Nanoporous monolithic microsphere arrays have anti-adhesive properties independent of humidity

Bioinspired artificial surfaces with tailored adhesive properties have attracted significant interest. While fibrillar adhesive pads mimicking gecko feet are optimized for strong reversible adhesion, monolithic microsphere arrays mimicking the slippery zone of the pitchers of carnivorous plants of the genus Nepenthes show anti-adhesive properties even against tacky counterpart surfaces. In contrast to the influence of topography, the influence of relative humidity (RH) on adhesion has been widely neglected. Some previous works deal with the influence of RH on the adhesive performance of fibrillar adhesive pads. Commonly, humidity-induced softening of the fibrils enhances adhesion. However, little is known on the influence of RH on solid anti-adhesive surfaces. We prepared polymeric nanoporous monolithic microsphere arrays (NMMAs) with microsphere diameters of a few 10 {\mu}m to test their anti-adhesive properties at RHs of 2 % and 90 %. Despite the presence of continuous nanopore systems through which the inner nanopore walls were accessible to humid air, the topography-induced anti-adhesive properties of NMMAs on tacky counterpart surfaces were retained even at RH = 90 %. This RH-independent robustness of the anti-adhesive properties of NMMAs significantly contrasts the adhesion enhancement by humidity-induced softening on nanoporous fibrillar adhesive pads made of the same material

Visualisation of Leishmania donovani Fluorescent Hybrids during Early Stage Development in the Sand Fly Vector

hybrids were produced by co-infecting sand flies with two strains carrying different drug resistance markers. However, the location and timing of hybridisation events in sand flies has not been described. strains carrying hygromycin or neomycin resistance genes and red or green fluorescent markers. Fed females were dissected at different times post bloodmeal (PBM) and examined by fluorescent microscopy or fluorescent activated cell sorting (FACS) followed by confocal microscopy. In mixed infections strains LEM3804 and Gebre-1 reached the cardia and stomodeal valves more rapidly than strains LEM4265 and LV9. Hybrids unequivocally expressing both red and green fluorescence were seen in single flies of both vectors tested, co-infected with LEM4265 and Gebre-1. The hybrids were present as short (procyclic) promastigotes 2 days PBM in the semi-digested blood in the endoperitrophic space. Recovery of a clearly co-expressing hybrid was also achieved by FACS. However, hybrids could not sustain growth in vitro. has profound epidemiological significance, because it facilitates the emergence and spread of new phenotypic traits

Eficiência de fungicidas para o controle da mancha-alvo, Corynespora cassiicola, na safra 2012/13: resultados sumarizados dos ensaios cooperativos.

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A clinical study to optimise a sand fly biting protocol for use in a controlled human infection model of cutaneous leishmaniasis (the FLYBITE study)

Background: Leishmaniasis is a globally important yet neglected parasitic disease transmitted by phlebotomine sand flies. With new candidate vaccines in or near the clinic, a controlled human challenge model (CHIM) using natural sand fly challenge would provide a method for early evaluation of prophylactic efficacy. Methods : We evaluated the biting frequency and adverse effects resulting from exposure of human volunteers to bites of either Phlebotomus papatasi or P. duboscqi, two natural vectors of Leishmania major. 12 healthy participants were recruited (mean age 40.2 ± 11.8 years) with no history of significant travel to regions where L. major-transmitting sand flies are prevalent. Participants were assigned to either vector by 1:1 allocation and exposed to five female sand flies for 30 minutes in a custom biting chamber. Bite frequency was recorded to confirm a bloodmeal was taken. Participant responses and safety outcomes were monitored using a visual analogue scale (VAS), clinical examination, and blood biochemistry. Focus groups were subsequently conducted to explore participant acceptability. Results: All participants had at least one successful sand fly bite with none reporting any serious adverse events, with median VAS scores of 0-1/10 out to day 21 post-sand fly bite. Corresponding assessment of sand flies confirmed that for each participant at least 1/5 sand flies had successfully taken a bloodmeal (overall mean 3.67±1.03 bites per participant). There was no significant difference between P. papatasi and P. duboscqi in the number of bites resulting from 5 sand flies applied to human participants (3.3±0.81 vs 3.00±1.27 bites per participant; p=0.56) .  In the two focus groups (n=5 per group), themes relating to positive participant-reported experiences of being bitten and the overall study, were identified. Conclusions: These results validate a protocol for achieving successful sand fly bites in humans that is safe, well-tolerated and acceptable for participants. Clinicaltrials.gov registration: NCT03999970 (27/06/2019)

Multilocus Microsatellite Typing (MLMT) of Strains from Turkey and Cyprus Reveals a Novel Monophyletic L. donovani Sensu Lato Group

In eastern Mediterranean, leishmaniasis represents a major public health problem with considerable impact on morbidity and potential to spread. Cutaneous leishmaniasis (CL) caused by L. major or L. tropica accounts for most cases in this region although visceral leishmaniasis (VL) caused by L. infantum is also common. New foci of human CL caused by L. donovani complex strains were recently described in Cyprus and Turkey. Herein we analyzed Turkish strains from human CL foci in Çukurova region (north of Cyprus) and a human VL case in Kuşadasi. These were compared to Cypriot strains that were previously typed by Multilocus Enzyme Electrophoresis (MLEE) as L. donovani MON-37. Nevertheless, they were found genetically distinct from MON-37 strains of other regions and therefore their origin remained enigmatic. A population study was performed by Multilocus Microsatellite Typing (MLMT) and the profile of the Turkish strains was compared to previously analyzed L. donovani complex strains. Our results revealed close genetic relationship between Turkish and Cypriot strains, which form a genetically distinct L. infantum monophyletic group, suggesting that Cypriot strains may originate from Turkey. Our analysis indicates that the epidemiology of leishmaniasis in this region is more complicated than originally thought

Eficiência de fungicidas para o controle da mancha-alvo, Corynespora cassiicola, na safra 2011/12: resultados sumarizados dos ensaios cooperativos.

