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Besides their ecological importance, epiphytic species of Orchidaceae play economic and social roles through their commercialization and some are at great risk of extinction. The objectives of this study were to characterize the leaf and root anatomy of fourteen epiphytic Orchidaceae species, which occur in indigenous territory in the Parque Estadual da Serra do Tabuleiro (P.E.S.T.), Santa Catarina, Brazil and to identify adaptive anatomical characteristics related to the epiphytic habit. The species are commercialized by the Guarani and were collected during interviews and guided tours with Guarani in the indigenous territory. The results reveal the species have many morphoanatomical structures that are useful during water shortages resulting from the epiphytic habit. Notable characteristics are related to reserving water (i.e., pseudobulbs and a hypodermis with water-storage cells) and resistance to desiccation in the leaf (i.e., conspicuous cuticle, suprastomatic chamber and extraxylary and pericyclic fibers) and root (i.e., tilosomes and/or exodermal thickening and cortex cells with phi thickenings or sclereids). Descriptions and the identification of adaptive characteristics of epiphytic plant species are useful for conservation and cultivation studies, especially for plants commercially used by the Guarani Indians.

Calcium is one of the most poorly reutilized nutrients. Its deficiencies cause various physiological disturbances and, consequently, reduce the quantity and quality of yields. Reduced content of Ca2+ ions in cells leads to development of, e.g., bitter pit in apples. Efficient and instantaneous mitigation of Ca2+ deficiencies is provided by foliar feeding. There are no detailed data on the effect of foliar feeding with various calcium forms on the cell structure or on the microanalysis and mapping of this element in apple fruit cells. Therefore, we carried out comparative studies of the ultrastructure of epidermis and hypodermis cells, to assess the content and distribution of calcium in the cell wall, cytoplasmic membrane, cytoplasm, and precipitates of Malus domestica Borkh. ‘Szampion’ fruit exposed to four Ca treatments, including the control with no additional Ca supplementation (I) and foliar applications of Ca(NO3)2 (II), CaCl2 (III), and Ca chelated with EDTA (IV). Light and transmission electron microscopy and an X-ray microanalyzer were used and showed a beneficial effect of calcium preparations on the ultrastructure of fruit epidermis and hypodermis cells, manifested in the presence of a normally developed cell wall with a regular middle lamella, preserved continuity of cytoplasmic membranes, and stabilized cell structure. In the selected elements of apical epidermis cells, the highest level of Ca2+ ions was detected in the middle lamella, cell wall, plasmalemma, and cytoplasm. The highest increase in the Ca2+ content in these cell constituents was recorded in treatment IV, whereas the lowest value of the parameters was noted in variant III.

Целью настоящего исследования явилось изучения данных литературы о возрастных изменениях кожи лица, а также сравнительный анализ показателей ультразвукового исследования эпидермиса, дермы и гиподермы в разных анатомических зонах лица у женщин молодого и старшего возраста. Материалы и методы исследования. Был выполнен систематический обзор отечественных и зарубежных исследований. Проведено измерение толщины разных слоев кожи (эпидермис, дерма, гиподерма) в области межбровной зоны, подбородка, носогубной складки и щек у 52 пациенток в возрасте от 15 до 75 лет (средний возраст 44,3 ± 11,9 лет) при помощи ультразвукового сканера Mindray DC-8. Исследование осуществлялось на базе кафедры фундаментальной медицины Балтийского федерального университета им. Канта. Статистическая обработка полученных результатов исследования проводилась в программном обеспечении Microsoft Excel 2017 с использованием средних значений ( М ), достоверность определялась при помощи критерия Стьюдента, а ее критический уровень в данном исследовании принимали равным 0,05. Результаты и их обсуждение. Описанные в обзоре литературы исследования отечественных и зарубежных авторов свидетельствуют об изменении с возрастом структуры и качества всех слоев кожи лица у женщин, особенно дремы. При проведении нашего исследования было выявлено утолщение кожного покрова различных анатомических зон лица у женщин после 40 лет. Установлено утолщение дермы у пациенток после 40 лет во всех анатомических зонах лица с достижением статистически значимого различия в области носогубных складок (p = 0,027) и щек ( p = 0,01). Заключение. До настоящего времени уделялось мало внимания изучению возрастных изменений кожи лица при помощи ультразвука, поэтому не существует стандартных параметров оценки ее толщины, что требует продолжения исследования., The purpose of current investigation was to study literature data on age-related changes in facial skin and a comparative analysis of the epidermis, dermis and hypodermis ultrasound parameters in different anatomical areas of the face in young and older women. Materials and methods. A systematic review of local and foreign studies was carried out. The thickness of different skin layers (epidermis, dermis, hypodermis) was measured in the area of the interbrow zone, chin, nasolabial folds and cheeks (salivary gland) in 52 patients aged 15 to 75 years (mean age 44.3±11.9 years) using an ultrasound scanner Mindray DC-8. The study was carried out on the basis of the Department of Fundamental Medicine of the Baltic Federal University named after Kant. Statistical processing of the results of the study was performed in Microsoft Excel 2017 using average values (M), the reliability was determined using Student's criterion, and its critical level in this study was taken to be 0.05. Results and its discussion. Literature review results indicate a change in the structure and quality of all layers of the facial skin in women, especially naps, with age. During our study, thickening of the skin of various anatomical areas of the face in women after 40 years was revealed. A thickening of the dermis was established in patients after 40 years in all anatomical areas of the face with the achievement of a statistically significant difference in the nasolabial folds (p = 0.027) and cheeks (p = 0.01). Conclusion. In present, not much attention has been put to the ultrasound investigation pf age-related changes in facial skin; therefore, there are no standard parameters for assessing its thickness, which requires further research.

