This newscast is updated weekdays at 6am Editor’s Note: Drew Hawkins is in the Netherlands as part of an International Health Study Fellowship with the Association of Health Care Journalists, supported by the Commonwealth Fund New Orleans is known across the world for Mardi Gras but many places also celebrate the Carnival season Take the small village of Schaijk in the Netherlands It’s about an hour and a half drive by car from Amsterdam in the southeastern region of the country The sprawling city and network of canals gives way to the “green deserts” of fields full of sheep and cattle Once you arrive in the village of about 7,000 residents The village ceremonially changes its name from Schaijk to “Moesland” — “moes” being the name of Dutch kale that is grown in the area The mayor gives over control of the village for the four days of the festivities to the “Prince of Moesland.” they’ve chosen a princess: Princess Nicole — her full title being "Thirsty Highness Princess Nicole the First.” Carnaval in the Netherlands traces its roots back to the 14th century but the modern day tradition started after World War II this is the first time a woman has been selected by the “Council of Eleven,” who meet each year to select the new royalty Other Carnaval associations in other villages in the region have chosen a princess before but Moesland has been considered more “traditional,” said Jeroen van Zuijlen chairman of the Carnaval Foundation De Moeslanden Locals said it was a sign that “times are changing.” But it’s a welcome change unique to the southeastern part of the Netherlands evolve to incorporate new traditions into old the Council of Eleven announced local resident Nicole Loeffen was selected to reign as princess And the theme for this year’s festivities would be “All You Need is Loeff.” who has been celebrating Carnaval for more than five decades said they were surprised by the announcement The nomination is teased in the local newspaper and kept a secret until the nominee accepts the crown “Yes of course it was a surprise,” Cornelissen said “But every year you are surprised who it is.” the village has been hard at work building “wagons” — parade floats — by hand many of whom have known each other since elementary school worked together every free moment to construct the elaborate floats Carnaval follows a full life cycle for villagers in Schaijk when elementary school children — with help from their parents and schoolteachers — build smaller floats for the "frotkot," the children’s parade that is held on Sunday For the “second part,” the groups of schoolchildren often stay together growing up and building floats for the adult parade on Monday And when they grow too old to build and pull the floats they move to the “third part” — they sing in the choir started by Cornelissen they can’t get me out,” Cornelissen said with a laugh “The ‘third part’ is the best,” Josje said The "frotkot" started Sunday afternoon as groups of children pulled their floats and handed out Dutch candies dressed in costumes unique to their group’s theme like Super Mario Brothers and Minecraft — “Moescraft” for the parade three-story float pushed by a tractor — much like floats during Mardi Gras in New Orleans — waving to the crowd as a brass band played Carnaval is seen as a strange tradition held by the southerners it’s a way of life that goes back centuries Former prince Joep Manders said he welcomed the change of having a princess rule the festivities “And it started actually when I was a prince And we made it a little bit more important since a couple of years in different areas they choose a princess as the leader of the town the villagers head into a giant tent built specially for Carnaval They grab beers and take seats around wooden tables The celebrations continue well into the night they’ll go back to regular life and wait for next year’s Carnaval The villagers said it’s a shame that half of the Netherlands doesn’t know Carnaval and they’re missing out on the festival of all festivals; a celebration for young and old which traditionally meant letting loose before the Catholic fasting period of Lent leading up to Easter “Even those who don’t understand carnival at all,” said Thirsty Highness Princess Nicole the First “[They] are always welcome to come and watch the parade.” Connecting decision makers to a dynamic network of information Bloomberg quickly and accurately delivers business and financial information The new order is stirring concerns over transparency and liquidity in an already fragmented trading landscape The high-frequency trading firms depicted in his 2014 bestseller worked almost unseen leveraging their technical prowess to mint a fortune in the shadows between stock buyers and sellers Which is what makes Oscar van Schaijk’s current schedule such a surprise the equestrian centre of former Dutch young rider Elisa Swanenberg Thomas and Sanne van Heel purchased the property and will renovate it to include a cross country track and new footing  They will relocate from Oijen to their new property in Schaijk renowned Dutch business entrepreneur and founder of Swanenberg Iron Group that now has businesses in real estate In 2013 the centre was completely renovated when daughter Elisa decided for a dressage career Hanoverian licensed stallion Akatschi (by (Akzent II x Wenzel I x Shogun xx) from Iron Spring Farm in the U.S.A Elisa and Akatschi made their international show debut in 2005 and they competed at Young Riders' level through 2008 with international starts in Hickstead She was trained with Coby van Baalen and Anky van Grunsven  In 2007 the family purchased Grand Prix horse Hiscox Karachi from Richard Davison's stable but he ended up being leased to Isreali Roy First Elisa quit competition at the end of the 2000s while Stable Swanenberg continued to exist as one of the leading equestrian venues in The Netherlands in her mid twenties with husband Frank van der Loop The now 34-year old works in her family's iron company as well as for her own business DGA Netwerk Zuid Elisa's mom Maria is an avid collector of perfume bottles and has created her own private perfume museum in Schaijk Akatschi spent his retirement at Coby van Baalen's yard Related LinksWhat's Happening: January 2009 - Part 1Hiscox Karachi Sold to The NetherlandsIron Spring Farm's Akatschi Sold to Dutch Young Rider Elisa Swanenberg Stalls for Rent at Durondeau Dressage in Peer, Belgium