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[
Indian J Exp Biol,
2015]
Advanced Glycation End products (AGE) generated in a non enzymatic protein glycation process are frequently associated with diabetes, aging and other chronic diseases. Here, we explored the protective effect of phlorotannins from brown algae Padina pavonica, Sargassum polycystum and Turbinaria ornata against AGEs formation. Phlorotannins were extracted from brown algae with methanol and its purity was analyzed by TLC and RP-HPLC-DAD. Twenty five grams of P. pavonica, S. polycystum, T. ornata yielded 27.6 +/- 0.8 g/ml, 37.7 g/ml and 37.1 +/- 0.74 g/ml of phloroglucinol equivalent of phlorotannins, respectively. Antioxidant potentials were examined through DPPH assay and their IC50 values were P. pavonica (30.12 +/- 0.99 g), S. polycystum (40.9 +/- 1.2 g) and T. ornata (22.9 +/- 1.3 g), which was comparatively lesser than the control ascorbic acid (46 +/- 0.2 g). Further, anti-AGE activity was examined in vitro by BSA-glucose assay with the extracted phlorotannins of brown algae (P. pavonica, 15.16 +/- 0.26 g/ml; S. polycystum, 35.245 +/- 2.3 g/ml; T. ornata, 22.7 +/- 0.3 g/ml), which revealed the required concentration to inhibit 50% of albumin glycation (IC50) were lower for extracts than controls (phloroglucinol, 222.33 +/- 4.9 g/ml; thiamine, 263 g/ml). Furthermore, brown algal extracts containing phlorotannins (100 l) exhibited protective effects against AGE formation in vivo in C. elegans with induced hyperglycemia.
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[
J Biol Chem,
2005]
Identification of cis-regulatory elements and their binding proteins constitutes an important part of understanding gene function and regulation. It is well accepted that co-expressed genes tend to share transcriptional elements. However, recent findings indicate that co-expression data show poor correlation with co-regulation data even in unicellular yeast. This motivates us to experimentally explore whether it is possible that co-expressed genes are subject to differential regulatory control using C. elegans'' excretory cell as an example. Excretory cell is a functional equivalent of human kidney. Transcriptional regulation of gene expression in the cell is largely unknown. We isolated a 10 bp excretory cell specific cis-element, Ex-1 from a
pgp-12 promoter. The significance of the element has been demonstrated by its capacity of converting an intestine specific promoter into an excretory cell-specific one. We also isolated a cDNA encoding an Ex-1 binding transcription factor, DCP-66 using a yeast one-hybrid screen. Role of the factor in regulation of
pgp-12 expression has been demonstrated both in vitro and in vivo. Search for occurrence of Ex-1 reveals that only a small portion of excretory cell-specific promoters contain Ex-1. Two other distinct cis-elements isolated from two different promoters can also dictate the excretory cell-specific expression but are independent of regulation by DCP-66. The results indicate that distinct regulatory elements are able to mediate the similar expression patterns.
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[
J Immunol,
2005]
ES-62 is a phosphorylcholine-containing glycoprotein secreted by filarial nematodes, which has previously been shown to possess a range of immunomodulatory capabilities. We now show, using a CD4+ transgenic TCR T cell adoptive transfer system, that ES-62 can modulate heterologous Ag (OVA)-specific responses in vivo. Thus, in contrast to the mixed IgG1-IgG2a response observed in control animals, ES-62-treated mice exhibited a Th2-biased IgG Ab response as evidenced by stable enhancement of anti-OVA IgG1 production and a profound inhibition of anti-OVA IgG2a. Consistent with this, Ag-specific IFN-gamma produced was suppressed by pre-exposure to ES-62 when T cells were rechallenged ex vivo. However, the response observed was not classical Th2, because although Ag-specific IL-5 production was enhanced by pre-exposure to ES-62, IL-13, and IL-4 were inhibited when T cells were rechallenged ex vivo. Moreover, such T cells produced lower levels of IL-2 and proliferated less upon Ag rechallenge ex vivo. Finally, pre-exposure to ES-62 inhibited the clonal expansion of the transferred Ag-specific CD4+ T cells and altered the functional response of such T cells in vivo, by modulating the kinetics and reducing the extent of their migration into B cell follicles.
