When Dutch prog rock legends Kayak released album number Seventeen longstanding keyboard player Ton Scherpenzeel looked back and pondered what might have been Dutch proggers Kayak came close to breaking America in the 70s Kayak’s last remaining original member Ton Scherpenzeel looked back over their career and discussed what went wrong his hopes for the future and his fear of flying… Nor should there be any, of course. Kayak’s history dates back to 1972 when the teenaged Scherpenzeel and drummer Pim Koopman became friends at a volleyball club in Hilversum The pair knew their eventual lead vocalist Max Werner as a fellow student at a local music conservatory completing the team with guitarist Johan Slager and bassist Cees van Leeuwen The band achieved huge success right off the bat with the following year’s debut album See See The Sun and its hits it was as good a start as anyone could have wished for.” Max Werner decided to abandon the mic and become a drummer The timing of such a switch could not have been any worse reach Number 55 on America’s Billboard chart and threaten to climb higher still Forced to seek a replacement singer and unable to tour there thanks to Scherpenzeel’s fear of flying Kayak’s American dream was abandoned to wither and die “Max hated his own voice – he had never wanted to be the lead singer when we started,” Scherpenzeel states ‘If you want me to remain in the band I will drum.’ He wasn’t the best drummer in the world we’d still have his voice along the drums America felt a lot further away than it does now Everything was done by letters and phone calls but without having to change things around something very big could have happened for Kayak there.” disappointment can still be heard in Scherpenzeel’s voice To have come within touching distance must have been especially heartbreaking the full significance didn’t really dawn on me,” he admits “We had a number one album in Holland and we were busy but it’s only afterwards when you see the paperwork and statistics when they’re out there on the internet Kayak could really have been something.’” Scherpenzeel used Kayak’s absence to join Camel from 1984 onwards Scherpenzeel was thrilled to hook up with Andy Latimer and company alternating between full-time band membership and touring muso status and appearing on Stationary Traveller It was the keyboardist’s problems with aviation that made it difficult for this relationship to continue Despite having sought various potential remedies things haven’t really changed for him “If it is not really necessary for me to fly Kayak were invited to appear on a popular Dutch TV show Things went so well that Scherpenzeel had no qualms with a permanent reunion though this wasn’t as simple as it sounds “I had remained in contact with Pim Koopman the other important member of the band who and we took things slowly at the start,” he relates Scherpenzeel and Koopman first discussed working together again in 1995 but it would take a further four years for Kayak to be reborn “We had a manager who forbade us to use the name,” Scherpenzeel explains It took his death in ’98 for us to receive the all-clear.” Regrettably, Koopman also died unexpectedly in 2009. In 2014, several line-ups later, a further obstacle arose when Kayak lost co-lead singers Edward Reekers and Cindy Oudshoorn before a tour for the rock opera Cleopatra – The Crown Of Isis a project that the group had spent two years preparing for Kayak’s current biography pointedly describes those exits as “unexpected and inconvenient” “I cannot say it any better than that,” comments Scherpenzeel through gritted teeth. Kayak replaced a man and woman with a solitary male – Bart Schwertmann – and their latest, all-new line-up also includes Kristoffer Gildenlöw, formerly of Pain Of Salvation though Scherpenzeel played most of the album’s bass parts himself So belated was Kayak’s tie-in with InsideOut that Scherpenzeel is thrilled by the possibilities of association with one of the biggest and most forward-thinking labels of the genre “This is the first time in 12 or 13 years that we’ve had a record label,” he chuckles but the experience taught us that to take things further you really do need extra support.” Kayak’s music is certainly blessed with commercial potential though at 11 and 10 mins respectively the grander pieces La Peregrina and Walk Through Fire really allow them to stretch out music must sound organic – it’s not a matter of how clever it is,” Scherpenzeel stresses “I love shorter songs just as much as the epic ones but the format of verse-chorus-verse-chorus is just as fulfilling Pop music has been a part of our repertoire: we are a two-sided band and that has sometimes confused our audience but it’s something that I cannot change.” Andy Latimer is a guest on Ripples On The Water a beautiful instrumental piece that Scherpenzeel wrote intentionally for the guitarist he plays one note and you know it’s him,” Scherpenzeel smiles “I’ve always loved Camel’s music Maybe I’ll go back to Camel someday: it’s up to Andy.” Kayak look set to be busy over the next couple of years they last performed in the UK back in March 1977 on a bill with Jan Akkerman and singer Kaz Lux who were promoting their concept album Eli “Jan didn’t want to play any Focus material which left the audiences disappointed so it was cut short,” Scherpenzeel recalls this was the band’s one and only foray to Britain to date of course Scherpenzeel is anxious to return under more favourable circumstances with just the English Channel between our two nations “European gigs are always possible – I did some two years ago with Camel Of course Kayak want to play wherever possible Dave LingSocial Links NavigationNews/Lives Editor Classic RockDave Ling was a co-founder of Classic Rock magazine His words have appeared in a variety of music publications Dave’s life was shaped in 1974 through the purchase of a copy of Sweet’s album ‘Sweet Fanny Adams’ along with early gig experiences from Status Quo As a lifelong season ticket holder of Crystal Palace FC he is completely incapable of uttering the word ‘Br***ton’ "Metal is the most positive thing in the world" “If you’re a Zep fan and really want to go see Zeppelin you might as well go and see one of the better tribute bands”: The epic life and career of John Paul Jones the heartbeat of Led Zeppelin and so much more Ghost's $130 Papa V Perpetua 'Ghildo' sells out in minutes Interview by Paul González-Morgan of Marine Strategy – Ben van Scherpenzeel Policy & Plans at the Port of Rotterdam is passionate about continuous improvement with a focus on ports and shipping in every aspect: nautical Ben’s past experience onboard vessels of Shell and Holland America Line together with the knowledge he has gained of the port since 2004 have been applied to initiate and develop many different projects that have had a positive impact on the port’s ecosystem and further afield across the shipping industry Container throughput at the Port of Rotterdam has risen significantly in Q1 2018 to 3.5 million TEU per quarter The continued growth in container throughput is a confirmation that Rotterdam is taking an increasingly important position in the maritime connection networks of large shipping company alliances We are seeing a somewhat more measured growth after the significant growth last year and this is entirely in accordance with our expectations What are the challenges of Marine Spatial Planning in the context of the Port of Rotterdam For traffic approaching the Port of Rotterdam we’ve worked hard together with our national authorities and all stakeholders at sea to create sufficient space for safe navigation and wind farms we’ve concentrated on deepening the river towards the Botlek area Overall it’s a challenge to use the existing infrastructure as efficient as possible by better planning – not only of ships but also related to maintenance like dredging Port call inefficiencies result in unnecessary delays costing billions in lost revenue and CO2 emissions How has the Port of Rotterdam optimised vessel calls to maximise efficiency This will be achieved by working on maximum cargo on board by online depth information This will mean taking into account “Just In Time” arrivals by exchange of ship planning – platform to exchange has become life: Pronto Fundamental for both data exchanges is to have standards that work for shipping port to port worldwide so parties do not build in unnecessary safety margins as they fully understand and trust the information provided Please talk to us about the importance of information exchange between all parties in the logistics chain: The departure time is the cornerstone to port logistics Most departure time dictate the arrival time of another vessel and that arrival time is key for planning resources for nautical services (pilots cargo services and vessel services (bunkers departure time depends on many parties: most of all terminal operations but certainly also bunker operations or any other critical service a vessel needs to complete before departure What are the three key elements that define productivity at a port terminal and avoiding idle time when commercial operations have finished Severe weather conditions can impact port operations How do you minimise the down-time at the terminal allowing ships to remain alongside safely under adverse weather conditions Is Blockchain a solution to increase port efficiency Last year the Municipality of Rotterdam and the Port of Rotterdam Authority jointly started BlockLab: a field lab for Blockchain Blockchain technology offers considerable potential to change the way we manage supply chains Its main contributions will be increased compliance and transparency and find more effective ways of resolving trust issues High-impact is therefore to be expected within the domains of supply chain finance and cross chain collaborations – especially when chains are decentralised and involve numerous stakeholders and transactions BlockLab puts Blockchain technology into practice The Lab develops the use of cases with alliances of engineers Is the port fully aligned with the demands of the ocean supply chain We’re working together with GS1 to connect to the supply chain standards allowing better resource planning in warehouses and factories How is the increasing size of ships shaping your plans for the port Ten years ago we already designed the Maasvlakte for the ships we have today We already applied more and stronger bollards based on the new IACS guidelines which will enter into force this year how do you see Brexit affecting the Port of Rotterdam in terms of capacity We all agree that Brexit will have its challenges however the Port of Rotterdam is treating this matter as a top priority and is fully committed in talking to all stakeholders (Dutch Government etc.) with a focus on contingency plans to ensure that the transition will go as efficient as possible What future plans do you have for the port The Dutch port infrastructure has been elected best port infrastructure in the world by the World Economic Forum for the sixth consecutive time A good infrastructure is essential for a port’s growth and development Billions have been and will be invested in the expansion of the intermodal network state-of-the-art terminals and ICT systems and the Offshore Center Rotterdam to name a few the digital infrastructure is of major importance By creating transparency across the chain and by sharing data we can further increase the efficiency and reliability of the logistics chain The Port of Rotterdam Authority invests 150 to 200 million euros per year in its port infrastructure Important investments for the coming year include the development of the Hartel Tank Terminal and the changes to the port railway via Thamesweg eliminating the clash between transport by rail and ocean-going vessels Find more Marine Strategy interviews here. Copyright (c) Marine Strategy Sign up for gCaptain’s newsletter and never miss an update and updates delivered daily straight to your inbox President Donald Trump's administration is considering softening its proposed fee on China-linked ships visiting U.S ports after a flood of negative feedback from industries that said the idea could be economically devastating By Dimitri Rhodes Nov 7 (Reuters) – Belgian oil tanker company CMB Tech says it will focus on the fast growing market in India as it reported third quarter results.. In a bold move amidst Russia’s intensified rocket attacks on civilian ships and Ukrainian ports Maersk has launched a new weekly container service into Ukraine signaling resilience in the country’s.. Subscribe to gCaptain Daily and stay informed with the latest global maritime and offshore news Stay informed with the latest maritime and offshore news For general inquiries and to contact us,please email: [email protected] To submit a story idea or contact our editors, please email: [email protected] For advertising opportunities contactEmail: [email protected]Phone: +1.805.704.2536 Essential news coupled with the finest maritime content sourced from across the globe Kayak’s 18th studio album ‘Out Of This World’ will be out on May 7th Legendary Dutch progressive rockers Kayak recently announced they will return with their 18th studio album ‘Out Of This World’ on May 7th Following the launch of the album’s first single in February they are now pleased to reveal the follow-up Ton Scherpenzeel comments: “Not the kind of song you’d expect from Kayak We get a little but disturbing peek inside the mind of someone who is overly obsessed by what he thinks is his love for another person.” The album is now also available for pre-order, arriving as a Limited CD Digipak, Gatefold 2LP + CD, and as Digital Album. An exclusive blood red vinyl edition is available directly from the band. Pre-order now here: https://kayak.lnk.to/OutOfThisWorld The full track-listing is as follows: spanning 70 minutes of energetic and incredibly diverse material- though still very much recognizable as Kayak It is clear that the new found energy has gained even more momentum since Hans Eijkenaar rejoined the group Although they were forced to cancel all tours because Ton Scherpenzeel suffered a heart attack at the end of 2019 and immediately after that because of the Covid pandemic The current line-up of the band is as follows: If you like what we do please support us on Ko-fi The Source for the Latest in Progressive Rock Interface announced this week it has reached a series of major sustainability milestones at its European manufacturing facility in Scherpenzeel As of January 2014 the plant has been operating with 100 percent renewable energy using virtually zero water in its manufacturing processes and has attained zero waste to landfill This is a key achievement for the facility and a significant step forward for the company as it strives towards Mission Zero — Interface’s pledge to eliminate any negative impact it has on the environment by 2020 and by doing so Interface announced this week it has reached a series of major sustainability milestones at its European manufacturing facility in Scherpenzeel, The Netherlands. As of January 2014 the plant has been operating with 100 percent renewable energy “2014 is a landmark year for us in many ways as we celebrate 20 years of Mission Zero and these figures from our European manufacturing operations show just how far we’ve come and what can be achieved with the right mind set and ambition," said Rob Boogaard while our achievements are to be celebrated this is certainly not a time to be complacent Mission Zero is not simply about reducing our own direct impacts — it’s also about taking full responsibility for the entire lifecycle of our products Our goal is to cut the umbilical cord to oil with the result that 44 percent of our raw materials in Europe are already recycled or bio-based We’ve come a long way but our Mission continues.” 