Exceptionally Well Located Equestrian Facility in Wellington, Florida Well-built Equestrian Estate With Multiple Business Opportunities in Sweden Stable Units for Rent at Lotje Schoots' Equestrian Center in Houten (NED) For Rent: Several Apartments and Stable Wing at High-End Equestrian Facility Stable Wing Available at Reiterhof Wensing on Dutch/German border Real Estate: Well-Appointed Country House with Extensive Equestrian Facility in the U.K. Rémi Blot Many of the coins featured military imagery When amateur treasure hunters discovered 107 ancient Roman coins on the banks of a river in the Netherlands at the end of 2017 Deducing that this practice persisted "was an important eureka moment in my academic career." Related: Photos: Roman-era silver jewelry and coins discovered in Scotland  found the coins with metal detectors along the Aa river in the village of Berlicum The stash included four silver denarii and 103 bronze sesterces (worth one-quarter of a denarius) After the van Schaijks reported the find to the Portable Antiquities of the Netherlands an agency that catalogues artifacts found in the country archaeologists found a bronze pendant from a horse harness and two more Roman coins at the site All of the coins were minted between 27 B.C The fact that these coins weren't overly valuable (there weren't any gold coins for instance); were scattered over a large area and not buried together in a "chest or amphora;" and were minted over a period spanning over 200 years suggested that the coins weren't hidden by one person or group as a single treasure trove but were likely "deposited by different people over a long period," Claes said A heavily corroded coin found by the Aa river with an image of the Roman Emperor Titus.(Image credit: Wim van Schaijk)Archaeologists dug a trench alongside the Aa river in the Netherlands.(Image credit: Cultural Heritage Agency of the Netherlands)Historian Liesbeth Claes and amateur treasure hunter Wim van Schaijk look at the corroded Roman coins.(Image credit: Tessa de Groot (T.de Groot&J.W.de Kort (ed.) A safe crossing: Research on a find of coins from the Roman period in the valley of the Aa near to Berlicum (in the municipality of Michielsgestel) Amersfoort 2021)During the team's analysis of the site they found a document dating from 1832 that noted a path cutting across the river where people could wade through the river," Claes said convinced us that travelers in Roman times brought offerings here for a safe crossing." Get the world’s most fascinating discoveries delivered straight to your inbox —Photos: Ancient 'Golden Horde' city revealed  —Photos: 2,000-year-old Roman road and coins discovered in Israel —Images: The largest stash of gold coins discovered in Israel  Even though the river isn't a rapidly flowing one it was important to be able to transport their goods safely to the other side," she said "And there's also the fact that in ancient times rivers always had some sacred connections," which would support the hypothesis that the coins were offerings Archaeologists have yet to find concrete evidence of a ford from Roman times at the site But they're hopeful that future excavations may unearth clues of its existence although the team commended the amateurs for discovering the coins going forward "It is advised not to allow metal detection in the advisory zone so that existing coins and other metal finds are not taken out of context without an archaeological investigation," they wrote in the translated report Her work has appeared in The New York Times She has won multiple awards from the Society of Professional Journalists and the Washington Newspaper Publishers Association for her reporting at a weekly newspaper near Seattle Laura holds a bachelor's degree in English literature and psychology from Washington University in St Louis and a master's degree in science writing from NYU 1,800-year-old warhorse cemetery held remains of a beloved horse — and a man considered an 'outsider' to Roman society Pompeii quiz: How much do you know about the Roman town destroyed by Mount Vesuvius Digital 'resurrection' of the Titanic sheds light on fateful night the ship tore apart Angry farmers from the Netherlands and a dozen other EU countries are heading to Brussels on the eve of the European elections They want to force an end to restrictive European climate and environmental regulations With approximately 25 tractors and truck tractors Dutch farmers left a meeting point in Schaijk They first headed to Someren to collect colleagues waiting there and then continued to the border crossing at Valkenswaard “We want to shake citizens awake before the elections so that they vote in the right direction,” said former dairy farmer Marcel Rijkers “We succeeded in that in the provisional elections and the parliamentary elections Rijkers had hoped that more farmers would participate but thanks to the drier weather in recent days farmers in some parts of the Netherlands can return to the fields “Farmers have also been discouraged by the government Several tractors had protest signs with “no farmers no future” and texts against the Green Deal The Demonstration starts at noon in the Atomium Park in Brussels Brussels is bracing itself since multiple farmers’ actions have recently gotten out of hand the farmers are not gathering around the EU administration buildings but on the outskirts of the city The protest is against the European Union’s agricultural policy Organizer Bart Dickens expects thousands of demonstrators with hundreds of tractors Dickens’ Belgian Farmers Defense Force (FDF) and other more radical farmers’ action groups will take the lead on Tuesday More moderate farmers’ organizations like LTO and the Belgian Farmers’ Union are not participating The organizers have determined “not to confront the police,” Dickens said Metrics details Current first-line treatments for uncomplicated falciparum malaria rapidly clear the asexual stages of the parasite but do not fully prevent parasite transmission by mosquitoes The standard membrane feeding assay (SMFA) is the biological gold standard assessment of transmission reducing activity (TRA) but its throughput is limited by the need to determine mosquito infection status by dissection and microscopy Here we present a novel dissection-free luminescence based SMFA format using a transgenic Plasmodium falciparum reporter parasite without resistance to known antimalarials