sells for $1.5 million during the fourth and final session of the OBS April Sale.OBS/VidHorse Photo this is the first time he has had two seven-figure offerings come out of his barn in the same week “Without my team we’re not able to do this they take just as much credit as I do,” Venosa said “We knew (the Tiz the Law colt) was one of the top horses in the sale you let the people evaluate them and that’s what they are worth The most important thing is I’m really excited about the home he’s going to.”Out of the winning stakes-placed Souper Speedy mare Georgian Dancer a half-sister to Canadian champion River Maid the Tiz the Law colt is one Venosa said has made an “amazing” transformation since he purchased him as a yearling the colt morphed into a sales topper with Finley prevailing as he bid alongside Spendthrift general manager Ned Toffey and Monique Delk executive director of racehorse development for St Elias.“Tiz the Law is a stallion where we’ve now bought three of them at the 2-year-old sales and he’s come forward like a whirlwind,” Finley said “I think he’s going to make an impact on the breed for a lot of years to come We’ve been doing business with Steven for about 30 years and bought our first really good horse out of Steve’s barn when he worked for J.J I have a lot of respect for the work that he does and this horse he really was a horse all three of us wanted.”Added Venosa “It seems like (the colt) gets better and better several people would come and look at him every day and every day he just really blossomed They’re showing up at 2-year-old sales To bring a horse like that of that size and work the way he did was very impressive.” for $1.4 million from the consignment of Kings Equine which sold two seven-figure horses during the week.“Very appreciative and glad a lot of adjectives that you can ultimately apply to the week,” said Tod Wojciechowski “The international buyers were here we had a broad spectrum of buyers from all over the world.” Grade 1-placed Langfuhr mare Fuhriously Kissed and is from the female family of Grade 1 winner On Fire Baby sold to Eclipse Thoroughbred Partners & Sabby Racing LLC for $560,000 providing Woods and his wife Angela a most appropriate sendoff.“It’s great It was fantastic how he sold,” an emotional Woods said and it was neat for a really nice horse to be my last horse and not some $5,000 horse And he’s going to a good spot.” and the debates surrounding trainer Bob Baffert Receive daily headlines, breaking news alerts, promotions, and much more! Rehabilitation for Musculoskeletal Conditions Volume 5 - 2024 | https://doi.org/10.3389/fresc.2024.1470002 Introduction: The rupture of the plantar fascia is a rare but significant injury that predominantly affects athletes and individuals engaged in high-impact activities and chronic degeneration from plantar fasciitis can predispose individuals to rupture It can involve a complete or partial tear of the plantar fascia fibers leading to a loss of structural integrity and functional support Spontaneous ruptures of the plantar fascia (occurring without any predisposing factors) are rarely observed in clinical practice No guidelines or other unequivocal recommendations are available for this pathological condition Method: A healthy 35-year-old male who works in an office setting and is a recreational cyclist with a silent clinical anamnesis experienced a spontaneous rupture of the plantar fascia of the left foot with no history of trauma He exhibited significant localized tenderness and swelling in the medial arch of the left foot with difficulty bearing weight on the affected foot An MRI confirmed a partial rupture of the medial cord of the plantar fascia accompanied by surrounding inflammation The patient underwent conservative treatment and transcutaneous electrical nerve stimulation) the patient reported a significant reduction in pain and marked improvement in functional mobility (as confirmed by VAS and Foot Function Index scores) and the patient could bear full weight on the foot without discomfort A follow-up ultrasound demonstrated complete resolution of the plantar fascia rupture and no residual inflammation Discussion: This case underscores the effectiveness of an integrated rehabilitative approach and provides a framework for managing similar cases in clinical practice This mechanism enhances the efficiency of the foot during push-off This case report details an integrated rehabilitative approach to a spontaneous plantar fascia rupture in a healthy 35-year-old male this report aims to provide insight into the effectiveness of non-surgical management strategies and contribute to the existing literature on this rare condition who works in an office setting and is a recreational cyclist (riding about 80 