Role of egg-laying behavior, virulence and local adaptation in a parasitoid's chances of reproducing in a new host.

Understanding the ability of parasitoid insects to succeed in new host populations is a relevant question for biological control and adaptive mechanisms. Cotesia typhae is an African parasitoid specialized on the moth Sesamiae nonagrioides, also called the Mediterranean corn borer. Two Kenyan strains of C. typhae differ in their virulence against a new host population from France. We explored behavioral and physiological hypotheses about this differentiation. Cotesia genus belongs to a group of Hymenoptera in which females inject a domesticated virus in their host to overcome its resistance. Since viral particles are injected along with eggs and since the strain with the higher virulence injects more eggs, we hypothesized that virulence could be explained by the quantity of virus injected. To test this assumption, we measured the injected quantities of eggs and viral particles (estimated by viral DNA segments) of each parasitoid strain along several ovipositions, to vary these quantities. Unexpectedly, results showed that virulence against the French host was not correlated to the injected quantities of eggs or viral segments, indicating that virulence differentiation is explained by other causes. The virulence against the respective natural hosts of the two C. typhae strains was also measured, and results suggest that local adaptation to a more resistant natural host may explain the pre-adaptation of one strain to the new host population. We also identified a differentiation of oviposition strategy and subsequent offspring number between the parasitoid strains, which is important in a biocontrol perspective.

Propiedades insecticidas del polvo de Laurelia sempervirens L. para el control de Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae) / Insecticidal properties of Laurelia sempervirens powder to Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae) control

Sitophilus zeamais is a key pest of stored cereals. The insecticidal properties of Laurelia sempervirens powder against S. zeamais were assessed in laboratory. The variable evaluated were mortality of adult insects by contact and fumigant toxicity, mortality of immature insects by contact toxicity, residual effect of powder stored under environmental or refrigerated conditions, adult insect emergence (F1), repellency and weight reduction and germination of maize. The concentrations evaluated were 0, 0.25, 0.5, 1.0, 2.0 and 4.0 percent and the experimental design was completely randomly. In contact toxicity bioassay all treatments showed mortality over 90 percent while fumigant activity did not exceed 40 percent. The F1 only was registered in treatment of 0.25 percent and control showing all other treatments 0 percent. In immature control bioassay all treatments exhibited statistical differences with control. The storage of powder under refrigerated conditions not improved the residual effect. All concentrations of powder assessed showed repellence. Germination and grain weight loss of maize seeds not showed significant differences with control. We concluded that L. sempervirens has promissory perspectives to stored grain pests control...

Sitophilus zeamais es plaga primaria de los cereales almacenados. Se evaluaron, en laboratorio, las propiedades insecticidas del polvo de follaje de Laurelia sempervirens para el control de S. zeamais. Las variables evaluadas fueron mortalidad por contacto y fumigante de insectos adultos, toxicidad por contacto sobre estados inmaduros, efecto residual del polvo almacenado en condiciones ambientales y de refrigeración, emergencia de insectos adultos (F1), repelencia y pérdida de peso y germinación del maíz. Las concentraciones evaluadas fueron 0; 0,25; 0,5; 1,0; 2,0 y 4,0 por ciento y el diseño experimental fue completamente al azar. En el bioensayo de toxicidad por contacto todos los tratamientos mostraron una mortalidad sobre el 90 por ciento, mientras que la actividad como fumigante no superó el 40 por ciento. La F1 sólo se registró en el tratamiento de 0,25 por ciento y en el testigo mostrando todos los otros tratamientos 0 por ciento. En el bioensayo de control de estados inmaduros todos los tratamientos exhibieron diferencias significativas con el testigo. El almacenamiento del polvo en condiciones de refrigeración no mejoró el efecto residual. Todas las concentraciones de polvo evaluadas registraron efecto repelente. La germinación y pérdida de peso de las semillas de maíz no mostraron diferencias significativas con el testigo. Se concluye que L. sempervirens tiene perspectivas promisorias para el control de plagas de los granos almacenados...

Azithromycin analogue CSY0073 attenuates lung inflammation induced by LPS challenge.

