serratus anterior compared with the push-up ascending phase on both of stable and unstable bases (P < 0.05). The lower serratus anterior showed increased activation on an unstable surface, which required more joint stability than did the stable base. Upper trapezius/upper serratus anterior ratio was significantly lower in the PUP than in the PUA phase with both stable and unstable bases of support (P < 0.05).Further studies are required to investigate the intramuscular variation in activation of the serratus anterior during exercises for rehabilitation.     

A new genus from the continental slope off Brazil and the discovery of the first males in the Hirsutiidae (Crustacea: Peracarida: Bochusacea)

Montucaris distincta gen. nov. sp. nov. , a new genus and species of hirsutiid, is described from the bathyal floor of the northern South Atlantic, off Brazil. It is easily distinguished from other members of the family by the reduced, one-segmented exopods of pereopods 5 and 6 and the array of stout triangular spines present on the anterior margin of the basis and proximal endopodal segments of the third pereopod. This discovery is notable as it includes the first record of males in the family, demonstrating that hirsutiids are gonochoristic. Developmental stages including mancas, preparatory females and juvenile males were recovered, in addition to numerous brooding females and two morphs of adult males. We infer that the new genus has a life cycle involving non-feeding terminal males with regressed mouthparts. These males display a bizarre cephalic configuration, with a clearly defined transverse dorsal suture on the dorsal cephalothoracic shield just behind the articulation of mandibles, a feature present also in syncarida and in some Thermosbaenacea and that we interpret as secondary. The peracaridan affinities of the Hirsutiidae are firmly established here after settling definitively the oostegitic nature of the setose plates present on the posteromedial margin of the pereopodal coxae of brooding females. The separate ordinal rank for hirsutiids – as the Bochusacea – with respect to the Mictacea, within which they had been previously classified, is favoured here. In addition, we highlight similarities between hirustiids and tanaidaceans that warrant further investigation. © 2006 The Linnean Society of London, Zoological Journal of the Linnean Society , 2006, 148 , 169–208.     

Saliva affects the antifungal activity of exogenously added histatin 3 towards Candida albicans

Antifungal activity of histatin 3 against two Candida albicans clinical isolates was determined in assays containing rabbit submandibular gland saliva. Histatin 3 inhibited the cell growth and germination of both isolates dose-dependently (10-100 microg ml(-1)) with maximum inhibition occurring after 60 min incubation. Adding fresh histatin 3 after 60 min caused further reduction in the viable cell count. Higher histatin 3 concentrations (50-100 microg ml(-1)) and prolonged exposure to peptide were required to inhibit germination. Histatin 3 was rapidly degraded in rabbit submandibular gland saliva and this may explain why fresh addition of histatin 3 increases candidacidal activity.     

Salivary gland branching morphogenesis

Salivary gland branching morphogenesis involves coordinated cell growth, proliferation, differentiation, migration, apoptosis, and interaction of epithelial, mesenchymal, endothelial, and neuronal cells. The ex vivo analysis of embryonic mouse submandibular glands, which branch so reproducibly and beautifully in culture, is a powerful tool to investigate the molecular mechanisms regulating epithelium-mesenchyme interactions during development. The more recent analysis of genetically modified mice provides insight into the genetic regulation of branching morphogenesis. The review begins, as did the field historically, focusing on the role of the extracellular matrix (ECM), and its components such as glycosaminoglycans, collagens, and laminins. Following sections describe the modification of the ECM by proteases and the role of cell-matrix and cell-cell receptors. The review then focuses on two major families of growth factors implicated in salivary gland development, the fibroblast growth factors (FGFs) and the epidermal growth factors (EGFs). The salivary gland phenotypes in mice with genetic modification of FGFs and their receptors highlight the central role of FGFs during salivary gland branching morphogenesis. A broader section mentions other molecules implicated from analysis of the phenotypes of genetically modified mice or organ culture experiments. The review concludes with speculation on some future areas of research.