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[
Heredity,
2001]
In Drosophila elegans, partial sexual isolation has developed between the brown and black morphs, which are distributed allopatrically. The present study aims to understand how they discriminate between potential mates. Mating experiments show that the females of the two morphs differ in sexual signal(s) and the males discriminate using these differences. Body colouration is not used as a sexual cue in this species. Between the females of the two morphs, a large difference was observed in the percentages of 7-pentacosene and 9-pentacosene on the cuticle. Genetical analysis using recombinant inbred lines supported the possibility that the concentration of these pentacosenes plays a role in mate discrimination of these two morphs. However, males did not respond to killed females at all, suggesting that cuticular hydrocarbons of females are not the only cue for the induction of male courtship behaviour. It may be that unknown signals or substances are essential to induce male courtship and pentacosenes modulate the attractiveness of females, positively in the black morph and negatively in the brown morph. Drosophila elegans F1 offspring had intermediate characteristics in mate discrimination and hydrocarbon composition between the parental brown and black morph strains. The number of loci responsible for the differences in the concentration of pentacosenes and the male and female components in the mate recognition between these two morphs is suggested to be more than one.
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[
BMC Syst Biol,
2012]
BACKGROUND: Synchronized bursting activity (SBA) is a remarkable dynamical behavior in both ex vivo and in vivo neural networks. Investigations of the underlying structural characteristics associated with SBA are crucial to understanding the system-level regulatory mechanism of neural network behaviors. RESULTS: In this study, artificial pulsed neural networks were established using spike response models to capture fundamental dynamics of large scale ex vivo cortical networks. Network simulations with synaptic parameter perturbations showed the following two findings. (i) In a network with an excitatory ratio (ER) of 80-90%, its connective ratio (CR) was within a range of 10-30% when the occurrence of SBA reached the highest expectation. This result was consistent with the experimental observation in ex vivo neuronal networks, which were reported to possess a matured inhibitory synaptic ratio of 10-20% and a CR of 10-30%. (ii) No SBA occurred when a network does not contain any all-positive-interaction feedback loop (APFL) motif. In a neural network containing APFLs, the number of APFLs presented an optimal range corresponding to the maximal occurrence of SBA, which was very similar to the optimal CR. CONCLUSIONS: In a neural network, the evolutionarily selected CR (10-30%) optimizes the occurrence of SBA, and APFL serves a pivotal network motif required to maximize the occurrence of SBA.
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[
Methods Cell Biol,
2012]
This chapter is an update of the previously published book chapter "Correlative Light and Electron Microscopy of Early C. elegans Embryos in Mitosis" (Muller-Reichert, Srayko, Hyman, O'Toole, & McDonald, 2007). Here, we have adapted and improved the protocol for the isolated meiotic embryos, which was necessary to meet the specific challenges a researcher faces while investigating the development of very early Caenorhabditis elegans embryos ex-utero. Due to the incompleteness of the eggshell assembly, the meiotic embryo is very fragile and much more susceptible to changes in the environmental conditions than the mitotic ones. To avoid phototoxicity associated with wide-field UV illumination, we stage the meiotic embryos primarily using transmitted visible light. Throughout the staging and high-pressure freezing, we incubate samples in an isotonic embryo buffer. The ex-utero approach allows precise tracking of the developmental events in isolated meiotic embryos, thus facilitating the comparison of structural features between wild-type and mutant or RNAi-treated samples.