km on asphalt twice a week) presented with a sudden onset of sharp pain in the arch of his left foot while walking (VAS score: 8) He reported no episodes of plantar fasciitis over the last years or previous trauma to the feet or lower extremities He denied any remarkable pre-existing or concomitant systemic illnesses and swimming recreationally only during his childhood and adolescence He routinary wears loafers when he's at work and sneakers in his recreational time He is not aware of previous pathological conditions of the plantar fascia or relevant genetic abnormalities in his family Foot Function Index (FFI) was administered at this time to measure the impact of this pathological condition in terms of pain and activity restriction (FFI score: 62%) T2-weighted MRI image showing the disruption of the medial cord of the plantar fascia at its proximal third with edema of the plantar fascia and surrounding soft tissue Physical therapy protocol–3 sessions/week started immediately after the diagnosis and performed for 4 weeks Achilles tendon and plantar fascia stretching exercises scheduled for the patient Timeline of our conservative plan for the spontaneous rupture of the plantar fascia the patient reported a significant reduction in pain (VAS score: 2) and marked improvement in functional mobility (FFI score: 14%) The patient had undergone physiotherapeutic sessions regularly (as confirmed by the therapist); NSAIDs have been taken only for 7 days after the diagnosis and the patient was able to bear full weight on the foot without discomfort No adverse events have been reported during this time interval He hadn't any limitations in practicing his routine sport activity The patient was advised to continue strengthening exercises and use supportive footwear during high-impact activities to prevent future injuries we elaborated and proposed a novel integrated rehabilitative approach This integrated rehabilitation approach was able to get a satisfying functional recovery in this case as demonstrated by a significant reduction in VAS and FFI scores The patient's recovery without surgical intervention underscores the potential benefits of non-invasive treatment strategies Our study has some limitations that should be addressed this report discusses only one instance of spontaneous rupture of the plantar fascia limiting the ability to generalize findings or draw broad conclusions A larger series of cases or a comparative study would provide more robust data the follow-up period in this case was relatively short; extended follow-up would provide a more comprehensive understanding of the prognosis and effectiveness of the treatment the patient in this case was a healthy adult male with a specific lifestyle (recreational cyclist) The findings may not be generalizable to different populations the assessment of pain and functional improvement relies heavily on patient-reported outcomes Objective measures such as functional tests could provide more reliable data the case report does not delve deeply into potential etiological factors that could contribute to spontaneous rupture in the absence of identifiable risk factors A discussion on possible underlying mechanisms such as genetic predisposition or microscopic degenerative changes the report highlights areas for future research and emphasizes the need for more extensive studies to better understand and manage spontaneous plantar fascia ruptures This case report aims to increase awareness of spontaneous plantar fascia rupture to keep in mind in the differential diagnosis This case demonstrates that an integrated rehabilitative program can be highly effective in treating this pathological condition and lead to full functional recovery without the need for surgical intervention offering a practical framework for clinicians managing similar cases Continued research and documentation of such cases are essential to further validate and refine conservative treatment protocols for plantar fascia ruptures The raw data supporting the conclusions of this article will be made available by the authors Ethical approval was not required for this study involving humans because an ethics committee approval is not necessary for a case report in our legislation This study was conducted in accordance with the local legislation and institutional requirements The participant provided his written informed consent to participate in this study Written informed consent was obtained from the participant/patient(s) for the publication of this case report GL: Writing – review & editing The author(s) declare that no financial support was received for the research The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher A constitutive model for the mechanical characterization of the plantar