and therefore unrestricted in its utility in compound screening Analyses of sixty-five compounds from the Medicines for Malaria Venture validation and malaria boxes identified 37 compounds with high levels of TRA (>80%); different assay modes allowed discrimination between gametocytocidal and downstream modes of action Comparison of SMFA data to published assay formats for predicting parasite infectivity indicated that individual in vitro screens show substantial numbers of false negatives These results highlight the importance of the SMFA in the screening pipeline for transmission reducing compounds and present a rapid and objective method In addition we present sixteen diverse chemical scaffolds from the malaria box that may serve as a starting point for further discovery and development of malaria transmission blocking drugs possibly modifying its response to other inhibitors by affecting parasite metabolism Here we report the development of a parasite line that constitutively expresses GFP-Luc and is free of limiting selection markers Our approach used frozen mosquitoes and machine homogenisation in a 96-well format to improve the efficiency of indiscriminate drug screening After validation and comparison of luciferase expression with microscopy based oocyst counts we used our luminescent SMFA to screen 47 marketed and experimental antimalarials and compared the results with data from the gametocytocidal lactate dehydrogenase (pLDH) activity assay and Pfs25 AlphaLISA assay the luciferase screen was used to evaluate the TRA of 18 compounds selected from the Medicines for Malaria Venture (MMV) malaria box specifically for their potential activity as gametocytocidals gametocytocidal or permanent inhibitory activity was confirmed by incubating gametocytes with compounds before ‘washing out’ the compound prior to mosquito feeding while specific activity against the parasite stages emerging in the mosquito gut was confirmed by adding compounds to the gametocyte culture immediately prior to mosquito feeding These assays identified novel transmission blocking small molecules and provide new insight into the predictive value of in vitro metabolic and gamete formation inhibition assays showing they can miss potent compounds and highlighting the advantages of the gold-standard SMFA for screening transmission blocking compounds our more scalable luciferase based read-out increases throughput of the assay and makes compound screening with the SMFA less resource intensive than when performed with microscopy To allow unrestrained evaluation of new compounds we developed a new parasite line, free of drug selection markers (full title: NF54-∆Pf47-5′hsp70-GFP-Luc; hereafter: NF54-HGL), that stably expresses a GFP-Luciferase fusion protein under control of the constitutive P. falciparum hsp70 promoter (Fig. 1A,B). (A) Schematic of the gene targeting strategy to insert a GFP-Luc reporter gene under control of the hsp70 promoter in the pfs47 locus showing the genomic locus of the wild-type (WT) and gene deletion mutants ∆pf47 containing the GFP-luciferase ORF before and after removal of the hdhfr resistance marker N (NcoI): restriction sites used for conformation of LR-PCR analysis; pf47: pf47 locus; hrp: histidine rich protein; cam: calmodulin; hsp: heat shock protein; hdhfr: human dihydrodrofolate reductase coding region; GFP-Luc: GFP-luciferase fusion protein; fcu: cytosine deaminase/uracil phosphoribosyl transferase; pbdt: P berghei dhfr transcription termination region; kb: kilo basepairs (B) Verification of the integration site by PCR and restriction digestion Genomic DNA fragments from the parental NF54 strain (WT) intermediate transgene NF54-HGL-hDHFR and final transgene NF54-HGL were amplified by PCR using primers MWV300 and MWV301 (P1 and P2) and analysed by DNA gel electrophoresis the PCR fragment was subjected to restriction digestion with XcmI/NcoI (X/N) (C) Luminescence intensity (relative light units The figure shows a comparison between uninfected red blood cells (RBC) mature gametocytes (Day 14 GCT) and asexual blood stage parasites (D–G) FLP-FRT mediated excision of the selection marker restores sensitivity to antifolates ∆Pf47GFP-luc-hdhfr and ∆Pf47GFP-luc to DHA (D) Pyrimethamine (F) and WR99210 (G) in a dose dependent asexual growth inhibition assay using pLDH enzyme activity as a readout Depicted data represents a triplicate measurement and was normalized to the MIN (10 μM DHA) and MAX (0.1% DMSO) controls Uninfected negative control mosquitoes showed low levels of luminescence (n = 617; mean RLU = 3.2 RLU; standard deviation = 1.9 RLU) Day 8 PI showed a clear separation of positive and negative mosquitoes and was chosen for subsequent assessments of infection intensity (i.e number of oocysts per midgut) and prevalence (i.e percentage of mosquitoes with oocyst stage infection) based on luminescence intensity Combining all mosquitoes from control feeds prevalence estimates for all mosquitoes from control feeds were 43.33% (208/480) for microscopy and 49.22% (350/711) for luminescence Standard Membrane Feeding Experiments for 7 compounds from the MMV validation box assessed both by microscopy and luciferase assay All compounds were tested in nine dilutions in duplicate The figure shows oocyst counts in individual mosquitoes and relative light units (RLU) from a distinct cohort of mosquitoes from the same cage The table shows IC50 values and cognate 95% confidence intervals (CI) determined from fitting the data to a logistic regression model by Maximum Likelihood Estimation NA: confidence interval not computed as the model did not converge (A) Single concentration (5 μM) rapid luminescent SMFA screen of 47 compound from MMV validation box assessed with the indirect and direct assay variants TRA% was calculated from relative differences between test and control feed luminescence intensity Error bars indicate 95% confidence intervals (B) Heat map comparing the SMFA data to published data from pLDH and Pfs25-based gametocyte viability assays The numbers indicate percentage inhibition at a compound concentration of 5 μM A color scale from green to red indicates low to high inhibition To provide more information on the mechanism of active compounds all compounds showing ≥ 80% TRA on the intensity scale in the indirect assay were re-tested this time adding compounds to the blood/gametocyte culture mix immediately prior to mosquito feeding (hereafter