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Gazolla MC, Almeida J, Lemes BL, Silva LM, Leal CM, Moura RO, da Silva Filho AA, Cajas RA, de Moraes J, Costa DS, Carvalho LSA
[
J Ethnopharmacol,
2023]
ETHNOPHARMACOLOGICAL RELEVANCE: Roots of Pothomorphe umbellata (L.) Miq. are used in traditional medicine of Africa and South America for the treatment of malaria and helminthiasis. However, neither P. umbellata nor its isolated compounds have been evaluated against Schistosoma species. AIMS OF THIS STUDY: To investigate the antischistosomal effects of P. umbellata root extracts and the isolated compound 4-nerolidylcatechol (4-NC) against Schistosoma mansoni ex vivo and in murine models of schistosomiasis. MATERIALS AND METHODS: The crude hydroalcoholic (PuE) and hexane (PuH) extracts of P. umbellata roots were prepared and initially submitted to an ex vivo phenotypic screening against adult S. mansoni. PuH was analyzed by HPLC-DAD, characterized by UHPLC-HRMS/MS, and submitted to chromatographic fractionation, leading to the isolation of 4-NC. The anthelmintic properties of 4-NC were assayed ex vivo against adult schistosomes and in murine models of schistosomiasis for both patent and prepatent S. mansoni infections. Praziquantel (PZQ) was used as a reference compound. RESULTS: of 2.9&#
x202f;&#
x3bc;M (0.91&#
x202f;&#
x3bc;g/mL) and a selectivity index higher than 68 against Vero mammalian cells, without affecting viability of nematode Caenorhabditis elegans. In patent S. mansoni infection, the oral treatment with 4-NC decreased worm burden and egg production in 52.1% and 52.3%, respectively, also reducing splenomegaly and hepatomegaly. 4-NC, unlike PZQ, showed in vivo efficacy against juvenile S. mansoni, decreasing worm burden in 52.4%. CONCLUSIONS: This study demonstrates that P. umbellata roots possess antischistosomal activity, giving support for the medicinal use of this plant against parasites. 4-NC was identified from P. umbellata roots as one of the effective in vitro and in vivo antischistosomal compound and as a potential lead for the development of novel anthelmintics.
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[
Lett Appl Microbiol,
2015]
The insecticidal activity of Bacillus thuringiensis is owing to the action of Cry and Cyt proteins. In addition to the synthesis of insecticidal proteins, some strains are able to synthesize -exotoxin, which is highly toxic to humans. In this regard, it is very important to have a simple method to detect -exotoxin in order to avoid the commercial production of this type of strains. In this work, we developed a simple and fast method, using the nematode Caenorhabditis elegans to detect indirectly the synthesis of -exotoxin by B. thuringiensis strain. Using this assay, we detected that ~ 60% of Mexican native strains (i.e. LBIT-471, 491, 492, 497, 507, 511, 515, 536, and 537) were toxic to the nematode (44 to 97% mortalities), and their -exotoxin (Ex(+) ) production, including a positive control (NRD-12), was confirmed by HPLC. In addition, the negative controls (Ex(-) ) LBIT-436 (HD-1) and LBIT-438 and also the native strains LBIT-499, 500, 521, 522, 533 and 542, did not show a detrimental effect against nematodes larvae, neither the synthesis of -exotoxin as determined by HPLC. Finally, we did not find a correlation between B. thuringiensis strains with similar plasmid patterns and the -exotoxin production. This article is protected by copyright. All rights reserved.
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[
Mycologia,
1972]
The hyphomycete Asteromyces cruciatus F. & Mme Moreau was described without a Latin diagnosis or a designated type. The taxon was validated by Hennebert. The known distribution of this monotypic genus has been limited. F. and Mme Moreau found the fungus in sand dunes at Point du Siege (under Psamma sp.) and between Franceville and Le Home (under Agropyrum sp.) on the Normandy coast of France. Brown found A. cruciatus in open sand in the intertidal zone at Studland, Dorset and Sandwich, Kent, England; and Nicot found it in sand dunes and beach samples at Malo-les-Bains on the North Sea coast of France.
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[
Biophys J,
2023]
Embryonic development requires the establishment of cell polarity to enable cell fate segregation and tissue morphogenesis. This process is regulated by Par complex proteins, which partition into polarized membrane domains and direct downstream polarized cell behaviors. The kinase aPKC (along with its cofactor Par6) is a key member of this network and can be recruited to the plasma membrane by either the small GTPase Cdc42 or the scaffolding protein Par3. Although in vitro interactions among these proteins are well established, much is still unknown about the complexes they form during development. Here, to enable the study of membrane-associated complexes ex vivo, we used a maleic acid copolymer to rapidly isolate membrane proteins from single C. elegans zygotes into lipid nanodiscs. We show that native lipid nanodisc formation enables detection of endogenous complexes involving Cdc42, which are undetectable when cells are lysed in detergent. We found that Cdc42 interacts more strongly with aPKC/Par6 during polarity maintenance than polarity establishment, two developmental stages that are separated by only a few minutes. We further show that Cdc42 and Par3 do not bind aPKC/Par6 simultaneously, confirming recent in vitro findings in an ex vivo context. Our findings establish a new tool for studying membrane-associated signaling complexes and reveal an unexpected mode of polarity regulation via Cdc42.