Glycosaminoglycan polymerization may enable osmotically inactive Na+ storage in the skin

Osmotically inactive skin Na(+) storage is characterized by Na(+) accumulation without water accumulation in the skin. Negatively charged glycosaminoglycans (GAGs) may be important in skin Na(+) storage. We investigated changes in skin GAG content and key enzymes of GAG chain polymerization during osmotically inactive skin Na(+) storage. Female Sprague-Dawley rats were fed a 0.1% or 8% NaCl diet for 8 wk. Skin GAG content was measured by Western blot analysis. mRNA content of key dermatan sulfate polymerization enzymes was measured by real-time PCR. The Na(+) concentration in skin was determined by dry ashing. Skin Na(+) concentration during osmotically inactive Na(+) storage was 180-190 mmol/l. Increasing skin Na(+) coincided with increasing GAG content in cartilage and skin. Dietary NaCl loading coincided with increased chondroitin synthase mRNA content in the skin, whereas xylosyl transferase, biglycan, and decorin content were unchanged. We conclude that osmotically inactive skin Na(+) storage is an active process characterized by an increased GAG content in the reservoir tissue. Inhibition or disinhibition of GAG chain polymerization may regulate osmotically inactive Na(+) storage.     

Efficient Transformation of Oil Palm Protoplasts by PEG-Mediated Transfection and DNA Microinjection

Background

Genetic engineering remains a major challenge in oil palm (Elaeis guineensis) because particle bombardment and Agrobacterium-mediated transformation are laborious and/or inefficient in this species, often producing chimeric plants and escapes. Protoplasts are beneficial as a starting material for genetic engineering because they are totipotent, and chimeras are avoided by regenerating transgenic plants from single cells. Novel approaches for the transformation of oil palm protoplasts could therefore offer a new and efficient strategy for the development of transgenic oil palm plants.

Methodology/Principal Findings

We recently achieved the regeneration of healthy and fertile oil palms from protoplasts. Therefore, we focused on the development of a reliable PEG-mediated transformation protocol for oil palm protoplasts by establishing and validating optimal heat shock conditions, concentrations of DNA, PEG and magnesium chloride, and the transfection procedure. We also investigated the transformation of oil palm protoplasts by DNA microinjection and successfully regenerated transgenic microcalli expressing green fluorescent protein as a visible marker to determine the efficiency of transformation.

Conclusions/Significance

We have established the first successful protocols for the transformation of oil palm protoplasts by PEG-mediated transfection and DNA microinjection. These novel protocols allow the rapid and efficient generation of non-chimeric transgenic callus and represent a significant milestone in the use of protoplasts as a starting material for the development of genetically-engineered oil palm plants.     

The gut microenvironment of sediment-dwelling Chironomus plumosus larvae as characterised with O2, pH, and redox microsensors

We devised a set-up in which microsensors can be used for characterising the gut microenvironment of aquatic macrofauna. In a small flow cell, we measured microscale gradients through dissected guts (O₂, pH, redox potential [E _h ]), in the haemolymph (O₂), and towards the body surface (O₂) of Chironomus plumosus larvae. The gut microenvironment was compared with the chemical conditions in the lake sediment in which the animals reside and feed. When the dissected guts were incubated at the same nominal O₂ concentration as in haemolymph, the gut content was completely anoxic and had pH and E _h values slightly lower than in the ambient sediment. When the dissected guts were artificially oxygenated, the volumetric O₂-consumption rates of the gut content were at least 10× higher than in the sediment. Using these potential O₂-consumption rates in a cylindrical diffusion–reaction model, it was predicted that diffusion of O₂ from the haemolymph to the gut could not oxygenate the gut content under in vivo conditions. Additionally, the potential O₂-consumption rates were so high that the intake of dissolved O₂ along with feeding could be ruled out to oxygenate the gut content. We conclude that microorganisms present in the gut of C. plumosus cannot exhibit an aerobic metabolism. The presented microsensor technique and the data analysis are applicable to guts of other macrofauna species with cutaneous respiration.     

Age-related nonrandom chromosomal abnormalities in human low-grade astrocytomas

We report a cytogenetic investigation of 55 low-grade astrocytomas in 52 patients, 15 children and 37 adults. In addition to numerical aberrations such as trisomy 7 and gonosomal losses, we found structural and/or numerical aberrations of chromosome 1 in eight astrocytomas. There was a striking difference between the rearranged chromosomes in pediatric and adult patients. Whereas the pediatric tumors revealed monosomies 1p with accompanying trisomy 1q, the astrocytomas in adults showed partial or complete monosomies 1q.     

Molecular and phylogenetic characterization of the sieve element occlusion gene family in <Emphasis Type="Italic">Fabaceae</Emphasis> and non-<Emphasis Type="Italic">Fabaceae</Emphasis>plants

Background  

The phloem of dicotyledonous plants contains specialized P-proteins (phloem proteins) that accumulate during sieve element differentiation and remain parietally associated with the cisternae of the endoplasmic reticulum in mature sieve elements. Wounding causes P-protein filaments to accumulate at the sieve plates and block the translocation of photosynthate. Specialized, spindle-shaped P-proteins known as forisomes that undergo reversible calcium-dependent conformational changes have evolved exclusively in the Fabaceae. Recently, the molecular characterization of three genes encoding forisome components in the model legume Medicago truncatula (MtSEO1, MtSEO2 and MtSEO3; SEO = sieve element occlusion) was reported, but little is known about the molecular characteristics of P-proteins in non-Fabaceae.     

