The precise determination of the window of implantation significantly improves ART outcomes.

The human endometrium is receptive to the embryo for a specific period of time known as the window of implantation (WOI). During this period, the endometrium shows a specific gene expression profile suitable for endometrial function evaluation. ER Map is a molecular tool able to accurately predict endometrial receptivity status by transcriptomic analysis. In this retrospective study, including 2256 subfertile patients undergoing ART treatment, the clinical value of precise WOI determination is studied in detail. Results obtained when single embryo transfers (sET) were scheduled either within the WOI timeframe as established by ER Map, or deviating from this WOI, are assessed and compared. Data obtained showed that 34.18% (771/2256) of patients had a displaced WOI. Analysis of ART outcomes showed significantly higher pregnancy rates in transfers scheduled within the WOI predicted compared to transfers that deviated more than 12h from this WOI (44.35% vs 23.08%, p < 0.001). The deviation from the WOI had also an impact on the progression of pregnancy, with a significant increase in pregnancy loss (~ twofold) observed in transfers that deviated more than 12h from the WOI predicted. These results indicate that the precise determination of the WOI and personalised embryo transfer can significantly improve clinical outcomes.

Tos e infiltrados pulmonares. Un final inesperado / Pulmonary infiltrates and cough. An unexpected end

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Virulence of Metarhizium brunneum (Ascomycota: Hypocreales) Strains Against Stinkbugs Euschistus heros and Dichelops furcatus (Hemiptera: Pentatomidae).

Three strains of fungi belonging to the genus Metarhizium Sorokin (ARSEF 4556, ARSEF 3297, native strain) were assayed against adults and nymphs of the Neotropical brown stinkbug Euschistus heros (F.) and the green-belly stinkbug Dichelops furcatus (F.). The most virulent strain, ARSEF 4556, caused over 90% mortality. The average survival time of the second and fifth instar nymphs and adults following immersion in 1 × 108 conidia ml-1 was 4.8, 5.7, and 5.2 d, respectively. The second instar nymphs were more susceptible than the adults. The LC50 values and median survival times for second instar and adult E. heros were 1.6 × 107 and 3.1 × 107 conidia ml-1 and 6 and 8 d, respectively. Eggs of E. heros and the closely related stinkbug, D. furcatus, were highly susceptible to ARSEF 4556 with the mean mortality of eggs immersed in 1 × 108 conidia ml-1 being 77.4% and 89.7%, respectively. The strain 3297 showed also good aptitudes for stinkbugs control with mortalities higher than 80% against nymphs and adults and eggs mortalities of 75.5% for E. heros and 79.6% for D. furcatus. This study has shown that it is possible to have a two-pronged control strategy, targeting adults and to reduce oviposition and targeting egg clusters to prevent emergence and dispersal of nymphs. Besides early instars of nymphs have been shown to be more susceptible to the fungal strains than late instars and adults.

Soil drenching with entomopathogenic fungi for control of the soil-dwelling life stages and adults of the same generation of Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae).

Four Beauveria bassiana and three Metarhizium brunneum isolates were evaluated, as soil drenches, against Spodoptera littoralis prepupae. Treatment efficacy was determined by assessing total mortality during development from prepupae through to pupae and adults; mortality and sub-lethal effects on reproduction were also quantified for adults emerging from surviving prepupae/pupae. All isolates were pathogenic but overall mortality varied between 31.7 and 83.3% (0% for control); average survival time was 7.5-10.5 days (14.0 days for control). From 1.7-15.0% of adults emerging from surviving prepupae/pupae were deformed (0% in control). Contact with fungal suspensions as prepupae/pupae caused a significant reduction in fecundity of emerging adult females (15-58.9%), and a significant reduction in egg viability (6.8-28.4%) compared with controls. Two isolates were selected for virulence evaluation against S. littoralis prepupae. The LC50s were 1.7 × 107 and 1.8 × 107 conidia ml-1 and the median survival times were 7 and 6 days for isolates EAMa 01/58-Su and EAMb 09/01-Su, respectively. Destruxin A was present in pupae developing from prepupae treated with isolates EAMa 01/58-Su (0.010 ± 0.002 µg pupae-1) and EAMb 09/01-Su (0.015 ± 0.003 µg pupae-1). The use of entomopathogenic fungi as soil drenches could be a key component of S. littoralis IPM strategies due to direct reductions in the number of soil-dwelling life stages and, also, the significant reduction in reproductive potential of surviving adults.