Establishing species limits in clades in Croton using characters from external morphology and common molecular markers has proved cumbersome, especially in Croton section Cyclostigma, a group of 50 Neotropical arborescent species commonly known as dragon's blood. Given this, we explored leaf and shoot apex anatomical characters for their utility in distinguishing species or groups of species in this section. We analysed 90 specimens belonging to section Cyclostigma and 14 specimens from related groups (sections Adenophylli, Cupreati and Sampatik), recording and describing 45 qualitative leaf characters. These characters were assembled into a matrix and analysed using statistical clustering methods based on similarity. Our results show that trichomes are one of the most diverse and variable anatomical features among the studied specimens. Novel anatomical features for Croton include the presence of a hypodermis and two previously unreported types of non-glandular trichomes. Although we did not detect a single anatomical character uniting section Cyclostigma, combinations of anatomical characters were useful to establish species limits and taxonomic identities within this section., ACF thanks FAPEMIG for the scholarship in Brazil during her doctoral degree, the CAPES foundation through PDSE (99999.009518/2014-02) supporting her stay at the Real Jardín Botánico-CSIC (Madrid) and the Programa de Capacitação Institucional (MPEG/MCTI) for the current CNPq research grant. RMSAM was supported by CNPq (477867/2013–8). RR was supported by a Visiting Scholar Fellowship to Brazil funded by FAPEMIG (CRA-BPV-00043-14) and CNPq research grant (477867/2013–8). Additional support was provided by Centro de Ciências Ambientais Floresta Escola (SECTES/UNESCO/HidroEX/FAPEMIG). We are grateful to two reviewers and the associate editor for their valuable suggestions and critical comments to earlier versions of this manuscrip

Rhizophora mangle L. is one of the most distributed species of neotropical mangroves. The species exhibits great phenological variability that is associated with saline concentrations of the sediment where it grows. Among the organs that are most affected by interstitial and tidal water salinity concentrations are the leaves. Since the hypersalinity generates water deficiency, it changes photosynthetic and hydraulic processes of the plant. To understand the relationship between the variation in leaf blade parameters and the water stress generated by salinity in two growth stages, morphoanatomical functional traits were quantified in leaves of juveniles and adults of R. mangle in three structurally different mangrove forests with different ranges of natural salinity (Oligohaline: 5.8–11.7 practical salinity units (PSU), Euhaline: 9.2–35.6 PSU and 23.9–47.7 PSU). We hypothesized that water stress caused by salinity generates modification in conductivity, water-storage, and photosynthetic tissues. Our results showed a greater number of morphoanatomical traits affected by salinity in juveniles compared to adults, greater variability in the traits associated with water accumulation and transport. Adults and juveniles subjected to higher values of salinity had traits more tolerant of variability in this factor, allowing superior adaptation to environments with high water deficit than individuals originating in oligohaline environments. This difference in adaptability to salinity between populations of R. mangle may imply different responses to climate change, where populations of oligohaline origin will be more susceptible to hypersalinization resulting from this phenomenon, while populations of euhaline origin could more effectively tolerate the aquatic stresses caused, allowing a prolongation of their permanence and the provision of their ecosystem services over time.

Rhizophora mangle, one of the main neotropic mangrove species, has wide phenological variability associated with soil salinity. Since global warming is one of the main drivers of changes in salinity, understanding the influence of this variable at the species level would help improve the prediction of climate change in the ecological services provided by mangroves. To understand the physiological and/or anatomical responses to water stress generated by edaphic salinity and its relationship with phenological and structural diversity, we quantified the functional traits of leaf tissue subjected to a cross-seeding experiment between two forests with different ranges of natural salinity (0–18 PSU1 and 20 to 70 PSU). A total of 180 propagules, 90 native and 90 from the other forest, were planted in each forest. Every three months for a year, soil salinity and growth, adaptability, and survival of propagules that were transformed into seedlings were measured. The traits evaluated between the two saline regimes presented significant differences, as stated in the working hypothesis. Likewise, there were modifications in the hypodermis and the xylem vessels in the exchanged seedlings, tissues related to water storage, and conduction. These responses allowed native euhaline forest seedlings to grow in oligohaline. The opposite occurred with seedlings originating in low salinities that did not survive in high salinities. Differences in adaptability between populations of R. mangle subjected to ranges of contrasting salinity may imply changes at the structural level, zoning, and abundance of the species front to the climate change processes., Salinity; Hypodermis; Foliar mesophyll; Xylem vessels; Cavitation.

The intracellular level of fatty aldehydes is tightly regulated by aldehyde dehydrogenases to minimize the formation of toxic lipid and protein adducts. Importantly, the dysregulation of aldehyde dehydrogenases has been implicated in neurologic disorder and cancer in humans. However, cellular responses to unresolved, elevated fatty aldehyde levels are poorly understood. Here, we report that ALH-4 is a C. elegans aldehyde dehydrogenase that specifically associates with the endoplasmic reticulum, mitochondria and peroxisomes. Based on lipidomic and imaging analysis, we show that the loss of ALH-4 increases fatty aldehyde levels and reduces fat storage. ALH-4 deficiency in the intestine, cell-nonautonomously induces NHR-49/NHR-79-dependent hypodermal peroxisome proliferation. This is accompanied by the upregulation of catalases and fatty acid catabolic enzymes, as indicated by RNA sequencing. Such a response is required to counteract ALH-4 deficiency since alh-4; nhr-49 double mutant animals are sterile. Our work reveals unexpected inter-tissue communication of fatty aldehyde levels and suggests pharmacological modulation of peroxisome proliferation as a therapeutic strategy to tackle pathology related to excess fatty aldehydes., Author summary Fatty aldehydes are generated during the turnover of membrane lipids and when cells are under oxidative stress. Because excess fatty aldehydes form toxic adducts with proteins and lipids, their levels are tightly controlled by a family of aldehyde dehydrogenases whose dysfunction has been implicated in genetic disease and cancer in humans. Here, we characterize mutant C. elegans that lack a conserved, membrane-associated aldehyde dehydrogenase ALH-4. Despite elevated levels of fatty aldehydes, these mutant worms survive by increasing the abundance of peroxisomes, which are important organelles for lipid metabolism. Such peroxisome proliferative response depends on the activation of transcription factors NHR-49 and NHR-79, via putative endocrine signals. Accordingly, the fertility of alh-4 mutant worms relies on NHR-49. Our work suggests a latent mechanism that may be activated during aldehyde dehydrogenase deficiency.