fascia Regional differences in the mechanical properties of the plantar aponeurosis Plantar fasciitis and the windlass mechanism: a biomechanical link to clinical practice PubMed Abstract | Google Scholar Effects of plantar fascia stiffness on the biomechanical responses of the ankle-foot complex The role of arch compression and metatarsophalangeal joint dynamics in modulating plantar fascia strain in running Elastic energy within the human plantar aponeurosis contributes to arch shortening during the push-off phase of running Risk factors for plantar fasciitis: a matched case-control study PubMed Abstract | Crossref Full Text | Google Scholar Risk factors affecting chronic rupture of the plantar fascia Ruptures of the plantar fascia: a systematic review of the literature Rupture of plantar fascia: current standard of therapy: a systematic literature review An ossifying bridge—on the structural continuity between the achilles tendon and the plantar fascia Plantar fascia anatomy and its relationship with achilles tendon and paratenon Plantar fascia rupture: diagnosis and treatment PubMed Abstract | Crossref Full Text | Google Scholar Sequential bilateral rupture of the plantar fascia in a tennis player PubMed Abstract | Crossref Full Text | Google Scholar PubMed Abstract | Crossref Full Text | Google Scholar Ultrasound evaluation of a spontaneous plantar fascia rupture Acute rupture of the plantar fascia in a soccer player Ultrasound-guided collagen injections for treatment of plantar fasciopathy in runners: a pilot study and case series Vitale L and Logroscino G (2024) Spontaneous rupture of the plantar fascia: a case report Received: 24 July 2024; Accepted: 15 August 2024;Published: 27 August 2024 Copyright: © 2024 Venosa, Romanini, Vitale and Logroscino. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) distribution or reproduction in other forums is permitted provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited in accordance with accepted academic practice distribution or reproduction is permitted which does not comply with these terms *Correspondence: Lorenzo Vitale, bG9yZW56by52aXRhbGVAZ3JhZHVhdGUudW5pdmFxLml0 Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher 94% of researchers rate our articles as excellent or goodLearn more about the work of our research integrity team to safeguard the quality of each article we publish Metrics details The carnivorous gastropod Rapana venosa (Valenciennes 1846) is one of the most notorious ecological invaders worldwide we present the first high-quality chromosome-scale reference R venosa genome obtained via PacBio sequencing and high-throughput chromosome conformation capture scaffolding The assembled genome has a size of 2.30 Gb 77.22% of which were functionally annotated Given its high heterozygosity (1.41%) and large proportion of repeat sequences (57.72%) it is one of the most complex genome assemblies This chromosome-level genome assembly of R venosa is an important resource for understanding molluscan evolutionary adaption and provides a genetic basis for its biological invasion control which limits our understanding of the internal phylogeny and evolutionary adaption of this important clade such studies are hampered by the lack of related genomic resources From inner to outer layers: photograph of R Hi-C assembly of chromosome interactive heat map Abscissa and ordinate represent order of each bin on corresponding chromosome group Color block illuminates intensity of interaction from white (low) to red (high) Taxon-annotated GC-coverage plot (BlobPlot) of the contigs used for R plotted relative to its base coverage and GC proportion Circle diameter is proportional the size of the contig it represents Circles are colored according to their assigned taxon at the phylum level (see legend) Histograms show the distribution of the total assembly length along each axis The data analyses used standard bioinformatic tools specified in the methods Towards a phylogeny of gastropod molluscs: an analysis using morphological characters Molecular phylogenetics of Caenogastropoda (Gastropoda: Mollusca) A molecular phylogenetic framework for the Muricidae a diverse family of carnivorous gastropods The first Conus genome assembly reveals a primary genetic central dogma of conopeptides in C The mitochondrial genome of the venomous cone snail Conus consors Salinity tolerance of larval Rapana venosa: implications for dispersal and establishment of an invading predatory gastropod on the North American Atlantic coast Expression and activity of critical digestive enzymes during early larval development of the veined rapa whelk Observations on the biology of the Veined Rapa whelk 1846) (Mollusca: Muricidae): A new gastropod in South Atlantic waters Veined