Phytopathogenic and antagonistic potentialities of fungi associated with pistachio bark beetle, Chaetoptelius vestitus (Coleoptera, Curculionidae), infesting pistachio (Pistacia vera) in Tunisia.

AIM: This study investigated the biodiversity of fungi associated with the pistachio bark beetle, Chaetoptelius vestitus, in Tunisia. We evaluated the phytopathogenic activities and tested antagonistic potentialities with respect to phytopathogens. METHODS AND RESULTS: A total of 41 fungal isolates were randomly isolated from C. vestitus adults and galleries. We identified 28 species belonging to 13 genera using ITS sequences of the ribosomal RNA operons. Pathogenicity assays performed using the excised shoot method revealed that isolates Aal_io_1 (Alternaria alternata), Feq_io_1 (Fusarium equiseti), Fgra_io_1 (Fusarium graminearum), Fve_io_1 (Fusarium verticilloides), Tro_io_1 (Trichothecium roseum) and Nqu_io_1 (Nothophoma quercina) displayed a high pathogenic activity on pistachio stems. Estimation of the antagonistic potentialities of isolated fungi against several phytopathogenic isolates as tested using a dual-culture method showed that isolates Tpi_io_1 (Talaromyces pinophilus), Pbi_io_2 (Penicillium bilaiae), Asc_io_1 (Aspergillus sclerotiorum) and Gla_io_1 (Geosmithia lavendula) displayed a broad range of antagonistic activities. CONCLUSION: Fungi associated with C. vestitus had a variable range of pathogenic activity on pistachio stem. Phytopathogenic fungi were antagonized by different fungal isolates which could be promising in pistachio protection against phytopathogenic fungi. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first that investigated the diversity of fungi associated with C. vestitus and evaluated both their phytopathogenic activity and antagonistic potential against fungal phytopathogens.

Which came first: The disease or the pest? Is there a host mediated spread of Beauveria bassiana (Ascomycota: Hypocreales) by invasive palm pests?

The red palm weevil (RPW) Rhynchophorus ferrugineus (Olivier) (Coleoptera: Curculionidae) is threatening the palm family worldwide, causing important economic losses. Current tactics to manage the weevil are largely based on chemical control, although the use of pesticides is hampered by several environmental constraints. Since the first introduction of RPW in Spain in 1996 and during its progressive spread around the Mediterranean basin, the number of reports of natural infection of RPW populations by entomopathogenic fungi (EPF) has been rising for 15 years, and this rise could support a pest-mediated EPF spread. To challenge this hypothesis, we assessed the usefulness of the region of elongation factor 1-alpha (EF1-α), Bloc nuclear intergenic region (Bloc) and inter simple sequence repeat (ISSR) markers, alone or in combination, to infer the relationships among Mediterranean Beauveria and Metarhizium strains isolated from the RPW. Second, the effect of abiotic factors, such as temperature, humidity and UV-B radiation, on the germination and growth of these EPFs strains as a function of their genealogy and geographic origin were determined. Finally, the pathogenicity of strains from different genetic clades was evaluated against larvae and adults of R. ferrugineus. The phylogenetic analysis based on the EF-1α gene identified eight different sequences among 24 fungal isolates of four fungal species. Similar clades were clustered when Bloc and ISSR analyses were performed. The results showed that strains of different origins were clustered in the same clade, and this outcome could be explained by an RPW-mediated EPF spread that was also influenced by time, geographical and other RPW related factors. Neither the response to abiotic factors nor virulence to RPW larvae and adults were related to the sequence type, with all B. bassiana strains well adapted to Mediterraneam climatic conditions. Taken together, these findings may help to select the best strain for RPW management.

Quantification of fungal growth and destruxin A during infection of Galleria mellonella larvae by Metarhizium brunneum.

