Novel Gallate Triphenylphosphonium Derivatives with Potent Antichagasic Activity

Chagas disease is one of the most neglected tropical diseases in the world, affecting nearly 15 million people, primarily in Latin America. Only two drugs are used for the treatment of this disease, nifurtimox and benznidazole. These drugs have limited efficacy and frequently induce adverse effects, limiting their usefulness. Consequently, new drugs must be found. In this study, we demonstrated the in vitro trypanocidal effects of a series of four gallic acid derivatives characterized by a gallate group linked to a triphenylphosphonium (TPP⁺) moiety (a delocalized cation) via a hydrocarbon chain of 8, 10, 11, or 12 atoms (TPP⁺-C₈, TPP⁺-C₁₀, TPP⁺-C₁₁, and TPP⁺-C₁₂, respectively). We analyzed parasite viability in isolated parasites (by MTT reduction and flow cytometry) and infected mammalian cells using T. cruzi Y strain trypomastigotes. Among the four derivatives, TPP⁺-C₁₀ and TPP⁺-C₁₂ were the most potent in both models, with EC₅₀ values (in isolated parasites) of 1.0 ± 0.6 and 1.0 ± 0.7 μM, respectively, and were significantly more potent than nifurtimox (EC₅₀ = 4.1 ± 0.6 μM). At 1 μM, TPP⁺-C₁₀ and TPP⁺-C₁₂ induced markers of cell death, such as phosphatidylserine exposure and propidium iodide permeabilization. In addition, at 1 μM, TPP⁺-C₁₀ and TPP⁺-C₁₂ significantly decreased the number of intracellular amastigotes (TPP⁺-C₁₀: 24.3%, TPP⁺-C₁₂: 19.0% of control measurements, as measured by DAPI staining) and the parasite’s DNA load (C₁₀: 10%, C₁₂: 13% of control measurements, as measured by qPCR). Based on the previous mode of action described for these compounds in cancer cells, we explored their mitochondrial effects in isolated trypomastigotes. TPP⁺-C₁₀ and TPP⁺-C₁₂ were the most potent compounds, significantly altering mitochondrial membrane potential at 1 μM (measured by JC-1 fluorescence) and inducing mitochondrial transition pore opening at 5 μM. Taken together, these results indicate that the TPP⁺-C₁₀ and TPP⁺-C₁₂ derivatives of gallic acid are promising trypanocidal agents with mitochondrial activity.     

Effect of the red imported fire ant on cotton aphid population density and predation of bollworm and beet armyworm eggs

The effects of the red imported fire ant, Solenopsis invicta Buren (Hymenoptera: Formicidae), on cotton aphid, Aphis gossypii Glover, populations and its predation of bollworm, Helicoverpa zea (Boddie), and beet armyworm, Spodoptera exigua (Hübner), (both Lepidoptera: Noctuidae) eggs were evaluated in cotton under field conditions during 2001 and 2002 in central and northern Texas. In central Texas, cotton aphid populations were approximately 5.5 times greater and predation of sentinel bollworm eggs 2 times greater in the presence of S. invicta versus in its absence, although aphid populations did not reach economic levels. Most predation of beet armyworm egg masses, measured via direct nocturnal observations, was due to S. invicta (68%) and cotton fleahopper, Pseudatomoscelis seriatus (Reuter) (21%), where S. invicta was present, and by the mite Abrolophus sp. (52%), spiders (13%), and minute pirate bug (Orius sp.) (13%) where S. invicta was absent. Predation of sentinel bollworm eggs and beet armyworm egg masses was approximately 1.5 and 4.1 times greater, respectively, in the presence of S. invicta versus in their absence. In the presence of S. invicta, the relative frequencies of minute pirate bug and cotton fleahopper were higher, and of S. invicta and native ants lower in beat bucket samples compared with their relative frequencies in nocturnal observations of predation upon beet armyworm egg masses. In the absence of S. invicta seven of eight predators sampled were similarly represented in beat bucket samples and nocturnal observations of beet armyworm egg mass predation, whereas minute pirate bug occurred at a higher frequency in beat bucket samples relative to nocturnal observations. These observations suggested that the relative frequencies of minute pirate bug, cotton fleahopper, S. invicta and native ants in beat bucket samples do not closely reflect the frequency with which these predators prey on noctuid eggs. Overall, the results of this study show that although S. invicta may promote aphid populations early in the growing season, it is an important predator of bollworm and beet armyworm eggs later in the season.     