BACKGROUND AND PURPOSE: Azithromycin is a macrolide antibiotic with anti-inflammatory and immunomodulating effects. Long-term azithromycin therapy in patients with chronic lung diseases such as cystic fibrosis has been associated with increased antimicrobial resistance, emergence of hypermutable strains, ototoxicity and cardiac toxicity. The aim of this study was to assess the anti-inflammatory effects of the non-antibiotic azithromycin derivative CSY0073. EXPERIMENTAL APPROACH: We compared the effects of CSY0073 with those of azithromycin in experiments on bacterial cultures, Pseudomonas aeruginosa biofilm, lung cells and mice challenged intranasally with P. aeruginosa LPS. KEY RESULTS: In contrast to azithromycin, CSY0073 did not inhibit the growth of P. aeruginosa, Staphylococcus aureus or Haemophilus influenzae and had no effect on an established P. aeruginosa biofilm. Bronchoalveolar lavage (BAL) fluids and lung homogenates collected after the LPS challenge in mice showed that CSY0073 and azithromycin (200 mg·kg(-1), i.p.) decreased neutrophil counts at 24 h and TNF-α, CXCL1 and CXCL2 levels in the BAL fluid after 3 h and IL-6, CXCL2 and IL-1ß levels in the lung after 3 h compared with the vehicle. However, only azithromycin reduced IL-1ß levels in the lung 24 h post LPS challenge. CSY0073 and azithromycin similarly diminished the production of pro-inflammatory cytokines by macrophages, but not lung epithelial cells, exposed to P. aeruginosa LPS. CONCLUSIONS AND IMPLICATIONS: Unlike azithromycin, CSY0073 had no antibacterial effects but it did have a similar anti-inflammatory profile to that of azithromycin. Hence, CSY0073 may have potential as a long-term treatment for patients with chronic lung diseases.

Effect of pps disruption and constitutive expression of srfA on surfactin productivity, spreading and antagonistic properties of Bacillus subtilis 168 derivatives.

AIMS: To analyse the effects of plipastatin operon disruption and constitutive expression of surfactin operon in Bacillus subtilis 168 on surfactin productivity, in vitro invasive growth and antagonism against fungi. METHODS AND RESULTS: The srfA native promoter was replaced by the constitutive promoter P(repU) in B. subtilis 168 after integration of a functional sfp gene. Moreover, the plipastatin synthesis was further disrupted in the B. subtilis 168 derivatives. In liquid media, an earlier and higher expression of P(repU), than that found with P(srfA), led to a specific surfactin production fivefold higher after 6 h of culture. On solid media, not only the invasive growth and the haemolytic activity but also the antifungal activity of the constitutive strains were improved when compared to the parental strain BBG111. As expected, the disruption of the plipastatin operon strongly reduced in vitro antifungal properties but, interestingly, enhanced specific surfactin production (1.47 g g(-1) of biomass), spreading behaviour and haemolytic activity of the strains. CONCLUSIONS: This work demonstrates for the first time the interdependency of surfactin and plipastatin regarding their biosynthesis as well as their influence on the biological activities of the producing strain. SIGNIFICANCE AND IMPACT OF THE STUDY: The constitutive overproduction of surfactin enhances the invasive growth and the in vitro antagonistic activity of the mutant strain. Both properties are known to play an important role in the biocontrol of plant diseases. Plipastatin operon disruption increases the surfactin productivity of mutant strains. These mutants are interesting for use in continuous bioprocesses for surfactin production or in bioremediation.

Respiration activity monitoring system (RAMOS), an efficient tool to study the influence of the oxygen transfer rate on the synthesis of lipopeptide by Bacillus subtilis ATCC6633.

The effect of oxygen transfer rate (OTR) on the synthesis of mycosubtilin, a non ribosomal lipopeptide antifungal biosurfactant, was investigated in the respiration activity monitoring system (RAMOS) for two Bacillus subtilis strains. These cultures were performed under definite oxygen-limited conditions without the adding of any anti-foam in the culture medium. By using four different filling volumes (FV) in the shaken bioreactors, different levels (20, 14, 9 and 7 mmol O(2)l(-1)h(-1)) of oxygen-limited growth could be obtained. A 25-fold increase of the specific productivity of mycosubtilin was observed for B. subtilis ATCC6633 in the case of the most severe oxygen limitation. But nearly no effect could be found with strain BBG100 carrying the constitutive P(repU) promoter instead of the natural P(myc) promoter. Transcript analysis of the fenF gene belonging to the myc operon indicated that the P(myc) promoter regulation could be slightly oxygen sensitive. Additionally, different patterns of the synthetised mycosubtilin homologues were obtained for different level of oxygen-limited growths. At the present state of investigation, oxygen regulation was thus shown to act at different levels suggesting the existence of a complex regulatory system of NRPS lipopeptide synthesis in the natural B. subtilis ATCC6633 strain.