Destruxin A is among the major secondary metabolites produced by the entomopathogenic ascomycete Metarhizium sp., and the lack of studies concerning production of destruxin A by the fungus is most likely the biggest obstacle for the registration of new fungal strains. Although several studies focus on the production of destruxin A in culture media, few studies examine destruxin A in vivo during host infection. In the current work, Galleria mellonella was used as an insect model to develop for the first time in vivo real-time PCR- and HPLC-MS-based quantification of fungal growth and metabolite production, respectively, during infection by two strains of M. brunneum. Total mortality of sixth instar G. mellonella larvae that were immersed in a suspension of 1.0×108conidiamL-1 of M. brunneum EAMa 01/58-Su or BIPESCO5 strains reached 85.5% and 78.8%, respectively, and the percentage of cadavers with fungal outgrowth was low at 12.2% and 4.4%, respectively. The average survival time of treated larvae was 5.5days for both fungal strains. Using EAMa 01/58-Su and BIPESCO5 specific primer set, real-time PCR showed that the patterns of fungal growth were different for the two strains, whereas no significant differences were detected in the number of fungal sequence copies recovered from the infected larvae. EAMa 01/58-Su and BIPESCO5 strains secreted destruxin A from days 2 to 6 and from days 2 to 5 post treatment, respectively. For EAMa 01/58-Su and BIPESCO5, the maximum titer of destruxin A in the host was on day 4 at 0.369 and 0.06µg/larva, respectively, and throughout the pathogenic process, the total production was 0.6 and 0.09µg/larva, respectively. These results demonstrated that the strains pose a low hazard, if any, to humans and the environment. The methods used in this study to quantify fungal growth and metabolite production provided valuable data to better understand the role of destruxin A during the growth of M. brunneum in the host larvae and to monitor the fate of destruxin A in food chains.

Transient endophytic colonizations of plants improve the outcome of foliar applications of mycoinsecticides against chewing insects.

The current work reports how spray application of entomopathogenic fungi on alfalfa, tomato and melon plants may cause an additional Spodoptera littoralis larvae mortality due to a temporal colonization of the leaves and subsequent ingestion of those leaves by the larvae. Most entomopathogenic fungi (EF) (Ascomycota: Hypocreales) endophytes seem to colonize their host plants in a non-systemic pattern, in which case at least a transient endophytic establishment of the fungus should be expected in treated areas after spray application. In this work, all strains were able to endophytically colonize roots, stems and leaves during the first 96h after inoculation. Whilst the treatment of S. littoralis larvae with a 10(8)ml(-1) conidial suspension resulted in moderate to high mortality rates for the Metarhizium brunneum EAMb 09/01-Su (41.7-50.0%) and Beauveria bassiana EABb 01/33-Su (66.7-76.6%) strains, respectively, an additive effect was detected when these larvae were also fed endophytically colonized alfalfa, tomato, and melon leaves, with mortality rates varying from 25.0% to 46.7% as a function of the host plant and total mortality rates in the combined treatment of 75-80% and 33-60% for B. bassiana and M. brunneum, respectively. Fungal outgrowth was not detected in any of the dead larvae feeding on colonized leaves, whereas traces of destruxin A were detected in 11% of the insects fed tomato discs endophytically colonized by M. brunneum. The combined effects of the fungal spray with the mortality caused by the feeding of insects on transient EF-colonized leaves have to be considered to estimate the real acute impact of field sprays with entomopathogenic fungi on chewing insects.

Stock management in hospital pharmacy using chance-constrained model predictive control.

One of the most important problems in the pharmacy department of a hospital is stock management. The clinical need for drugs must be satisfied with limited work labor while minimizing the use of economic resources. The complexity of the problem resides in the random nature of the drug demand and the multiple constraints that must be taken into account in every decision. In this article, chance-constrained model predictive control is proposed to deal with this problem. The flexibility of model predictive control allows taking into account explicitly the different objectives and constraints involved in the problem while the use of chance constraints provides a trade-off between conservativeness and efficiency. The solution proposed is assessed to study its implementation in two Spanish hospitals.

Development of a QuEChERS-based extraction method for the determination of destruxins in potato plants by UHPLC-MS/MS.

Ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) with electrospray ionization has been proposed for the determination of fifteen natural destruxins (A, B, C, D, E, Ed, Ed1, A2, B2, D2, E2, Cl, DesmA, DesmB, and DH-A), secondary metabolites with insecticidal and phytotoxic activities produced by Metarhizium species fungus, which are being studied as biological agents in pest control. Therefore, procedures to control them in the food chain are required, starting with crops. As a consequence, in this study, a simple QuEChERS-based destruxin (dtx) extraction procedure has been developed and validated in four different parts of potato plant (tuber, root, stem and leaves) for the first time. For dtx A, the limits of detection obtained, ranged between 0.5 and 1.3 µg/kg, and for quantification, ranged between 1.7 and 4.2 µg/kg. Precision values were below 8.5%; and in all cases, recoveries were higher than 91%. Finally, the method has been applied in potato samples inoculated by EAMa 01/58-Su strain, where dtxs A and B were detected and quantified. In all cases, dtx B concentration was higher than dtx A.