Objective High frequency ultrasound allows visualization of epidermis, dermis and hypodermis, precise measurement of skin thickness, as well as assessment of skin oedema, fibrosis and atrophy. The aim of this pilot cross-sectional observational study was to assess the performance and multiobserver variability of ultra-high-frequency (UHF) (50 MHz) ultrasound (US) in measuring skin thickness as well as the capacity of UHF-derived skin features to differentiate SSc patients from healthy controls. Methods Twenty-one SSc patients (16 limited and five diffuse SSc) and six healthy controls were enrolled. All subjects underwent US evaluation by three experts at three anatomical sites (forearm, hand and finger). Dermal thickness was measured and two rectangular regions of interest, one in dermis and one in hypodermis, were established for texture feature analysis. Results UHF-US allowed a precise identification and measurement of the thickness of the dermis. The dermal thickness in the finger was significantly higher in patients than in controls (P < 0.05), while in the forearm it was significantly lower in patients than in controls (P < 0.001). Interobserver variability for dermal thickness was good to excellent [forearm intraclass correlation coefficient (ICC) = 0.754; finger ICC = 0.699; hand ICC = 0.602]. Texture computed analysis of dermis and hypodermis was able to discriminate between SSc and healthy subjects (area under the curve >0.7). Conclusion These preliminary data show that skin UHF-US allows a very detailed imaging of skin layers, a reliable measurement of dermal thickness, and a discriminative capacity between dermis and hypodermis texture features in SSc and healthy subjects.

The rhamnose biosynthetic pathway, which is highly conserved across nematode species, was characterized in the nematode Caenorhabditis elegans. The pathway is up-regulated before each larval molt, suggesting that rhamnose biosynthesis plays a role in cuticle or surface coat synthesis., L-Rhamnose is a common component of cell-wall polysaccharides, glycoproteins and some natural products in bacteria and plants, but is rare in fungi and animals. In the present study, we identify and characterize a biosynthetic pathway for dTDP-rhamnose in Caenorhabditis elegans that is highly conserved across nematode species. We show that RML-1 activates glucose 1-phosphate (Glc-1-P) in the presence of either dTTP or UTP to yield dTDP-glucose or UDP-glucose, respectively. RML-2 is a dTDP-glucose 4,6-dehydratase, converting dTDP-glucose into dTDP-4-keto-6-deoxyglucose. Using mass spectrometry and NMR spectroscopy, we demonstrate that coincubation of dTDP-4-keto-6-deoxyglucose with RML-3 (3,5-epimerase) and RML-4 (4-keto-reductase) produces dTDP-rhamnose. RML-4 could only be expressed and purified in an active form through co-expression with a co-regulated protein, RML-5, which forms a complex with RML-4. Analysis of the sugar nucleotide pool in C. elegans established the presence of dTDP-rhamnose in vivo. Targeting the expression of the rhamnose biosynthetic genes by RNAi resulted in significant reductions in dTDP-rhamnose, but had no effect on the biosynthesis of a closely related sugar, ascarylose, found in the ascaroside pheromones. Therefore, the rhamnose and ascarylose biosynthetic pathways are distinct. We also show that transcriptional reporters for the rhamnose biosynthetic genes are expressed highly in the embryo, in the hypodermis during molting cycles and in the hypodermal seam cells specifically before the molt to the stress-resistant dauer larval stage. These expression patterns suggest that rhamnose biosynthesis may play an important role in hypodermal development or the production of the cuticle or surface coat during molting.

