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The aim of this study was to examine data demonstrating that Scarpa's fascia, a superficial fascia of the anterior abdominal wall, is a vascularized tissue. Specimens of the fascia of seven volunteers undergoing abdominoplasty surgical procedures at the Plastic Surgery Unit of the University of Padova Medical Center were collected. Fractal analysis and quantitative assessment of the vascular network of the fascia was carried out, exploiting the presence of blood in the vessels. Each sample was divided and processed for histological/immunohistochemical analysis (into 5 micron-paraffin embedded sections and cryo-sectioned free-floating samples) as well as for electron microscopy study. A rich vascular pattern forming a fine, dense meshwork with an area percentage of 6.20±2.10% von Willebrand factor stained vessels was noted in all the specimens of the fascia examined; the area percentage of the αSMA-stained vessels was 2.93 ± 1.80 %. The diameters of the vessels fell between the 13 - 65 μm range; the network was composed of arteries, veins, capillaries and lymphatic segments. Topological results showed that the vascular network within Scarpa's fascia is well-branched (segments: 6615 ± 3070 and 8.40 ± 3.40 per mm

The superficial fascia has only recently been recognized as a specific anatomical structure. Furthermore, whereas it is actually recognized that the innervation of the deep/muscular fascia plays a key role in proprioception and nociception, there are very few studies that have analyzed these characteristics in the superficial fascia. In this work, our group analyzed two different anatomical districts (abdomen and thigh), from volunteer patients, undergoing surgery procedures. Each sample was processed for histological analysis by Hematoxylin&Eosin, and by immunohistochemistry stainings (in 5-micron-paraffin embedded section and in cryosectioned free floating samples), with antibodies specific for nerve fibers: S100 antibody for myelinating and non-myelinating Schwann cells, PGP9.5 antibody as pan-neuronal marker, tyrosine hydroxylase for autonomic innervation. The results revealed a huge innervation: the nervous structures were found above all around blood vessels and close to adipocytes, but they penetrated also in the connective tissue itself and are found in the midst of fibro-adipose tissue. The tissue is pervaded by both thin (mean diameter of 4.8 ± 2.6 μm) and large nerve fiber bundles of greater diameter (21.1 ± 12.2 μm). The ratio S100/TH positivity was equal to 2.96, with a relative percentage of autonomic innervation with of 33.82%. In the light of these findings is evident that the superficial fasciae have a clear and distinct anatomical identity and a specific innervation, which should be considered to better understand their role in thermoregulation, exteroception and pain perception. The knowledge of the superficial fascia may improve grading and developing of different manual approach for treatments of fascial dysfunctions, and the understanding of how some factors like temperature or manual therapies can have an impact on sensitivity of the fascia.

A small number of peptide growth factor ligands are used repeatedly in development and homeostasis to drive programs of cell differentiation and function. Cells and tissues must integrate inputs from these diverse signals correctly, while failure to do so leads to pathology, reduced fitness, or death. Previous work using the nematode C. elegans identified an interaction between the bone morphogenetic protein (BMP) and insulin/IGF-1-like signaling (IIS) pathways in the regulation of lipid homeostasis. The molecular components required for this interaction, however, were not fully understood. Here we report that INS-4, one of 40 insulin-like peptides (ILPs), is regulated by BMP signaling to modulate fat accumulation. Furthermore, we find that the IIS transcription factor DAF-16/FoxO, but not SKN-1/Nrf, acts downstream of BMP signaling in lipid homeostasis. Interestingly, BMP activity alters sensitivity of these two transcription factors to IIS-promoted cytoplasmic retention in opposite ways. Finally, we probe the extent of BMP and IIS interactions by testing additional IIS functions including dauer formation, aging, and autophagy induction. Coupled with our previous work and that of other groups, we conclude that BMP and IIS pathways have at least three modes of interaction: independent, epistatic, and antagonistic. The molecular interactions we identify provide new insight into mechanisms of signaling crosstalk and potential therapeutic targets for IIS-related pathologies such as diabetes and metabolic syndrome., Author summary Systemic homeostasis depends on the coordinated action of different cells and tissues throughout the body in response to internal and external cues. This coordination is achieved in part by cellular communication through the production, release, and response to molecular signals in a process known as cell signaling. These signals and their response pathways are highly conserved among animal species; therefore, we are able to use genetic tools in a simple animal model to identify broadly relevant mechanisms of function. Here, we demonstrate that two conserved signaling pathways, the bone morphogenetic protein (BMP) and insulin pathways, crosstalk to control lipid storage. We identify molecular components that mediate this regulation and determine their sites of action. Furthermore, we show that these two signaling pathways have multiple modes of interaction in a variety of developmental and physiological contexts. Due to the devastating human diseases caused by misregulation of these pathways, including cancer, cardiovascular disease, diabetes, and metabolic syndrome, understanding the mechanisms of crosstalk is critical.