Setting up and modelling of overflowing fed-batch cultures of Bacillus subtilis for the production and continuous removal of lipopeptides.

This work is related to the set-up of overflowing exponential fed-batch cultures (O-EFBC) derived from carbon limited EFBC dedicated to the production of mycosubtilin, an antifungal lipopeptide belonging to the iturin family. O-EFBC permits the continuous removal of the product from the bioreactor achieving a complete extraction of mycosubtilin. This paper also provides a dynamical Monod-based growth model of this process that is accurate enough to simulate the evolution of the specific growth rate and to correlate it to the mycosubtilin specific productivity. Two particular and dependant phenomena related to the foam overflow are taken into account by the model: the outgoing flow rate of a broth volume and the loss of biomass. Interestingly, the biomass concentration in the foam was found to be lower than the biomass concentration in the bioreactor relating this process to a recycling one. Parameters of this model are the growth yield on substrate and the maximal specific growth rate estimated from experiments led at feed rates of 0.062, 0.071 and 0.086h(-1). The model was extrapolated to five additional experiments carried out at feed rates of 0.008, 0.022, 0.040, 0.042 and 0.062h(-1) enabling the correlation of the mean specific growth rates with productivity results. Finally, a feed rate of 0.086h(-1) corresponding to a mean specific growth rate of 0.070h(-1) allowed a specific productivity of 1.27mg of mycosubtiling(-1) of dried biomassh(-1).

Real time in situ microscopy for animal cell-concentration monitoring during high density culture in bioreactor.

An in situ microscope (ISM) device is utilised in this study to monitor hybridoma cells concentration in a stirred bioreactor. It generates images by using pulsed illumination of the liquid broth synchronised with the camera frame generation to avoid blur from the cell's motion. An appropriate image processing isolates the sharp objects from the blurred ones that are far from the focal plane. As image processing involves several parameters, this paper focuses on the robustness of the results of the cells counting. This stage determines the applicability of the measuring device and has seldom been tackled in the presentations of ISM devices. Calibration is secondly performed for assessing the cell-concentration from the cell automated numeration provided by the ISM. Flow cytometry and hemacytometer chamber were used as reference analytical methods. These measures and the output of the image processing allow estimating a single calibration parameter: the reference volume per image equal to 1.08 x 10(-6) mL. In these conditions, the correlation coefficient between both reference and ISM data sets becomes equal to 0.99. A saturation of this system during an ultrasonic wave perfusion phase that deeply changes the culture conditions is observed and discussed. Principal component analysis (PCA) is used to undergo the robustness study and the ISM calibration step.

Effects of divided attention on encoding and retrieval processes: assessment of attentional costs and a componential analysis.

Recent research has shown that divided attention at retrieval, in contrast to encoding, affected memory performance only minimally. This immunity at retrieval was associated with a significant secondary task cost. In this article the authors further investigated these effects employing a cued-recall task and a multimeasure approach with accuracy, latency, overall attentional costs, and the temporal distribution of attentional costs associated with the encoding and retrieval of low- and high-frequency words. The results of 2 experiments yielded a complex pattern of both similarities and differences between encoding and retrieval. Simultaneous inspection of the different measures of performance was instrumental in identifying 3 major types of retrieval (unsuccessful, slow, and fast), as well as different phases of the retrieval process, each of which was characterized by a different demand for attentional resources.

A human immunodeficiency virus type 1 Env-granulocyte-macrophage colony-stimulating factor fusion protein enhances the cellular immune response to Env in a vaccinia virus-based vaccine.

