Revision on the effects of an entomopathogen bacteria Bacillus thuringiensis on the immune response of Red Palm Weevil (Rhynchophorus ferrugineus), is it a question of gender?

1 Università degli Studi di Palermo, Italia; 2 Università degli Studi dell’Insubria, Italia Il punteruolo rosso, Rhynchophorus ferrugineus (Olivier 1790) (Coleoptera: Curculionidae) fitofago infestante delle palme e Galleria mellonella L. (Lepidoptera: Pyralidae) parassita delle arnie, sono stati utilizzati come modelli per approfondire le conoscenze della relazione ospite-nematode entomopatogeno. Gli effetti del nematode entomopatogeno Steinernema carpocapsae (Nematoda: Steinernematidae) sull'immuno attività delle larve di R. ferrugineus e di G. mellonella, sono stati messi a confronto. E’ stata anche valutata la capacità di completare il ciclo vitale di S. carpocapsae nei due insetti. Sebbene S. carpocapsae sia efficace nel controllo biologico del punteruolo rosso, non è tuttavia in grado di completare il suo ciclo all’interno dell’ospite, mentre lo stesso nematode si riproduce con successo in G. mellonella. Sulla base di precedenti dati di laboratorio e sulla revisione sistematica della letteratura, gli effetti indotti dal nematode sulla risposta immunitaria del Coleottero e del Lepidottero, sono stati comparati. In particolare sono stati analizzati: la popolazione emocitaria immunocompetente (numero e tipi cellulari), l’attività del sistema profenolossidasi-fenolossidasi (proPO) e infine è stata valutata l’inibizione e proliferazione del batterio simbionte Gram negativo Xenorhabdus nematophila (Enterobacteraceae).Since their discovery, bioinsecticides based on Bacillus thuringiensis (Bt) have demonstrated targeted insect control with minimal environmental impact. Understanding the biology of insect pests, as well as interac- tions with pathogens and immune systems, can help to enhance their effect within these interactions. On the basis of previous laboratory data and a systematic literature review we analyzed the effects induced by the entomopathogen bacteria Bt, on red palm weevil (RPW), an highly invasive pest of palms that had a significant economic, environmental and social impact when introduced into Italy. The bacteria-induced effects on physiology, immune response and stress answers in the RPW. Rhynchophorus ferrugineus Olivier (Coleoptera: Curculionidae) were considered. Previously a Bt strain, commercialized against another family of Coleopters, was found to be a potential pathogen of RPW. Data showed that Bt interacts negatively with the growth and the vitality of RPW moreover with hemocytes of R. ferrugineus whose numbers decreased drastically in the hemolymph both in total number and in type. In particular, we focused on the Bt stress- induced infections considering the genders and the instars. The interaction between Bt and RPW hemocytes was analyzed by evaluating the expression heat shock proteins (HSPs) particularly HSP70 in the supernatant of the hemocyte lysate obtained from larvae and adults. HSPs are rapidly synthesized in the cells after stress exposition including infection by pathogens. The western blot analysis, showed that the HSP70 expression was modulated in the time (3h, 6h, 12h, 19, 24h) in the response to Bt treatment, highlighting that Bt is a stress factor for the larvae but also for adults. The protein expression was increased approximately seven times after 3 hours from treatment and after 6 hours it returning to control value. Data show difference in how males and females invest in Bt infection answer. Our data on immunity support the theory that males choose investment in reproduction over investment in immunity. In fact, B. thuringiensis-induced mortality data show the effect that females mount a stronger immune response and they also suffer lower mortality

Elevated cortisol modulates hsp70 and hsp90 gene expression and protein in sea bass head kidney and isolated leukocytes