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.

The disproportionate length of an axon makes its structural and functional maintenance a major task for a neuron. The heterochronic gene lin-14 has previously been implicated in regulating the timing of key developmental events in the nematode C. elegans. Here, we report that LIN-14 is critical for maintaining neuronal integrity. Animals lacking lin-14 display axonal degeneration and guidance errors in both sensory and motor neurons. We demonstrate that LIN-14 functions both cell autonomously within the neuron and non-cell autonomously in the surrounding tissue, and we show that interaction between the axon and its surrounding tissue is essential for the preservation of axonal structure. Furthermore, we demonstrate that lin-14 expression is only required during a short period early in development in order to promote axonal maintenance throughout the animal's life. Our results identify a crucial role for LIN-14 in preventing axonal degeneration and in maintaining correct interaction between an axon and its surrounding tissue.

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.

The biology and behavior of adults differ substantially from those of developing animals, and cell-specific information is critical for deciphering the biology of multicellular animals. Thus, adult tissue-specific transcriptomic data are critical for understanding molecular mechanisms that control their phenotypes. We used adult cell-specific isolation to identify the transcriptomes of C. elegans’ four major tissues (or “tissue-ome”), identifying ubiquitously expressed and tissue-specific “enriched” genes. These data newly reveal the hypodermis’ metabolic character, suggest potential worm-human tissue orthologies, and identify tissue-specific changes in the Insulin/IGF-1 signaling pathway. Tissue-specific alternative splicing analysis identified a large set of collagen isoforms. Finally, we developed a machine learning-based prediction tool for 76 sub-tissue cell types, which we used to predict cellular expression differences in IIS/FOXO signaling, stage-specific TGF-β activity, and basal vs. memory-induced CREB transcription. Together, these data provide a rich resource for understanding the biology governing multicellular adult animals., Author summary C. elegans is the simplest multi-cellular model system, with only 959 somatic cells in the fully-developed adult. This work describes the isolation and RNA-seq analysis of the worm’s major adult tissues. Previously, the isolation of adult tissues has been hampered by the worm’s tough outer cuticle, but identification of the transcriptomes of adult tissues is necessary to understand the biology of adults, which differs substantially from that of embryonic and larval cells. We recently developed a method to isolate and RNA-sequence adult tissues, and applied it here to characterize the muscle, neuron, intestine, and epidermis adult transcriptomes and isoform profiles. The data reveal interesting new characteristics for adult tissues, particularly the hypodermis’ metabolic function, which we have functionally tested. The tissue transcriptomes were also used to identify relevant human tissue orthologs in an unbiased manner. Finally, we present a new prediction tool for gene expression in up to 76 tissues and cell types, and we demonstrate its utility not only in predicting cell-specific gene expression, but in diagnosing expression changes in different genetic pathways and contexts.

The development of treatments using stem cells has drawn the attention of researchers to fat deposits given the fact they represent an almost unlimited reservoir of such cells, which can be accessed through minimally invasive techniques. However, the standardization of these studies has been made difficult because of the controversies of nomenclature regarding the many components of adipose tissue. Despite their distinct and independent structures with different metabolic responses, the terms hypodermis and subcutaneous adipose tissue are many times used as synonyms. However, the correct distinction between these two layers, the knowledge of their behavior and an uniformity of these terminologies are of utmost importance. Thus, the purpose of this study was to make a bibliographic review on the theme, aiming to show the anatomical, histological and metabolic differences between these two tissues and standardize their nomenclature.