Vaccinia virus (VV) infection induces protective T- and B-cell responses, making recombinants based on VV good candidates for the development of effective vaccines to other viruses. VV recombinants expressing the human immunodeficiency virus (HIV) envelope protein (Env) have been generated in several laboratories and shown to induce anti-HIV cellular and humoral immune responses in vaccinated humans and in chimpanzees. To increase the immunogenicity of the Env antigen, a VV recombinant was generated that expresses a chimeric antigen consisting of the Env protein fused to an immunostimulatory cytokine, granulocyte-macrophage colony-stimulating factor (GM-CSF). The chimeric protein retained GM-CSF biological activity when expressed by this recombinant virus (VV-GM-gp120) in cells infected in vitro. Infection of BALB/c mice with VV-GM-gp120 triggered a higher HIV-specific cellular immune response, as measured by interferon-gamma production, than that induced by a VV recombinant expressing the native Env protein. Moreover, although anti-gp120 antibody titres were similar in sera from mice inoculated with either of the VV recombinants, immunization with the recombinant expressing the fusion protein elicited antibodies against a broader spectrum of Env epitopes. These results indicate that HIV Env antigen fusion to GM-CSF provides a means to improve the anti-HIV immune response.

Effects of divided attention on encoding and retrieval processes in human memory: further support for an asymmetry.

Despite a tradition in cognitive psychology that views encoding and retrieval processes in human memory as being similar, F. I. M. Craik, R. Govoni, M. Naveh-Benjamin and N. D. Anderson (1996) have recently shown that notable differences exist between the 2 when divided-attention manipulations are used. In this article, the authors further examined this asymmetry by using several manipulations that changed task demands at encoding and retrieval. The authors also used a secondary-task methodology that allowed a microlevel analysis of the secondary-task costs associated with encoding and retrieval. The results illustrated the resiliency of retrieval processes to manipulations involving different task demands. They also indicated different loci of attention demands at encoding and retrieval. The authors contend that whereas encoding processes are controlled, retrieval processes are obligatory but do require attentional resources for their execution.

Functional assessment of macular hole surgery by scanning laser ophthalmoscopy.

OBJECTIVE: This study aimed to evaluate more precisely the benefit of macular hole surgery. DESIGN: The design was a prospective study of 40 eyes in 40 patients examined before and after surgery of full-thickness macular holes with a scanning laser ophthalmoscope (SLO). PARTICIPANTS: Forty patients with full-thickness macular holes participated. Three of the holes were stage 2, 23 were stage 3, and 14 were stage 4. INTERVENTION: The SLO examination consisted of macular imaging and assessment of macular function using tests produced by an acousto-optical modulator. MAIN OUTCOME MEASURES: Preferred retinal locus (PRL) was determined, visual acuity was measured, and scotoma was detected by microscotometry and by the line test (modified Watzke-Allen test). RESULTS: Anatomic success was achieved in 32 of 40 cases as assessed biomicroscopically. On SLO examination before surgery, macular holes were seen as a central bright round disc outlined by a thin dark edge surrounded by a dark ring and a less dark area with ill-defined limits. In all cases, the PRL was located on the upper edge of the hole, a scotoma was always detected inside the hole, and the line was seen as broken in 26 of 32 cases. After surgery, the hole closed completely in 25 of 32 eyes; it disappeared from 14 of these 25 eyes and was replaced by a dark or clear disc in 11. In the other seven successful cases, its size shrank and its edge flattened but remained faintly visible. The hole remained unchanged in eight cases. Eccentric PRL became central in 28 of 32 cases. The scotoma disappeared in 23 of 32 cases. The line was seen as continuous in 24 of 32 cases. Complete anatomic and functional successes were achieved in 19 of the 32 cases of macular hole closure. CONCLUSION: The SLO examination allows accurate assessment of the anatomic and functional results of macular hole surgery. Various degrees of functional success were recorded, depending on the test used.

Scanning laser ophthalmoscope imaging of fluorescein-labelled blood cells.