2014 7pm EST / 4pm PST / 12am GMT / 1am CET Metrics details The V-ATPase is the main regulator of intra-organellar acidification Assembly of this complex has extensively been studied in yeast We identified 11 male patients with hemizygous missense mutations in ATP6AP1 encoding accessory protein Ac45 of the V-ATPase Homology detection at the level of sequence profiles indicated Ac45 as the long-sought human homologue of yeast V-ATPase assembly factor Voa1 restored V-ATPase-dependent growth in Voa1 mutant yeast Patients display an immunodeficiency phenotype associated with hypogammaglobulinemia hepatopathy and a spectrum of neurocognitive abnormalities Ac45 in human brain is present as the common Our work unmasks Ac45 as the functional ortholog of yeast V-ATPase assembly factor Voa1 and reveals a novel link of tissue-specific V-ATPase assembly with immunoglobulin production and cognitive function we describe a novel ATP6AP1-linked immunodeficiency and identified disease mutations in ATP6AP1 in 11 male patients with abnormal protein glycosylation Yeast V-ATPase assembly factor Voa1 was predicted to be homologous to Ac45 which was confirmed by functional complementation of Voa1 mutant yeast with the processed C-terminal domain of Ac45 Identification of different Ac45 protein isoforms in human brain liver and B cells indicated the presence of tissue-specific regulation of organelle acidification The dominating clinical symptoms displayed by the present cohort of patients with various ATP6AP1 mutations (Table 1) include hepatopathy and immune abnormalities Recurrent bacterial infections were associated with hypogammaglobulinemia ranging from plantar abscesses and gastrointestinal infections in family 1 to multiple episodes of childhood pneumonia and purulent otitis media in families 2–6 Several patients were successfully treated with intravenous immunoglobulins some of the patients responded very poorly to childhood vaccinations Hepatopathy ranged from mild hypertransaminasemia to cirrhosis and end-stage liver failure gastric problems were noted in the majority of patients and laboratory abnormalities included leukopenia The two brothers from family 4 displayed high-normal levels of IgD+/CD27− naïve B cells and lowered levels of IgD+/CD27+ intermediate and switched memory B cells suggesting a problem in B-cell differentiation Patients with the p.E346K substitution (families 3–5) showed in addition splenomegaly abnormal hepatic histology and neurological symptoms such as epilepsy mild intellectual disability and behaviour abnormalities Muscle weakness with mildly elevated serum creatine kinase (CK) was demonstrated in a few patients the presence of the p.E346K mutation predicted a more severe phenotype within the ATP6AP1 disease spectrum which is in accordance with the early death of two patients (3.2 and 5.1) due to liver failure 2 and 6 display a milder disease course and are currently doing well without cognitive impairment radiography showed no signs of osteopetrosis as observed in ATP6V0A3 deficiency There was no apparent renal phenotype with no signs of metabolic acidosis Hypokalemia was reported for patients 3.1 and 5.1 no signs for renal tubulopathy as observed in ATP6V0A4 deficiency were seen (a) A hepatocyte is surrounded by fat globules of variable size having the typical aspect of triglycerides (b) Hepatocyte showing relative translucency due to proliferated SER Arrows point to lipofuscin bodies (lysosomes) (c) Higher magnification showing an atypical lipofuscin body with a central reticulate content (black arrow) White arrow: section through a Golgi apparatus (d) A mitochondrion is engulfed in an autophagosome (arrow) cleaved-Ac45-KKNN function is comparable to that of Voa1 While Ac45 protein levels were lower than Voa1 levels for mutated Ac45 proteins were unchanged compared with non-mutated Ac45 protein reduced V-ATPase function observed for the Y313C or E346K mutation cannot be ascribed to decreased protein abundance Though GFP-tagging slightly reduced fitness of processed Ac45-KKNN on restrictive medium (data not shown) dilysine-dependent ER localization was verified these results indicate that the human and yeast proteins function about equally well in V0 V-ATPase assembly in the ER likely at least some of the symptoms are related to V-ATPase dysfunction which is supported by our studies on V-ATPase restricting growth conditions in yeast The question is why these systems are affected Possible explanations could include the tissue-specific processing of Ac45 or the existence of additional functions of Ac45 beyond pH regulation via its effect on the V-ATPase Our studies in human hepatocytes show that Ac45 in contrast to what was observed in neuroendocrine cells mostly localizes to the early secretory pathway This is in agreement with the presence of mostly intact-Ac45 carrying non-processed high-mannose glycans Further studies are needed to elucidate the mechanisms driving differential Ac45 glycosylation and processing in brain since tissue-specific forms of Ac45 could suggest a possible mechanism for the tissue-restricted disease symptoms in Ac45-deficient patients A glycosylation defect therefore may affect B-cell activation and thus antibody production or pH regulation are mainly affecting antibody production by a defective accessory subunit of the V-ATPase combined with its observed steady-state localization in the early secretory pathway (ER suggests that differential proteolytic processing might represent such a mechanism the identification of tissue-specific proteolytic processing of Ac45 and the availability of Voa1 mutant yeast as valuable model to further dissect the individual functions of Ac45 will facilitate future research to understand the functional roles and isoforms of Ac45 in the immune system muscle and brain and its relation to the V-ATPase in human Screening for abnormal protein glycosylation in plasma of patients with hepatopathy and immune dysfunction with or without neurological symptoms provides a rapid way to identify additional individuals with ATP6AP1 deficiency Genomic DNA was extracted from fibroblast pellets or white blood cells from 10 patients and available family members. Primers (Supplementary Table 4) were designed to amplify the 10 exons of ATP6AP1 (GenBank accession number NM_001183.4) including at least 50 bp of the flanking intronic regions Standard PCR reactions were based on 1 μl DNA and 0.2 μl Platinum Taq polymerase (Invitrogen) in a total volume of 25 μl Standard reaction conditions were 10 min at 95 °C The reaction was completed with a final elongation of 7 min at 72 °C For the sequencing of the resulting PCR product the BigDye Terminator Ready reaction cycle sequencing kit v.3.1 (Applied Biosystems) was used Analysis of the results was performed on an ABI3100 Avant (Applied Biosystems) Total RNA from different human adult and fetal tissues was ordered from Stratagene Europe (Amsterdam All fetal tissues are from 20- or 21-week-old embryos after gestation RNA was isolated using the NucleoSpin RNA II kit (Macherey-Nagel Germany) according to the manufacturer’s protocols the RNA was treated with DNase I (Invitrogen The Netherlands) while bound to the RNA binding column concentration and purity of the RNA were assessed using agarose gel electrophoresis and spectrometry 5 μg of total RNA was transcribed into cDNA by using the iScript cDNA synthesis kit (Bio-Rad Laboratories USA) according to the manufacturer’s protocol cDNA was purified by using the NucleoSpin extract II kit (Macherey-Nagel) according to the manufacturer’s protocol qPCR data were analysed by using comparative quantitation and the relative Q-values of the genes of interest calculated by equalizing the lowest Ct value to 1 The normalization factor for the reference genes was determined using the GeNORM program (medgen.ugent.be/genorm) and used to normalize the Q-values Individual experiments were performed in triplicate who gave written informed consent for scientific use of the buffy coats Peripheral blood mononuclear cells were isolated by density gradient centrifugation (Lymphoprep; Nycomed Pharma CD19+ B cells were positively selected using anti-CD19 magnetic microbeads (Miltenyi Biotec The isolated human donor B cells were lysed in cold lysis buffer (50 mM Hepes pH 7.4 0.1% deoxycholate supplemented with complete protease inhibitor mix (Roche Diagnostics)) to a final concentration of around nine million B cells per 20 μl lysisbuffer Tissue samples were powdered using a vessel and liquid nitrogen and subsequently lysed with cold lysis buffer to a final concentration of 100 mg tissue sample per 500 μl lysis buffer All samples were incubated on ice for 15 min and repeatedly shortly vortexed during the incubation the homogenates were centrifuged at 14,000 RCF for 10 min after which the supernatants were collected Subsequent PNGase F treatment was done as follows: supernatant containing 21 μg of protein was incubated for 4–6 h at 37 °C with 2.5 μl of 500,000 U ml−1 of PNGase F (New England Biolabs) in a final volume of 34 μl containing 50 mM sodium phosphate buffer pH 7.5 (G7 buffer from PNGase F kit New England Biolabs) and complete protease inhibitor mix (Roche Diagnostics) A second incubation was performed overnight at 37 °C after adding an additional 1 μl of PNGase F and the samples were boiled for 5 min at 99 °C Samples were separated on 10% SDS–PAGE (7 μg of protein per lane) and the proteins were transferred to a 0.2-μm polyvinylidene difluoride membrane After blocking in 5% milk in PBS+1% Tween-20 the membrane was incubated overnight at 4 °C with primary antibody rabbit anti-mouse Ac45 polyclonal #49 antiserum (directed towards A271-T283 and L443-I457 of mouse Ac45 Belgium) at a dilution of 1:5,000 in blocking buffer Goat-anti rabbit-HRP secondary antibody (Dako P0448) at a dilution of 1:5,000 in 2.5% milk in PBS-1% Tween-20 were used for ECL detection the membrane was incubated with mouse anti-GapdH monoclonal antibody (Ab8245-100 Abcam) at a dilution of 1:2,000 in 3% BSA in PBS/0.1% Tween-20 for 1 h at room temperature Incubation with the secondary antibody (Goat-anti-mouse-HRP Dako P0447) was for 1 h at room temperature at a dilution of 1:5,000 in 2.5% milk in PBS-1% Tween-20 Chemoluminescent signals was detected using ECL (Pierce) IHH55 were cultured in gelatin-coated culture flasks in Williams medium E supplemented with 10% FCS 0.022 U ml−1 insulin and 0.045 μg ml−1 dexametasone Culturing was done at 37 °C under an atmosphere of 5% CO2 IHH cell cultures were tested negative for mycoplasm IHH cells were transfected with a hsAc45/pcDNA3 construct using Lipofectamine LTX (Invitrogen) according to manufactures’ guidelines cells were cultured on gelatin-coated cover slips for 3 days and fixed for 1 h by 4% paraformaldehyde in PBS at room temperature After blocking of residual parafomaldehyde with 50 mM NH4Cl in PBS cells were permeabilized using 0.1% Triton-X100 in PBS (PBS-T) and incubated with anti-Ac45 antibody (1:1,000) and mouse-anti-EEA1 (BD Biosciences mouse-anti-GM130 (BD Transduction Laboratories 1:100) antibodies in 1% BSA in PBS-T (blocking buffer) for 12 h at 4 °C cells were incubated for 45 min at room temperature with secondary antibodies Goat-anti-rabbit-Alexa488 and Goat-anti-mouse-Alexa568 or Donkey-anti-rabbit-Alexa488 combined with Donkey-anti-goat-Alexa568 at a dilution of 1:200 in blocking buffer MilliQ water and dehydrated using methanol and subsequently mounted in Mowiol containing 2.5 μM DAPI Imaging was performed using an Olympus FV1000 confocal laser scanning microscope using a × 63 oil objective All images were captured with an aspect ratio of 1,200 × 1,200 using the FluoView version 4.1 software at a scanning speed of 12.5 μs per pixel Image analysis was performed using Fiji software Relative fluorescence intensities were calculated over a 10-μm cross section by setting the highest measured value to 100 IHH cells were seeded into gelatin-coated 12-well plates at a density of 2 × 105 cells per well Cells were washed using PBS and starved in starvation medium (DMEM without methione and cysteine 0.022 U ml−1 insulin and 0.045 μg ml−1 dexametasone) for 1 h cells were incubated for a 30-min pulse period in starvation medium containing 0.1 mCi ml−1 EasyTag EXPRESS 35S protein labelling Mix (PerkinElmer) For analysis of the N-glycosylation of Ac45 the 30-min pulse was carried out in the presence of tunicamycin cells were washed with PBS and lysed in lysis buffer lysates were incubated with anti-Ac45 antibodies (1:500) in lysis buffer supplemented with 0.8% SDS for 12 h at 4 °C Immune complexes were precipitated by Protein A-sepharose (GE Healthcare Life Sciences) and resolved on 10% SDS–PAGE EndoH (New England Biolabs) and PNGaseF (New England Biolabs) treatment was performed on immunoprecipitated Ac45 protein according to manufacturer’s guidelines Radiolabelled proteins were visualized by fluorography The tissue cylinder was immediately immersed in cold 2.5% glutaraldehyde in 0.1 M Cacodylate buffer (pH 7.4) for 2 h. post fixed in 1% osmium tetroxide for 1 h. dehydrated through ethanol series and embedded in epoxy resin Semi-thin sections (1 μm) stained with 1% toluidine blue were used for orientation cell identification and gross histopathological changes (that is Ultrathin sections (60 nm) were cut with diamond knives and either left unstained or stained with uranyl acetate and lead citrate Sections from three blocks were placed on 300-mesh copper grids and viewed and photographed with JEOL JEM 100SX and 100CX electron microscopes coding sequence for Voa1 signal peptide (M1–A24) and the first Voa1 amino acid after signal cleavage (D25) followed by a single HA tag or HA-GFP Sequence coding for C-terminal KKNN of Voa1 was added to Ac45 where indicated All pMR plasmids have 245 bp of VOA1 3′ UTR flanking sequence Inverse PCR with primers encoding K262Q and K263Q mutations and pMR092 template produced pMR1312 pMR1303 and pMR1304 were made from pMR1213 and pMR1214 by inserting GFP immediately after the HA tag E346K and M428I mutations were made in pMR1303 resulting in pMR1303E346K and pMR1303M428I Coding sequence region in all plasmid constructs was verified by DNA sequencing Liquid cultures of yeast strains were grown overnight at 30 °C in synthetic minimal medium plus dextrose (SD) supplemented to select for plasmid (-Ura) diluted to 0.4 OD600 per ml in rich medium (YEPD) buffered to pH 5 with 50 mM succinate/phosphate and grown until densities reached 1 OD per ml Cells were pelleted and suspended to 0.8 OD per ml in H2O This suspension and serial 1:8 dilutions were spotted onto YEPD pH 5 and restrictive medium (100 mM CaCl2 in pH-unadjusted YEPD or 60 mM CaCl2 in YEPD buffered to pH 7.5 with 50 mM HEPES) Growth was recorded after 46 h incubation at 30 °C Yeast membrane proteins were prepared from glass-bead lysates by centrifugation at 13,000g as described24 The pelleted membrane fraction was solubilized in sample buffer (8 M urea 0.01% bromophenol blue) to 100 OD600 equivalents per ml and 0.5 OD equivalents separated by SDS–PAGE Proteins were transferred to nitrocellulose membrane and analysed by western blot using a 1:300 dilution of monoclonal anti-HA primary antibody (BioLegend) and a 1:15,000 dilution of goat anti-mouse secondary antibody labelled with IRDye 800CW (LI-COR) Blocking of non-specific binding and incubation with antibody was done in 5% non-fat dry milk Wash steps after antibody incubation were in the same buffer but without milk a final wash was done in 10 mM Tris-HCl pH 7.5 The authors declare that all data supporting the findings of this study are available within the article and its Supplementary Information files ATP6AP1 deficiency causes immunodeficiency with hepatopathy cognitive impairment and abnormal protein glycosylation Regulation and isoform function of the V-ATPases A phenocopy of CAII deficiency: a novel genetic explanation for inherited infantile osteopetrosis with distal renal tubular acidosis Mutations in the gene encoding B1 subunit of H+-ATPase cause renal tubular acidosis with sensorineural deafness encoding a new kidney vacuolar proton pump 116-kD subunit cause recessive distal renal tubular acidosis with preserved hearing Defects in TCIRG1 subunit of the vacuolar proton pump are responsible for a subset of human autosomal recessive osteopetrosis Atp6i-deficient mice exhibit severe osteopetrosis due to loss of osteoclast-mediated extracellular acidification Impaired glycosylation and cutis laxa caused by mutations in the vesicular H+-ATPase subunit ATP6V0A2 Novel insights into V-ATPase functioning: distinct roles for its accessory subunits ATP6AP1/Ac45 and ATP6AP2/(pro) renin receptor Cytoplasmic terminus of vacuolar type proton pump accessory subunit Ac45 is required for proper interaction with V(0) domain subunits and efficient osteoclastic bone resorption ATP6S1 elicits potent humoral responses associated with immune-mediated tumor destruction Biosynthesis of the vacuolar H+-ATPase accessory subunit Ac45 in Xenopus pituitary A novel accessory subunit for vacuolar H(+)-ATPase from chromaffin granules V-ATPase-mediated granular acidification is regulated by the V-ATPase accessory subunit Ac45 in POMC-producing cells Accessory subunit Ac45 controls the V-ATPase in the regulated secretory pathway Versatile roles of V-ATPases accessory subunit Ac45 in osteoclast formation and function V-ATPase subunit ATP6AP1 (Ac45) regulates osteoclast differentiation and protease exocytosis in osteoclast-mediated bone resorption Identification of domains within the V-ATPase accessory