The study was focused on assessing the presence of arabinogalactan proteins (AGPs) and pectins within the cell walls as well as prenyl lipids, sodium and chlorine content in leaves of Tilia x euchlora trees. The leaves that were analyzed were collected from trees with and without signs of damage that were all growing in the same salt stress conditions. The reason for undertaking these investigations was the observations over many years that indicated that there are trees that present a healthy appearance and trees that have visible symptoms of decay in the same habitat. Leaf samples were collected from trees growing in the median strip between roadways that have been intensively salted during the winter season for many years. The sodium content was determined using atomic spectrophotometry, chloride using potentiometric titration and poly-isoprenoids using HPLC/UV. AGPs and pectins were determined using immunohistochemistry methods. The immunohistochemical analysis showed that rhamnogalacturonans I (RG-I) and homogalacturonans were differentially distributed in leaves from healthy trees in contrast to leaves from injured trees. In the case of AGPs, the most visible difference was the presence of the JIM16 epitope. Chemical analyses of sodium and chloride showed that in the leaves from injured trees, the level of these ions was higher than in the leaves from healthy trees. Based on chromatographic analysis, four poly-isoprenoid alcohols were identified in the leaves of T. x euchlora. The levels of these lipids were higher in the leaves from healthy trees. The results suggest that the differences that were detected in the apoplast and symplasm may be part of the defensive strategy of T. x euchlora trees to salt stress, which rely on changes in the chemical composition of the cell wall with respect to the pectic and AGP epitopes and an increased synthesis of prenyl lipids.

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.

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.

Studies on the anatomy of Piper leaves demonstrate the presence of a subepidermal tissue distinct from the adjacent epidermis, which cells show thin walls and hyaline contents. Some authors consider such cells a hypodermal tissue, while others refer to them as components of a multiple epidermis. In this study, the nature of this subepidermal tissue was investigated through the analysis of leaf ontogeny in three Piper species. The analysis showed that the referred tissue originates from the ground meristem and, thus, should be considered a hypodermis. The studied species suggests that the role of the hypodermis would be to protect the photosynthetic apparatus from excess light, regulating the intensity of light reaching the chlorophyll parenchyma. Os estudos de anatomia foliar de espécies de Piper revelam a presença de um tecido subepidérmico distinto da epiderme e cujas células apresentam paredes finas e conteúdo hialino. Estas células são referenciadas por alguns autores como um tecido hipodérmico e por outros, como sendo constituintes de uma epiderme múltipla. Nesse estudo verificou-se a natureza deste tecido subepidérmico a partir da análise da ontogênese foliar de três espécies de Piper. A análise revelou que o referido tecido tem origem do meristema fundamental e, portanto, deve ser denominado de hipoderme. Para as espécies avaliadas, sugere-se que a hipoderme teria função de, proteger o aparato fotossintético do excesso de luminosidade, regulando a intensidade luminosa que atinge o parênquima clorofiliano.

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.

The rigid crustacean exoskeleton, the cuticle, is composed of the polysaccharide chitin, structural proteins and mineral deposits. It is periodically replaced to enable growth and its construction is an energy-demanding process. Ecdysis, the shedding event of the old cuticle is preceded by a preparatory phase, termed premolt, in which the present cuticle is partially degraded and a new one is formed underneath it. Procambarus clarkii (Girard), an astacid crustacean, was used here to comprehensively examine the changing patterns of gene expression in the hypodermis underlying the cuticle of the carapace at seven time points along ~14 premolt days. Next generation sequencing was used to construct a multi-tissue P. clarkii transcript sequence assembly to be generally used in a variety of transcriptomic studies. An aimed reference transcriptome was created here for the performance of a digital transcript expression analysis, determining the gene expression profiles in each of the examined premolt stages. The analysis revealed a cascade of sequential expression events of molt-related genes involved in chitin degradation, synthesis and modification, as well as synthesis of collagen and four groups of cuticular structural genes. The novel description of major transcriptional events during premolt and determination of their timing provide temporal markers for future studies of molt progress and regulation. The peaks of expression of the molt-related genes were preceded by expression peaks of cytoskeletal genes hypothesized to be essential for premolt progress by regulating protein synthesis and/or transport probably by remodeling the cytoskeletal structure.

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.

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 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.