PURPOSE: To demonstrate the feasibility of a technique for the visualization by scanning laser ophthalmoscope (SLO) of fluorescein-labelled autologous leukocytes and platelets in retinal vessels. METHOD: Individual blood samples from rats and rabbits were centrifuged to isolate platelets and leukocytes, then passively labelled with fluorescein and reinjected into the same animal. An SLO was used to visualize and record cell displacement in the retinal circulation. Labelled platelets were analysed by flow cytometry. RESULTS: By SLO, platelets appeared as a heterogeneous particle flow, and individual leukocytes appearing as brighter spots could easily be traced. Flow cytometry showed that after labelling platelets were well individualized and their size was slightly increased. CONCLUSION: Circulating blood cells can be visualized in retinal vessels by a simple method consisting of passive labelling of autologous platelets and leukocytes by fluorescein. No platelet toxicity was detected. This method could be applied to the study of blood cell movement in human retinal vascular diseases.

Autologous platelet concentrate as an adjunct in macular hole healing: a pilot study.

PURPOSE: A pilot study was undertaken to assess the efficacy of autologous platelets in macular hole healing. PATIENTS AND METHODS: Eight eyes of eight patients with stage 3 or 4 macular holes, two of which had failed to heal after previous vitrectomy and gas tamponade, were included. The procedure consisted of pars plana vitrectomy with removal of posterior cortical vitreous, stripping of associated epimacular membranes, 15% perfluoroethane-air tamponade, and instillation of autologous platelet concentrate onto the posterior pole. Strict postoperative facedown positioning was observed for 12 days. Postoperative evaluation included visual acuity measurement, biomicroscopic macular appearance and scanning laser ophthalmoscope examination. The follow-up period ranged from 3 to 13 months (mean, 7 months). RESULTS: Of eight eyes, flattening of the surrounding retina and closure of the hole were achieved in seven (87.5%). Visual acuity improved two lines or more in four eyes (50%) Four eyes (50%) reached a postoperative visual acuity of 20/50 or more. Increased nuclear sclerosis was observed in six eyes (75%), and retinal detachment occurred in two eyes (25%). CONCLUSIONS: Autologous platelet concentrate administered peroperatively in full-thickness macular holes seems to be a safe and effective adjunct to vitrectomy with removal of posterior hyaloid and gas tamponade. A larger multicenter randomized prospective study is underway to verify these encouraging results before advocating the use of autologous platelets in macular hole surgery.

Electrophysiological exploration of visual function in mitochondrial diseases.

In a group of 10 children (ranging from 5 months to 15 years old) affected by diseases with mitochondrial dysfunction, 4 suffered from mitochondrial myopathy, 4 from mitochondrial encephalopathy and 2 from Friedreich's ataxia. The clinically detectable visual impairment consisted of 3 grey ocular fundi (the other 7 were normal) associated, in 2 subjects, with a mild nystagmus. Electrophysiological assessment, consisting of ERGs and flash VEPs, was systematically performed. The normal ERGs in all subjects confirmed the normal functioning of retinal electrogenesis. In contrast, the VEPs of 6 out of 10 subjects were modified: in 2 of the 4 subjects with mitochondrial myopathy, the VEPs had a hyperamplitude; in the 2 subjects with Friedreich's ataxia, the implicit time of the principal VEP peaks was increased, together with a hyperamplitude in 1 case; lastly, in 2 of the 4 subjects with mitochondrial encephalopathy, the VEPs were altered. These modifications reflected visual pathway conduction disorders with no clinical expression. Various underlying pathophysiological mechanisms possibly responsible for these modifications are discussed.

Autologous platelet concentrate for the treatment of full-thickness macular holes.

BACKGROUND: To improve the anatomic success rate in the surgery of full-thickness macular holes, we tested, in a prospective pilot study, the effects of autologous platelet concentrate deposited on the macula at the end of surgery. METHODS: Two consecutive groups of patients were compared. Twenty eyes (group 1, mean symptom duration 11 months) were operated on with injection of an autologous platelet concentrate on the macula after fluid-gas exchange. Another 20 eyes (group 2, mean symptom duration 11 months) were subsequently operated on without autologous platelet concentrate. For all stage 3 holes, posterior hyaloid was detached en bloc at the level of the optic disc. The patient was left supine for 24 h after surgery, and then remained face down for 10 days. RESULTS: In group 1, 19 cases were an anatomic success, i.e. there was flattening of the retina surrounding the hole and reattachment of the edge of the hole to the retinal pigment epithelium; in 9 cases the hole was even undetectable. Final visual acuity was 0.5 or more in 9 eyes, and 0.4 or more in 14. Visual acuity improved by two lines or more in 17 of the 19 successfully operated eyes. In group 2, only 13 cases were an anatomic success. The functional results for the successfully operated eyes were identical to those of group 1. CONCLUSION: These results strongly suggested that autologous platelet concentrate could significantly improve the success rate in macular hole surgery and led us to begin a comparative, prospective, randomized trial.

