Compromising the plasma membrane as a secondary target in photodynamic therapy-induced necrosis

Photodynamic therapy (PDT) is a non-invasive treatment widely applied to different cancers. The goal of PDT is the photo-induced destruction of cancer cells by the activation of different cell death mechanisms, including apoptosis and/or necrosis. Recent efforts focusing on understanding the mechanisms of cell death activated by PDT find that it depends on the type of photosensitizer (PS), targeted organelles, and nature of the light used. It is generally accepted that very short incubation times are required to direct the PS to the plasma membrane (PM), while longer periods result in the accumulation of the PS in internal compartments such as the endoplasmic reticulum or mitochondria. Glycosylation of the PS targets cancer via saccharide receptors on the cell surface, and is generally assumed that these compounds rapidly internalize and accumulate, e.g. in the endoplasmic reticulum. Herein we demonstrate that a minor fraction of a glycosylated chlorin compound residing at the PM of cancer cells can activate necrosis upon illumination by compromising the PM independently of the length of the incubation period. The results presented here show that the PM can also be targeted by glycosylated PS designed to accumulate in internal organelles. PS activation to induce necrosis by compromising the plasma membrane has the benefits of fast cell death and shorter irradiation times. The findings described here expand our understanding of the cellular damage induced by phototherapies, presenting the possibility of activating another cell death mechanism based on the incubation time and type of light used.     

The many faces of Peridinium cinctum (Peridiniaceae,Peridiniales): morphological and molecular variability in a common dinophyte

Peridinium cinctum is a common freshwater dinophyte with a long history of research. Erich Lindemann was the first to assess intraspecific variability in this species focusing on plate pattern variation. Since then, this issue has been neglected but with the application of DNA sequence diagnostics, a combination of morphological and molecular characters may enable taxonomic delimitations. Our aim was to identify distinct morphotypes using plate pattern as the main characteristic and then compare them to the geographic occurrence of particular ribotypes (as inferred from sequences of the Internal Transcribed Spacer: ITS) in samples from Central Europe. Approximately 200 observations were carried out under the inverse light microscope for each of a total of 15 strains. We observed two main variations from the abundant plate pattern in P. cinctum, namely an unusual position of the 2a plate and the irregular shape of the 1a plate. In 88 (predominantly clonal) strains, we identified five different ribotypes (submitted as 71 new GenBank entries) which had no clear correlation to the defined morphotypes and/or spatial occurrences. In four cases, we detected two distinct ribotypes at the same locality. However, samples collected south of the Danube River presented a different predominant morphotype from the rest of the samples, thus implying a potential biogeographic signal as inferred from morphology. In general, there is morphological and molecular variability in P. cinctum, which is under-studied and which may uncover geographic or ecological correlations or even the existence of cryptic species.     

4-N-Alkanoyl and 4-N-alkyl gemcitabine analogues with NOTA chelators for 68-gallium labelling

The conjugation of 4-N-(3-aminopropanyl)-2′-deoxy-2′,2′-difluorocytidine with 2-(p-isothiocyanatobenzyl)-1,4,7-triazacyclononane-1,4,7-triacetic acid (SCN-Bn-NOTA) ligand in 0.1?M Na₂CO₃ buffer (pH 11) at ambient temperature provided 4-N-alkylgemcitabine-NOTA chelator. Incubation of latter with excess of gallium(III) chloride (GaCl₃) (0.6?N AcONa/H₂O, pH?=?9.3) over 15?min gave gallium 4-N-alkylgemcitabine-NOTA complex which was characterized by HRMS. Analogous [⁶⁸Ga]-complexation of 4-N-alkylgemcitabine-NOTA conjugate proceeded with high labeling efficiency (94%–96%) with the radioligand almost exclusively found in the aqueous layer (～95%). The high polarity of the gallium 4-N-alkylgemctiabine-NOTA complex resulted in rapid renal clearance of the ⁶⁸Ga-labelled radioligand in BALB/c mice.     