rapa whelk (Rapana venosa) range extensions in the Virginia waters of Chesapeake Bay Preliminary estimates of consumption rates of Rapana venosa (Gastropoda Muricidae); a new threat to mollusk biodiversity in the Rio de la Plata Occurrence of imposex and seasonal patterns of gametogenesis in the invading veined rapa whelk Rapana venosa from Chesapeake Bay Comparison of predation signatures left by Atlantic oyster drills (Urosalpinx cinerea Say Muricidae) and veined rapa whelks (Rapana venosa Valenciennes The alien mollusc Rapana venosa (Valenciennes Muricidae) in the northern Adriatic Sea: population structure and shell morphology Molecular response and developmental speculations in metamorphosis of the veined rapa whelk Symbiotic microbiome and metabolism profiles reveal the effects of induction by oysters on the metamorphosis of the carnivorous gastropod Rapana venosa Integrated mRNA and miRNA transcriptomic analysis reveals the response of Rapana venosa to the metamorphic inducer (juvenile oysters) Real-time DNA sequencing from single polymerase molecules A 3D map of the human genome at kilobase resolution reveals principles of chromatin looping Fast and accurate long-read assembly with wtdbg2 Pilon: an integrated tool for comprehensive microbial variant detection and genome assembly improvement Genome survey on invasive veined rapa whelk (Rapana venosa) and development of microsatellite loci on large scale The oyster genome reveals stress adaptation and complexity of shell formation Compatibility between snails and schistosomes: insights from new genetic resources The genome of the golden apple snail Pomacea canaliculata provides insight into stress tolerance and invasive adaptation Giant African snail genomes provide insights into molluscan whole-genome duplication and aquatic-terrestrial transition Genome and transcriptome mechanisms driving cephalopod evolution BUSCO applications from quality assessments to gene prediction and phylogenomics HiC-Pro: an optimized and flexible pipeline for Hi-C data processing Fast and accurate long-read alignment with Burrows-Wheeler transform HiCUP: pipeline for mapping and processing Hi-C data Juicer provides a one-click system for analyzing loop-resolution Hi-C experiments De novo assembly of the Aedes aegypti genome using Hi-C yields chromosome-length scaffolds Juicebox provides a visualization system for Hi-C contact maps with unlimited zoom RepeatModeler2 for automated genomic discovery of transposable element families LTR_FINDER: an efficient tool for the prediction of full-length LTR retrotransposons De novo identification of repeat families in large genomes Using RepeatMasker to identify repetitive elements in genomic sequences a database of repetitive elements in eukaryotic genomes Tandem repeats finder: a program to analyze DNA sequences Insights into bilaterian evolution from three spiralian genomes Complete DNA sequence of the mitochondrial genome of the sea-slug Aplysia californica: conservation of the gene order in Euthyneura High speed BLASTn: an accelerated MegaBLAST search tool TRNAscan-SE On-line: integrating search and context for analysis of transfer RNA genes Infernal 1.1: 100-fold faster RNA homology searches AUGUSTUS: ab initio prediction of alternative transcripts Systematic identification of novel protein domain families associated with nuclear functions HISAT: a fast spliced aligner with low memory requirements StringTie enables improved reconstruction of a transcriptome from RNA-seq reads NCBI sequence read archive. https://identifiers.org/ncbi/insdc.sra:SRR22889214 (2022) NCBI sequence read archive. https://identifiers.org/ncbi/insdc.sra:SRR23517974 (2022) NCBI sequence read archive. https://identifiers.org/ncbi/insdc.sra:SRR23501451 (2022) NCBI sequence read archive. https://identifiers.org/ncbi/insdc.sra:SRR23501452 (2022) NCBI sequence read archive. https://identifiers.org/ncbi/insdc.sra:SRR23501453 (2022) NCBI sequence read archive. https://identifiers.org/ncbi/insdc.sra:SRR23501454 (2022) Yang, M., Song, H. & Zhang, T. Rapana venosa breed wild species isolate MY-2022, whole genome shotgun sequencing project. GenBank https://identifiers.org/ncbi/insdc:JAQIHA000000000 (2023) Song, H. Annotations of Rapana venosa genome. Figshare. https://doi.org/10.6084/m9.figshare.22362598.v1 (2023) BlobTools: Interrogation of genome assemblies The genome of the venomous snail Lautoconus ventricosus sheds light on the origin of conotoxin diversity Improved Pairwise Alignment of Genomic DNA TreeFam: a curated database of phylogenetic trees of animal gene families MUSCLE: multiple sequence alignment with high accuracy and high throughput New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0 RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies