Hollow Microneedle Arrays for Intradermal Drug Delivery and DNA Electroporation

The association of microneedles with electric pulses causing electroporation could result in an efficient and less painful delivery of drugs and DNA into the skin. Hollow conductive microneedles were used for (1) needle-free intradermal injection and (2) electric pulse application in order to achieve electric field in the superficial layers of the skin sufficient for electroporation. Microneedle array was used in combination with a vibratory inserter to disrupt the stratum corneum, thus piercing the skin. Effective injection of proteins into the skin was achieved, resulting in an immune response directed to the model antigen ovalbumin. However, when used both as microneedles to inject and as electrodes to apply the electric pulses, the setup showed several limitations for DNA electrotransfer. This could be due to the distribution of the electric field in the skin as shown by numerical calculations and/or the low dose of DNA injected. Further investigation of these parameters is needed in order to optimize minimally invasive DNA electrotransfer in the skin.     

Resistance to Brown Ring Disease in the Manila clam, Ruditapes philippinarum: A study of selected stocks showing a recovery process by shell repair

European stocks of the Manila clam Ruditapes philippinarum are affected by the Brown Ring Disease (BRD), which is caused by Vibrio tapetis. BRD is characterized by an accumulation of a brown organic matrix on the inner face of the shell. Clams that recover from BRD develop a white mineralized layer covering the brown matrix. Stocks of clams that showed resistance to BRD development, as enhanced recovery, have been monitored since 2000. We have examined two selected stocks: a Low Susceptibility (LS) stock and a High Susceptibility stock (HS), over three generations. The LS stock showed less evidence of the BRD symptoms, and more evidence of total shell repair, both in the field and following experimental challenge with V. tapetis, indicating that some clams may be less vulnerable to a V. tapetis attack than others. The inner face of the valves of the LS and HS clams of the two last generations were analysed with scanning electron microscopy. Examination of shells from BRD-affected clams showed that during the repair process, calcium crystals were progressively laid down until the affected zone was entirely covered. By the end of the shell repair process, a final organic layer covered the calcium crystal mounds. This layer seemed essential in the recovery process. The results indicate that the shell repair capability of the clams is the principal mechanism implicated in the development of BRD resistance in the Manila clam stocks. However, this resistance did not increase with generation because the broodstock was maintained at a site where selection pressure was low, due to a low prevalence of V. tapetis.     

Cavin proteins: New players in the caveolae field

Caveolae are specialized lipid microdomains, forming small invaginations in the plasma membrane, known to be implicated in multiple functions including lipid storage, cell signaling and endocytosis. Formation of these wide flask-shaped invaginations is dependent on the expression of a caveolar coat protein, namely caveolin. Until now, the accepted paradigm was that caveolin was the sole and only structural protein of caveolae since its expression was necessary and sufficient to drive caveolae biogenesis. The recent characterizations of PTRF/cavin-1 and subsequently other cavin family members in caveolae formation have highlighted additional levels of complexity in the biogenesis of these plasma membrane invaginations. In this review, recent advances on the role of the different cavin family members in the regulation of caveolae structures as well as potential new functions will be discussed.     

Lipid body accumulation alters calcium signaling dynamics in immune cells

There is well-established variability in the numbers of lipid bodies (LB) in macrophages, eosinophils, and neutrophils. Similarly to the steatosis observed in adipocytes and hepatocytes during hyperinsulinemia and nutrient overload, immune cell LB hyper-accumulate in response to bacterial and parasitic infection and inflammatory presentations. Recently we described that hyperinsulinemia, both in vitro and in vivo, drives steatosis and phenotypic changes in primary and transformed mast cells and basophils. LB reach high numbers in these steatotic cytosols, and here we propose that they could dramatically impact the transcytoplasmic signaling pathways. We compared calcium release and influx responses at the population and single cell level in normal and steatotic model mast cells. At the population level, all aspects of Fc?RI-dependent calcium mobilization, as well as activation of calcium-dependent downstream signaling targets such as NFATC1 phosphorylation are suppressed. At the single cell level, we demonstrate that LB are both sources and sinks of calcium following Fc?RI cross-linking. Unbiased analysis of the impact of the presence of LB on the rate of trans-cytoplasmic calcium signals suggest that LB enrichment accelerates calcium propagation, which may reflect a Bernoulli effect. LB abundance thus impacts this fundamental signaling pathway and its downstream targets.     