In fish, interactions between Hsps and cortisol are involved in stress modulated physiological processes including innate immune responses. Cortisol exerts a role in the regulation of Hsps synthesis. Fish head kidney is a lymphomieloid and endocrine organ releasing cortisol, and it is the central organ for immune-endocrine interactions. In sea bass, cortisol intraperitoneal injection and in vitro treatment of head kidney cells show that inducible Hsp70 and Hsp90 are modulated by this hormone. However, an inverse relationship between mRNA expression (real-time PCR) and Hsp70 and Hsp90 protein levels (densitometric band analysis) was found. Time-course assays indicate a cortisol-mediated regulation. Furthermore, Hsp70 gene modulation appears to be more susceptible to the cortisol action and the mRNA was transcribed within 3. h post-injection. The restoration of the homeostatic conditions was observed at a week p.i., when plasma cortisol baseline was reached. Although fish manipulation and injection exerted stressing effects as indicated by serological parameters, differences between cortisol treated specimens compared to untreated or sham fish are statistically significant. Similar results were found by examining in vitro total cells and isolated leukocytes from head kidney cultured for 3. h with increasing cortisol concentration. Finally, MTT test and DNA fragmentation experiments showed that the apoptotic effect expected in cortisol-treated cells could be counteracted by high Hsp70 intracellular levels

Cadmium, Copper and Tributyltin effects on fertilization of Paracentrotus lividus (Echinodermata)

Marine environments are continuously being threatened by a large number of xenobi- otics from anthropogenic sources. The effect of chemical pollution on living organisms are numerous and may impair reproductive success of adults species of marine invertebrate and vertebrate through effects on gamete quality. Echinoderms are characterized by external fertilizzation and gametes, free of any type of protection, may be in contact with toxic substances so the reproductive success depends largely on the environment conditions. The purpose of this work is to assess the effects on the in vitro fertiliza- tion of exposure of sea urchin Paracentrotus lividus gametes to xenobiotic substances as CuSO4, CdCl2 and TBTCl. The effect of contaminant were assessed by two experimental set in which gametes were treated with different concentration (0, 10-3, 10-5 10-7 10-9 M) of different substances as CdCl2, CuSO4 and TBTCl. The effects were evaluated as percentage of fertilization. The results showed that the gametes exposure to xenobiotic decreased the percentage of fertilization and that more sensitive to treatment were the sperm cells, propably because the toxic effect affected the motility of the sperm. In conclusion, the absence of fertilization (spermiotoxicity) may submit the toxic effects of these substances to the level of body and may candidate the sea urchis as biosensors for the evaluation of environmental quality

In vitro effect of cadmium and copper on separated blood leukocytes of Dicentrarchus labrax.

The immunotoxic effects of heavy metals on blood leukocytes of sea bass (Dicentrarchus labrax) were examined. The cells, separated by a discontinuous Percoll-gradients, were exposed in vitro to various sublethal concentrations of cadmium and copper (10(-7) M, 10(-5) M, and 10(-3) M) and their immunotoxic effect was then evaluated by measuring neutral red uptake, MU assay, DNA fragmentation and Hsp70 gene expression. First of all, we demonstrated that the cells treated in vitro could incorporate Cd and Cu. A relationship between heavy metal exposure and dose-time-dependent alterations in responses of leukocytes from blood was found for both metals, but copper was more immunotoxic than cadmium in all assays performed. A significant reduction in the cells' ability to uptake neutral red and viability by MU assay was recorded, indicating that both cadmium and copper could change the membrane permeability, inducing cellular apoptosis when the concentration of metals reached 10(-3) M. The apoptotic effect may also explain the high level of cytotoxicity found when the leukocytes were exposed to higher concentration of metals. These results demonstrated that toxic effect of copper and cadmium affect on the mechanisms of cell-mediated immunity reducing the immune defences of the organism. (C) 2014 Elsevier Inc. All rights reserved

Expression of a glucocorticoid receptor (D1GR1) in several tissues of the teleost fish Dicentrarchus labrax

Since glucocorticoids have a role in maintaining the homeostatic status in fish, in the present paper mRNA expression (in situ hybridization) and tissue immunohistochemical localization of a glucocorticoid receptor (DlGR1) in several Dicentrarchus labrax organs are reported. Riboprobe and specific antibodies were prepared by using the DlGR1 that has been previously cloned and sequenced from peritoneal cavity leukocytes. Both mRNA and receptor were identified in head kidney, spleen, gills, intestine, heart and liver tissues. The functional roles of DlGR1 localization are discussed

The language of the mobile phone manual: Reading challenges to serve the technology