Differentiation of secretory cells leads to sharp increases in protein synthesis, challenging endoplasmic reticulum (ER) proteostasis. Anticipatory activation of the unfolded protein response (UPR) prepares cells for the onset of secretory function by expanding the ER size and folding capacity. How cells ensure that the repertoire of induced chaperones matches their postdifferentiation folding needs is not well understood. We find that during differentiation of stem-like seam cells, a typical UPR target, the Caenorhabditis elegans immunoglobulin heavy chain-binding protein (BiP) homologue Heat-Shock Protein 4 (HSP-4), is selectively induced in alae-secreting daughter cells but is repressed in hypodermal daughter cells. Surprisingly, this lineage-dependent induction bypasses the requirement for UPR signaling. Instead, its induction in alae-secreting cells is controlled by a specific developmental program, while its repression in the hypodermal-fated cells requires a transcriptional regulator B-Lymphocyte–Induced Maturation Protein 1 (BLMP-1/BLIMP1), involved in differentiation of mammalian secretory cells. The HSP-4 induction is anticipatory and is required for the integrity of secreted alae. Thus, differentiation programs can directly control a broad-specificity chaperone that is normally stress dependent to ensure the integrity of secreted proteins., A study in the nematode Caenorhabditis elegans shows that dedicated developmental programs can bypass the requirements for the unfolded protein response during the differentiation of secretory cells, anticipating their future high folding needs., Author summary During differentiation, cells that specialize in secretion of proteins, such as antibody-secreting B cells, prepare for the onset of secretory function by expanding the size of the major secretory organelle, the endoplasmic reticulum (ER), and by increasing the expression of molecular chaperones and folding enzymes. This pre-emptive expansion of the ER depends on activation of the ER stress response pathways and is required for the secretory phenotype. In addition, cells may also need to up-regulate a selected subset of chaperones because different secreted proteins may require different chaperones for their folding and secretion. Except in specialized cases, how this selective up-regulation is achieved, and whether it depends on the ER stress pathways, is not well understood. Using Caenorhabditis elegans, we find that a chaperone BiP/HSP-4, which is usually induced in most cells by stress, is selectively induced during differentiation of stem cells into the alae-secreting cells while being repressed in their sister lineage, the hypodermal cells. We find that induction of this chaperone is independent of the known ER stress pathways, while its repression requires a known regulator of development in mammals, BLIMP1/BLMP-1. The pre-emptive induction of BiP/HSP-4 is important for the integrity of secreted alae and cuticle, suggesting that a general molecular chaperone that is a canonical target of ER stress pathways can be selectively regulated by development to ensure the quality of secreted proteome and functionality of the cells postdifferentiation.

Variability of eight morpho-anatomical characters of 2-year-old needles of the Bosnian pine (Pinus heldreichii Christ.), collected from natural populations in Montenegro (Lovćen, Zeletin and Bjelasica) and Serbia (Zlatibor-Pešter) were investigated. The needles have 0–13 resin ducts, predominantly of the medial type (settled in mesophyll tissue). The average values were as follows: 7.86 cm (needle length), 1.44 mm (needle width), 0.85 mm (needle thickness), 20.82 μm (cuticle+epidermis thickness), 21.08 μm (height of hypodermal cells), 3.31 (number of hypodermis layers), 4 (number of resin ducts) (Formula presented.) 57.05 μm (resin duct diameter). Needle width and thickness were the most variable characters. Principal component analysis (Formula presented.) separation of the geographically most distant populations Lovćen and Zlatibor-Pešter from themselves and from other populations. Cluster analysis suggests biggest similarities between populations Lovćen and Bjelasica and biggest differences between Lovćen and Zlatibor-Pešter. Plant Biosystems - An International Journal Dealing with all Aspects of Plant Biology (2016), 150(2): 254-263

Heme-containing peroxidases are important components of innate immunity. Many of them functionally associate with NADPH oxidase (NOX)/dual oxidase (DUOX) enzymes by using the hydrogen peroxide they generate in downstream reactions. Caenorhabditis elegans encodes for several heme peroxidases, and in a previous study we identified the ShkT-containing peroxidase, SKPO-1, as necessary for pathogen resistance. Here, we demonstrated that another peroxidase, HPX-2 (Heme-PeroXidase 2), is required for resistance against some, but not all pathogens. Tissue specific RNA interference (RNAi) revealed that HPX-2 functionally localizes to the hypodermis of the worm. In congruence with this observation, hpx-2 mutant animals possessed a weaker cuticle structure, indicated by higher permeability to a DNA dye, but exhibited no obvious morphological defects. In addition, fluorescent labeling of HPX-2 revealed its expression in the pharynx, an organ in which BLI-3 is also present. Interestingly, loss of HPX-2 increased intestinal colonization of E. faecalis, suggesting its role in the pharynx may limit intestinal colonization. Moreover, disruption of a catalytic residue in the peroxidase domain of HPX-2 resulted in decreased survival on E. faecalis, indicating its peroxidase activity is required for pathogen resistance. Finally, RNA-seq analysis of an hpx-2 mutant revealed changes in genes encoding for cuticle structural components under the non-pathogenic conditions. Under pathogenic conditions, genes involved in infection response were differentially regulated to a greater degree, likely due to increased microbial burden. In conclusion, the characterization of the heme-peroxidase, HPX-2, revealed that it contributes to C. elegans pathogen resistance through a role in generating cuticle material in the hypodermis and pharynx., Author summary Reactive oxygen species (ROS) production by the host tissues is one of the first lines of defense when microbial infection occurs. ROS has been shown to be involved in multiple protective pathways in innate immunity. However, given the complexity of mammalian systems, the exact manner in which ROS are used for host defense remains incompletely understood. In this study, we use Caenorhabditis elegans as a simplified model system to decipher the protective functions of ROS in innate immunity. We describe a peroxidase, HPX-2, that protects C. elegans from multiple infectious microbes by strengthening barrier tissue. This finding brings insight into the mechanisms by which peroxidases utilizes ROS to contribute to innate immunity. With infectious diseases being one of the most important causes of morbidity and mortality around the world, understanding ROS production and its function in pathogen resistance will provide us with important information in developing new therapies against pathogens.