MrBayes: bayesian inference of phylogenetic trees PAML 4: Phylogenetic analysis by maximum likelihood Download references This research was supported by the National Natural Science Foundation of China (Grant No the China Agriculture Research System of MOF and MARA and the Creative Team Project of the Laboratory for Marine Ecology and Environmental Science Qingdao National for Marine Science and Technology (Grant No Hao Song was supported by the Young Elite Scientists Sponsorship Program by CAST (Grant No 2021QNRC001) and Youth Innovation Promotion Association by CAS The funders had no role in the study design These authors contributed equally: Hao Song CAS Key Laboratory of Marine Ecology and Environmental Sciences Laboratory for Marine Ecology and Environmental Science Qingdao National Laboratory for Marine Science and Technology Research and Development Center for Efficient Utilization of Coastal Bioresources The authors’ contributions to specific working groups are indicated below Li: data processing and statistical analysis All authors have read and approved the final manuscript The authors declare no competing interests 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/s41597-023-02459-7 Anyone you share the following link with will be able to read this content: a shareable link is not currently available for this article Sign up for the Nature Briefing: Translational Research newsletter — top stories in biotechnology The Twirling Candy colt consigned as Hip 1062 in the ring at the OBS Spring Sale Steven Venosa of S G V Thoroughbreds ended the Ocala Breeders' Sales Spring 2-Year-Olds in Training Sale strong April 23 with a pair of six-figure juveniles topping off an overall successful week in Central Florida.  the colt was bred by George Bolton and Barry Lipman in Kentucky and purchased by pinhooking partnership Turning Hawk B.S.S for $165,000 from Eaton Sales at the Keeneland September Yearling Sale.  He was the seventh horse purchased by Red Baron's Barn and Rancho Temescal throughout the week Gross receipts for their purchases came to $1,915,000.  "A group of us selected the horse and we all liked the horse the colt was purchased by Vekuma Holdings for $350,000 He worked an eighth-mile in :20 3/5 during the under tack show.  Sign up for BloodHorse Daily Venosa said that Hip 996 held a special place in his consignment as he signed for the colt as a yearling for $40,000 from the Abbie Road Farm consignment to the Ocala Breeders' Sales October Yearlings Sale Venosa signed the ticket in the name of Big D stable in whose name he pinhooks one horse every year.  "Anytime I lead a horse up here and sell one how could you not be happy?" said Venosa "It was the only horse I came here to buy My son's name is Dylan and every year I buy a horse for him When I go home and do the cards at night he sits there and I tell him the numbers That horse was Hip 996 and I would read the cards and say 'That's my horse!' and his horse had the most shows Dad!' I called him when it sold and he said 'That's my horse!' "  Photo: Joe DiOrioSteve Venosa (left) and George Weaver at the OBS Spring Sale Venosa said he has been pleased to see an overall increase in market depth with horses being traded at all levels of the game.  Obviously we sold him (Hip 1062) really well but we also sold one for $15,000 It's good to see that there are people there The other horses are kind of struggling a little bit but it seems like it's starting to get better The middle market—there are some people there It's nice that the racetracks are starting to let fans back in has completed its first bunkering operations with Eastern Pacific Shipping’s 210,000 dwt bulker Mount Tai the event also marked a milestone for Eastern Pacific Shipping since the bunkering is 100th LNG bunkering operation in Singapore Like all EPS-managed LNG dual fuel vessels which is chartered on consecutive voyages with BHP is equipped with ME-GI two-stroke engines that have negligible levels of methane slip The vessel will be a part of EPS’ low-carbon emission fleet transporting iron ore along the green corridor from West Australia to Northeast Asia “Working with like-minded customers like EPS is crucial in accelerating the energy transition Shipping companies can meet decarbonisation targets until around 2030 to 2035 Their progress toward net zero will be further improved when bio-LNG and ultimately renewable synthetic LNG are available for blending,” Tahir Faruqui head of ShelldownstreamLNG said FueLNG Venossa recently finished its inaugural LNG loading operations through ship-to-ship (STS) transfer FueLNG christened its newest LNG bunker vessel FueLNG Venosa at a ceremony held at Hyundai Mipo Dockyard in Ulsan Chartered from Korea Line LNG, the LNG bunker