Foliar C,N, and P stoichiometry characterize successful plant ecological strategies in the Sonoran Desert

Ecological processes are centered to water availability in drylands; however, less known nutrient stoichiometry can help explain much of their structure and ecological interactions. Here we look to the foliar stoichiometry of carbon (C), nitrogen (N), and phosphorus (P) of 38 dominant plant species from the Sonoran Desert, grouped in four different functional types to describe ecological characteristics and processes. We found that foliar N, C:N, C:P, and N:P stoichiometric ratios, but not P, were higher than those known to most other ecosystems and indicate P but not N limitations in leaves. Biological N fixers (BNF) had even higher leaf N concentrations, but bio-elemental concentrations and stoichiometry ratios were not different to other non-N-fixing legume species which underscores the need to understand the physiological mechanisms for high N, and to how costly BNF can succeed in P-limiting drylands environments. Stoichiometry ratios, and to lesser extent elemental concentrations, were able to characterize BNF and colonizing strategies in the Sonoran Desert, as well as explain leaf attribute differences, ecological processes, and biogeochemical niches in this dryland ecosystem, even when no direct reference is made to other water-limitation strategies.     

Where can switchgrass production be more profitable than corn and soybean? An integrated subfield assessment in Iowa,USA

Perennial bioenergy crops are considered an important feedstock for a growing bioeconomy. However, in the USA, production of biofuel from these dedicated, nonfood crops is lagging behind federal mandates and markets have yet to develop. Most studies on the economic potential of perennial biofuel crops have concluded that even high‐yielding bioenergy grasses are unprofitable compared to corn/soybeans, the prevailing crops in the United States Corn Belt. However, they did not account for opportunities precision agriculture presents to integrate perennials into agronomically and economically underperforming parts of corn/soybean fields. Using publicly available subfield data and market projections, we identified an upper bound to the areas in Iowa, United States, where the conversion from corn/soybean cropland to an herbaceous bioenergy crop, switchgrass, could be economically viable under different price, land tenancy, and yield scenarios. Assuming owned land, medium crop prices, and a biomass price of US$ 55 Mg^?1, we showed that 4.3% of corn/soybean cropland could break even when converted to switchgrass yielding up to 10.08 Mg ha^?1. The annualized change in net present value on each converted subfield patch ranged from just above US$ 0 ha^?1 to 692 ha^?1. In the three counties of highest economic opportunity, total annualized producer benefits from converting corn/soybean to switchgrass summed to US$ 2.6 million, 3.4 million, and 7.6 million, respectively. This is the first study to quantify an upper bound to the potential private economic benefits from targeted conversion of unfavorable corn/soybean cropland to switchgrass, leaving arable land already under perennial cover unchanged. Broadly, we conclude that areas with high within‐field yield variation provide highest economic opportunities for switchgrass conversion. Our results are relevant for policy design intended to improve the sustainability of agricultural production. While focused on Iowa, this approach is applicable to other intensively farmed regions globally with similar data availability.     

Delivery of ALX-0171 by inhalation greatly reduces respiratory syncytial virus disease in newborn lambs

Respiratory syncytial virus (RSV) is a common cause of acute lower respiratory disease in infants and young children worldwide. Currently, treatment is supportive and no vaccines are available. The use of newborn lambs to model hRSV infection in human infants may provide a valuable tool to assess safety and efficacy of new antiviral drugs and vaccines. ALX-0171 is a trivalent Nanobody targeting the hRSV fusion (F) protein and its therapeutic potential was evaluated in newborn lambs infected with a human strain of RSV followed by daily ALX-0171 nebulization for 3 or 5 consecutive days.

Colostrum-deprived newborn lambs were infected with hRSV-M37 before being treated by daily nebulization with either ALX-0171 or placebo. Two different treatment regimens were examined: day 1–5 or day 3–5 post-infection. Lambs were monitored daily for general well-being and clinical parameters. Respiratory tissues and bronchoalveolar lavage fluid were collected at day 6 post-inoculation for the quantification of viral lesions, lung viral titers, viral antigen and lung histopathology.