subunit Ac45 involved in V-ATPase transport and Ca2+-dependent exocytosis Vma21p is a yeast membrane protein retained in the endoplasmic reticulum by a di-lysine motif and is required for the assembly of the vacuolar H(+)-ATPase complex Vma22p is a novel endoplasmic reticulum-associated protein required for assembly of the yeast vacuolar H(+)-ATPase complex VMA12 is essential for assembly of the vacuolar H(+)-ATPase subunits onto the vacuolar membrane in Saccharomyces cerevisiae VMA12 encodes a yeast endoplasmic reticulum protein required for vacuolar H+-ATPase assembly Role of Vma21p in assembly and transport of the yeast vacuolar ATPase Voa1p functions in V-ATPase assembly in the yeast endoplasmic reticulum Structural gene organization and evolutionary aspects of the V-ATPase accessory subunit Ac45 function and regulation of the vacuolar (H+)-ATPase Vma9p need not be associated with the yeast V-ATPase for fully-coupled proton pumping activity in vitro Gene identification in the congenital disorders of glycosylation type I by whole-exome sequencing VMA21 deficiency causes an autophagic myopathy by compromising V-ATPase activity and lysosomal acidification Role of furin in granular acidification in the endocrine pancreas: identification of the V-ATPase subunit Ac45 as a candidate substrate The fate of newly synthesized V-ATPase accessory subunit Ac45 in the secretory pathway Iterative orthology prediction uncovers new mitochondrial proteins and identifies C12orf62 as the human ortholog of COX14 a protein involved in the assembly of cytochrome c oxidase Rot1p of Saccharomyces cerevisiae is a putative membrane protein required for normal levels of the cell wall 1,6-beta-glucan HMMER web server: interactive sequence similarity searching Mapping the Arabidopsis organelle proteome ORFeome cloning and global analysis of protein localization in the fission yeast Schizosaccharomyces pombe and how of organelle acidification by the yeast vacuolar H+-ATPase A genome-wide enhancer screen implicates sphingolipid composition in vacuolar ATPase function in Saccharomyces cerevisiae CCDC115 deficiency causes a disorder of Golgi homeostasis with abnormal protein glycosylation TMEM199 deficiency is a disorder of Golgi homeostasis characterized by elevated aminotransferases and cholesterol and abnormal glycosylation V-type ATPase proton pump expression during enamel formation The vacuolar-type H(+)-ATPase at a glance - more than a proton pump RNASEK Is a V-ATPase-associated factor required for endocytosis and the replication of rhinovirus RILP regulates vacuolar ATPase through interaction with the V1G1 subunit Regulation of B cell differentiation by intracellular membrane-associated proteins and microRNAs: role in the antibody response process and present antigens: a crucial role for cell polarity Antigen presentation by B lymphocytes: how receptor signaling directs membrane trafficking Core fucosylation of IgG B cell receptor is required for antigen recognition and antibody production Transferrin and apolipoprotein C-III isofocusing are complementary in the diagnosis of N- and O-glycan biosynthesis defects Plasma N-glycan profiling by mass spectrometry for congenital disorders of glycosylation type II Diagnostic serum glycosylation profile in patients with intellectual disability as a result of MAN1B1 deficiency Next-generation sequencing of a 40 Mb linkage interval reveals TSPAN12 mutations in patients with familial exudative vitreoretinopathy Immortalized human hepatocytes as a tool for the study of hepatocytic (de-)differentiation Fast and accurate automatic structure prediction with HHpred in Molecular Cloning: A Laboratory Manual 3rd edn Cold Spring Harbor Laboratory Press (2001) Protein sorting in yeast: mutants defective in vacuole biogenesis mislocalize vacuolar proteins into the late secretory pathway A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae sequence- and ligation-independent PCR cloning method Mutations that suppress the thermosensitivity of green fluorescent protein Glycoproteomics analysis of human liver tissue by combination of multiple enzyme digestion and hydrazide chemistry Predicting transmembrane protein topology with a hidden Markov model: application to complete genomes Download references We dedicate this paper to the memory of Maciej Adamowicz who sadly passed away during the preparation of this manuscript Loes de Vries and Max van Essen for excellent technical assistance Virginie Steenveldt-Verkade and Özlem Seyrani (Translational Metabolic Laboratory The Netherlands) for culturing of patient fibroblasts Dr Arjan de Brouwer (Department of Human Genetics The Netherlands) for human tissue cDNA samples Belgium) for the anti-mouse Ac45 antiserum This work was financially supported by the Institute of Genetic and Metabolic Disease (IGMD) (grant to D.J.L. by grants from the Dutch Organisation for Scientific Research ZONMW-NWO (Medium Investment grant 40-00506-98-9001 and VIDI grant 91713359 to D.J.L.) and by National Institutes of Health grant GM38006 to T.H.S Sharita Timal and Margret Ryan: These authors contributed equally to this work Centre for Neuroscience and Radboud Institute for Molecular Life Sciences Radboud Institute for Molecular Life Sciences Radboud Institute for Molecular Life Sciences and Donders Centre for Neuroscience Department of Laboratory Medicine and Pathology Nijmegen Centre for Mitochondrial Disorders (NCMD) Temple Street Children’s University Hospital Personalized Genomic Medicine Pediatric Genetics and Metabolism Goryeb Children’s Hospital Department of Pediatrics and Adolescent Medicine Julia Vodopiutz & Susanne Greber-Platzer Centre for Molecular and Biomolecular Informatics designed and performed experiments and analysed data performed clinical studies and analysed clinical data All authors contributed to writing the paper The authors declare no competing financial interests Supplementary Tables 1-5 and Supplementary References (PDF 2265 kb) Download citation Anyone you share the following link with will be able to read this content: a shareable link is not currently available for this article Molecular and Cellular Biochemistry (2024) Sign up for the Nature Briefing newsletter — what matters in science Ensuring a smooth and comfortable experience for passengers has long been the key for major airports looking to differentiate themselves Ross Davies talks to Marianne van Scherpenzeel of Amsterdam Airport Schiphol – long considered a front runner in the field – to learn more about the initiatives it has in place to slake the demands of the modern traveller It might be the length of the queue at the check-in desk the number of boutiques or even the softness of the seat in the departure lounge but the gamut of metrics that constitute passenger satisfaction at the world’s busiest airports is steadily becoming more complex The days when an airport simply represented a transient transportation hub are long gone By dint of the burgeoning airport city concept – identified by TIME magazine in 2011 as one of the "ten ideas that will change the world" – today’s terminals are just as likely to house offices hotels and high-end restaurants as they are standard currency exchange and car rental desks Today’s customer-service-savvy passengers are also a lot more exacting when it comes to expectations of what a desirable airport experience should entail While a swift check-in and shorter wait at the baggage carousel remain paramount comfort and leisure facilities are arguably just as highly valued by passengers particularly as a means of offsetting the potential itchy feet and lengthy waiting times synonymous with air travel with a turnover of approximately 50 million passengers each year Amsterdam Airport Schiphol has long been considered something of a pioneer: it is set on shifting the image of a modern airport away from a necessary evil and towards a situation in which passengers are actually happy to spend prolonged periods of time it has been declared Europe’s best airport at both the Skytrax World Airport Awards and ACI Europe Best Airport Awards Manager policy and projects Marianne van Scherpenzeel argues that these accolades have come about as a result of ongoing efforts to improve passenger processes and experience such as the revamp of one of its departure lounges and the roll-out of self-service technology in its check-in areas "We have done several things over the latest period," she says "One of those was to update lounge three The addition of new shops and refurbishments has completely transformed the waiting experience We have also installed self-service drop-off points for several airlines so passengers can check in their baggage independently." Don’t let policy changes catch you off guard Stay proactive with real-time data and expert analysis Self-service at airports is a trend that has gathered speed in recent years the usage of which has grown steadily in recent years due to their manifest popularity with passengers particularly to the baggage drop-off machines," she claims "As well as adding speed and mitigating waiting times the advantage of self-service is that passengers can remain in control of their own baggage processes Schiphol has long valued technology as an indispensable asset in facilitating various passenger procedures it became one of the first airports in the world to introduce millimetre-wave security scanners – eschewing arduous metal detection gates – while it has also made use of iris scan technology at its border passage since 2001 "It’s really important to pursue innovation especially if it helps us to add more speed comfort and control to passenger processes such as checking in and baggage drop-off," says van Scherpenzeel "We are still looking to develop even more self-service processes as well as improving the ‘no-queue’ concept which is an automated border control passage developed in collaboration with the government We also have a mobile app through which passengers can check their flight information." airports such as Schiphol have also ramped up their interaction with passengers in order to gauge overall satisfaction levels the airport is appreciative of the rising influence of social media as a forum where passengers propagate opinion and word of mouth – both positive and unfavourable "We still conduct a bi-monthly survey using a broad passenger questionnaire on a number of quality items in order to gauge their overall perception of the airport," explains van Scherpenzeel "But we also monitor Twitter and Facebook to see what is being said about us and how we can improve our quality." Schiphol will undoubtedly have been buoyed by the aforementioned accolades that have been bestowed upon it this year – "it’s an indication that we are on the right track and motivates us to continue making passengers feel welcome," confirms van Scherpenzeel A major contributing factor to the airport’s passenger experience offering remains its impressive folio of amenities – in 2011 it also opened a 200m² park which serves as a waiting area and is complemented by a connecting rooftop terrace "The area represents somewhere nice where passengers can relax during their stay especially if they have a longer transfer time between flights," says van Scherpenzeel "Adding more comfort is certainly a major driver for us." tourist-friendly facilities – it also has a library and a Dutch-themed museum – make it a stand-alone airport in many ways they clearly cannot come at the expense of efficiency while essential to the smooth running of operations Airports are still defined and represented by the staff that work across their array of public service areas training is required to bolster both hard and soft skills it is really dependent on the situation," she says a security agent will receive a different kind of training from one of our floor managers as it’s a different kind of job hospitality remains the central focus for our staff a number of masterclasses are provided for them." Despite being the fourth-busiest airport in Europe Schiphol currently has the highest peak-hour capacity on the continent owing to its status as a major transit hub And with passenger volumes expected to increase in the future there will be an onus on the airport to maintain the high levels of satisfaction to which passengers have become accustomed will necessitate a creative and far-sighted approach "We will try and stay as innovative as possible," she says "This will require us to keep abreast of what passengers want and continually improve the overall process we hope to remain Europe’s preferred airport." Given the developments over the last year – recognised by passengers and the industry alike – this goal would appear to be more than achievable Give your business an edge with our leading industry insights View all newsletters from across the GlobalData Media network We focus on the greatest sustainability challenges across all sectors Offering unparalleled depth and breadth of expertise In this follow up to our most recent edition of the SustainAbility Institute by ERM's annual Sustainability Trends Report we explore the sustainability issues and trends shaping the business agenda in 2022 from the perspective of the technology sector The supplement focuses on the five environmental and governance (ESG) related trends we anticipate will most impact the industry: The ESG risks and opportunities facing the tech sector are striking While for many sectors COVID-19 has posed significant challenges for technology it has also opened numerous new opportunities the sector has experienced significant growth as people and businesses were forced to go online and rapidly embrace digital solutions to continue economic and welfare activities when lockdowns were introduced across the world Pandemic-initiated growth is likely to continue as remote working and many other uses of technology to connect people in more innovative ways are likely to stick long term The pervasiveness and ubiquitous nature of technology also means it faces regulatory and civil society scrutiny in areas like data ethics Edelman’s 2021 Trust Barometer shows that public trust in the sector is waning; in fact while trust in business overall is growing trust in the technology sector has consistently declined over the past decade.2 The sector has high-profile environmental challenges as well the sustainable sourcing of critical minerals The technology sector has great potential to help society and other industries manage the transition to a sustainable economy especially considering the sector’s exponential growth Tech companies already provide technological solutions to other sectors to support their transitions to sustainable operations If the sector can build and maintain public trust regarding data privacy and digital innovation – so that more digitally enabled solutions can be embraced in everyday lives – then the low-carbon transition can be a digitally enabled one Learn more about these five sustainability trends and what companies might expect to result from the trend as the decade advances Download the report supplement 2022 edition of our annual report in which we explore the sustainability issues and trends that will shape the business agenda in 2022 This report a tool for businesses to use on the road to circularity built on ERM’s expertise and experience working with clients over five decades This blog explores how businesses are creating and re-creating additional new normals in response to new information and shifting expectations related to each of our 10 sustainability trends for 2021 This website is using a security service to protect itself from online attacks The action you just performed triggered the security solution There are several actions that could trigger this block including submitting a certain word or phrase You can email the site owner to let them know you were blocked Please include what you were doing when this page came up and the Cloudflare Ray ID found at the bottom of this page Interview with Dick Wittenberg (Dutch journalist and writer) about his career and ambitions Dick Wittenberg (1953) is a Dutch journalist and writer His ambition to write started when he was about seven years old and the urge never stopped after working for 30 years as a journalist for several newspapers he decided to focus more on writing non-fiction books This resulted in three books: Prikkeldraad (Barbed wire) 2015 Het gezicht van armoede (The face of poverty) 2015 The last book was crowned with the Bob den Uyl prize and the Dirk Scherpenzeel prize He now works as correspondent ‘worldsavers’ at the online journalistic platform The Correspondent It was early in life that Wittenberg knew he wanted to become a writer Although his dad passed away when he was still young Wittenberg remembers sitting beside him in the living room watching him write From that moment on he wanted to become a writer too As his dad was an accountant it seems more likely that he was noting numbers instead of writing stories but these domestic scenes resulted in Wittenbergs desire to become a writer In the assumption that writers and journalists were the same thing he left for Utrecht to study journalism The educational innovations of the seventies that implied democratization of education did not work well for Wittenberg the project based subjects and excessive freedom did not give him the descent foundation he would have liked What did help to gain writing skills was his range of correspondence friends that made him write six to seven letters a day But even though his study didn’t provide him with a good set of skills he never worried about getting a job He just wanted to get started and learn the profession by experience He returned to Eindhoven and applied for the first job he could find from there on he applied for other jobs Wittenberg ended up at the Institute of social research for Surinamese Maybe this is where his affection with minorities started but because of an incident with corrupt management this job ended A while after the resignation he started at NRC Handelsblad as a correspondent Eindhoven It seems characteristic for the unruly attitude of Wittenberg that he moved to the sports department when he wanted something else In the time the department was highly unpopular and conservative Because he wasn’t interested in just reporting games he shone a new light on the department by publishing background stories of sportsmen and analysing sports behaviour instead of just reporting a match Due to the NRC Handelsblad he also ended up as a reporter in London In the contrary of his preference to be situated in a place in the shadow he was now staged in the spotlights This pressure weighed heavy on him; besides long working hours and a baby at home he felt the responsibility towards his paper because of the enormous financial outlay for his correspondence.  