vessel has a total capacity of 18,000 cbm and is expected to bring significant economies of scale. The ship is designed to facilitate safe and quick turnaround of vessels carrying out simultaneous cargo handling and bunkering operations; and is capable of bunkering different types of LNG fuel tanks. Besides bunkering operations, FueLNG Venosa will provide gas-up and cool-down services to LNG carriers and LNG-fueled vessels after dry docking in Singapore or en route to loading operations. “Combining Singapore’s strategic location and supportive infrastructure with our expertise and partnerships, we are working towards making Singapore a prominent LNG bunkering hub,” FueLNG highlighted. Daily news and in-depth stories in your inbox Ingersoll Rand Engineering Project Solutions At Ingersoll Rand’s Engineering Project Solutions, we have been managing and implementing engineered to-order air packages for complex technical requirements for over 60 years. We provide specialized custom compressed air and gas compressors as well as nitrogen generation packages to international EPC contractors and engineering companies across a range of […] Volume 7 - 2020 | https://doi.org/10.3389/fmed.2020.606462 This article is part of the Research TopicFrom Development to Senescence, Bridging the Gap in Lung FibrosisView all 8 articles chronic pulmonary pathologies represent the third leading cause of death in the elderly population Evidence-based projections suggest that >65 (years old) individuals will account for approximately a quarter of the world population before the turn of the century are described as the nine “hallmarks” that govern cellular fitness Any deviation from the normal pattern initiates a complex cascade of events culminating to a disease state originally employed to describe aberrant changes in cancer cells can be also used to describe aging and fibrosis Pulmonary fibrosis (PF) is the result of a progressive decline in injury resolution processes stemming from endogenous (physiological decline or somatic mutations) or exogenous stress dietary or occupational exposure accelerates the pathogenesis of a senescent phenotype based on (1) window of exposure; (2) dose recurrence; and (3) cells type being targeted the threshold to generate an irreversibly senescent phenotype is lowered we do not have sufficient knowledge to make accurate predictions we provide an assessment of the literature that interrogates lung epithelial and immune senescence at the intersection of aging environmental exposure and pulmonary fibrosis As the medical field battles the COVID-19 global pandemic, scientists and clinicians have come to terms with the notion that we know very little of the mechanisms mediating lung injury and resolution in the aged and susceptible respiratory system (14–16) this manuscript summarizes the evidence linking lung injury and remodeling in the context of biological aging and chemical exposure The nine “hallmarks” of cell fitness Healthy cells rely on a balance between survival checkpoints and aberrant hallmarks A total of nine factors have been defined: genomic instability and altered intercellular communication (represented by circles in matching colors) Skewing of this equilibrium generates a progressively senescent phenotype as a result of age-related checkpoint dysfunction driving cellular toxicity and heavy metals exposure disrupts unfolded protein response Nutrient utilization is then shifted toward glycolysis leading to: (1) Enhanced AMPK signaling driven by excess AMP resulting in proliferative latency and pRB/p16 pathway activation (2) Excess pyruvate enters mitochondria for conversion into AcCoA; this leads to mitochondria ROS generation and toxic cell acidification (3) Oxidative stress from mitochondria damages cytoplasmic proteins and DNA; AcCoA functions as a acetyl donor for localized histone hyperacetylation while histone chaperones Asf1 and HIRA greatly remodels chromatin independently of replicative state of the cell Gene expression resulting from chromatin reprogram results in senescent associated secretory phenotype (SASP) In light of this evidence, any therapeutic that effectively reduces numbers or activity of senescent cells has significant healthcare potential. While technological advancements produced a broad arsenal of pharmacological moieties, their efficacy against age- and fibrotic-related senescence has been hindered by the paucity of models that mimic the clinical course of disease (42–44) clinically relevant models of chemical induced fibrosis are limited by intrinsic differences among the thousands of environmental or occupational stressors the lung comes into contact daily To overcome these limitations and provide adequate prediction of how do environmental exposures that