Fourteen compounds had > 80% TRA in the indirect assay but not in the direct assay suggesting the absence of strong activity against the parasite life stages occurring in the mosquito gut BC1 inhibitor atovaquone and protein synthesis inhibitor cycloheximide had comparable TRA in the indirect and direct SMFA formats indicating activity against mosquito stage parasites but not excluding the possibility of concurrent gametocytocidal action the pLDH and gametogenesis assays identified 6/47 and 13/47 compounds respectively as having gametocytocidal activity The activity of all but one of the six compounds identified by the pLDH assay (napthoquine) was confirmed by the gametogenesis assay while all hits from either viability assay were confirmed in the indirect luminescent SMFA high TRA was observed for a number of compounds where the inhibition assays showed low activity indicating that some compounds had activity only identifiable with transmission as the definitive readout Total false negative rates for the pLDH and gametogenesis assays were 32% (15/47) and 17% (8/47) Excluding compounds that blocked transmission in the direct luminescent SMFA which we presumed to have activity downstream of the gametocyte the SMFA still identified a larger number of transmission-blocking compounds than either gametocytocidal assay with false negativity rates of 25% (10/40) in the pLDH assay and 10% (4/40) in the gametogenesis assay This is not surprising since the SMFA covesr a higher biological content than either of the other two methodologies Dose response experiments for 18 compounds from the MMV malaria box assessed in the luminescent SMFA in a wash-out assay variant All compounds were tested in duplicate in two independent experiments The figure shows oocyst intensities relative to vehicle control (0.1% DMSO) as a function of concentration for the compounds indicated at the top of the panels The firefly luciferase expressing parasite line NF54-HGL enables the SMFA to be performed for indiscriminate compound screening with a semi-automated luciferase based read-out The results of the current study demonstrate the versatility of the SMFA for screening compounds with activity against different transmission stages of the malaria parasite and validate the luminescent assay format in combination with microtiter plate-based processing of mosquitoes for improving the efficiency of the biological gold-standard for assessing Plasmodium transmission in controlled experiments luminescence measurements provide continuous data that reflect actual numbers of developing sporozoites which may be more directly relatable to mosquito infectivity for which oocyst counts are an accepted proxy An added advantage of the luminescent assay is that it is less subjective than microscopy based assessments of oocyst numbers We cannot exclude the possibility that in heavily infected mosquitoes with higher oocyst intensities than those reported here nutrient and space competition may lead to lower per capita sporozoite production resulting in a non-linear relationship between luminescence and oocyst intensity and lower luminescence based TRA estimates than would be determined from oocyst number alone Though we advise caution in extrapolating actual oocyst number from luminescence values we believe luminescence based TRA estimates may be more accurate than those made using oocyst data The possibility that some compounds may effect mosquito metabolism and by proxy parasite developmental success Readouts based on oocyst number may be insensitive to such subtle effects while the more quantitative luminescence readout may not be Additional experiments will be required to elucidate the interaction of mosquito and parasite fitness in relation to compound exposure An experimental and computational strategy to achieve such an assessment has been implemented in our laboratory and will be described elsewhere Potent transmission-blocking molecules The depicted compounds showed an IC50 < 100 nM in the luminescent SMFA The results of the current study demonstrate the sensitivity and flexibility of the SMFA for determining the effect of small molecules on P falciparum gametocyte transmission to mosquitoes while validating a method that increases the assays scalability and ease of use Comparison to gametocyte inhibition assays indicates that parasite development in the mosquito may be required to ensure assay sensitivity to transmission-blocking small molecules Of the 18 compounds selected in this analysis based on the results of previous gametocyte assays our data indicate that 3 compounds have greater activity on transmission stage parasites than asexual stage parasites These molecules may be of particular interest for further drug discovery and elucidation of novel drug targets An AgeI restriction site was introduced immediately upstream of the start codon by introducing a PCR fragment amplified with primers MWV252 and MWV253 through KpnI/HpaI restriction digestion and ligation The promoter region of the hsp70 gene (PF3D7_0818900) was placed upstream of GFP through sacII/AgeI restriction digestion and ligation after amplification with primers MWV198 and MWV200 and subcloning in pEGFP-N1 (Clontech laboratories CA) through BamHI/SacII digestion and ligation The 3′ transcription termination region from the hsp86 gene (PF3D7_0708400) was introduced following PCR amplification using primers MWV268 and MWV271 and XbaI/ApaI restriction digestion and ligation Finally the reporter cassette was introduced in the gene targeting plasmid by KpnI/NgoMIV restriction digestion and ligation yielding plasmid pMV163 5′hsp70 and 3′hsp86 fragments were amplified by PCR amplification (Phusion falciparum DNA (NF54 strain) and all PCR fragments were sequenced after TOPO TA (Invitrogen) sub-cloning Parasites in positive wells were transferred to the semi-automated culture system and cultured for further phenotype and genotype analyses Integrity of the transgenes was analysed by PCR using primers MWV300 and MWV301 and XcmI/NcoI restriction digestion Assays were performed in triplicate (three serial dilutions for each compound) and all plates included triplicate controls to assess minimum (MIN control To compare the three different strains data were normalized and expressed as the percentage reduction in replication relative to the blocking and non-blocking controls The midguts of mosquitoes fed previously with NF54-HGL