In Cyperaceae, leaf anatomical characters, in particular the presence of a hypodermis or of a multiple epidermis, have contributed in taxonomic and phylogenetic studies. In this family, the leaf epidermis is often described as uniseriate, and the cells of the subepidermal layers having no chloroplasts are treated as hypodermis. Both tissues have a different ontogenetic origin and hence are not homologous. The aim of the present work was to verify the origin of the subepidermal layers in eight species belonging to Cyperoideae. All species studied presented multiple epidermal layers that were confirmed by leaf ontogeny. In Fimbristylis complanata, F. dichotoma, Pycreus flavescens and P. polystachyos the mature leaves present multiple epidermal layers with cells of the distinct layers similar in shape and size; in the other species studied these cells are different. Especially in the latter case, a multiple epidermis is easily interpreted erroneously as a hypodermis, possibly leading to erroneous evolutionary conclusions. Making correctly distinction between a hypodermis and a multiple epidermis, and hence in case of doubt investigating the origin of the questioned tissue, is compulsory in order to use both characters in a phylogenetic context. Though in the past often called ‘hypodermis’, our leaf ontogenetical observations show that in all species studied, the subepidermical layers constitute a multiple epidermis, originating from the protodermis. Fil: Martins, Shirley. Universidade Estadual Paulista Julio de Mesquita Filho; Brasil Fil: Pilatti, Vanesa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina Fil: Vegetti, Abelardo Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina Fil: Scatena, Vera Lucia. Instituto de Biociencias; Brasil

Across organisms, manipulation of biosynthetic capacity arrests development early in life, but can increase health- and lifespan post-developmentally. Here we demonstrate that this developmental arrest is not sickness but rather a regulated survival program responding to reduced cellular performance. We inhibited protein synthesis by reducing ribosome biogenesis (rps-11/RPS11 RNAi), translation initiation (ifg-1/EIF3G mutation and egl-45/EIF3A RNAi), or ribosome progression (cycloheximide treatment), all of which result in a specific arrest at larval stage 2 of C. elegans development. This quiescent state can last for weeks—beyond the normal C. elegans adult lifespan—and is reversible, as animals can resume reproduction and live a normal lifespan once released from the source of protein synthesis inhibition. The arrest state affords resistance to thermal, oxidative, and heavy metal stress exposure. In addition to cell-autonomous responses, reducing biosynthetic capacity only in the hypodermis was sufficient to drive organism-level developmental arrest and stress resistance phenotypes. Among the cell non-autonomous responses to protein synthesis inhibition is reduced pharyngeal pumping that is dependent upon AMPK-mediated signaling. The reduced pharyngeal pumping in response to protein synthesis inhibition is recapitulated by exposure to microbes that generate protein synthesis-inhibiting xenobiotics, which may mechanistically reduce ingestion of pathogen and toxin. These data define the existence of a transient arrest-survival state in response to protein synthesis inhibition and provide an evolutionary foundation for the conserved enhancement of healthy aging observed in post-developmental animals with reduced biosynthetic capacity., Author summary Protein synthesis is an essential cellular process, but post-developmental reduction of protein synthesis across multiple species leads to improved health- and lifespan. To better understand the physiological responses to impaired protein synthesis, we characterize a novel developmental arrest state that occurs when reducing protein synthesis during C. elegans development. Arrested animals have multiple survival-promoting phenotypes that are all dependent on the cellular energy sensor, AMP kinase. This survival response acts through the hypodermis and causes a reduction in pharyngeal pumping, indicating that the animal is responding to a perceived external threat, even in adults. Furthermore, exposing animals to pathogens, or xenobiotics they produce, can recapitulate these phenotypes, providing a potential evolutionary explanation for how a beneficial response in adults could evolve through the inhibition of an essential biological process such as protein synthesis.

We have identified Conserved Non-coding Elements (CNEs) in the regulatory region of Caenorhabditis elegans and Caenorhabditis briggsae mab-9, a T-box gene known to be important for cell fate specification in the developing C. elegans hindgut. Two adjacent CNEs (a region 78 bp in length) are both necessary and sufficient to drive reporter gene expression in posterior hypodermal cells. The failure of a genomic mab-9∷gfp construct lacking this region to express in posterior hypodermis correlates with the inability of this construct to completely rescue the mab-9 mutant phenotype. Transgenic males carrying this construct in a mab-9 mutant background exhibit tail abnormalities including morphogenetic defects, altered tail autofluorescence and abnormal lectin-binding properties. Hermaphrodites display reduced susceptibility to the C. elegans pathogen Microbacterium nematophilum. This comparative genomics approach has therefore revealed a previously unknown role for mab-9 in hypodermal function and we suggest that MAB-9 is required for the secretion and/or modification of posterior cuticle.

Cadherins are one of the major families of adhesion molecules with diverse functions during embryonic development. Fat-like cadherins form an evolutionarily conserved subgroup characterized by an unusually large number of cadherin repeats in the extracellular domain. Here we describe the role of the Fat-like cadherin CDH-4 in Caenorhabditis elegans development. Cdh-4 mutants are characterized by hypodermal defects leading to incompletely penetrant embryonic or larval lethality with variable morphogenetic defects. Independently of the morphogenetic defects cdh-4 mutant animals also exhibit fasciculation defects in the ventral and dorsal cord, the major longitudinal axon tracts, as well as migration defects of the Q neuroblasts. In addition CDH-4 is essential for establishing and maintaining the attachment between the buccal cavity and the pharynx. Cdh-4 is expressed widely in most affected cells and tissues during embryogenesis suggesting that CDH-4 functions to ensure that proper cell contacts are made and maintained during development.

Epithelial differentiation is a very early event during development of most species. The nematode Caenorhabditis elegans, with its well-defined and invariant lineage, offers the possibility to link cell lineage, cell fate specification and gene regulation during epithelial differentiation. Here, we focus on the regulation of the gene lin-26, which is required for proper differentiation of epithelial cells in the ectoderm and mesoderm (somatic gonad). lin-26 expression starts in early embryos and remains on throughout development, in many cell types originating from different sublineages. Using GFP reporters and mutant rescue assays, we performed a molecular dissection of the lin-26 promoter and could identify almost all elements required to establish its complex spatial and temporal expression. Most of these elements act redundantly, or synergistically once combined, to drive expression in cells related by function. We also show that lin-26 promoter elements mediate activation in the epidermis (hypodermis) by the GATA factor ELT-1, or repression in the foregut (pharynx) by the FoxA protein PHA-4. Taken together, our data indicate that lin-26 regulation is achieved to a large extent through tissue-specific cis-regulatory elements.