With unstable politics and the death of princess Dianna it were tempestuous times for England and consequently also for Wittenberg Publishing everyday worked addictive and ego flattering but eventually also caused a burnout ‘Publishing everyday worked addictive and ego flattering but eventually also caused a burnout.’ When he returned to the Netherlands he could start as an editor on the foreign department focussing mainly on Africa Although he never thought of it himself, it appeared to be spot on In this position he could fly to Africa four times a year write background stories while not being obligated to bring the daily news. Obviously he mainly visited countries nobody was originally interested in, and reported stories that would otherwise not be told But the times are over that newspapers had the financial resources to fly reporters all over the world several times a year. As we come to talk about contemporary journalism and how digitalization influenced journalism does Wittenberg make clear that he doesn’t see a substantial difference between pressed or digital journalism But he does observe an increasing level of snack-content in articles I'm not sure if he is joking when he talks about some contemporary journalist as ‘modern vampires’ By this term he means journalists who are reporting gossip and celebrity news instead of covering the role of a trustable guide The ideas for his books were born during his last years of employment at the NRC Because of a shrinking budget there was no money available for thorough research and long background stories But Wittenberg stayed occupied with his ideas and decided to quit his job and become a freelancer With some rough ideas for his books he found a publisher and raised the necessary funds This way he created a particular certainty for himself and enabled himself to become a book writer after all Simultaneously Rob Wijnberg started the online platform The Correspondent and Wittenberg decided to join on a freelance basis.  The research for his books turned out to be a fantastic experience for him In the contrary to writing for a newspaper he experienced the freedom to follow side-tracks and didn’t feel the pressure of tight deadlines For his book Prikkeldraad he visited the place in America where barbed wire was invented and ended up in an elderly home that was funded by money made in the barbed wire industry This home fascinated him so much that he stayed there for almost his whole stay His eyes twinkle when we talk about his ‘barbed wire journey’ Wittenberg wants to create decent and enjoyable pieces of writing and by doing so he wants to contribute to society  Later it becomes clear that twinkling eyes are the highest profit you get from publishing non-fictional books about subjects most people walk past. His previous books were on the Malawian village Dickisoni that Wittenberg follows for over 10 years The daily poverty that is normal to half of the world’s population The connection between this village and Wittenberg is strong and he tries to return every year  He sais that it keeps him grounded and reminds him to all that is good in life It’s one of the greatest privileges of his career that he was able to travel so much for his work and is accepted in a culture that is so different from our own When I ask about the origin of his engagement to minority groups and subjects Wittenberg refers to his youth Because of the choices and support of his mom he was able to escape this milieu but never forgot where he grew up He always stayed connected to minorities in the shadow of society But this is not the goal of his writership Wittenberg wants to create decent and enjoyable pieces of writing and by doing so he wants to contribute to society  Additionally he emphasizes that he doesn’t want to write about celebrities and superficial media events He prefers to write background stories that the reader doesn’t read every day and in doing so enlighten people and events that are usually left aside In his writing Wittenberg doesn’t necessarily looks out for stories; he comes across subjects that interest him and keeps an archive (A small part of this archive is displayed on the title photo.) He later states that he doesn’t see himself as a journalist but more as an archivist Now he becomes older he also likes to share his knowledge with young journalists and likes to mentor writers with ambition.  After his books he decided to put his writing on the back burner but if it’s up to him he keeps writing and collecting stories till the end Our newsletter arrives every Monday with fresh reviews Formed in the Nethrlands back in 1972 by keyboard player Ton Scherpenzeel progressive rock band Kayak have run a course of highs an dlows over a career that spans 45 years After a three year break they’re back with an almost complete line up and their 17th album – Seventeen We had the chance to chat with founder  the and one constant of the band LOUDER THAN WAR: First question from me is that having been active band for 45 years I’m ashamed to say that the new album is the first of yours I’ve heard How come you’ve managed to steer under my radar for so long LTW: It’s been quite a journey with all manner of ups and downs but can we focus on a couple of stops on the journey  Signing with a major label like EMI in the early days must have been a massive boost when you’re twenty and a beginner in this business it is all just as exciting as it is ‘normal’ to get a major record company like that interested You have no idea what you’re up against the sacrifices that have to be made and the chance of really becoming a success with something you started because it was your passion That’s great when you’re twenty and naive as hell and you sort of expect the world to like you LTW: What was behind the reunion  in 2000 after the split in 1992 Or was it a ‘split’ or just a break… TS: We always made the joke about ‘we have waited till the kids grew up and moved out’ but really we did not think about doing anything with Kayak for over fifteen years since 1981 Until Pim and I found each other again in a creative way We wouldn’t have used the name while he was still alive- not that he had any rights but he was just crazy enough not to accept Kayak going on without him Still it took a couple of years before it actually happened as no major record company was really interested to begin with you must have seen a few changes in the music industry – which do you see as the biggest changes and what are the challenges in making music in 2018 TS: In all sorts of ways I would say the transition from analog to digital made the biggest difference but on the other hand it is much cheaper to make an album It is difficult to make some money in the business while on the other hand the number of acts bands and artist that all want a piece of the cake has grown immensely In the past you would hear or read the odd review reviews from all over the world are pouring in One fart in Holland and they smell it in Australia The big challenge I would say is to stand out amidst that all LTW: You’ve recorded a real variety of music  from rock operas to female vocalists which area do you think you work best and think I have the ability to write both shorter songs as well as longer epics or music that belongs in a certain framework and story like rock operas the theatre-like stuff as well as intimate ballads or out-of-the-box music The problem with rock operas is usually about the money if you want to do it properly LTW: You seem to have been dogged with all sorts of struggles deaths and departures but resolutely carried on – how do you feel these issues have affected the band TS: Sure how could it not affect you or a band Ultimately it lead to a band that has me as the sole survivor Life is all about changes and Kayak is no exception LTW: And although you have these guys in the band so what sort of input do they have when it comes to recording LTW: About Andy Latimer’s contribution to Ripples On The Water Did you have him in mind to actually play on that track from the beginning I knew he would have to play it And I am very fortunate that he wanted to do it recognizable like only the real great players are He plays one note and you instantly know it is him We have kept in touch throughout the years For some mysterious reason it never happened LTW: How did the association with Inside Out come about I know Thomas Waber has his finger on the prog pulse…. TS: We already offered Cleopatra to InsideOut Arjen Lucassen gave me the tip to contact Thomas But they weren’t really into this sort of thing maybe it was too much musical-like for their taste When our manager Rob Palmen sent them the demos for Seventeen it is much more of a rock band now compared to the rock opera idea of Cleopatra I guess you’re always enthused about new material but how do you think the new album sits with in the Kayak catalogue it is too fresh still and I could not say where it fits in People say it’s the band going back to the seventies LTW: And finally – Record World in the USA made you ‘Most Promising Band of the Year’ in 1977…forty years on If the promise was that we’d conquer the world (or the US at least) and sell many albums If the promise was to stay true to ourselves Often things went wrong at the wrong moment If your singer decides he does not want to sing while a US tour is in the making If two of your singers decide to leave while you’ve got a rock opera written with them in mind co-producer and co-writer dies in the middle of a tour I have always been able to make a living out of my music outside Kayak The Kayak website Interview by Mike Ainscoe. You can find more of Mike’s writing on Louder Than War at his author’s archive. He can be found on Facebook and his website is www.michaelainscoephotography.co.uk Louder Than War is run by a small but dedicated independent team and we rely on the small amount of money we generate to keep the site running smoothly Any money we do get is not lining the pockets of oligarchs or mad-cap billionaires dictating what our journalists are allowed to think and write and we want to continue bringing you news on the most interesting releases the latest gigs and anything else that tickles our fancy just pure enthusiasm for a scene that each and every one of us is passionate about We do not charge artists for the exposure we give them and to many Subscribing to one of our paid tiers takes just a minute helping to keep the flame of independent music burning PLEASE SUBSCRIBE TO LTW and website in this browser for the next time I comment Δdocument.getElementById( "ak_js_1" ).setAttribute( "value" This site uses Akismet to reduce spam. Learn how your comment data is processed. Following the conclusion of the José Ignacio Domecq Cup and then the Barón de Strabrock Cup the third and final cup of the Iberian Polo Tour de Primavera (Spring) will begin this afternoon The XXI Jerez Polo Cup will be held on the fields of Santa Maria Polo Club Six teams will compete in this cup with a handicap of up to eight goals with each organization fielding two qualifying teams The top two will then advance directly to the final (held on Sunday while the remaining teams will fill out the two subsidiary cups It should be noted that the Quinto Elemento team won the previous two tournaments and will seek to secure a perfect performance a member of Camel and the founder of prog/jazz band Nathan Mahl has succumbed to kidney cancer at the age of 54 LeBlanc's passing was confirmed by the official Camel website "It is with a hugely heavy heart that we announce the gentle passing of our friend and keyboard player We are unable to adequately express the sorrow we are experiencing at this time Joining the band in 2000 as a replacement for departing keyboardist Ton Scherpenzeel LeBlanc toured with Camel and was featured on their 2002 release A Nod and a Wink He was forced to take a leave of absence prior to the group's 2003 farewell tour but was back in the lineup for their Retirement Sucks tour in 2013 and appeared on 2014's live DVD In From the Cold Camel's renewed activity came as LeBlanc was battling the cancer that would ultimately claim his life and he was forced to sit out the band's 2014 live dates manage to complete work on a new Nathan Mahl album According to LeBlanc's Facebook page he passed "peacefully at home in Kemptville with his wife and daughters," leaving behind "a legacy of beautiful and inspiring music – music that tells a story and each story exposes a search within for the answers to life and art See Other Rockers We've Lost This Year Rock and Roll Hall of Fame's Worst Snubs Keyboard player Guy LeBlanc, a member of Camel and the founder of prog/jazz band Nathan Mahl, has succumbed to kidney cancer at the age of 54.\nRead More LeBlanc's passing was confirmed by the official Camel website According to LeBlanc's Facebook page In 2021 Kayak released their 18th album Out Of This World they didn't realise it would be their final farewell There haven’t been too many reasons to be cheerful for musicians in the 2020s so far. But Ton Scherpenzeel, founder member of Dutch prog mainstays Kayak That’s not the result of enforced isolation Rather it’s an upbeat attitude that’s been reinforced through the relentlessly turbulent history of a band that have released their 18th studio album and are approaching 50 years since their original formation in 1972 The most recent tribulation to confront them could have spelled the end not just of Kayak but of their driving force Keyboardist and songwriter Scherpenzeel suffered a heart attack in 2019 which forced him to re-evaluate his lifestyle as well as his attitude to his art.  “It made me realise how precious life is,” he tells Prog fair-haired 68-year-old looks sprightly enough on the other end of a Zoom call I’m not going to make another 20 albums So now I’m approaching every album as if it’s the last I’m as dedicated to this one as I was with the first one I’ve learned not to trust my body anymore I’m not an excessive drinker or smoker So I’m walking every day for an hour I like playing piano and normally that’s enough movement But my attitude has changed because I still want to go on for a little while yet…”  It’s arguably the best they’ve made since Scherpenzeel reformed the band at the turn of the millennium after their initial 1982 split And although the current line-up sees no fewer than four members take the lead mic at certain points Kayak have long since moved beyond being a band of one voice is a group whose lead singer has regularly switched amid personnel changes resulting in no fewer than 19 former members being listed on their Wikipedia page Kayak forged a role at the forefront of the Dutch prog scene in the early 70s after teenage friends Scherpenzeel and drummer Pim Koopman put a band together with Max Werner on vocals Johan Slager on guitar and Cees van Leeuwen on bass.  