Insecticidal Activity of a Destruxin-Containing Extract of Metarhizium brunneum Against Ceratitis capitata (Diptera: Tephritidae).

Tephritid fruit flies are major pests that limit fruit production around the world; they cause important damages, increasing directly and indirectly annual costs, and their management is predominately based on the use of chemical insecticides. This research investigated the insecticidal activity of the crude extract obtained of Metarhizium brunneum Petch EAMb 09/01-Su strain and its capacity to secrete secondary metabolites including destruxins (dtx). Dtx A and A2 had insecticidal activity against Ceratitis capitata (Wiedemann) when administered per os. The crude extract of seven Metarhizium and one Beauveria isolates were evaluated per os against medfly adults. The crude extracts of the isolate EAMb 09/01-Su resulted in mortality ranging between 95 and 100% at 48 h. The high-pressure liquid chromatography profile showed two active peaks (F5B and F6 subfractions) related with dtx A2 and dtx A, which caused 70 and 100% mortality on C. capitata at 48 h postfeeding, respectively. The LC50 was 104.92 ppm of dtx A, contained in the F6 subfraction, and the LT50 was 4.16 h at a concentration of 400 ppm of dtx A contained in the F6 subfraction. Moreover, the average survival time of adults exposed to this subfraction was 12.6 h with only 1 h of exposure. The insecticide metabolites of the F6 subfraction of the EAMb 09/01-Su isolate retained >90% of its insecticidal activity after exposure to 60°C for 2 h and 120°C for 20 min. These results highlight the potential of this strain as a source of new insecticidal compounds of natural origin for fruit fly control.

Establishment of fungal entomopathogens Beauveria bassiana and Bionectria ochroleuca (Ascomycota: Hypocreales) as endophytes on artichoke Cynara scolymus.

Entomopathogenic fungi (EPF) are commonly found in diverse habitats and are known to cause mycoses in many different taxa of arthropods. Various unexpected roles have been recently reported for fungal entomopathogens, including their presence as fungal endophytes, plant disease antagonists, rhizosphere colonizers and plant growth promoting fungi. In Tunisia, a wide range of indigenous EPF isolates from different species, such as Beauveria bassiana and Bionectria ochroleuca, were found to occur in the soil, and to be pathogenic against the artichoke aphid Capitophorus elaeagni (Hemiptera: Aphididae). Since endophytic fungi are recently regarded as plant-defending mutualists and their presence in internal plant tissue has been discussed as an adaptive protection against insects, we were interested on elucidating the possible endophytic behavior of B. bassiana and B. ochroleuca on artichoke, Cynara scolymus, after foliar spraying tehcnique. The leaf spray inoculation method was effective in introducing the inoculated fungi into the plant tissues and showed, then, an endophytic activity on artichoke even 10 days later. According S-N-K test, there was significant differences between the two fungal treatments, B. ochroleuca (84% a) and B. bassiana (78% a), and controls (0% b). Likewise, the inoculated entomopathogenic fungi were also isolated from new leaves even though with significant differences respectively between controls (0% c), B. bassiana (56% b) and B. ochroleuca (78% a). These results reveals significant new data on the interaction of inoculated fungi with artichoke plant as ecological roles that can be exploited for the protection of plants.

One Metarhizium brunneum Strain, Two Uses to Control Ceratitis capitata (Diptera: Tephritidae).

We determined the virulence and insecticidal activity of the hypocrealean fungus Metarhizium brunneum (Petch) strain EAMb 09/01-Su and its crude extract against Mediterranean fruit fly Ceratitis capitata Wiedemann (Diptera: Tephritidae) and we evaluated the combined use of the fungus with its crude extract. We also determined the effect of fermentation time, temperature, and initial pH of the M. brunneum culture medium on the insecticidal activity of the crude extract. When C. capitata adults were sprayed with a conidial suspension, the strain EAMb 09/01-Su caused 100% mortality with a mean lethal time (LT50) of 5.6 d and mean lethal concentration (LC50) of 2.84 f#x2013; 10(7) conidia per milliliter. Fermentation time significantly affected the lethality of the crude extract when it was provided to C. capitata per os. The highest level of mortality (73.3%) and the shortest median survival time (25.5 h) was obtained from 15-d-old cultures. The crude extract was demonstrated to be thermostable, given that the mortality was >50% at 48 h when the extract had been heated to 100°C for 3 h. Lastly, the optimum initial pH for maximum crude extract activity in terms of mortality ranged between 7 and 9. Additivity was observed for all M. brunneum EAMb 09/01-Su strain crude extract combinations tested, indicating compatibility between products. We concluded that the M. brunneum EAMb 09/01-Su strain is a promising tool for medfly control alone or in combination with its crude extract.