Eye fixations near corners: evidence for a centre of gravity calculation based on contrast, rather than luminance or curvature.

Subjects scanned line drawings of polygons in order to count the number of corners. The positions their eyes fixated were studied as a function of the size of the angle and whether the apex of the angle was present or absent. The results showed that the eyes tended to land at a position near the centre of gravity of the corner configurations. The observed landing positions were coherent with the hypothesis that the centre of gravity was calculated within an attentional spotlight centered on the apex of the corners, and that the calculation was based not on the total luminance distribution, nor on the distribution of energy in a neurophysiologically motivated curvature detector, but simply on the basis of a contrast detector.

Contribution of scanning laser ophthalmoscopy to the functional investigation of subjects with macular holes.

A study was designed to validate a functional investigation performed with the scanning laser ophthalmoscope before surgery for macular holes in 12 eyes: The assessment included fundus examination, a functional examination resulting in evaluation of the preferred retinal lows, visual acuity and recording of visual evoked potentials. The preferred retinal locus was evaluated by presenting a small square area, and visual acuity was determined by means of calibrated figures. The visual evoked potentials were evoked by three alternating checkerboards (check size, 30', 2 Hz) centered over the hole and seen at an angle of 6.5 x 6.5 degrees, 2.5 x 2.5 degrees and 6.5 x 6.5 degrees with central exclusion of 2.5 x 2.5 degrees. The appearance of the fundus visualized by scanning laser ophthalmoscopy consisted of a clear central disk corresponding to the hole, surrounded by a very dark ring, associated with a second, less dark ring with unclear margins. Fixation was unstable in one case with a visual acuity of 20/70. In 11 cases, fixation was localized to the superior retina with a visual acuity superior to 20/70. The visual evoked potentials evoked by 6.5 x 6.5 degrees were discernible in all 12 eyes; visual evoked potential by annular stimuli were discernible in 11 cases. The 2.5 x 2.5 degrees stimulus evoked no response in eight cases, proving the area of the hole was nonfunctional. A response was recorded in the four other cases, where the dimension of the holes was less than 2 degrees. The results of this scanning laser ophthalmoscopic assessment demonstrated a precise evaluation of the residual macular function in the cases of full-thickness macular holes.

Focal visual evoked potentials generated by scanning laser ophthalmoscope in patients with age-related macular degeneration treated by perifoveal photocoagulation.

Perifoveal laser photocoagulation has been proposed for the treatment of subfoveal neovascular membranes in age-related macular degeneration. We evaluated residual function in seven eyes of six treated patients by means of transient focal visual potentials evoked with a scanning laser ophthalmoscope. The site of the preferred retinal locus was determined. The modulation of the helium-neon laser beam generated three tests (a homogeneous 6 x 6 degrees square--offset and onset--and two alternating pattern checkerboards 6 x 6 degrees and 2.5 x 2.5 degrees 60', 2 Hz) projected onto the preferred retinal locus. The focal visual evoked potentials were recorded. One eye had an unstable fixation with no discernible focal visual evoked potentials. The other six eyes had a stable fixation located in the superior retina, temporally for the right eyes and nasally for the left eyes. The homogeneous 6 x 6 degrees square evoked discernible responses in all six patients. The two checkerboards evoked discernible responses in five of six patients. These results were compared with those recorded in four controls in whom the three tests were projected onto the same retinal areas as in the patients. Evoked responses were more often recorded in the preferred retinal locus of the treated patients with age-related macular degeneration than in the corresponding retinal areas of the controls. The scanning laser ophthalmoscope allowed us to control the site of stimulation in the patients' and controls' retinas. These preliminary results suggest that there may be a functional plasticity of the visual system after therapeutic laser-induced central scotoma.