This article presents the results of a research study undertaken with different mobile phone users, focusing on the manual reading effects and impacts on the readers and on the use of this device. By relying on genre theory (Volochinov, 1988 [1929]; Bakhtin, 2000; Bazerman, 2006) and on the Sociodiscursive Interaction (Bronckart, 1999; Schneuwly, 2004), the aspects related to the manners of apprehension, constitution and circulation of the genre mobile phone user manual were analyzed. Despite the fact that the mobile phone users faced some difficulties during the manual reading – a fact that compromises its communicative purpose the analysis shows that the dominant language capacity required for this genre is the one which describe and prescribe actions. The study of the language used in this genre helped to identify its general characteristics and focus on the major problems present in its elaboration and apprehension by different levels of knowledge users. Key words: textual genre, language, manual, technology

An intronic SNP in the thyroid hormone receptor β gene is associated with pituitary cell-specific over-expression of a mutant thyroid hormone receptor β2 (R338W) in the index case of pituitary-selective resistance to thyroid hormone

<p>Abstract</p> <p>Background</p> <p>The syndrome of resistance to thyroid hormone (RTH) is caused by mutations in the thyroid hormone receptor β gene (<it>THRB</it>). The syndrome varies from asymptomatic to diffuse hypothyroidism, to pituitary-selective resistance with predominance of hyperthyroid signs and symptoms. The wide spectrum of clinical presentation is not completely attributable to specific <it>THRB </it>mutations. The <it>THRB </it>gene encodes two main isoforms, TR β1 which is widely distributed, and TR β2, whose expression is limited to the cochlea, retina, hypothalamus, and pituitary. Recent data demonstrated that in mice an intron enhancer region plays a critical role in the pituitary expression of the β2 isoform of the receptor. We thus hypothesized that polymorphisms in the human homologous region could modulate the pituitary expression of the mutated gene contributing to the clinical presentation of RTH.</p> <p>Methods</p> <p>Screening and <it>in vitro </it>characterization of polymorphisms of the intron enhancer region of the <it>THRB </it>gene in the index case of pituitary-selective RTH.</p> <p>Results</p> <p>The index case of pituitary-selective resistance is characterized by the missense R338W exon 9 mutation in <it>cis </it>with two common SNPs, rs2596623T and rs2596622C, located in the intron enhancer region of the <it>THRB </it>gene. Reporter gene assay experiments in GH3 pituitary-derived cells indicate that rs2596623T generates an increased pituitary cell-specific activity of the TR β2 promoter suggesting that rs2596623T leads to pituitary over-expression of the mutant allele.</p> <p>Conclusions</p> <p>The combined coding mutation and non-coding SNP therefore generate a tissue-specific dominant-negative condition recapitulating the patient's peculiar phenotype. This case illustrates the role of regulatory regions in modifying the clinical presentation of genetic diseases.</p

Cytotoxic Potential of the Coelomic Fluid Extracted from the Sea Cucumber Holothuria tubulosa against Triple-Negative MDA-MB231 Breast Cancer Cells

Growing evidence has demonstrated that the extracts of dierent holothurian species exert beneficial effects on human health. Triple negative breast cancers (TNBC) are highly malignant tumors that present a poor prognosis due to the lack of effective targeted therapies. In the attempt to identify novel compounds that might counteract TNBC cell growth, we studied the effect of the exposure of the TNBC cell line MDA-MB231 to total and filtered aqueous extracts of the coelomic fluid obtained from the sea cucumber Holoturia tubulosa, a widespread species in the Mediterranean Sea. In particular, we examined cell viability and proliferative behaviour, cell cycle distribution, apoptosis, autophagy, and mitochondrial metabolic/cell redox state. The results obtained indicate that both total and fractionated extracts are potent inhibitors of TNBC cell viability and growth, acting through both an impairment of cell cycle progression and mitochondrial transmembrane potential and a stimulation of cellular autophagy, as demonstrated by the increase of the acidic vesicular organelles and of the intracellular protein markers beclin-1, and total LC3 and LC3-II upon early exposure to the preparations. Identification of the water-soluble bioactive component(s) present in the extract merit further investigation aiming to develop novel prevention and/or treatment agents efficacious against highly metastatic breast carcinoma