Background Previously we demonstrated the micro- and macrofilaricidal properties of imatinib in vitro. Here we use electron and multiphoton microscopy to define the target of imatinib in the adult and microfilarial stages of Brugia malayi and assess the effects of pharmacologically relevant levels of imatinib on the adult parasites. Methods After fixation of adult B. malayi males and females, sections were stained with polyclonal rabbit anti-c-Abl antibody (or isotype control) and imaged with multiphoton fluorescent microscopy. Microfilariae were fixed and labeled with rabbit anti-c-Abl IgG primary antibody followed by anti-rabbit gold conjugated secondary antibody and imaged using transmission electron microscopy (TEM; immunoEM). In addition, adult B. malayi males and females were exposed to 0 or 10μM of imatinib for 7 days following which they were prepared for transmission electron microscopy (TEM) to assess the drug’s effect on filarial ultrastructure. Results Fluorescent localization of anti-c-Abl antibody demonstrated widespread uptake in the adult filariae, but the most intense signal was seen in the reproductive organs, muscle, and intestine of both male and female worms. Fluorescence was significantly more intense in the early microfilarial stage (i.e. early morula) compared with later development stages (i.e. pretzel). Anti-c-Abl antibody in the microfilariae localized to the nuclei. Based on TEM assessment following imatinib exposure, imatinib appeared to be detrimental to embryogenesis in the adult female B. malayi. Conclusions At pharmacologically achievable concentrations of imatinib, embryogenesis is impaired and possibly halted in adult filariae. Imatinib is likely a slow microfilaricide due to interference in intra-nuclear processes, which are slowly detrimental to the parasite and not immediately lethal, and thus may be used to lower the levels of L. loa microfilariae before they are treated within the context of conventional mass drug administration., Author summary While Loa loa, or the “African eye worm”, does not commonly cause clinical disease, infection with high blood levels of microfilariae from this helminth infection is problematic in those who receive mass drug administration (MDA) in the eradication efforts of lymphatic filariasis and onchocerciasis. Ivermectin, the drug of choice for both MDA programs, causes a rapid killing of Loa loa microfilariae and in those with high pre-treatment levels, a severe inflammatory reaction can result in encephalopathy, and rarely death. Using the filariae Brugia malayi as an in vitro surrogate model for any of the blood-borne filarial parasites (including Loa loa) we have previously shown that imatinib can act as a slow microfilaricide. Understanding imatinib’s targets in adult and microfilariae may predict the in vivo effects on Loa loa microfilarial loads, and anticipate potential side-effects for future clinical trials. In this study, we localized c-Abl, the target of imatinib, to the reproductive structures of adult B. malayi, and to the nuclei of the microfilariae. Pharmacologically achievable levels of imatinib most strikingly caused necrosis of developing microfilariae in adult female B. malayi. These data further support pursuing clinical trials in the safety and efficacy of imatinib for Loa. loa and other filarial co-infections.

Background Regulation of organ and body size is a fundamental biological phenomenon, requiring tight coordination between multiple tissues to ensure accurate proportional growth. In C. elegans, a TGF-β pathway is the major regulator of body size and also plays a role in the development of the male tail, and is thus referred to as the TGF-β/Sma/Mab (for small and male abnormal) pathway. Mutations in components of this pathway result in decreased growth of animals during larval stages, with Sma mutant adults of the core pathway as small as ~60-70% the length of normal animals. The currently accepted model suggests that TGF-β/Sma/Mab pathway signaling in the C. elegans hypodermis is both necessary and sufficient to control body length. However, components of this signaling pathway are expressed in other organs, such as the intestine and pharynx, raising the question of what the function of the pathway is in these organs. Results Here we show that TGF-β/Sma/Mab signaling is required for the normal growth of the pharynx. We further extend the current model and show that the TGF-β/Sma/Mab pathway can function in multiple tissues to regulate body and organ length. Specifically, we find that pharyngeal expression of the SMAD protein SMA-3 partially rescues both pharynx length and body length of sma-3 mutants. Conclusions Overall, our results support a model in which the TGF-β/Sma/Mab signaling pathway can act in multiple tissues, activating one or more downstream secreted signals that act non cell-autonomously to regulate overall body length in C. elegans. Electronic supplementary material The online version of this article (doi:10.1186/s12861-014-0043-8) contains supplementary material, which is available to authorized users.

There are several transforming growth factor-beta (TGF-beta) pathways in the nematode Caenorhabditis elegans. One of these pathways regulates body length and is composed of the ligand DBL-1, serine/threonine protein kinase receptors SMA-6 and DAF-4, and cytoplasmic signaling components SMA-2, SMA-3, and SMA-4. To further examine the molecular mechanisms of body-length regulation in the nematode by the TGF-beta pathway, we examined the regional requirement for the type-I receptor SMA-6. Using a SMA-6::GFP (green fluorescent protein) reporter gene, sma-6 was highly expressed in the hypodermis, unlike the type-II receptor DAF-4, which is reported to be ubiquitously expressed. We then examined the ability of SMA-6 expression in different regions of the C. elegans body to rescue the sma-6 phenotype (small) and found that hypodermal expression of SMA-6 is necessary and sufficient for the growth and maintenance of body length. We also demonstrate that GATA sequences in the sma-6 promoter contribute to the hypodermal expression of sma-6.