Koopman left in 1976 suffering from respiratory problems their sound had evolved into a noticeably more radio-friendly AOR style always a reluctant singer and troubled by stage fright despite his striking tenor voice pulled a rare move known in the trade as “a reverse Phil Collins” by switching from lead vocals back to the drum kit that was his first love (Werner would return to the mic briefly as part of the 1999 reunion before having to leave after the first tour due to health problems).  That left centre-stage open for Kayak fan Edward Reekers to join the band and for casual observers he’s probably the most recognisable face by virtue of arriving just in time to front Kayak’s biggest hit The group split in 1982, and Camel fans may remember Scherpenzeel’s subsequent tenure alongside Andrew Latimer and co in that band but after a one-off live show for a Dutch TV show in 1999 Kayak decided to reform with former Vandenberg man Bert Heerink replacing the ailing Werner up front a few months later only for Reekers to rejoin again on vocals in 2005.  who injected a new dimension into the Kayak sound as an additional lead singer from 2003.  This second coming of Kayak saw them increasingly explore their progressive roots with Scherpenzeel’s classically influenced keyboards and fascination for myths and legends returning to the fore on albums such as 2003’s Merlin – Bard Of The Unseen (a more successful proggier reworking of 1981’s Merlin album with new tracks added) and the multivoice rock opera Nostradamus: The Fate Of Man in 2005 Readers should expect the unexpected in this band co-founder and key songwriter Pim Koopman died suddenly of a heart attack it was really up to me whether the band would continue or not,” says Scherpenzeel “because we started the band together And in the end what kept me going – still keeps me going – was the endless drive to create It’s not like I want a band because it’s so cosy and nice playing with people It’s a drive to create something that wasn’t there before.”  crowdfunded rock opera Cleopatra – The Crown Of Isis suggested the post-Koopman Kayak still had plenty of creative potential Scherpenzeel was stunned to learn that both the band’s singers we had troubles in the band,” he says “It took me two years to get myself together and convince myself we should continue He ended up recruiting a whole new line-up in 2017, who would make the following year’s Seventeen album the highest placing they’d achieved since 1980 The only way was surely up… then came not only the aforementioned heart attack but a global pandemic sweeping touring plans off the table and giving him further pause for reflection with actions and consequences that touch us all.” he’d already bounced back from his near-death experience and written much of the new album including the title track of Out Of This World a storming prog tour-de-force geared around a brilliantly baroque quasi-classical synth motif Its lyrics would soon take on extra layers of resonance as much of the planet locked itself down.  ‘Why don’t we get out of this world and fly to a place we could live in?’ Kayak’s (relatively) new frontman Bart Schwertmann sings It speaks of a wanderlust and the search of relief from a screwed-up planet “It’s a theme that’s been in the back of my mind for a long time,” Scherpenzeel says "In fact it actually quotes a song of ours from 1975 ‘Remember Noah wrought his ship a long long time ago?’ It’s about a man who takes off in a spaceship with a selected few but it’s the same idea with this new song the desire to escape the world’s problems Corona hits and it seems to fit even better to the current times…” Elsewhere on the new album the styles diversify considerably: the short is funk-inflected soft rock that has a distinctly 80s feel Cary is irresistible accordion-led folk-pop and Kaja is a transcendent instrumental passage that’s a sublime showcase for the emotive melodic strokes of new guitarist Marcel Singor the rich banks of harmonies characterising Mystery recall Asia or Yes at their most accessible the irresistibly bombastic Under A Scar feels like an emotional high point of a rock opera yet to be written slow-burning nine-minute fable A Writer’s Tale is imbued with bewitching pomp rock melodrama and Critical Mass adopts an breathless time signature to hopscotch around another beguiling Scherpenzeel piano riff and gnarly what strikes the listener right away is the immediacy of the songs – with hooks riffs and instrumental flourishes demanding attention at the first time of asking.  “It’s natural to me,” says Scherpenzeel I have to grab the listener quickly.’ I’m always trying to make every song stand out But those come out naturally in the way I write.” Another stand-out upbeat track is Traitor’s Gate a gutsy rocker geared around a stabbing central riff and a theme of romantic treachery Given that it’s preceded by a bittersweet ballad are we to deduce that Scherpenzeel has had to deal with heartbreak as well as a heart condition?  thankfully I’ve been married 45 years this year so it’s not really personal,” he smiles it all stemmed from coming up with that title where I noticed the Traitor’s Gate sign I had the title and then I put myself into the role of the person in the song.” Traitor’s Gate was co-written and sung by top session drummer Hans Eijkenaar (who departed with the Reekers/Oudshoorn-led line-up and later rejoined) Singor and Swedish former Pain Of Salvation bassist Kristoffer Gildenlöw Scherpenzeel now feels galvanised by his band.  “I’m lucky to have such fantastic players and singers around me,” he says “I’m so fortunate to know they are willing to work weeks and months on this album and do their thing but then you have to find the right people who want to play it.” “It was inspired by losing David Bowie then every year it gets more as everybody’s ageing Later in the song I wanted to make the point that it’s gonna happen to us all it doesn’t matter if you’re famous or not We can’t expect our heroes to be any different Assuming Scherpenzeel’s conservative estimate is right and he’s not likely to make another 20 albums before he joins the great rock’n’roll hall of fame in the sky we can only hope he sticks around long enough to make a few more records as good as Out Of This World he might have the same band alongside him too But maybe that’s asking a little too much.  This article originally appeared in issue 120 of Prog Magazine Johnny is a highly experienced and versatile music writer whose tastes range from prog and hard rock to R’n’B He has written about music professionally for 30 years surviving the Britpop wars at the NME in the 90s (under the hard-to-shake teenage nickname Johnny Cigarettes) before branching out to newspapers such as The Guardian and The Independent and magazines such as Uncut “A handy surrogate for anyone craving the feel of Yes’ glory days… and how did they beat Jon Anderson to the title?” The Flower Kings’ Love is a fully-achieved ambition “I’m super-proud of everything Rush accomplished I’m excited to be one of the folks in Envy Of None”: Alex Lifeson loves working alone in his home studio – and he’s even brought the guitar solos back Metrics details Wnt-induced β-catenin-mediated transcription is a driving force for stem cell self-renewal during adult tissue homeostasis Enhanced Wnt receptor expression due to mutational inactivation of the ubiquitin ligases RNF43/ZNRF3 recently emerged as a leading cause for cancer development targeting canonical Wnt receptors such as LRP5/6 holds great promise for treatment of such cancer subsets we employ CIS display technology to identify single-domain antibody fragments (VHH) that bind the LRP6 P3E3P4E4 region with nanomolar affinity and strongly inhibit Wnt3/3a-induced β-catenin-mediated transcription in cells Structural analysis reveal that individual VHHs variably employ divergent antigen-binding regions to bind a similar surface in the third β-propeller of LRP5/6 sterically interfering with Wnt3/3a binding anti-LRP5/6 VHHs block the growth of Wnt-hypersensitive Rnf43/Znrf3-mutant intestinal organoids through stem cell exhaustion and collective terminal differentiation VHH-mediated targeting of LRP5/6 provides a promising differentiation-inducing strategy for treatment of Wnt-hypersensitive tumors These Wnt-enhancing properties complicate the application of LRP6-targeting mAbs in vivo we selected three highly potent VHHs that bind LRP6 with nanomolar affinity and efficiently block Wnt3/3a-dependent β-catenin signaling Structural analysis revealed that these VHHs all bind a surface of the third propeller domain of LRP6 that is likely involved in Wnt3 binding treatment with anti-LRP6 VHHs induces strong growth inhibition of Wnt-hypersensitive intestinal organoids by driving collective terminal differentiation we identify a highly potent set of VHHs that target Wnt-hypersensitive tumors VHHs targeting LRP6P3E3P4E4 block cellular responses to Wnt3a The P3E3P4E4 module of the extracellular domain was used to generate anti-LRP6 VHHs LRP6P3E3; blue/orange and LRP6P4E4; green/orange b Wnt luciferase reporter assay performed in LRP6-overexpressing HEK293T cells stimulated with Wnt3a-conditioned medium and treated with 10 μM of the indicated anti-LRP6P3E3P4E4 VHHs c Wnt luciferase reporter assay performed in HEK293T cells stimulated with Wnt3a-conditioned medium and treated with 10 μM of the indicated anti-LRP6P3E3P4E4 VHHs Graphs show average (bars) and range (dots) of luciferase activity in duplicate cell cultures transfected in parallel Anti-LRP6P3E3P4E4 VHHs show nM affinities b Luciferase activities normalized to unstimulated control are represented as a heat map Anti-LRP6P3E3P4E4 VHHs cause a dose-dependent decrease of Wnt signaling in both HEK293T cells overexpressing LRP6 (a) or untransfected HEK293T cells (b) c Binding affinities of anti-LRP6P3E3P4E4 VHHs as revealed by ITC measurements One representative experimental titration curve of each VHH binding to LRP6P3E3P4E4 is shown Structural analysis of L-P2-B10 and L-P2-D07 anti-LRP6 binding mode a Alignment of the LRP6P3E3P4E4 complexes with L-P2-B10 and LRP6–L-P2-D07 in two orthogonal views Cartoon trace with transparent surface representation of LRP6P3E3 (blue) b Interface of LRP6–L-P2-B10 with on the left-hand side LRP6P3 (gray) and CDR2 and CDR3 loops of L-P2-B10 (green and red Interacting side chains are shown as sticks surface presentation of L-P2-B10 (orange) with interacting residues of LRP6P3 shown in sticks; numbers represent the six blades of β-propeller 3 with in addition the CDR1 loop in blue (left) and the surface of L-P2-D07 in red (right) d Binding-surface areas of L-P2-B10 (orange) L-P2-D07 (red) and DKK-C31 (magenta) on LRP6P3E3P4E4 (gray) the VHHs act as a competitive inhibitor for Wnt3 ligands by sterically blocking access of these ligands to LRP6P3 Characterization of anti-LRP5/6 VHHs with highest potency a IC50 calculations of inhibition of cellular responses to Wnt3a by titration of the indicated anti-LRP6P3 VHHs in a luciferase reporter assay b IC50 calculations by combination of Wnt3a and Rspo1 c Activation of Wnt signaling mediated by CHIR-99021 is not affected by anti-LRP6P3 VHHs Wnt luciferase reporter assay performed in HEK293T cells treated with 5 µM CHIR-99021 and 10 μM of the indicated anti-LRP6P3 VHHs Graph shows average (bars) and range (dots) of luciferase activity in duplicate cell cultures transfected in parallel d Wnt luciferase reporter assay performed in LRP6-/- HEK293T cells Wnt pathway activation achieved upon LRP5 overexpression is counteracted by anti-LRP5/6P3 VHHs e Western blot showing inhibition of Wnt3a-induced LRP6 phosphorylation (pLRP6) by the different anti-LRP5/6P3 VHHs f anti-LRP5/6P3 VHHs do not inhibit Wnt1-mediated cellular responses Wnt luciferase reporter assay in HEK293T cells overexpressing Wnt1 and treated with 10 μM of the indicated anti-LRP5/6P3 VHHs g Top panels: Schematic representation of the growth conditions associated phenotypes and experimental set up Bottom panels: anti-LRP5/6P3 VHHs induce death of Wnt3a-treated wild type (WT) intestinal organoids Organoids were cultured in WENR and treated with 10 μM of the indicated anti-LRP5/6P3 VHHs for 4 d (Scale bar Red asterisks indicate cell death; green arrows indicate organoids showing villi and crypts; blue arrows indicate cystic organoids Anti-LRP5/6 VHHs drive collective differentiation of R/Z-mutant tumor organoids a Anti-LRP5/6P3 VHHs block tumorigenic R/Z mutant organoid growth Organoids were cultured in EN and treated with 10 μM of the indicated anti-LRP5/6P3 VHH for 4 d (Scale bar Red asterisks indicate cell death; green arrows indicate organoids showing villi and crypts; orange arrows indicate organoids showing a mixed phenotype of cell death and villi crypts structures b Anti-LRP5/6P3 VHHs strongly diminish cell viability of tumorigenic R/Z mutant organoids Organoids were cultured in EN and treated with 10 μM of the indicated anti-LRP5/6P3 VHHs for 4 d Graph represents relative cell viability normalized to PBS treatment c Schematic representation of the cell types present in the intestine and the role of Wnt signaling in maintaining intestinal physiology d qRT-PCR showing anti-LRP5/6P3 VHH treatment (3 d) strongly inhibits the expression of Wnt target genes stem cell-associated genes and Paneth cell markers while inducing a transcriptional program for differentiation in tumorigenic R/Z mutant organoids Graph represents fold change in gene expression as compared to PBS treatment ( = 1) e Anti-LRP5/6P3 VHH treatment (3 d) attenuates proliferation as shown by a decrease in Ki67 staining (left) Anti-LRP5/6P3 VHH treatment decreases the number of Paneth cells (Lys) while increasing the number of differentiated enteroendocrine cells (ChgA) (right) Phalloidin staining was used to mark the organoids (Scale bar anti-LRP5/6P3 VHHs effectively target Wnt-dependent tumorigenic organoids by removing an essential pathway for renewal of stem-like tumor cells and promoting their terminal differentiation From an initial panel of 33 LRP6P3E3P4E4-binding VHHs we selected three VHHs that most efficiently inhibited cellular responses to Wnt3a at low nanomolar IC50 values These VHHs efficiently blocked the growth of murine tumorigenic R/Z-mutant organoids We show that L-P2-B10 employs a particularly long CDR3 loop that interacts with the VHH framework to create a stable flat paratope for recognition of a plane epitope at the center of the LRP6 P3 domain while CDR1 does not participate in the interaction L-P2-D07 employs a much shorter CDR3 loop that interacts with the same relatively flat epitope This recognition motif is absent in the CDR sequence of our LRP6P3-binding VHHs in line with their lack of interference with Wnt1-mediated signaling and apparent specificity for inhibition of Wnt3/3a-mediated cellular responses due to conservation of the VHH-binding epitope on LRP5P3 and LRP6P3 allowing for selective blockade of Wnt3/Rspo-mediated cellular responses while leaving non-blocked Wnts unaffected The broad species cross reactivity of these VHHs as anticipated based on in silico modeling of the contact surfaces (rodent non-human primate) should facilitate both therapeutic proof-of-concept experiments as well as toxicology studies Human embryonic kidney (HEK)293T cells (ATCC CRL-3216) were cultured in RPMI medium (Thermo Scientific) supplemented with 10% Fetal Calf Serum (Bodinco B.V.) 100 units/mL penicillin and 100 μg/mL streptomycin (Thermo Scientific) Mouse L-cells were cultured in DMEM containing 1 g/L glucose (Thermo Scientific) with the same supplements L-cells stably expressing Wnt3a were used to generate Wnt3a-conditioned medium (Wnt3a-CM) R-spondin1-CM and Noggin-CM were produced using HEK293T cells stably transfected with human Rspo1-V5 or after transient transfection with mouse Noggin-Fc expression vector CHIR-99021 (Tocris bioscience) was used at 5 μM for 5 h Organoids were cultured in matrigel droplets (Corning) Products were cloned into pGEM-T Easy vector system I (Promega) and subsequently sequenced using a T7 sequencing primer Western blotting was performed using standard procedures with Immobilon-FL (Milipore) PVDF membranes For blocking the Odyssey blocking buffer (LI-COR) was used and immunoblot analysis was performed using the LI-COR Odyssey Infrared Imaging System For Immunoblotting: mouse anti-α-Tubulin (Sigma Aldrich 1:10,000); mouse anti-actin (MP Biomedicals (CLONE C4) 1:10,000); rabbit