gametocyte culture were mounted on glass slides in a droplet of PBS and GFP expression was visualized and photographed on a Leica fluorescence microscope with digital camera or processed in the luciferase activity assay on day 8 PI For the individual mosquitoes 45 μl of homogenised material was transferred to wells in a Krystal 2000 black & white 96-well plate (Porvair Next 45 μl of Bright-Glo luciferase assay substrate (Promega WI) was added to each well using a Cybi Selma dispensing station (Cybio Germany) and the plate was incubated for 3 minutes at room temperature Luciferase activity was measured using a Synergy 2 multi-purpose plate reader (Biotek Background was determined by measuring uninfected mosquitoes that had undergone the same treatment Assays were performed as described previously13 Statistical analysis was conducted using R (Foundation for Statistical Computing USA) and GraphPad Prism 5.0 (GraphPad Software Inc. data were expressed relative to the negative (vehicle) controls for oocyst density and oocyst prevalence (i.e IC50 values were calculated by applying a four-parameter logistic regression model using a Maximum Likelihood Estimation algorithm to find the best fit A semi-automated luminescence based standard membrane feeding assay identifies novel small molecules that inhibit transmission of malaria parasites by mosquitoes The decline in paediatric malaria admissions on the coast of Kenya Effect of Antimalarial Drugs on Plasmodium falciparum Gametocytes Artesunate combinations for treatment of malaria: meta-analysis Reduction of transmission from malaria patients by artemisinin combination therapies: a pooled analysis of six randomized trials Revisiting the circulation time of Plasmodium falciparum gametocytes: molecular detection methods to estimate the duration of gametocyte carriage and the effect of gametocytocidal drugs Strategic use of antimalarial drugs that block falciparum malaria parasite transmission to mosquitoes to achieve local malaria elimination Primaquine to prevent transmission of falciparum malaria Primaquine or other 8-aminoquinoline for reducing P Rationale for short course primaquine in Africa to interrupt malaria transmission Male and Female Plasmodium falciparum Mature Gametocytes Show Different Responses to Antimalarial Drugs A Male and Female Gametocyte Functional Viability Assay To Identify Biologically Relevant Malaria Transmission-Blocking Drugs A combination of new screening assays for prioritization of transmission-blocking antimalarials reveals distinct dynamics of marketed and experimental drugs Anopheles gambiae Circumsporozoite Protein–Binding Protein Facilitates Plasmodium Infection of Mosquito Salivary Glands A scalable assessment of Plasmodium falciparum transmission in the standard membrane-feeding assay using transgenic parasites expressing green fluorescent protein-luciferase A transgenic Plasmodium falciparum NF54 strain that expresses GFP–luciferase throughout the parasite life cycle falciparum in vitro killing rates allow to discriminate between different antimalarial mode-of-action Comparative whole genome transcriptome analysis of three Plasmodium falciparum strains A Plasmodium falciparum screening assay for anti-gametocyte drugs based on parasite lactate dehydrogenase detection A malaria gametocytocidal assay using oxidoreduction indicator Activity of clinically relevant antimalarial drugs on Plasmodium falciparum mature gametocytes in an ATP bioluminescence “transmission blocking” assay Identification of MMV Malaria Box Inhibitors of Plasmodium falciparum Early-Stage Gametocytes Using a Luciferase-Based High-Throughput Assay A high-throughput assay for the identification of malarial transmission-blocking drugs and vaccines Antiapicoplast and gametocytocidal screening to identify the mechanisms of action of compounds within the malaria box Identification of inhibitors of Plasmodium falciparum gametocyte development Gametocytocidal screen identifies novel chemical classes with Plasmodium falciparum transmission blocking activity Isolation and functional characterization of two distinct sexual-stage-specific promoters of the human malaria parasite Plasmodium falciparum Identifying genomic changes associated with insecticide resistance in the dengue mosquito Aedes aegypti by deep targeted sequencing standard membrane feeding assay for assessing the transmission blocking activity of vaccines and drugs against Plasmodium falciparum Population dynamics of sporogony for Plasmodium vivax parasites from western Thailand developing within three species of colonized Anopheles mosquitoes The relevance and applicability of oocyst prevalence as a read-out for mosquito feeding assays The number of sporozoites produced by individual malaria oocysts KRIBB11 inhibits HSP70 synthesis through inhibition of heat shock factor 1 function by impairing the recruitment of positive transcription elongation factor b to the hsp70 promoter Luciferase inhibition by a novel naphthoquinone Measuring the blockade of malaria transmission—An analysis of the Standard Membrane Feeding Assay Malaria: some considerations regarding parasite productivity Substantial contribution of submicroscopical Plasmodium falciparum gametocyte carriage to the infectious reservoir in an area of seasonal transmission Epidemiology and Infectivity of Plasmodium falciparum and Plasmodium vivax Gametocytes in Relation to Malaria Control and Elimination The role of anti-malarial drugs in eliminating malaria Nowhere to hide: interrogating different metabolic parameters of Plasmodium falciparum gametocytes in a transmission blocking drug discovery pipeline towards malaria elimination The Activities of Current Antimalarial Drugs on the Life Cycle Stages of Plasmodium: A Comparative Study with Human and Rodent Parasites Antimalarial drugs and the mosquito transmission of Plasmodium Persistence of atovaquone in human sera following treatment: inhibition of P falciparum development in vivo and in vitro Molecular basis of differential resistance to cycloguanil and pyrimethamine in Plasmodium falciparum malaria Lead clinical and preclinical antimalarial drugs can significantly reduce sporozoite transmission to vertebrate populations Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings The open access malaria box: a drug discovery catalyst for neglected diseases Targeting Plasmodium PI(4)K to eliminate malaria A long-duration dihydroorotate