Objective: To define the relationship between the long saphenous vein and the connective framework of the subcutaneous tissue (hypodermis) of the lower limb. Methods: The connective skeleton of the hypodermis was studied by anatomical dissection, stereomicroscopy of cross-sectioned specimens and ultrasound imaging in 88 lower extremities. Results: The long saphenous vein runs for most of its length in a narrow compartment delineated deeply by the muscular fascia and superficially by a connective tissue lamina descending from the inguinal ligament in the anteromedial part of the thigh and medial aspect of the calf. These two fascia fuse at the boundaries of the compartment. The long saphenous vein adventitia is anchored to both fasciase by thick connective tissue strands. Conclusion: The anatomical relationship between the long saphenous vein and the connective framework of the hypodermis suggests that: (1) only the vein running within the deep compartment of the hypodermis should be considered as the ‘true’ long saphenous vein; (2) the other subcutaneous veins running outside the compartment should be considered as collaterals of the long saphenous vein; (3) the connective sheath surrounding the long saphenous vein could oppose dilatation of this vessel should valvular incompetence develop; and (4) thigh muscle contraction could modify the calibre of the long saphenous vein as happens in the deep veins. Finally, the authors propose to term the deep compartment of the medial thigh and the leg hypodermis the ‘long saphenous vein compartment’ and consequently the hypodermic connective lamina, by which it is superficially delimited, as the ‘long saphenous vein fascia’.

Comparative morphological and anatomical investigations on the normal and cristate forms of Echinocereus pectinatus (Scheidw.) Eng. stem (Cactaceae A.L Juss.) from Fomin’s Botanical Garden collections have been conducted. There were found a few characters identical for the stem structure in both forms. But also were found morphological and anatomical differences between normal and cristate forms. The most obvious sign of fasciation is a change of cross stem sections from circular to elliptical shape. The plants are also distinguished by the size and the form of transversal projections of the parenchymatous cells and by shape and location of the inclusions.

Plants that are adapted to waterlogged conditions develop aerenchyma in roots for ventilation. Some wetland plant species also form an apoplastic barrier at the outer cell layers of roots that reduces radial oxygen loss (ROL) from the aerenchyma and to prevent toxic compounds from entering the root. The composition of the apoplastic barrier is not well understood. One potential component is suberin, which accumulates at the hypodermal/exodermal cell layers of the roots under waterlogged soil conditions or in response to other environmental stimuli. However, differences in suberin content and composition between plant species make it difficult to evaluate whether suberin has a role in preventing ROL. In this article, we summarize recent advances in understanding apoplastic barrier formation in roots and, between various plant species, compare the chemical compositions of the apoplastic barriers in relation to their permeability to oxygen. Moreover, the relationship between suberin accumulation and the barrier to ROL is discussed.

Full thickness wounds require a dermal component to achieve functional permanent skin restoration. Currently available tissue-engineered skin substitutes lack a subcutaneous fat layer that would functionally contribute some of the mechanical and thermoregulatory properties of normal skin. To generate a trilayer engineered skin equivalent, we included bone marrow mesenchymal (BM-MSC) or adipose tissue-derived (ASC) stromal cells in a human plasma hydrogel exposed to adipogenic clues for three weeks. Approximately half of the cells differentiated under these conditions into mature adipocytes that survived for two years in culture with minimal medium change. In vitro generation of bona fide fully differentiated adipocytes was assessed by leptin secretion and ultrastructurally demonstrated through semithin to ultrathin sectioning and lipid staining with osmium tetroxide. Furthermore, presence of BM-MSCs or ASCs within the subcutaneous layer contributed to the epidermal differentiation program, with more proliferating basal cells depositing basal membrane proteins and differentiating into mature keratinocytes that were able to generate a pluristratified epithelium. In conclusion, we engineered a fully differentiated human skin trilayer that could present multiple applications such as use for in vitro drug absorption tests and regenerative therapies. Copyright (c) 2012 John Wiley & Sons, Ltd.

Current control strategies for onchocerciasis and lymphatic filariasis (LF) rely on prolonged yearly or twice-yearly mass administration of microfilaricidal drugs. Prospects for near-term elimination or eradication of these diseases would be improved by availability of a macrofilaricide that is highly effective in a short regimen. Flubendazole (FLBZ), a benzimidazole anthelmintic registered for control of human gastrointestinal nematode infections, is a potential candidate for this role. FLBZ has profound and potent macrofilaricidal effects in many experimental animal models of filariases and in one human trial for onchocerciasis after parental administration. Unfortunately, the marketed formulation of FLBZ provides very limited oral bioavailability and parenteral administration is required for macrofilaricidal efficacy. A new formulation that provided sufficient oral bioavailability could advance FLBZ as an effective treatment for onchocerciasis and LF. Short-term in vitro culture experiments in adult filariae have shown that FLBZ damages tissues required for reproduction and survival at pharmacologically relevant concentrations. The current study characterized the long-term effects of FLBZ on adult Brugia malayi by maintaining parasites in jirds for up to eight weeks following brief drug exposure (6–24 hr) to pharmacologically relevant concentrations (100 nM—10 μM) in culture. Morphological damage following exposure to FLBZ was observed prominently in developing embryos and was accompanied by a decrease in microfilarial output at 4 weeks post-exposure. Although FLBZ exposure clearly damaged the parasites, exposed worms recovered and were viable 8 weeks after treatment., Author Summary Onchocerciasis and lymphatic filariasis are debilitating diseases caused by infections with filarial nematodes. The World Health Organization aims to eliminate these infections as public health problems. Despite prolonged control efforts, including chemotherapy through mass drug administration (MDA), transmission and infections persist. Addition of a microfilaricide that is efficacious in a short regimen would enhance prospects for achieving elimination goals. We investigated the long-term effects of the macrofilaricidal drug, flubendazole (FLBZ), on Brugia malayi. Adult parasites were exposed in culture to FLBZ at pharmacologically relevant concentrations (100 nM—10 μM) for up to 24 hr prior to implantation into the abdominal cavity of a jird for long-term maintenance. The greatest drug effect was on embryogenesis; morphological damage was most evident in early developmental stages. There was also a decrease in the release of microfilaria (mf) from the adult. Interestingly, no damage was observed to fully formed mf. Although further studies are required to determine to what extent these findings can be extrapolated to a field setting, an exposure profile which may produce similar effects in vivo has been defined.