No primeiro ano de realização dos ensaios cooperativos para o controle da mancha-alvo observou-se que há necessidade de ajustes na metodologia, como, por exemplo, não iniciar as aplicações no estádio vegetativo nos ensaios cooperativos. Como as aplicações se iniciaram muito cedo nos ensaios, sendo limitadas a três aplicações, a última aplicação foi realizada entre os estádios R3 e R5, comprometendo a proteção das plantas no final do ciclo da cultura e a porcentagem de controle final dos tratamentos. No entanto, pôde-se observar que há diferença entre os produtos, objetivo principal dos ensaios. Os fungicidas do grupo dos benzimidazóis, amplamente utilizados no campo para controle da mancha-alvo, apresentaram eficiência inferior aos outros produtos. Os produtos com maior eficiência nos ensaios não possuem registro no MAPA até o momento. Outra variável que deve ser analisada com maior rigor é o potencial de dano da mancha-alvo. Para os ensaios cooperativos foram utilizadas cultivares relatadas como suscetíveis no campo e, mesmo assim, a média de redução de produtividade para a testemunha sem controle foi baixa, de 10,3%. Conhecer a reação das cultivares pode auxiliar na definição da necessidade de controle químico específico para essa doença.bitstream/item/69328/1/CT94.pd

Salivary Gland Transcriptomes and Proteomes of Phlebotomus tobbi and Phlebotomus sergenti, Vectors of Leishmaniasis

Phlebotomine female sand flies require a blood meal for egg development, and it is during the blood feeding that pathogens can be transmitted to a host. Leishmania parasites are among these pathogens and can cause disfiguring cutaneous or even possibly fatal visceral disease. The Leishmania parasites are deposited into the bite wound along with the sand fly saliva. The components of the saliva have many pharmacologic and immune functions important in blood feeding and disease establishment. In this article, the authors identify and investigate the protein components of saliva of two important vectors of leishmaniasis, Phlebotomus tobbi and P. sergenti, by sequencing the transcriptomes of the salivary glands. We then compared the predicted protein sequences of these salivary proteins to those of other bloodsucking insects to elucidate the similarity in composition, structure, and enzymatic activity. Finally, this descriptive analysis of P. tobbi and P. sergenti transcriptomes can aid future research in identifying molecules for epidemiologic assays and in investigating sand fly-host interactions

Comparison of 16 national methods in the life cycle assessment of carbon storage in wood products in a reference building

Wood and bio-based construction products are perceived as a way to use renewable resources, to save energy and to mitigate greenhouse gas (GHG)-emissions during production and to store carbon during the entire service life of the building. This article compares the carbon footprint per kilogram of wood products (softwood beams, plywood, oriented strand board panel, and fibre board) from the perspective of the life cycle assessment methodology for greenhouse gas (GHG) emissions of practitioners from 16 countries participating in the IEA Annex 72. These materials are used in PAL6 softwood structure multi-residential building. This article aims at comparing the carbon footprint accounting methods from 16 countries for PAL6 multi-residential building. Each national team applied the reference study period (RSP), life cycle modules covered, modelling rules, the geographical scope of inventory data as well as the LCA database according to its specific national method. The results show that there are three types of methodology to assess a building with biogenic content (0/0, -1/+1, -1/+1*). The results were more variable plywood, oriented strand board, and fibreboard than the softwood beams due to the variability in the wood transformation processes among the countries. A net negative carbon balance was obtained for the softwood beam for the countries using -1/+1* with a clear assumption of the fraction of the carbon permanently stored at the end-of-life (EoL). The carbon storage is only possible if it is secured at the EoL. Participating countries apply different definitions of permanence and EoL scenarios. Guideline on assessing, monitoring, and legally reporting carbon storage at the EoL are needed, based on concertation between standard, life cycle assessment, wood industry, and climate expertspublishedVersio

Systematic functional analysis of Leishmania protein kinases identifies regulators of differentiation or survival

Differentiation between distinct stages is fundamental for the life cycle of intracellular protozoan parasites and for transmission between hosts, requiring stringent spatial and temporal regulation. Here, we apply kinome-wide gene deletion and gene tagging in Leishmania mexicana promastigotes to define protein kinases with life cycle transition roles. Whilst 162 are dispensable, 44 protein kinase genes are refractory to deletion in promastigotes and are likely core genes required for parasite replication. Phenotyping of pooled gene deletion mutants using bar-seq and projection pursuit clustering reveal functional phenotypic groups of protein kinases involved in differentiation from metacyclic promastigote to amastigote, growth and survival in macrophages and mice, colonisation of the sand fly and motility. This unbiased interrogation of protein kinase function in Leishmania allows targeted investigation of organelle-associated signalling pathways required for successful intracellular parasitism