dehydrogenase inhibitor (DSM265) for prevention and treatment of malaria A novel multiple-stage antimalarial agent that inhibits protein synthesis First-in-man safety and pharmacokinetics of synthetic ozonide OZ439 demonstrates an improved exposure profile relative to other peroxide antimalarials Designing the next generation of medicines for malaria control and eradication Complete in vitro maturation of Plasmodium falciparum gametocytes Infectivity of cultured Plasmodium falciparum gametocytes to mosquitoes Cultivation of fertile Plasmodium falciparum gametocytes in semi-automated systems Negative selection using yeast cytosine deaminase/uracil phosphoribosyl transferase in Plasmodium falciparum for targeted gene deletion by double crossover recombination Negative selection of Plasmodium falciparum reveals targeted gene deletion by double crossover recombination Transformation with human dihydrofolate reductase renders malaria parasites insensitive to WR99210 but does not affect the intrinsic activity of proguanil Removal of Heterologous Sequences from Plasmodium falciparum Mutants Using FLPe-Recombinase High efficiency transfection of Plasmodium berghei facilitates novel selection procedures In Application of genetic engineering to research on tropical disease pathogens with special reference to Plasmodia (ed S A set of independent selectable markers for transfection of the human malaria parasite Plasmodium falciparum Generalized linear mixed models: a practical guide for ecology and evolution Download references We want to thank Chris Janse and Shahid Kahn for providing us with plasmid PL1063 Astrid Pouwelsen and Jolanda Klaassen for mosquito breeding and for their assistance during SMFA evaluation Hans-Peter Duerr is gratefully acknowledged for statistical analyses and Paul Willis for selection of compounds for SMFA evaluation This work was supported by a grant from the Bill and Melinda Gates Foundation (OPP1040394) are supported by PATH Malaria Vaccine Initiative and the European Community’s Seventh Framework Programme (SIGNAL; Marie Curie Career Integration Grant PCIG12-GA-2012-333936) Radboud University Nijmegen Medical Center selected compounds and performed data analyses KJD and RWS own stock in TropIQ Health Sciences All other authors have no competing financial interests to 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My Profile Logout Slovak banjoist Richard Ciferský, founder of innovative music accessory company Banjolit and banjo man with Bill Faster in the Slovak Republic, has released a unique and interesting project called The Adventures of Willo the Dwarf with a children’s book he has written the package opens to reveal a 20-page book with charming illustrations by Kateřina Hruschková that follows the story of Willo and his search for the pot of gold at the end of the rainbow Each section of the book’s text corresponds with an audio track on the CD and the reader is prompted to listen to the song at the end of each brief chapter One can easily see a young child or grandchild wanting to “read” the story over and over insisting on also hearing the songs that complete the tale Richard wrote this story in his native Slovak language and then translated it into English with the assistance of Becky Buller Willo’s adventure brings him into contact with all sorts of things youngsters love and the moral of the story supports belief in our dreams along with the values of love and friendship Performing on the CD are Michal Vavro on guitar It’s hard to imagine a young child or early reader who wouldn’t enjoy The Adventures of Willo the Dwarf It can be purchased for $19 from the Banjolit web site. emailAbout the AuthorJohn LawlessJohn had served as primary author and editor for The Bluegrass Blog from its launch in 2004 until being folded into Bluegrass Today in September of 2011 managing a strong team of columnists and correspondents Latest Event ListingsSearch Events powered by AhSo © Bluegrass Today 2025powered by AhSo NEW ORLEANS (AP) — The good times and parades still rolled in New Orleans for Mardi Gras Day celebrations early Tuesday thunderstorms and tornado warnings in the afternoon Carnival Season's final parades in Louisiana's most populous city which is world-famous for its Mardi Gras bash were moved to earlier time slots — with masked riders on elaborate floats and costumed revelers determined to beat the bad weather and partake in the conclusion of the pre-Lenten jollification people donning skeleton masks and antlers on their heads gathered in New Orleans' historic Treme neighborhood Members of the Northside Skull and Bones Gang woke up the city with drums and chants “Mardi Gras is all about having these moments This city is like nowhere else in the country right now,” said Kendall Calyen marveling at the 200-year-old tradition unfolding before his eyes The city’s two biggest parades — hosted by the social clubs Krewe of Zulu and Krewe of Rex — started earlier shortened their routes and are going without marching bands New Orleans Police Superintendent Anne Kirkpatrick said The parades wrapped up by the afternoon to avoid worsening weather The National Weather Service forecasted thunderstorms and winds up to 60 mph (100 kph) and had issued tornado watches for the area New Orleans Mayor LaToya Cantrell told the Associated Press that despite weather challenges the city was able modify the parades to ensure that there were still traditional Mardi Gras Day festivities “My favorite part is the authenticity of the city of New Orleans on full display for the world to see which is woven and embedded in our culture that’s through our people,” Cantrell said family and friends gathered wearing Mardi Gras purple Impromptu barbecues wafted smoke from street corners and hip-hop blared Jeremy Speed, a 40-year-old Krewe of Zulu member who grew up watching the parade, rode a float and tossed his hand-painted coconut “throws ” to onlookers He was disappointed that the route had been shortened due to weather but was grateful that the parades weren't canceled altogether They gave us a little something,” Speed said Two other parades which had been scheduled to meander through the city later on Tuesday with nearly 200 truck floats were postponed to Sunday At the start of the year, some seemed skeptical whether Carnival celebrations would take place as normal, following the devastating Jan. 1 truck attack that killed 14 people on Bourbon