Recombinant artificial forisomes provide ample quantities of smart biomaterials for use in technical devices

Forisomes are mechanoproteins that undergo ATP-independent contraction–expansion cycles triggered by divalent cations, pH changes, and electrical stimuli. Although native forisomes from Medicago truncatula comprise a number of subunits encoded by separate genes, here we show that at least two of those subunits (MtSEO1 and MtSEO4) can assemble into homomeric forisome bodies that are functionally similar to their native, multimeric counterparts. We expressed these subunits in plants and yeast, resulting in the purification of large quantities of artificial forisomes with unique characteristics depending on the expression platform. These artificial forisomes were able to contract and expand in vitro like native forisomes and could respond to electrical stimulation when immobilized between interdigital transducer electrodes. These results indicate that recombinant artificial forisomes with specific characteristics can be prepared in large amounts and used as components of microscale and nanoscale devices.     

Fringes of the empire: Diet and cultural change at the Roman to post‐Roman transition in NW Iberia

A growing number of paleodiet investigations over recent years have begun to reveal the stark dietary differences that existed between regions of the Roman Empire, as well as significant changes in subsistence strategies after its fall. The present study explores the dietary changes at the Roman to post‐Roman (Germanic) transition in the Northwest Iberian Peninsula, in order to improve our understanding of the changes that occurred at end of the Roman Empire in different regions across Europe and to also consider the influence of climate had on them. The carbon and nitrogen stable isotope investigation in bone collagen from A Lanzada, NW Spain (100–700 AD), which was an important commercial, coastal settlement has been presented. A human sample of 59 individuals, 6 of them subadults, is compared with 31 faunal specimens, which include a number of marine fish. Isotope data for the terrestrial fauna reveal the influence of the sea on the local isotope baseline. Analysis of the human samples indicates a mixed marine‐terrestrial diet. A shift in mean human δ¹³C values from ?16.7‰ to ?14.3‰ provides clear evidence for a significant change in diet in the post‐Roman period, probably through the intensification of both marine resources exploitation and C₄‐plant consumption (presumably millet). A deterioration of paleoenvironmental conditions, together with a poor socioeconomic situation and the arrival of new people, the Sueves, who brought a new political and socioeconomic system have been discussed as the main causes for the dietary modification in post‐Roman times.     

Effects of simulated microgravity on thyroid carcinoma cells

We aimed to investigate whether simulated microgravity on thyroid carcinoma cells could help to perform in vitro cancer studies such as antitumor drug tests more reliable and to spare animal experiments. We cultured cancer cells at 0 g to enable formation of three-dimensional multicellular tumor spheroids (MCTS), which will resemble the originating tumors. Under microgravity human follicular cells (ML-1 cell line) keep floating with-out stirring so that initial cell-cell interactions required for spheroid formation will be induced by forces due to biochemical components actually expressed on surfaces of cells, whereas gravity related push- or shear events will not influence MCTS formation. Within 12 hours of clinorotation the monolayer turned spontaneously into MCTS with remarkable features: An increase of extracellular matrix proteins and TGF-beta 1. Thyroglobulin, ft3 and ft4 secretion were markedly reduced. These data are in agreement with the observation that astronauts show low thyroid hormone levels after spaceflight.     

Cysteamine, the natural metabolite of pantetheinase, shows specific activity against Plasmodium

In mice, loss of pantetheinase activity causes susceptibility to infection with Plasmodium chabaudi AS. Treatment of mice with the pantetheinase metabolite cysteamine reduces blood-stage replication of P. chabaudi and significantly increases survival. Similarly, a short exposure of Plasmodium to cysteamine ex vivo is sufficient to suppress parasite infectivity in vivo. This effect of cysteamine is specific and not observed with a related thiol (dimercaptosuccinic acid) or with the pantethine precursor of cysteamine. Also, cysteamine does not protect against infection with the parasite Trypanosoma cruzi or the fungal pathogen Candida albicans, suggesting cysteamine acts directly against the parasite and does not modulate host inflammatory response. Cysteamine exposure also blocks replication of P. falciparum in vitro; moreover, these treated parasites show higher levels of intact hemoglobin. This study highlights the in vivo action of cysteamine against Plasmodium and provides further evidence for the involvement of pantetheinase in host response to this infection.     

Spatial and temporal regulation of the forisome gene <Emphasis Type="Italic">for1</Emphasis> in the phloem during plant development

Forisomes are protein aggregates found uniquely in the sieve elements of Fabaceaen plants. Upon wounding they undergo a reversible, calcium-dependent conformational switch which enables them to act as cellular stopcocks. Forisomes begin to form in young sieve elements at an early stage of metaphloem differentiation. Genes encoding forisome components could therefore be useful as markers of early sieve element development. Here we present a comprehensive analysis of the developmental expression profile of for1, which encodes such a forisome component. The for1 gene is highly conserved among Fabaceaen species and appears to be unique to this phylogenetic lineage since no orthologous genes have been found in other plants, including Arabidopsis and rice. Even so, transgenic tobacco plants expressing reporter genes under the control of the for1 promoter display reporter activity exclusively in immature sieve elements. This suggests that the regulation of sieve element development is highly conserved even in plants where mature forisomes have not been detected. The promoter system could therefore provide a powerful tool for the detailed analysis of differentiation in metaphloem sieve elements in an unexpectedly broad range of plant species. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.