anti-LRP6 (Cell Signaling (C5C7) 1:1000); rabbit anti-Phospho-LRP6 (Ser1490) (Cell Signaling All goat secondary antibodies were conjugated with either Alexa Fluorphores (Life Technologies For immunofluorescence: rabbit anti-Ki67 (Abcam As secondary antibodies goat anti-rabbit IgG-488 (Life Technologies 1:300) and goat anti-mouse-IgG-568 (Life Technologies B-2261) was used at 1:500 and Phalloidin (Ph647) (Cell Signaling Transfection efficiency was controlled and normalized by including 1 ng of TK-Renilla reporter plasmid in all transfections All transfections were performed with FuGENE 6 transfection reagent (Promega) according to the manufacturer’s protocol cells were incubated with 10 μM of VHH and either control conditioned medium or Wnt3a-CM overnight Luciferase activity was measured using the Dual Luciferase Reporter Kit (Promega) according to the manufacturer’s protocol in a Centro XS 960 microplate luminometer (Berthold) Values were normalized to untreated controls For bright field microscopy organoids were grown in 10 μL matrigel droplets and images were captured with an EVOS (Thermo Scientific) R/Z dKO organoids were grown in 10 μL matrigel in slide angiogenesis chambers (Ibidi) in 40 μl EN and treated with 10 μM of IWP-2 (R&D systems) or 10 μM of each VHH for 3 d After treatment organoids were fixed in paraformaldehyde (4% diluted in 0.1 M Na-Pi) for 1 h after which paraformaldehyde was removed and 20 mM NH4Cl (in PBS) was added for 10 min Organoids were permeabilized in PBD 0.2 T buffer (1% BSA 0.2% TX-100 in PBS) for 2 h at room temperature (RT) Organoids were incubated overnight with primary antibody at 4 °C and 3 h with secondary antibody including HOECHST at RT in PBD 0.2 T Organoids were mounted in Ibidi mounting medium and images were acquired with a Zeiss LSM700 confocal microscope Organoids were trypsinized in the presence of Y-27632 (Tocris) and 1000 single cells per well were seeded in a 96-well plate in Basement Membrane Extract (BME) (Cultrex) the BME was polymerized for 20 min at 37 °C Organoids were cultured for the indicated times in the absence or presence of the indicated VHHs The Cell titer Glo Luminiscent Cell viability assay (Promega) was used to assess viability of the organoids in a Centro XS 960 microplate luminometer Human LRP6 containing domains P3E3P4E4 (Uniprot residues 639–1244) was cloned in the expression vector pUPE107.03 (U-Protein Express BV The Netherlands) with an N-terminal cystatin secretion signal and a C-terminal His6-tag LRP6P3E3P4E4 was transiently co-expressed in HEK293 deficient in N-acetylglucoaminyltransferase I (GnTI-) cells with human MESD Six days after transfection the conditioned medium was harvested by centrifugation for 20 min at 1000 × g LRP6 was purified by the addition of Ni-Sepharose excel resin (GE Healthcare) and incubated overnight at 4 °C A packed column was washed using immobilized metal ion affinity chromatography (IMAC) A buffer (25 mM HEPES pH 8.0 Proteins were eluted in IMAC B (25 mM HEPES pH 8.0 followed by a purification step with size-exclusion chromatography (SEC) using a Superdex 200 HiLoad 16 60 column (GE Healthcare) in 15 mM HEPES pH 7.4 and 150 mM NaCl and concentrated to 13 mg/mL To produce biotinylated human LRP6P3E3P4E4 it was cloned in an expression vector (U-Protein Express BV The Netherlands) with a C-terminal BAP-tag followed by a His6-tag LRP6P3E3P4E4 was transiently co-expressed in HEK293 GnTI-cells with human MESD and Bir-A Five days after transfection 0.2 μM Biotin (Sigma Aldrich) was added to the medium and incubated overnight to obtain biotinylated LRP6P3E3P4E4 Purification was performed as described for the non-biotinylated LRP6P3E3P4E4 After five rounds of selection, the recovered DNA was PCR-amplified with Phusion Green HF (Thermo Scientific) using primers TAC6 and NOTIRECREV (Supplementary Table 3) to amplify the VHH segment and introduce a C-terminal V5-tag and NotI site downstream of the VHH coding sequence digested with NcoI and NotI (New England Biolabs) and ligated into a pET vector containing additional polyHis- and FLAG- tags C-terminal of the insert UK) and plated on 2YT media plates containing 50 µg/mL kanamycin (Sigma Aldrich) Individual colonies were picked and grown for soluble expression of VHH induction of 1 mL cultures was started using 1 mM IPTG (Sigma Aldrich) in cultures grown to OD600 ~1 followed by further overnight incubation at 30 °C Cells were lysed by snap-freeze and thaw followed by treatment with Bugbuster Master Mix (Merck) according to the manufacturer’s instructions soluble supernatant fraction was directly used in ELISA screening ELISA screening was carried out on NUNC Maxisorp plates coated with 250 ng per well of streptavidin followed either by overnight incubation at 4 °C with 100 ng/well of biotinylated LRP6P3E3P4E4 (target wells) or PBS buffer (negative control wells) Horseradish peroxidase-conjugated anti-V5 secondary antibody (Abcam) diluted 1: 3000 in 2% BSA in PBS was used to detect VHH/anti-V5 binding Binding was detected using SureBlue TMB peroxidase substrate (Insight Biotechnology) Clones showing an OD > 0.4 on LRP6 and specificity over the negative control wells (LRP6/Streptavidin ration > 5) were selected for a confirmatory ELISA Clones with confirmed binding were sequenced Sequences were aligned and correlated to values from the confirmatory ELISA (LRP6 signal/streptavidin signal) to select the best unique sequence hit clones coli BL21 cells (BL21-Gold Competent Cells Agilent) grown in Terrific Broth (TB) (Sigma Aldrich) containing 50 µg/mL kanamycin Induction and lysis was performed as described above C-terminal V5-FLAG3-His6-tagged VHH material was purified from cell lysates by IMAC either using His MultiTrap HP plates or HisTrap 1 and 5 mL columns (GE Healthcare Life) in case of purifications in 96 well format from 1 mL cultures or from large scale cultures Protein-containing fractions were pooled and imidazole was removed by either buffer exchange (PD Multitrap G-25) or extensive dialysis (snakeskin dialysis tubes MWC 10 kDa The final purification step (using PBS) was performed with size exclusion chromatography (SEC) using a Superdex 75 HiLoad 16 60 column (GE Healthcare Life Sciences) Cellular debris was removed by centrifugation for 30 min at 8000 × g and filtered with a glass fiber prefilter (Ministart Imidazole was added to a final concentration of 15 mM VHHs were purified using Ni-Sepharose 6 Fast Flow beads (GE Healthcare) Fractions containing protein after elution with 300 mM imidazole were concentrated and further purified by SEC using a Superdex 75 Hiload 16 60 column in 15 mM HEPES pH 7.4 and 150 mM NaCl and concentrated to 10 mg/mL Isothermal titration calorimetry (ITC) was performed using a MicroCal Auto ITC200 (Malvern Instruments Ltd) A 10 µM solution of LRP6P3E3P4E4 in DPBS was placed into the 200 μL sample cell at 25 °C Titration was performed with 2 µL injections (first injection 0.5 µL) of VHHs in DPBS at a concentration of 100 µM every 120 s (26 injections total) Data were fitted using the MicroCal PEAQ-ITC Analysis Software (Malvern Instruments Ltd) according to standard procedures Each ligand test was performed in triplicate and values for n Quartz crystal microbalance (QCM) technology was used to analyze the affinities of the VHHs towards LRP6 The experiments were performed using an Attana A200 C-Fast system (Attana AB) LRP6P3E3P4E4 was immobilized on a low-non-specific binding chip (LNB) via amine coupling according to the manufacturer’s protocol Two sensor chips were docked in the instrument and the temperature was set to 22 °C HBS-T was passed over the surfaces at a flow rate of 100 µL/min until the baseline was stabilized (frequency change ≤ 0.2 Hz over 600 s) After reduction of the flow rate to 10 µL/min a freshly prepared activation solution (0.2 M 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide 0.05 M sulfo-N-hydroxysuccinimide) was injected for 300 s on both surfaces Then 50 µg/mL of LRP6P3E3P4E4 in sodium acetate buffer a de-activation solution consisting of 1 M ethanolamine Immobilization of LRP6P3E3P4E4 caused a frequency shift of 200 Hz in channel A The flow was set to 25 µL/min and the experiment was initiated after baseline stabilization VHHs diluted in HBS-T at four concentrations (0.25–0.13–0.06–0.03 µg/mL) were injected sequentially for 84 s in parallel on both surfaces In a single-cycle-kinetics approach starting at the lowest concentration each injection was followed by 30 s dissociation and followed by injection of the next higher concentration Finally dissociation was observed by injection of HBS-T for 300 s After each cycle the surface was regenerated with an 84 s pulse of 10 mM glycine All VHHs were injected in duplicate in two independent experiments Data were collected using Attester software (Attana AB) and analyzed with Evaluation software (Attana AB) and Clamp XP (University of Utah) The signal obtained from the reference surface was subtracted from the sensograms obtained for the LRP6P3E3P4E4– immobilized surface To calculate the kinetic parameters (ka and kd) and affinity constant (KD) the experimental data were fitted using a homogeneous ligand model (1:1 fitting model) Purified LRP6P3E3P4E4 was mixed with purified L-P2-B10 (C-terminal His6-tag) or VHH L-P2-D07 (C-terminal V5-FLAG3-His6-tag) respectively in a 1:1.05 receptor:VHH ratio and incubated for 2–4 h at 4 °C The final concentration of the mixed receptor:VHH complexes was 131 and 145 μM Crystals of the LRP6P3E3P4E4:L-P2-B10 complex grew at 18 °C as long needles by sitting drop vapor diffusion technique equilibrating the protein mixture with a reservoir solution containing 0.1 M sodium citrate pH 5.5 and 10% PEG w/v 4000 in a 1:1 protein:mother liquor ratio Crystals were cryo-protected by briefly taking them up in mother liquor supplemented with 25% (v/v) ethylene glycol before flash-freezing in liquid nitrogen Crystals of the LRP6P3E3P4E4:L-P2-D07 complex grew by sitting drop vapor diffusion in a 1:1 protein:mother liquor ratio with a reservoir solution containing 0.1 M MES Crystals were transferred into cryo-protectant solution of mother liquor supplemented with 20% (v/v) glycerol and flash frozen in liquid nitrogen Figures were generated with PyMOL (The PyMOL Molecular Graphics System Further information on experimental design is available in the Nature Research Reporting Summary linked to this Article Targeting Wnt signaling in colorectal cancer A review in the theme: cell signaling: proteins Surrogate Wnt agonists that phenocopy canonical Wnt and beta-catenin signalling Molecular regulation and pharmacological targeting of the beta-catenin destruction complex Tumour suppressor RNF43 is a stem-cell E3 ligase that induces endocytosis of Wnt receptors ZNRF3 promotes Wnt receptor turnover in an R-spondin-sensitive manner The R-spondin/Lgr5/Rnf43 module: regulator of Wnt signal strength Stem cells marked by the R-spondin receptor LGR5 APC mutations occur early during colorectal tumorigenesis Comprehensive molecular characterization of gastric adenocarcinoma Whole-exome sequencing of neoplastic cysts of the pancreas reveals recurrent mutations in components of ubiquitin-dependent pathways Targeting PTPRK-RSPO3 colon tumours promotes differentiation and loss of stem-cell function Identification of novel candidate genes for early-onset colorectal cancer susceptibility Porcupine inhibitor suppresses paracrine Wnt-driven growth of Rnf43;Znrf3-mutant neoplasia Chemical modulation of WNT signaling in cancer WNT signaling in bone development and homeostasis The complex world of WNT receptor signalling Dissecting molecular differences between Wnt coreceptors LRP5 and LRP6 Reconstitution of a frizzled8.Wnt3a.LRP6 signaling complex reveals multiple Wnt and Dkk1 binding sites on LRP6 Wnt isoform-specific interactions with coreceptor specify inhibition or potentiation of signaling by LRP6 antibodies a context-dependent activator and inhibitor of Wnt signalling LDL-receptor-related protein 6 is a receptor for Dickkopf proteins Essential requirement for Wnt signaling in proliferation of adult small intestine and colon revealed by adenoviral expression of Dickkopf-1 Sclerostin expression and functions beyond the osteocyte Structural basis of Wnt signaling inhibition by Dickkopf binding to LRP5/6 Crystal structures of the extracellular domain of LRP6 and its complex with DKK1 Wnt antagonists bind through a short peptide to the first beta-propeller domain of LRP5/6 Epigenetic inactivation of the Wnt antagonist DICKKOPF-1 (DKK-1) gene in human colorectal cancer Inhibition of tumorigenesis driven by different Wnt proteins requires blockade of distinct ligand-binding regions by LRP6 antibodies Novel bispecific domain antibody to LRP6 inhibits wnt and r-spondin ligand-induced Wnt signaling and tumor growth The first propeller domain of LRP6 regulates sensitivity to DKK1 Detection of endogenous LRP6 expressed on human cells by monoclonal antibodies specific for the native conformation Nanobodies as versatile tools to understand antibody fragment libraries built using automated hexamer codon additions and validated through next generation sequencing CIS display: in vitro selection of peptides from libraries of protein-DNA complexes Inhibition of HER3 activation and tumor growth with a human antibody binding to a conserved epitope formed by domain III and IV Antagonistic TNF receptor one-specific antibody (ATROSAB): receptor binding and in vitro bioactivity Single-cycle kinetic analysis of ternary DNA complexes by surface plasmon resonance on a decaying surface Structural and functional studies of LRP6 ectodomain reveal a platform for Wnt signaling Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche Interplay between metabolic identities in the intestinal crypt supports stem cell function Redundant sources of Wnt regulate intestinal stem cells and promote formation of Paneth cells Adult intestinal stem cells: critical drivers of epithelial homeostasis and regeneration Identification of stem cells in small intestine and colon by marker gene Lgr5 A colorectal tumor organoid library demonstrates progressive loss of niche factor requirements during tumorigenesis Small molecule-mediated disruption of Wnt-dependent signaling in tissue regeneration and cancer Niche-independent high-purity cultures of Lgr5 + intestinal stem cells and their progeny Control of Wnt receptor turnover by R-spondin-ZNRF3/RNF43 signaling module and its dysregulation in cancer Molecular basis for the preferential cleft recognition by dromedary heavy-chain antibodies Crystal structure of a camel single-domain VH antibody fragment in complex with lysozyme Reduced affinity to and inhibition by DKK1 form a common mechanism by which high bone mass-associated missense mutations in LRP5 affect canonical Wnt signaling Wnt-mediated self-renewal of neural stem/progenitor cells Regulation of Wnt/LRP signaling by distinct domains of Dickkopf proteins Kremen2 modulates Dickkopf2 activity during Wnt/LRP6 signaling Paneth cells constitute the niche for Lgr5 stem cells in intestinal crypts Apc restoration promotes cellular differentiation and reestablishes crypt homeostasis in colorectal Wnt addiction of genetically defined cancers reversed by PORCN inhibition Nanobodies and nanobody-based human heavy chain antibodies as antitumor therapeutics Nanobody-based cancer therapy of solid tumors Simultaneous paralogue knockout using a CRISPR-concatemer in mouse small intestinal organoids Loss of the tumor suppressor CYLD enhances Wnt/beta-catenin signaling through K63-linked ubiquitination of Dvl Constitutive transcriptional activation by a beta-catenin-Tcf complex in APC-/- colon carcinoma Analyzing real-time PCR data by the comparative C(T) method An in vitro selection strategy for conferring protease resistance to ligand binding peptides Selection of a high-affinity WW domain against the extracellular region of VEGF receptor isoform-2 from a combinatorial library using CIS display Protein production by auto-induction in high density shaking cultures DIALS: implementation and evaluation of a new integration package How good are my data and what is the resolution Overview of the CCP4 suite and current developments Structural evaluation of EGFR inhibition mechanisms for nanobodies/VHH domains Towards automated crystallographic structure refinement with phenix.refine Coot: model-building tools for molecular graphics MolProbity: all-atom structure validation for macromolecular crystallography Download references We thank members of the laboratory of M.M.M We thank the European Synchrotron Radiation Facility (ESRF) for the provision of synchrotron radiation facilities; and beamline scientists of the ESRF for assistance We thank