Street in New Orleans The Department of Homeland Security upgraded Mardi Gras to its highest risk rating leading to an influx of law enforcement agents A zigzagging layout of barriers paralleled the main parade route to guard against fast-moving vehicles and armored trucks; SWAT teams and helicopters were present Amid the weather threat and security measures Along with colorful floats and paradegoers with their necks hunched from a hefty haul of shimmery beads, were renowned Black masking Indians whose spectacular beaded and bejeweled costumes are topped with feathered headdresses Such groups have taken to the streets to represent a central part of African American culture in the city dating back to the 1800s wore an intricate blue beaded outfit that she spent all year threading beads onto using dental floss The member of the Wild Magnolias quickly put together a headband the night before Fat Tuesday as a back-up in case her massive feathered headdress flew off in the wind Batiste will be preparing for next year: “For real The Krewe of Zulu parade is known for hundreds of marchers tossing trinkets including highly coveted decorated coconuts the Krewe of Rex — a social club led by the honorary king of Carnival season — bears the traditional colors of purple The weeks of Carnival culminate with feasting on Fat Tuesday, which precedes Ash Wednesday, the start of Lent This year’s Mardi Gras Day fell unusually late in the year But the outsized culture of street parties extravagant balls and spirited parades has evolved into a decidedly secular spectacle in the Big Easy Sign up for our daily Newsletter and stay up to date with all the latest news You are receiving this pop-up because this is the first time you are visiting our site You are using software which is blocking our advertisements (adblocker) we are relying on revenues from our banners So please disable your adblocker and reload the page to continue using this site.Thanks GEGE Machinebouw introduced waterjet cutting lines for products This innovation means using a moving waterjet blade to process leeks has become a common practice among leek growers there was always a bottleneck when positioning the leeks because they had to be placed at a fixed distance and you lose volume and limit shelf life," begins this Dutch engineering company's director His company has now solved this issue thanks to an Artificial intelligence (AI)-based camera system These vegetables come in all lengths and thicknesses as much as 70% of the leeks had to be handled by people afterward Linking this camera system means the machine now recognizes where the root begins so we can direct the waterjet blade to the exact right position up to 95% precision slicing is sometimes possible Those parts of the leeks the cameras can't see are still cut the end products have a better quality because cutting the root base in the correct place ensures maximum shelf life and minimum product loss." “The moving waterjet blade slices the vegetables efficiently pressurized jet of water cuts the product perfectly in the exact right place Traditional cutting systems blunt the blades quickly The advantage of water cutting is that the blade doesn't become blunt “That results in a very sharp cutting edge Water shoots out of the nozzle at 600 meters per second as it prevents fungi and germs from being carried away from the blade and neither does the machine damage the products Capacity remains the same: 160 leeks per minute Research shows that this technique improves leeks shelf life by quite a few days." Demo modelThis new camera system was developed last year and GEGE expects to deliver the first machines this year making half our revenue in the Netherlands and the rest overseas We've also sold waterjet cutters in places like Canada "You cannot use waterjet cutting for all products certain types of hard cheese and boiled eggs did not yield the desired results It proved suitable for fruits and vegetables such as leeks "You can discuss this technique's usefulness to no end but the best way is to test it in practice That's why we have a demo model so companies can test this method on their products." "The integrated camera system completes the circle you can have a fantastic automated machine you still won't achieve the desired result It's not too hard to cut correctly; the issue is positioning them correctly We manage to do that by integrating vibrators our cameras don't have a standard Vision system; they use AI That promises something for the future since the system only gets smarter the more accurate the system becomes," Bart concludes FreshPublishers © 2005-2025 FreshPlaza.com Stibbe is advising Barça Produccions and Barcelona Football Club alongside Spanish law firm Perez Llorca in relation to a business combination agreement with Mountain & Co – a special acquisition company (SPAC) listed on NASDAQ – which will result in Barça Produccions becoming a company listed on NASDAQ The transaction values the pro-forma company at approximately €1 billion will give Barça Produccions access to the US capital markets to create a stronger platform to accelerate its multi-pronged growth strategy FC Barcelona has the largest fan base among all professional football clubs and an exceptional brand recognition Barça Produccions intends to capitalise on these factors and leverage strong market tailwinds to produce original content that stimulates fan engagement and drives long-term profitable growth FC Barcelona expects Barça Produccions to serve as an important source of revenue in the coming years and has assigned significant resources to support the business Get quality reporting directly into your inbox The suspects were members of a “prolific organized crime gang flooding Europe with millions of counterfeit cigarettes,” and are believed to have been involved in violent robberies conducted in the Netherlands at the end of March police dismantled two illegal tobacco factories in the cities of Schaijk and Heerlen Europol pointed out that the capacity of the dismantled illicit factories was more than a million cigarettes a day equating to a “tax loss of over 243,000 euro (US$294,000) per day per factory.” Dutch police arrested 21 Polish and Ukrainian nationals working in the factories and seized the 5.4 million counterfeit cigarettes 40 tonnes of raw tobacco and 800 kilograms of hookah tobacco The second phase of the operation was carried out last week in