lead to cellular senescence lead to different lung pathologies it is absolutely necessary to test chemical exposure in aged from the aforementioned TERT and SP-C mutant mice In-depth analysis of the fibrogenic effects of environmental exposure on the senescent lung is critical to advance the field and better address the needs of susceptible populations At least three aspects need to be considered: window of exposure; dose duration and recurrence of exposure/injury; cell type-specific responses Modeling chemical exposure on the fibrotic phenotype Depiction of possible outcomes resulting from aging and chemical exposure and their relationship to fibrogenesis ) may drive rapid and possibly lethal fibrosis after a single exposure aging may lead to different disease profiles based on factor such as genetic instability (i.e. an individual presenting somatic mutations may be predisposed to develop a fibrotic phenotype without toxic challenge (2a Similar responses can be observed following mild/moderate repeated exposure ) passing the “fibrogenic threshold" ) or never reach that threshold depending on factor such as genetic susceptibility aging may be associated with fibrotic and non-fibrotic outcomes depending on individual biological clocks While to date is still unclear what effects environmental exposure elicits on such susceptible parenchyma work from our group and others intends to fill this knowledge gap In what seems like a prohibitive task (investigating the effects of hundreds of chemically diverse moieties on dozens of cell types) technological advancements in single cell sequencing analysis and multi-omics approaches significantly eased these challenges The next section will summarize epithelial mesenchymal and immune cell senescence induced by aging or chemical exposure Modeling cell specific responses to chemical exposure Simplified depiction of the divergent effects of particular matter exposure in immune epithelial (inclusive of alveolar and mucus producing goblet cells) and mesenchymal (fibroblasts and myofibroblasts) The paucity of data that comprehensively examines the responses of a dysfunctional lung epithelium to environmental challenge blurs our ability to determine the mechanistic overlap preceding fibrogenesis chemical challenge has been predominantly examined in juvenile/healthy lungs While these studies helped defining senescent-like phenotype following exposure Addressing each of these questions may advance out understanding of senescence By combining this observational evidence with the currently available pulmonary disease models the next decade of pulmonary research has important clinical implications to our pursuit of mechanistic answers of sublethal toxicity in the susceptible lung Justification of these differences may provide important insights to our understanding of both cellular conditions It is fundamental to recognize that the behavior of immune cell subtypes is unique and often dependent on the surrounding environment The failure of broad-spectrum immunomodulatory therapy in PF is a reminder of this The next section summarizes the current knowledge of age- and chemical-induced senescence on a cell by cell basis Phenotypic characterization of nine prototypical macrophage populations including M1 the authors bring up significant points related to the differential responses of lung resident AMs and MoAMs to a second challenge This (and other) work could reshape how we view macrophage biology A more complete understanding of monocyte biology in the context of aging and fibrosis could help identify unique signatures to define monocyte-derived alveolar macrophages and even achieve selective targeting during the fibrogenic process Phenotypic characterization of peripheral and tissue resident lymphocyte populations including CD4 and unconventional T cells (invariant natural killer T cells iNKT); CD8αα+ cells; mucosal-associated invariant T cells MAIT; γδ T cells; and intestinal intraepithelial lymphocytes While identification of these tissue resident lymphocytes is progressing fairly rapidly thanks to single-cell techniques analysis of their roles in specific disease states still at its infancy further functional examination of these cells in the context of a senescent state (aging and fibrosis) represents a promising strategy to advance the field and perhaps therapy This review exposes the wealth of evidence that pertains to aging While demonstrating some degree of mechanistic redundancy across the spectrum of senescence it also highlights a number of knowledge gaps that need to be addressed to impact human health (i.e. the next cycle of research questions should test: (1) whether senescence of the remodeled/fibrotic lung occurs faster and through the same mechanisms as