U-protein Express for the provision of protein expression facilities We thank Guangyun Yu and Ronald Pieters (Utrecht University Department of Pharmaceutical sciences - Molecular Pharmacy for providing access to the MicroCal Auto ITC200 which is partly financed by the Dutch Cancer Society This work was supported by European Research Council Starting Grant 242958 the Netherlands Organization for Scientific Research NWO VICI Grant 91815604 European Union Grant FP7 Marie Curie ITN 608180 “WntsApp” (to M.M.M.) Fondazione Michelangelo for the Advancement of the Study and Treatment of Cancer (to N.F.) BioStruct-X Grant 283570 and NWO Spinoza Grant 01.80.104.00 (to P.G.) These authors contributed equally: Nicola Fenderico Oncode Institute and Department of Cell Biology Gottfried Schatz Research Center for Cell Signaling A patent application covering this work has been filed by UMCU and Isogenica The remaining authors declare no competing interests Journal peer review information: Nature Communications thanks Wenqing Xu and the other anonymous reviewers for their contribution to the peer review of this work Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations Download citation DOI: https://doi.org/10.1038/s41467-018-08172-z 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 They decided to take up this challenge after their existing clientele expressed a clear need for an additional screen supplier Ridder started to explore the opportunities and we soon realized these existed." a start was made at the time to develop an extensive portfolio “At that moment the portfolio was still far from complete but we did have a number of good products to enter the market with It was a good time to introduce a new supplier.” Ridder started with twelve screens - all available from stock and in three different sizes Now seventeen types of screens are supplied from stock and the complete assortment consists of more than thirty types including screens produced entirely in the Netherlands Co-creationLast year Ridder started with "co-creation" In addition to the already existing screens operational director: “Together with the customers we develop a variant that meets their specific wishes and needs This way we contribute to the solution to the challenge that our customers face in their project supported by the existing knowledge and experience within the Ridder Group A great development that we are proud of.” In addition to the introduction of new and co-created screens Ridder is constantly looking to optimize the current screens we are investigating how we can achieve higher energy savings with our energy screens for example by increasing the thickness of the foil But we always keep looking at the net return When we compare the additional costs of such a new product with the advantages we still have to conclude that we have the ideal mix with the current RES 10 FR screen That is also an outcome of a development.” Mario ScherpenzeelNiels and Johan tell about the knowledge within Ridder they would not be able to realize new developments The foundation of this knowledge was laid by Mario Scherpenzeel Johan: "The professional and stable base that he created within our department ensures that there is now an incredibly good team that is ready to go through." Niels also agrees that this foundation is very important to Ridder: "Thanks to the foundation that Mario laid the team can now continue to develop with enthusiasm and energy." Present situation Ridder has already delivered more than 4000 ha of climate screens Last year Ridder reached a milestone by supplying 1500 ha in the Netherlands There are now four knitting machines at the production location in Maasdijk there is no intention to bring the total production to the Netherlands “The most important thing is that production in the Netherlands is going to provide us with a lot of knowledge and experience This makes us less dependent on our partners in Asia It also ensures that with our production we are closer to the customer “It is now also possible for our customers to come and view the production of their screen,” says Niels “Our department now consists of eighteen employees Most people will know my colleagues Joachim Keus And of course we can always use new people Bright futureWith the celebration of the fifth anniversary “We have now positioned ourselves well in the market We have made progress and are developing new screens ourselves,” says Johan Ridder would like to make the difference in the current market over the next five years Five years plus one“The invitations had already been printed and we had made many arrangements,” Johan laughs But then he says: “Delay does not mean cancellation If the current circumstances do not make it possible to celebrate this year For more information:  Ridderwww.ridder.com[email protected] FreshPublishers © 2005-2025 HortiDaily.com a dedicated photographer from the Netherlands has been able to wholeheartedly document the early stages of Sultanate of Oman’s renaissance He recently offered his exceptional collection as a special gift to Oman based on his deep love for the country and on his keenness to enrich its national archives his collection illuminates this pivotal era of Oman’s modern history which he collated with the earlier one under the title “Oman Scherpenzeel said that over the past four years he spent in Oman in the early 70s he was able to document a large number of scenes scenery and landmarks of Omani governorates that reflected the social cultural and human heritage of the country Oman has seen a comprehensive transformation thanks to the country’s ability to keep pace with the events of the age as they unfolded Scherpenzeel said that photos are more truthful in projecting reality at a time the rhythm of life is changing at a rapid pace crisscrossing with all aspects of social interactions He affirmed that he was already in Oman by June 1970 three weeks prior to late Sultan Qaboos Bin Said’s accession to power in the country “Life in Oman was marvelous and Omani people were gentle sociable and truthful in their friendship,” said Scherpenzeel noting that one could move freely in the fascinating landscape and that one could even sleep safely on the golden dunes Scherpenzeel said he could still remember his visit to Sur and the long trip to the easterly wilayat from Muscat by 4-wheel on the rough roads he could observe Omani people’s devotion to the Islamic religion and sound Arabian habits and traditions of hospitality Oman’s Ambassador to the Netherlands said that these sincere reflections represent a model of the Sultanate of Oman’s soft power that distinguishes Omani people among nations of the world when he visited Scherpenzeel at his home in the Netherlands he was astonished at the man’s amicable reaction “It was as if he had known me for a very long time His warm welcome unveiled his deep love to Oman and his thorough knowledge of the country the reputation of its people and the grandeur of its leadership.” Many things struck me as I entered Scherpenzeel’s home: Every corner is adorned with an image from Oman his bedroom is bedecked with historical paintings What baffled me more was the fact that Scherpenzeel made accounts of Omani villages and countryside in such details that gave me the impression that I was listening to an academic in Omani history and civilization The ambassador explained that he visited Scherpenzeel to thank him for the gift he presented to the Sultanate of Oman and to express gratitude for such a humanitarian gesture For more information, please read Privacy policy Kayak have announced their farewell tour which is set to take place in April & May 2022 These shows will see them perform tracks from across their career Band lead Ton Scherpenzeel comments “It’s been a great ride but we’ve come to the end of the line As the other band members would have liked to continue But it takes a lot more than just that to make a good tour and circumstances for bands like us are not getting any easier In that respect I feel I’ve reached the limit of my capabilities I’d rather stop too early than go on too long.” https://www.kayakonline.info/ https://www.facebook.com/kayakmusicband https://www.instagram.com/kayak.band/ TotalRock’s beginnings go way back to 1997 – founded as ‘Rock Radio Network’ by the voice of metal Tommy Vance and his trusty producer of the BBC Friday Rock Show along with the walking encyclopaedia of metal Netherlands-based meat business Vion Food Group is investing in a plant in the south of the country as part of a consolidation process Netherlands-based meat business Vion Food Group is investing in its plant in the town of Boxtel as part of a consolidation process Vion plans to spend EUR35m (US$39.6m) to modernise the facility in “order to further increase its competitiveness on the national and international meat markets” according to a statement posted on its website.  Some of the fresh meat production carried out at the company’s site in the town of Scherpenzeel located in the Dutch province of Gelderland will be transferred to Boxtel in the south of the country Vion said the “majority” of the 120 workers employed in the fresh division at  Scherpenzeel will be relocated to its other factories in Boxtel or “will leave by means of natural labour turnover” Around 20 workers may not find relocation a suitable possibility It expects to complete the transfer by the middle of 2021.  Vion noted the operational transfer “will shorten the supply chain which will result in an improvement in the quality and freshness of the meat and more efficient production”.   Nominations are now open for the prestigious Just Food Excellence Awards - one of the industry's most recognised programmes celebrating innovation This is your chance to showcase your achievements Don't miss the opportunity to be honoured among the best - submit your nomination today View all newsletters from across the GlobalData Media network Experts stressed the importance of standardisation, data sharing and transparency at Riviera’s webinar, Voyage and vessel optimisation when making port calls Icelandic maritime energy management provider Marorka’s director of service International Taskforce Port Call Optimization chairman Ben van Scherpenzeel PortXchange international business consultant - port call optimisation Captain Abhishek Nair and Shell general manager shipping & maritime for Europe & Africa Captain Kaia Bjerre van de Ven discussed issues related to the future of data sharing and digitalisation in port operations Mr Björnsson said real-time optimisation was made possible when vessels were able to receive real-time weather and sea-state forecasts and terminal data. While improvements in connectivity and modelling have helped Marorka provide customers with efficient speed profiles for voyages Mr Björnsson said the question of data sharing remains an issue He suggested the industry needs to have standards in place for data sharing and must incentivise companies to share operational data Mr van Scherpenzeel said data sharing is burdened by the differing standards and formats present throughout the industry While standardisation will require co-operation investments in implementation and cultural change Any data sharing process has to be applicable geographically as well as across operational types and profiles Mr van Scherpenzeel explained berth planning in the context of vessels and ports that are not linked by a central database Most ports are run by a public body while the terminals are managed by private organisations “Berth planning is dictated by a terminal The access to the terminal is through a public waterway normally organised by the public authority with the help of a service provider for nautical services That collaboration is key to our customers who arrive at the port," he said terminals and service providers to realise one planning system that allows ships to arrive at port at a safe Citing a practical case of a mariner receiving conflicting navigational data Captain Nair showed how this could lead to efficiency losses throughout the supply chain the data [a mariner] has on [a] chart or publication does not match the information [he or she is] getting from the port authorities or the agents just a difference of 10 cm between a chart and the port’s data can mean a loading difference of 2,000 tonnes on a VLCC And as a master you will probably err on the side of safety," he said Noting the delays and costs incurred due to a lack of data sharing, Captain van de Ven said "shipping is not linear; sharing data helps us react at any time" “We still see container vessels up to 20,000 TEU waiting for bunker barges that are delayed This is about being aware of how different parts in the chain are active,” she said Captain van de Ven said she expects standardisation will be driven by IMO and supported by industry bodies like BIMCO and Intertanko The experts also pointed out it is important to incentivise change in addition to creating a standard Noting that “incentives work better than regulations” Mr van Scherpenzeel said by modifying contracts masters can be incentivised to share data without fear of breaching the contract Captain Nair said the process involves changing work culture and that PortXchange is conducting an ongoing pilot programme with chemical clusters in Houston Recognising that companies have widely varying levels of digital maturity he said “You have to work with them slowly find what works best for each user and finally bring them towards one common standard.” Mr Bjönsson said Marorka places emphasis on a practical approach He said performance management has benefited from increased transparency between owners charterers and managers and is expected to be useful in vessel optimisation standards alone cannot improve performance Decisions made on bad data have the potential to be worse than decisions made on gut feeling." Captain Nair said existing data can be used for optimisation stressing the need for more willingness within shipping to adapt to new technology We want things to work right before we step into it Let’s try to make this digitalisation journey together." Some 74% of webinar attendees felt that current regulations encourage digitalisation and 43% agreed with our panel that willingness is the biggest barrier to further digitalisation in the industry 73% of attendees said their company has a clear idea of digitalisation in the coming years with 39% suggesting there is a clear roadmap for the next five years A strong majority at 80% said they expect to see full IoT connected vessels between 2025 and 2030 You can view this webinar and all of the webinars from Riviera’s Vessel Optimisation Webinar Week, in full, in our webinar library And you can sign up to attend our upcoming webinars on our events page. Panellists (left to right): PortXchange’s international business consultant for port call optimisation Captain Abhishek Nair, International Taskforce port call optimisation chairman Ben van Scherpenzeel, Icelandic maritime energy management provider Marorka’s director of service, Haraldur Orri Björnsson and Shell general manager shipping & maritime for Europe & Africa, Captain Kaia Bjerre van de Ven Newsletter Search View more... SCHERPENZEEL – A ground-breaking project which recycles discarded fishing nets into nylon yarn to produce carpet tiles is to be rolled out to the Lake Ossa region of Cameroon, in a project led by Dutch carpet tile manufacturer Interface, and the Zoological Society of London. Explore all our independent journalism from as little as 30p per day. Less than an espresso - but more stimulating. Create an accountNewsletterRead our top news stories delivered straight to your inbox. Search View more... Help to support the purpose-led work of our team of journalists and partners – and expose your product or service to our global readership Request a media pack which details the benefits of advertising on ETN including our rates and details of our distribution please fill out the form below: YOUR DETAILS – See our privacy policy here for details of how we take your privacy very seriously Advertising Terms and Conditions Subscribe to our magazine today and receive exclusive online content '+scriptOptions._localizedStrings.webview_notification_text+' "+scriptOptions._localizedStrings.redirect_overlay_title+" "+scriptOptions._localizedStrings.redirect_overlay_text+" Muscat – Dutch photographer Eric van Scherpenzeel who documented the beginning of Omani renaissance from 1970 to 1974 recently presented his photos as a gift to the sultanate The photos captured an important stage of modern Omani history for posterity Scherpenzeel spoke of he great love for Oman and its the beauty and hence his decision