Anti-Gb3 Monoclonal Antibody Inhibits Angiogenesis and Tumor Development

Inhibiting the growth of tumor vasculature represents one of the relevant strategies against tumor progression. Between all the different pro-angiogenic molecular targets, plasma membrane glycosphingolipids have been under-investigated. In this present study, we explore the anti-angiogenic therapeutic advantage of a tumor immunotherapy targeting the globotriaosylceramide Gb3. In this purpose, a monoclonal antibody against Gb3, named 3E2 was developed and characterized. We first demonstrate that Gb3 is over-expressed in proliferative endothelial cells relative to quiescent cells. Then, we demonstrate that 3E2 inhibits endothelial cell proliferation in vitro by slowing endothelial cell proliferation and by increasing mitosis duration. Antibody 3E2 is further effective in inhibiting ex vivo angiogenesis in aorta ring assays. Moreover, 3E2 treatment inhibits NXS2 neuroblastoma development and liver metastases spreading in A/J mice. Immunohistology examination of the NXS2 metastases shows that only endothelial cells, but not cancer cells express Gb3. Finally, 3E2 treatment diminishes tumor vessels density, proving a specific therapeutic action of our monoclonal antibody to tumor vasculature. Our study demonstrates that Gb3 is a viable alternative target for immunotherapy and angiogenesis inhibition.     

Correction to: Imaging plant responses to water deficit using electrical resistivity tomography

Plant and Soil - The original version of this article unfortunately contained a mistake. The authors would like to add Andreas Kemna as the fifth author of this paper. The updated list of authors...     

Comparative proteomic analysis of a sea urchin (Heliocidaris erythrogramma) antibacterial response revealed the involvement of apextrin and calreticulin

Echinoderms evolved early in the deuterostome lineage, and as such constitute model organisms for comparative physiology and immunology. The sea urchin genome sequence (Strongylocentrotus purpuratus) revealed a complex repertoire of genes with similarities to the immune response genes of other species. To complement these genomic data, we investigated the responses of sea urchins to the injection of bacteria using a comparative proteomics approach on a closely related species. In the sea urchin, Heliocidaris erythrogramma, the relative abundance of many proteins was altered in response to the injection of both bacteria and saline, suggesting their involvement in wounding responses, while others were differentially altered in response to bacteria only. The identities of 15 proteins that differed in relative abundance were determined by mass spectrometry. These proteins revealed a significant modification in energy metabolism in coelomocytes towards the consumption of glutamate and the production of NADPH after injection, as well as an increased concentration of cell signalling molecules, such as heterotrimeric guanine nucleotide-binding protein. The injection of bacteria specifically increased the abundance of apextrin and calreticulin, suggesting that these two proteins are involved in the sequestration or inactivation of bacteria.     

Imaging plant responses to water deficit using electrical resistivity tomography

Background and aims

Monitoring root water uptake dynamics under water deficit (WD) conditions in fields are crucial to assess plant drought tolerance. In this study, we investigate the ability of Electrical Resistivity Tomography (ERT) to capture specific soil water depletion induced by root water uptake.

Methods

A combination of surface and depth electrodes with a high spatial resolution (10 cm) was used to map 2-D changes of bulk soil electrical conductivity (EC) in an agronomic trial with different herbaceous species. A synthetic experiment was performed with a mechanistic model to assess the ability of the electrode configuration to discriminate abstraction patterns due to roots. The impact of root segments was incorporated in the forward electrical model using the power-law mixing model.

Results

The time-lapse analysis of the synthetic ERT experiment shows that different root water uptake patterns can be delineated for measurements collected under WD conditions but not under wet conditions. Three indices were found (depletion amount, maximum depth, and spread), which allow capturing plant-specific water signatures based moisture profile changes derived from EC profiles. When root electrical properties were incorporated in the synthetic experiments, it led to the wrong estimation of the amount of water depletion, but a correct ranking of plants depletion depth. When applied to the filed data, our indices showed that Cocksfoot and Ryegrass had shallower soil water depletion zones than white clover and white clover combined with Ryegrass. However, in terms of water depletion amount, Cocksfoot consumed the largest amount of water, followed by White Clover, Ryegrass+White Clover mixture, and Ryegrass.

Conclusion

ERT is a well-suited method for phenotyping root water uptake ability in field trials under WD conditions.