Beauveria bassiana (Ascomycota: Hypocreales) wound dressing for the control of Euzophera pinguis (Lepidoptera: Pyralidae).

Injury to olive tree trunks and branches because of biotic and abiotic factors, such as pruning and mechanical harvesting, attracts the olive pyralid moth Euzophera pinguis Haworth (Lepidoptera: Pyralidae). This moth has become increasingly important in the Mediterranean region during recent years. The use of an entomopathogenic fungus for wound dressing for pest control is reported for the first time in this study. Beauveria bassiana (Ascomycota: Hypocreales) strain EABb 08/04-Ep was originally obtained from a diseased E. pinguis larva and has shown effective E. pinguis control in an olive crop in Jaén, Andalusia, Spain, under field conditions during the spring and fall of 2008 and 2009 and the spring of 2011. Experimental artificial 30 by 30-mm square wound cages were large enough to allow the E. pinguis females to oviposit. Approximately 80 and 40-60% of the control wounds contained live larvae in the experiments that occurred during the spring and fall, respectively. The B. hassiana wound dressing gave similar results as the chlorpyrifos wound dressing throughout the experiment, with efficacies reaching 80-85% in the spring and 90-95% in the autumn. The B. bassiana fungus was recovered from 60-90% of the wounds at the completion of the experiments and after 60 d of treatment. These data indicate that strain EABb 08/04-Ep applied to the pruning wounds can be an effective tool for the microbial control of E. pinguis in olive crops. Moreover, B. bassiana may be used within integrated pest management strategies to minimize chemicals, depending on the population density of the pyralid moth.

Biocontrol of Bactrocera oleae (Diptera: Tephritidae) with Metarhizium brunneum and its extracts.

The susceptibility of preimaginal and adult olive fruit fly, Bactrocera oleae (Gmelin) (Diptera: Tephritidae), to a strain of the mitosporic ascomycete Metarhizium brunneum (Petch) (Hypocreales: Clavicipitaceae) and the insecticidal activity of its crude extract to olive fruit fly adults were investigated. Strain EAMb 09/01-Su caused 60% mortality to B. oleae adults, with average survival time (AST) of 8.8 d. In soil treatments against pupariating third-instar larvae, preimaginal B. oleae mortality reached 82.3%, whereas preimaginal mortality targeting puparia was 33.3%. The crude extract of EAMb 09/01-Su strain caused 80.0% adult mortality when administered per os, with AST of 27.7 h. The crude extract was demonstrated to be quite thermostable and photoresistant. These results indicate that M. brunneum EAMb 09/01-Su strain and its crude extract show potential to be used in an integrated pest management olive fruit fly management strategy targeting both adults and preimaginals.

Telangiectasia macular eruptiva Perstans y síndrome de Conn / Telangiectasia macularis eruptiva Perstans and Conn syndrome

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Embolismo gaseoso cerebral por accidente de buceo / Cerebral gas embolism as a result of diving accident

La patología relacionada con los accidentes de buceo es quizás poco conocida por su baja prevalencia. Se clasifica en no disbárica o disbárica, en función de su relación con cambios de presión. Presentamos el caso de un buceador que, después de realizar una inmersión de 30 minutos a 22 metros de profundidad, presentó bradipsiquia y afasia transitoria. En la tomografía axial computarizada craneal se objetivó un infarto silviano izquierdo. Se realizó tratamiento en cámara hiperbárica con buena evolución clínica. En relación con el caso, revisamos las distintas formas de presentación de los accidentes disbáricos, así como la utilidad de la cámara hiperbárica en el tratamiento

Clinical alterations related to diving accidents are little known perhaps because of its low prevalence. They can be classified as dysbarism-related and non-dysbarism-related according to its relation with pressure changes. We present the case of a diver that showed bradypsychia and transitory aphasia after carrying out an immersion of 30 minutes in duration and 22 meters deep. A left sylvian cerebral infarct was demostrated in cranial computerized axial tomography. Treatment was carried out in a hyperbaric chamber with good clinical evolution. We review with regard to this case the different forms of clinical presentation of dysbarism-related accidents, as well as the usefulness of the hyperbaric chamber in their treatment