The novel gene<Emphasis Type="Italic"> CpEdi-9</Emphasis> from the resurrection plant<Emphasis Type="Italic"> C. plantagineum</Emphasis> encodes a hydrophilic protein and is expressed in mature seeds as well as in response to dehydration in leaf phloem tissues

The resurrection plant Craterostigma plantagineum Hochst. is used as an experimental system to investigate desiccation tolerance in higher plants. A search for genes activated during early stages of dehydration identified the gene CpEdi-9, which is expressed in mature seeds and in response to dehydration in the phloem cells of vascular tissues of leaves. Elements for the tissue-specific expression pattern reside in the isolated promoter of the CpEdi-9 gene, as shown through the analysis of transgenic plants. The CpEdi-9 promoter could be a suitable tool for expressing genes in the vascular system of dehydrated plants. CpEdi-9 encodes a small (10 kDa) hydrophilic protein, which does not have significant sequence homologies to known genes. The predicted protein CpEDI-9 shares some physicochemical features with LEA proteins from plants and a nematode. Based on the unique expression pattern and on the nucleotide sequence we propose that CpEdi-9 defines a new class of hydrophilic proteins that are supposed to contribute to cellular protection during dehydration. This group of proteins may have evolved because desiccation tolerance requires the abundant expression of protective proteins during early stages of dehydration in all tissues.Abbreviations ABA Abscisic acid - ABRE ABA-responsive element - Edi Early dehydration induced - GUS Glucuronidase - LEA Late embryogenesis abundant - MU Methylumbelliferone This article is dedicated to Prof. Dr. Francesco Salamini on the occasion of his 65th birthday and his departure from the Max Planck Institute in Köln     

Death is the major fate of medial edge epithelial cells and the cause of basal lamina degradation during palatogenesis

During mammalian development, a pair of shelves fuses to form the secondary palate, a process that requires the adhesion of the medial edge epithelial tissue (MEE) of each shelf and the degeneration of the resulting medial epithelial seam (MES). It has been reported that epithelial-mesenchymal transformation (EMT) occurs during shelf fusion and is considered a fundamental process for MES degeneration. We recently found that cell death is a necessary process for shelf fusion. These findings uncovered the relevance of cell death in MES degeneration; however, they do not discard the participation of other processes. In the present work, we focus on the evaluation of the processes that could contribute to palate shelf fusion. We tested EMT by traditional labeling of MEE cells with a dye, by infection of MEE with an adenovirus carrying the lacZ gene, and by fusing wild-type shelves with the ones from EGFP-expressing mouse embryos. Fate of MEE labeled cells was followed by culturing whole palates, or by a novel slice culture system that allows individual cells to be followed during the fusion process. Very few labeled cells were found in the mesenchyme compartment, and almost all were undergoing cell death. Inhibition of metalloproteinases prevented basal lamina degradation without affecting MES degeneration and MEE cell death. Remarkably, independently of shelf fusion, activation of cell death promoted the degradation of the basal lamina underlying the MEE ('cataptosis'). Finally, by specific labeling of periderm cells (i.e. the superficial cells that cover the basal epithelium), we observed that epithelial triangles at oral and nasal ends of the epithelial seam do not appear to result from MEE cell migration but rather from periderm cell migration. Inhibition of migration or removal of these periderm cells suggests that they have a transient function controlling MEE cell adhesion and survival, and ultimately die within the epithelial triangles. We conclude that MES degeneration occurs almost uniquely by cell death, and for the first time we show that this process can activate basal lamina degradation during a developmental process.     

Trophic relationships in the nearshore zone of Martel Inlet (King George Island,Antarctica): δ<Superscript>13</Superscript>C stable-isotope analysis

Carbon isotopic composition was used to assess the linkage between three different potential sources of energy and the community in the shallow coastal zone of Martel Inlet. Stable ¹³C ratios ranged from –28.7 for the zooplankton plus phytoplankton to –14.4 for the grazer Nacella concinna. Microphytobenthos (–16.7) was considerably more enriched in ¹³C than were suspended particulate matter (SPM) (–25.6) and macroalgal fragments (–23.6 and –21.1), indicating that stable carbon isotope analysis might be used to discern the relative contribution of these sources of primary production. There is a benthic-pelagic coupling between plankton, benthic suspensivores, the ophiuroid Ophionotus victoriae and the icefish Chaenocephalus aceratus. Benthic grazers such as N. concinna, deposit feeders such as Yoldia eightsi and the nematodes showed a tight coupling with the microphytobenthos and the sediment. Some omnivorous/depositivorous polychaetes, echinoids, amphipods and the fish Notothenia coriiceps showed values close to the ratios of the macroalgal fragments. Benthic carnivores and/or scavengers were generally enriched over suspensivores and depleted in relation to microphytobenthos grazers, showing a considerable overlap in ¹³C values throughout the food web, without any clear coupling with the primary sources of organic matter. The trophic web in the shallow zone of high benthic production and under seasonal ice cover in the Antarctic is more complex than it is in shelf areas, where SPM is the main food source. The soft-bottom community in the shallow zone of Martel Inlet is enriched in ¹³C due to the significant input of carbon from the microphytobenthos and macroalgal fragments.     