Poland and targeted “the criminal masterminds running this gang,” Europol said Police raided a dozen of objects across the country arrested nine people and seized 54 tons of tobacco as well as machinery used for the production of cigarettes Polish officers also “seized firearms and weapons alongside clothing resembling police uniforms radio communication devices and signal jammers,” the statement said Europol said that initial estimates are that the loss to the Polish budget will exceed 52 million Polish zloty ($13.72 million) Investigators are trying to find “potential links to other European countries,” the European police agency said Support from readers like you helps OCCRP expose organized crime and corruption around the world you’ll be directly supporting investigative journalism as a public good You’ll also gain access to exclusive insights and benefits German and Hungarian authorities arrested the alleged kingpins of a Hungarian gang of traffickers for sexual exploitation,.. Authorities in Italy and Spain have crushed a sophisticated drug trafficking and money laundering network that is believed.. Spanish police arrested one of Europe’s biggest money launderers linked to an Irish criminal clan and dismantled the Spanish.. Pearl GTL Technology Manager and Vice-President QSRTC; and other officials at the agreement signing ceremony Doha: University of Doha for Science and Technology (UDST) has initiated a collaboration agreement with Qatar Shell Research and Technology Center (QSRTC) to support Engineering Technology students with their capstone projects Pearl GTL Technology Manager and Vice-President QSRTC signed the agreement in the presence of a number of University officials and a delegation from the energy company The agreement sets a joint strategy to help students identify their suitable capstone project and get industry insights throughout the process A capstone project is usually the final assignment at university with students carrying out research to develop an innovative solution to a real-world challenge A Qatar Shell engineer will guide the students provide feedback and help find adequate resources to complete all requirements for their project UDST will dedicate technical supervision and will allocate the needed space for these projects this collaboration brings forward many opportunities for the students to enhance their work and develop new skills and competencies President of UDST said: “At UDST we believe that applied education leads to academic excellence Our students learn by applying the knowledge they gained to real-world settings and this type of learning can only be achieved with the support of our partners from different industries Qatar Shell is one of our long-standing partners who has been helping us for years to nurture and guide young ambitious minds It is our pleasure to be partnering once again on this particular initiative and we look forward great outcomes.” said: “Qatar Shell is an active supporter of Qatar’s journey towards a knowledge-based economy We are proud to share our skills and expertise with the future leaders of Qatar so that they can continue to innovate the technologies of tomorrow and tackle the challenges that lay ahead of us Partnering with UDST on this initiative will help to equip students with some of the tools they need to benefit the technical and economic development of Qatar.” two amateur archaeologists from Brabant discovered over a hundred Roman coins near to Berlicum in the north of the province Another interesting fact is that the coins offered were chosen very precisely ‘This was an important eureka moment in my academic career.' Claes: ‘You can determine from different characteristics that this was not a hoard of valuable treasure that people deliberately buried for later These coins were lying spread over a large area and were just loose in the earth they turned out not to be particularly valuable coins What we found were mainly bronze coins; there were no gold ones That's not what you'd expect if the find were a treasure trove the coins were from a very broad time period: the oldest coins date from the Republic [up to 27 BC Ed.] and the most recent from the time of Emperor Marcus Aurelius [up to 180 AD] This indicates that the coins must have been deposited by different people over a long period.'’ ‘In the 1832 entry in the land registry - one of the Netherlands' oldest detailed maps - my colleagues noticed that at that time there was a path cutting across the river convinced us that travellers in Roman times brought offerings here for a safe crossing It may not have been a particularly rapidly flowing river but for traders in particular it was important to be able to transport their goods safely to the other side. And there's also the fact that in ancient times rivers always had some sacred connections.'   ‘I was mainly involved in studying the coins themselves I looked at whether the travellers selected particular coins for their offerings It seems that they did: coins with military imagery seemed to be popular offerings because what does a military image have to do with a safe crossing But then I realised that this was also quite common before Roman times: we regularly find offerings of axes swords and helmets in other rivers and stretches of water This could be a pre-Roman custom that continued in the Roman period but in a different way That was an important eureka moment in my academic career.'  ‘Definitely. We stayed in contact with Nico and Wim throughout the research. They also helped with additional field research at the find site. For me, this is a good example of how amateur archaeologists and academics can complement one another. That starts with amateur archaeologists reporting their finds, as Wim and Nico did, to Portable Antiquities of the Netherlands To show our thanks, we recently presented them with the first copy of our report.’ Brothers Nico and Wim van Schaijk were responsible for finding the coins. In late 2017 they detected the coins using a metal detector and excavated them. In this report the brothers take us back to the find site in a field beside the river Aa The Unification of the Mediterranean World 400BC-400AD Numismatics in Leiden: more than two sides to the same coin Reevaluating Conceptions of Imperial Monetary Flow: New Methodologies and Frameworks Coin streams within the Roman West (AD 83-138) Financial flows to/from Soviet Central Asia