Administration by inhalation of ALX-0171 was well-tolerated in these hRSV-infected newborn lambs. Robust antiviral effects and positive effects on hRSV-induced lung lesions and reduction in symptoms of illness were noted. These effects were still apparent when treatment start was delayed and coincided with peak viral loads (day 3 post-infection) and at a time point when signs of RSV disease were apparent. The latter design is expected to have high translational value for planned clinical trials. These results are indicative of the therapeutic potential of ALX-0171 in infants.     

Gracilaria-Mytilus interaction on a commercial algal farm in Chile

Mytilus chilensis is an invertebrate that competes for space with the alga Gracilaria chilensis in farmed areas in Chile and, for this reason, is considered a contaminant organism. Mussel beds are considered to play a role in the regeneration of nitrogen and, as a consequence, they could be an important source of ammonium for the algae. In this study, we manipulated the mussel cover in experimental plots, creating replicated areas with 0%, 30% and 60% coverage. In half of the plots the mussels were killed with a gas torch so their effect would be mechanical, without nutrient regeneration. After 15 days, each plot was planted with 12 G. chilensis bundles (100 g each) per square meter. Ammonium concentrations increased significantly in the waters around the mussel bed in contrast to areas without mussels or dead mussels. Mussel cover had a significant negative effect on the length of the G. chilensis bundles planted in the experimental plots. However, no significant differences were detected between experimental quadrats with live mussels and those with dead mussels during a 5 month period. These results indicate that the mechanical effect of the mussel can, to some extent, be responsible for the decline in G. chilensis abundance in farms where mussel beds have been established.     

New World Podostemaceae: Ecological and evolutionary enigmas

The largely tropical Podostemaceae occur in a unique habitat for angiosperms: attached to rocks in river rapids and waterfalls. Little taxonomic work has been published on New World Podostemaceae since Van Royen's monograph in the early 1950s. A paucity of complete herbarium material severely limits taxonomic study and precludes addressing a range of enigmatic ecological and evolutionary issues that relate to the family. The size, high level of endemism, and life history of the family are discussed. The distributions of two species in Mexico are used to illustrate the need for concerted collecting efforts. Recommendations for collection strategies are made that reflect the unusual habitat and life history of these plants.     

Comparative studies between freshly isolated and spontaneously immortalized bovine granulosa cells: Protein secretion,steroid metabolism,and responsiveness to growth factors

A bovine granulosa cell line (BGC-1) has been obtained by spontaneous immortalization of primary cultures. BGC-1 cells have retained some characteristics of primary cultures, such as the hormonal regulation of fibronectin biosynthesis. In the present study we have compaed BGC-1 cells and primary cultures of bovine granulosa cells in terms of protein secretion, steroid metabolism, and mitogenic responses to growth factors. The pattern of protein secretion in BGC-1 cells was qualitatively similar to that of primary cultures. The main differences were a higher proportion of fibronectin and the relative amounts of several other unidentified proteins. Progesterone levels in BGC-1 cultures were undetectable. When BGC-1 cells and primary cultures were incubated with [³H]-pregnenolone, the former showed a lower conversion rate to progesterone. In contrast, the conversion rate of [³H]-progesterone to 5α-reduced metabolites was markedly increased in BGC-1 cells. We also examined the effects of epidermal growth factor (EGF), insulin like growth factor-1 (IGF-I), and transforming growth factor-b? (TGF-b?) on DNA synthesis under serum-free conditions. Both primary cultures and BGC-1 cells exhibited a stimulatory response to EGF and IGF-I on [³H]-thymidine incorporation. Neither BGC-1 cells nor primary cultures showed a significant response to TGF-b? when added alone. However, in the presence of a combination of EGF and IGF-I, TGF-b? displayed an inhibitory effect on primary cultures while it stimulated DNA synthesis in BGC-1 cells even further. The addition of conditioned medium from BGC-1 cells (BGC-1-CM) stimulated DNA synthesis on primary cultures to a greater extent than the addition of conditioned medium from primary cultures. These results suggest that BGC-1 cells may be a useful model to study the regulation of granulosa cell function during the period previous to the preovulatory stage of follicular development. The differential responses of the immortalized cells to growth regulators may offer some clues on the mechanisms that control cell proliferation in normal tissues. © 1995 Wiley-Liss, Inc.