Solution structure of subunit F(6) from the peripheral stalk region of ATP synthase from bovine heart mitochondria

The ATP synthase enzyme structure includes two stalk assemblies, the central stalk and the peripheral stalk. Catalysis involves rotation of the central stalk assembly together with the membrane-embedded ring of c-subunits driven by the trans-membrane proton-motive force, while the alpha and beta-subunits of F(1) are prevented from co-rotating by their attachment to the peripheral stalk. In the absence of structures of either the intact peripheral stalk or larger complexes containing it, we are studying its individual components and their interactions to build up an overall picture of its structure. Here, we describe an NMR structural characterisation of F(6), which is a 76-residue protein located in the peripheral stalk of the bovine ATP synthase and is essential for coupling between the proton-motive force and catalysis. Isolated F(6) has a highly flexible structure comprising two helices packed together through a loose hydrophobic core and connected by an unstructured linker. Analysis of chemical shifts, (15)N relaxation and RDC measurements confirm that the F(6) structure is flexible on a wide range of timescales ranging from nanoseconds to seconds. The relationship between this structure for isolated F(6) and its role in the intact peripheral stalk is discussed.     

Carbohydrate, glycolipid, and lipid components from the photobiont (Scytonema sp.) of the lichen, Dictyomema glabratum

The photobiont of the lichen, Dictyonema glabratum (Scytonema sp.), was isolated and cultivated in a soil-extract medium and submitted to chemical analysis. Successive extractions with CHCl3-MeOH, aqueous MeOH, and H2O gave rise to solutions of lipids (25%), low-molecular-weight carbohydrates (22%), and polysaccharides (4%), respectively. TLC of the lipid extract showed the presence of glycolipids, which were further purified and examined by NMR spectroscopy and GC-MS. Monogalactosyldiacylglycerol (1%), digalactosyldiacylglycerol (0.8%), trigalactosyldiacylglycerol (0.4%), and sulfoquinovosyldiacylglycerol (0.5%) were identified. The most abundant fatty acid ester in each fraction was palmitic (C16:0), but a great variation of the ester composition from one to another was found. Others present were those of C12:0, C14:0, C15:0, C16:1, C17:0, C18:0, C18:1, C18:2, C18:3, C22:0, C22:2, and C24:0. The lipid extract was also subjected to acid methanolysis, which gave rise to dodecane, 2-Me-heptadecane, 2,6-Me2-octadecane, and 8-Me-octadecane, methyl esters of C14:0, C15:0, C16:0, C16:1, C17:0, C18:0, C18:1, C18:2, C20:0, and C24:0 fatty acids, and the dimethyl ester of decanedioic acid. The polysaccharide had mainly Glc, Gal, and Man, with small amounts of 3-O-methylrhamnose and 2-O-methylxylose, both found in plants, and unexpectedly, some of the units were beta-galactofuranose, typical of fungal, but not cyanobacterial polysaccharides. The low-molecular-weight carbohydrates showed mannose as the main free reducing sugar, which differs from Nostoc sp. and Trebouxia sp. photobionts.     

Peroxisomal proliferation protects from beta-amyloid neurodegeneration

Alzheimer disease is a neurodegenerative process that leads to severe cognitive impairment as a consequence of selective death of neuronal populations. The molecular pathogenesis of Alzheimer disease involves the participation of the beta-amyloid peptide (Abeta) and oxidative stress. We report here that peroxisomal proliferation attenuated Abeta-dependent toxicity in hippocampal neurons. Pretreatment with Wy-14.463 (Wy), a peroxisome proliferator, prevent the neuronal cell death and neuritic network loss induced by the Abeta peptide. Moreover, the hippocampal neurons treated with this compound, showed an increase in the number of peroxisomes, with a concomitant increase in catalase activity. Additionally, we evaluate the Wy protective effect on beta-catenin levels, production of intracellular reactive oxygen species, cytoplasmic calcium uptake, and mitochondrial potential in hippocampal neurons exposed to H(2) O(2) and Abeta peptide. Results show that the peroxisomal proliferation prevents beta-catenin degradation, reactive oxygen species production, cytoplasmic calcium increase, and changes in mitochondrial viability. Our data suggest, for the first time, a direct link between peroxisomal proliferation and neuroprotection from Abeta-induced degenerative changes.