Using a mixed-dye injection technique, we found a novel kind of muscle fiber with a lumen, established its precise location in the subcutaneous muscle layer along the acupuncture muscle of the bladder line, and determined its detailed ultrastructure. The channels with flowing liquid were a novel kind of muscle fibers with lumens and they were located in the subcutaneous muscle layer of rat. Their detection was realized by using chrome-hematoxylin and a mixture of fluorescent nanoparticles and commercial Pelikan ink. These acupuncture muscle channels were hidden among the neighboring skin skeletal muscle fibers and were barely distinguishable from them with light microscopes. Only with a transmission electron microscope were their characteristic features shown to be different from normal skin skeletal muscle. These features included undifferentiated muscle fibers that resembled immature myofibrils without Z-lines and reassembled telophase nuclei.

The seedling root system of the seagrass Posidonia oceanica consists of a primary root and up to four adventitious roots. Under culture, germination and early growth began with the emergence of the primary root in the first week. Then the two adventitious root primordia originally present in the seed emerged at 3 and 5 weeks respectively, followed successively by further adventitious roots. Primary roots reached 17 mm at 4 weeks, but then their growth decreased markedly. In contrast the adventitious roots showed a pattern of continued elongation. Anatomical observations of both primary and adventitious roots revealed a multilayered hypodermis of thick-walled cells enclosing a wide cortex (99% of the root transverse area) and narrow stele. A well-distinguished endodermis was only observed in the primary roots, while differentiated xylem elements were found solely in the adventitious roots, but it is unclear to what degree differences between the two root types are due to different root maturity or to their role in water and nutrient uptake. Overall, the P. oceanica seedling root system is composed of multiple, rapidly formed roots which are strong yet flexible due to a large proportion of cortical tissue and further strengthened by a multilayered hypodermis, characteristics which could potentially facilitate initial anchorage and establishment.

An illustrated glossary of terms used in the analysis of the shells of recent and fossil foraminifera supplemented by a rigorous selection of terms that facilitate an understanding of their biology and their use in ecology and biostratigraphy. The glossary includes some 650 entries illustrated by 83 - often composite – figures many of which are stereographs or 3D models. A taxonomic index lists the 140 taxa illustrated.

Caenorhabditis elegans PEB-1 is a novel DNA-binding protein expressed in most pharyngeal cell types and outside the pharynx in the hypodermis, hindgut, and vulva. Previous RNAi analyses indicated that PEB-1 is required for normal morphology of these tissues and growth; however, the peb-1 null phenotype was unknown. Here we describe the deletion mutant peb-1(cu9) that not only exhibits the morphological defects observed in peb-1(RNAi) animals, but also results in penetrant larval lethality characterized by defects in pharyngeal function and molting. Consistent with a function in molting, we found that PEB-1 was detectable in all hypodermal and hindgut cells underlying the cuticle. Comparison to molting-defective lrp-1(ku156) mutants revealed that the peb-1(cu9) mutants were particularly defective in shedding the pharyngeal cuticle, and this defect likely contributed to feeding defects and lethality. Most markers of pharyngeal cell differentiation examined were expressed normally in peb-1(cu9) mutants; however, g1 gland cell expression of a kel-1Colon, two colonsgfp reporter was reduced. As g1 gland cells have prominent functions during molting, we suggest defective gland cell differentiation contributes to peb-1(cu9) molting defects. In comparison, other peb-1 mutant phenotypes, including hindgut abnormalities, appeared independent of the molting defect. Similar phenotypes resulted from late loss of pha-4 function, suggesting that PEB-1 and PHA-4 have common functions in some tissues where they are co-expressed.

The relationships between the connective framework of the lower extremity hypodermis and the saphenous veins was studied by dissection, stereomicroscopy, ultrasonography and histology in 64 lower limbs. A fibroelastic lamina was evidenced in the hypodermis of the medial aspect of the thigh and leg and in the back face of the leg. This lamina, together with the underlying muscular fascia, fixed the boundaries of two compartments occupied by the saphenous veins and nerves. The adventitia of the saphenous veins was connected to the compartment walls by thick connective strands. The saphenous veins ran deeply in the hypodermis, closely ensheathed by a fibroelastic sleeve. As a consequence, they could no longer be considered as a truly superficial vein. This term seems to be appropriate only for their tributaries, which ran in a more superficial plane just below the dermis. The role of the saphenous vessels in blood return from the lower limbs may be greater than classically accepted. In fact, due to their close fascial ensheathing and adventitial anchoring, muscular contractions may enhance blood flow within these vessels as occurs in the intermuscular veins. Finally, dilative pathology of the saphenous vein may be resisted by the membranous lamina as a sort of a fibroelastic shield.