to gift these to the sultanate He visited Oman for the second time in the 1990s to photograph specific areas which he recently published as a comparative study in a book titled Oman’s Past and Present features and various details reflecting social and cultural aspects of Omani people across several governorates,” he told ONA three weeks prior to the Late Sultan Qaboos bin Said’s accession to power “Life in Oman was marvelous; the people were gentle sociable and truthful in their friendship,” said Scherpenzeel adding that visitors could travel in the fascinating landscape at will and sleep safely on sand dunes under stars in the desert visited Scherpenzeel at his home to thank him for the gift “It was as if he had known me for a very long time,” the ambassador said H E Dr Harthi was struck by what he saw in Scherpenzeel’s home his bedroom was bedecked with historical paintings mementos and even old rifles from Oman,” he said Muscat Daily is now the largest selling broadsheet newspaper in the Sultanate of Oman with 33,500 daily copies and 28,000 subscribers. Muscat Daily provides unrivalled national news coverage from Oman © 2021 Apex Press and Publishing. All Rights Reserved. Powered by Mesdac Broadband TV News January 20, 2016 11.08 Europe/London By Fiber Nederland (Fiber NL) a provider of triple play and smart home solutions is acquiring its competitor Stipte (formerly Scarlet Netherlands) which was founded three years ago by Martijn van Scherpenzeel and operates independently of network operators “Between powerhouses such as KPN and Ziggo Fiber NL is a relatively small player but in a consolidated market competition has generally more to lose than to gain service-oriented and able to react faster to trends.” Van Scherpenzeeland points out that for instance Fiber NL beat KPN in 2015 by being the first to offer an integrated smart home solution The company also took a majority stake in the technology company that offers the Smart Alarm home automation services Van Scherpenzeel added: “With the acquisition of Stipte known for its operational excellence and its organisation and brand fits well with Fiber NL we hope to expand our role as a challenger We are pleased to welcome Matthijs Kamp as a partner and collaborate with an enthusiastic group of professionals This acquisition has no impact on staffing and delivery of services to customers of Stipte.” Arnhem-based Robert covers the Benelux, France, Germany, Austria and Switzerland as well as IPTV, web TV, connected TV and OTT. Email Robert at rbriel@broadbandtvnews.com Today, consumers are increasingly using bandwidth-intensive and latency-sensitive workloads, such as 4K and 8K streaming, online gaming, and AR/VR applications. As a result, Internet Service Providers must update their networks and by extension Wi-Fi experiences and performance. … [Download the White Paper ...] Copyright © 2025 Broadband TV News LLP · Log in set a target for his business to have zero impact on the environment by 2020 many thought it an outrageous ambition But the company has made such great strides that its European operations are within touching distance At a time when very few companies were making public commitments to sustainability, "mission zero" was a bold vision Since then Interface's journey towards sustainability has seen a momentous shift in the way the company operates – and some outstanding results Interface's plants in Holland and Northern Ireland have been using around 90% less carbon and 95% less water than in 1996 All the gas used here is biogas produced from green waste that comes from a local fish processing company The gas is produced from the anaerobic digestion of fish waste which is filtered to the natural gas standard pumped into the grid and bought by Interface The switch to 100% biogas has eliminated all energy-related emissions at the plant which at the beginning of 2014 stood at 8,900 tonnes of CO2 a year The Dutch facility uses hardly any water in the manufacturing process – the only water it consumes is on the domestic front Interface's manufacturing facilities use 95% less water today than they did 18 years ago 95% of Interface's energy consumption comes from renewable sources a massive leap from 2012 when the figure stood at 33% This has shielded the company from energy price volatility and carbon-related price movements Interface's facility in Craigavon has used 100% green electricity since 2006 and has recycled all of its waste since 2008 In 2014, the company will reduce its scope 1 and 2 emissions in Europe by 90% from a 1996 baseline So how has Interface achieved such massive reductions in energy and water use Meeting the challenge of zero impact on the environment is not about making a few tweaks an aggressive energy efficiency programme and the technology to go with it It means an approach to sustainability that influences every aspect of the company's business inspiring Interface and its people to continually push at the boundaries This attitude was applauded by the Guardian judges who were impressed with Interface's "clear and determined pursuit of mission zero to realise a radical and transformative change" One of the innovations that helped Interface reduce its energy use is a new pre-coat manufacturing line Pre-coat is an important part of the backing that supports the look and feel of a carpet by anchoring the yarn in place – in Interface's case this layer is latex The new pre-coat line makes more efficient use of hot air through ultra-efficient ovens cutting gas consumption and dramatically reducing the company's dependence on fossil fuels the new pre-coat process has reduced energy use by 40% per square metre of carpet produced and cut the use of materials by 8% in the Scherpenzeel factory It is just one of a wide range of energy-saving measures introduced by the carpet tile manufacturer more sustainable materials and the installation of LED lighting many of Interface's savings have come from fundamental shifts in the way it makes its products The company has also changed the way it operates A new closed-loop piping system in its Scherpenzeel plant recycles all cooling and manufacturing water meaning the only water used by the factory is for domestic purposes These latest achievements bring Interface closer to achieving its ambitious goal in 2014 the company's European operations will achieve a 90% reduction in greenhouse gas emissions That exceeds the EU Commission's proposed 2030 carbon reduction target by more than 100% Interface is proud of these milestones on its "mission zero" journey It is the kind of progress that is beyond the imagination of many companies but for Interface it is by no means the end of the story "mission zero" is not just about its own operations – it is taking responsibility for the lifecycle of its products And that is where the carpet tile revolutionaries are going to focus their attention next Lorna Thorpe is part of the wordworks network The Guardian Sustainable Business Sustainability Case Studies contain articles on all the initiatives that met the criteria for the GSB Awards. ‘The waste processors want to visually check if there’s anything that doesn’t belong in the bags and you can’t do that with the opaque grey bags If we can’t see what the quality of the material is we can’t process it,’ Rudie Marsman of Van Scherpenzeel recycling firm told the broadcaster At Van Scherpenzeel two to three tonnes of plastic and metal packaging and cartons (PMD) end up being burnt unnecessarily every year Waste disposal companies told local councils as early as January that opaque bags containing such refuse are no longer welcome because ‘the assumption is they contain ordinary household waste’ locals authorities have failed to notify the public that opaque bags are banned and transparent bags are now the order of the day these were issued via packaging industry initiative Plastic Heroes but when local councils took over the collection of PMD waste in 2015 the bags were no longer available which have introduced containers for plastic or wheelie bins It should have been explained to the public that the transparent bag served a purpose,’ Marsman told NOS Waste processing companies are reportedly struggling to process the enormous amount of PMD waste: although more people are sorting their waste the amount of other refuse in among the PMD waste is growing too A report by Rijkswaterstaat showed that 21% of collected PMD waste contains items that sorting machines cannot handle, such as food waste, plastic toys, household implements and clothes, NOS writes. We could not provide the Dutch News service, and keep it free of charge, without the generous support of our readers. Your donations allow us to report on issues you tell us matter, and provide you with a summary of the most important Dutch news each day. Many thanks to everyone who has donated to DutchNews.nl in recent days! We could not provide this service without you. If you have not yet made a contribution, you can do so here. Please help us making DutchNews.nl a better read by taking part in a short survey. By EMN Horizon Forbidden West has been out for quite some time now and one thing is for certain we are going to have a sneak-peek into a collection of stunning art from the Horizon Forbidden West Art Team The developers uploaded a collection including early concept art of environment We will not be showing everything that the team has uploaded for obvious reasons. You can view them in all their glory here here are some of our favourites from the collection With a fleet of 200 trucks and  three hubs in Poland which also serve the Slovak and Bulgarian markets in addition to the main location in the Netherlands Mandersloot is one of the market leaders in conditioned groupage transport to Central Europe "The advantage of the various depots is that we are locally present and speak the language of the customer while a branch manager from the Netherlands is active in all countries we can immediately solve problems with a certain shipment and we offer on the spot services such as stickers documentation and quality controls," says Willem van Leuveren frozen goods and so-called high value cargo "We are a customer at almost all retailers in the mentioned Central European countries We can therefore apply very competitive rates for retail," says Willem van Leuveren "We often cross-dock the Dutch products in our logistics centre in Scherpenzeel the trucks must be on their way to our hubs so that the cargo can be delivered to the Eastern European the next day For this we have in-house transport from vans to LHVs because we are also active in urban distribution and can therefore also cover the last mile our activities go much further than transport from A to B the company took an important step in obtaining the IFS Logistics certificate this means not only a safety system for the transport and cross-docking of food-related products but it is a system in which we can guarantee the quality of every conditioned product on all fronts," says Willem "This has also changed things for the customer They used to be able to choose from various temperature ranges now they can only choose between four different set points for LTL transport We drive with these temperatures on a continuous basis so that there is less temperature fluctuation during transport Willem mentions the recruitment of qualified drivers as the main challenge because we always drive with drivers from the country of origin so from Poland with Polish drivers and from the Netherlands with Dutch drivers." He does not expect that road transport to these countries will be replaced by other modalities "The network in Central Europe is not suitable for this and the truck is still the fastest means of transport." Web portalAutomation is a spearhead for Mandersloot "Our web portal is available to all customers so that they can track their shipments and view documents 24 hours a day We also send our key accounts a performance report every week but we find it is important to communicate proactively to the customer There can always be something that slows down the freight in which I expect that most of the planning will take place automatically within a few years I do not think that the human factor will be superfluous but planning will become a different profession the planners knew all the postcode areas by heart Nowadays they have to think much more systematically That is why we also focus on the necessary education and training "Mandersloot remains a real family business In 1964 it was founded by Teuni and Wuf Mandersloot the management is led by Ronald and Marjon Mandersloot and the third generation is already involved in the company," says Willem "Sustainability is of paramount importance to us This year we have installed solar panels on the roof of our headquarters in Scherpenzeel and the purchase of LNG trucks is planned for next year." For more information:Willem van LeuverenMandersloot t Zwarte Land 103925 CK Scherpenzeel+ 3133-2776444[email protected]www.mandersloot.eu FreshPublishers © 2005-2025 FreshPlaza.com Mandersloot will be the first logistics service provider in Europe with 3 LHVs (Longer Heavier Vehicles) on an international route to supply the Czech Republic specialized in conditioned transport of mainly food will use the LHVs mainly for the line-haul of groupage goods between the parent company in Scherpenzeel (gld) and the subsidiary in Prague Sustainability is of paramount importance at Mandersloot is equipped with solar panels and in addition several LHVs are already being used for inland transport sees the deployment of LHVs over long distances as an important next step in the process to reduce the company’s carbon footprint In addition to its head office in the Netherlands Mandersloot has several modern premises in Central Europe Mandersloot is able to supply all major retail organizations in Poland With a fleet of more than 200 vehicles and a customer-oriented focus authorities and employees who made this fantastic milestone possible For more information:Willem van Leuveren / Ronald Marchal Mandersloot’s Expeditiebedrijf b.v.Tel: +31 33 277644Email: [email protected] www.mandersloot.eu  Kim Carter, a young Dutch gymnast of Maltese descent, yet again played a prominent role in her club's success in the national Rhythmical Gymnastics finals, held recently. On April 13, Carter and her colleagues had represented their club - GTV de Badhoeve - in the last qualifying round ahead of the main finals. A fine performance on that occasion earned GTV de Badhoeve the right to compete in the finals last month in Scherpenzeel. Under the heading 'GTV de Badhoeve - the best Rhythmical Gymnastic Club in the Holland', regional newspaper Het Witte Weekblad reported that Carter and her team-mates produced an excellent performance in the finals, winning three gold medals and a first placing in the overall rankings. In Category 1B, GTV de Badhoeve was very successful. In the 'hoop exercise', Sarah Vogel, Tessa Herngreen, Daphne van Delden, Kim Carter, Dominique Corver, Stephanie Klassen and Nikki Wesdorp were the season's favourites for the Dutch title. Expectations were eventually met as the team obtained 18.392 points to become national champions. There was stiffer competition in the ribbon exercise but a second and third placing in the qualifying rounds secured a place in these finals for GTV de Badhoeve. Encouraged by the other successes on the day, the team carried out their exercise almost to perfection and with a total of 15.599 points, they clinched another gold. Thanks to these fine performances, valuable points were obtained towards the overall standings and rankings. In 2001, GTV de Badhoeve were third but this time around they dethroned former champions Pax Hoofdorp. GTV de Badhoeve were crowned Dutch champions with 1,753 points, enjoying a lead of 120 points over their nearest rivals. GTV de Badhoeve are planning a visit to Malta. About 20 to 30 girls will be coming over in October to give a training demonstration at a local club. Carter is currently spending her summer holidays on the islands. To view comments, please register for free or log in to your account. 1 ContributorKeep On Missing You LyricsFeelings of confusionSpinning through my mindReliving every momentKnowing it still hurtsAllies in all mattersPartners in all crimesNo one came between usTogether we fooled the worldWe were like hand and gloveYou’re the last one I wanted to looseI still keep on missing youWith all my heart I know I doWe laughed To learn more, check out our transcription guide or visit our transcribers forum Find answers to frequently asked questions about the song and explore its deeper meaning 2 ContributorsQueen Of Hearts LyricsThe curtains drawn