an architecturally pristine one (2) Since it is understood that age-related dysfunction lowers the threshold necessary to trigger an irreversible senescent phenotype can we model and accurately predict such levels (3a) Can we identify mutual factors involved in senescence across the spectrum of chemical exposure (3b) Can we identify shared and exclusive factors that drive senescence across the lung disease spectrum how does PF senescence compare to that observed in COPD (4) Can we modulate/reprogram the behavior and communication of specific cell types (5) Can senolytics modulate seemingly irreversible changes in the fibrotic (and aging) lung or their imbalance during menopause and andropause support or protects the lung from exogenous stressors and disease is not well-understood This is likely the most important puzzle piece to understand clinical datasets there is a canyon-sized knowledge gap in front of us Although we are a long way away from getting all the answers it is comforting to see an increasingly collaborative scientific community and frequent technological advancements that help us comprehensively study cell biology The successes of the next decade of research lies in good hands This work was supported by the University of Utah Center on Aging Pilot Grant The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest CrossRef Full Text | Google Scholar PubMed Abstract | CrossRef Full Text | Google Scholar Idiopathic pulmonary fibrosis: pathogenesis and management Idiopathic pulmonary fibrosis in US Medicare beneficiaries 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This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) *Correspondence: Alessandro Venosa, YWxlc3NhbmRyby52ZW5vc2FAcGhhcm0udXRhaC5lZHU= Metrics details The present study evaluated the structural and ultrastructural characteristics of Rapana venosa egg capsules starting from observations of their antifouling activity and mechanical resistance to water currents in mid-shore habitats and electron microscopy were used to evaluate the surface and structure of the R These measurements revealed an internal multilamellar structure of the capsule wall with in-plane distributions of layers with various orientations It was found that the walls contained vacuolar structures in the median layer which provided the particular characteristics viscoelastic and swelling measurements were also carried out This study revealed the presence and distribution of chitosan in the capsule of R Chitosan identification in the egg capsule wall structure was carried out through SEM–EDX measurements The biopolymer presence in the capsule walls may explain the properties of their surfaces as well as the mechanical resistance of the capsule and its resistance to chemical variations in the living environment Independent evolution also results in adaptive solutions being often dissimilar among various phylogroups knowledge concerning the mechanisms of adaptation in one phylogroup cannot be simply extrapolated to other groups This explains our interest in the particular adaptations of the egg capsule structure and composition in R venosa egg capsules were analysed starting from observations of their antifouling activity and mechanical resistance to currents in mid-shore areas venosa capsules with the aim of better understanding their structural and compositional adaptations This information can shed light on the reproductive biology and remarkable hardiness of this invasive species The novelty and importance of this study also arises from elucidating the ultrastructural composition of the capsules which impacts the taxonomy and the evolution of mollusc species or of biomaterials models (a) General appearance of the egg capsules; scale bar = 2 mm; (b) The capsule surface features; the small image shows the observed autofluorescence; scale bar = 10 μm; (c) The edge of the transverse section stained with acridine orange; scale bar = 5 μm (d) Lamellar layer (Lm) array in the section line; scale bar = 1 μm Autofluorescence was observed when microscopic studies at an excitation wavelength of 450–480 nm with an emission wavelength of 515–520 nm (green filter) were carried out to evaluate the structure (Fig. 1b) Rapana venosa egg capsule transverse section areas and structural characteristics analysed using various staining procedures; (a) PAS staining; scale bar = 10 µm vacuoles (Vc); scale bar = 10 µm; (c) H&E staining; scale bar = 10 µm (Lm 1 = superficial external layer; Lm 2 = internal layer) The observed exfoliated superficial material on the outer side of the capsule surface (Fig. 2c) can be explained by the lamellar structure of the superficial layer By combining these staining techniques, the thickness of the layers and the changes in the ratio between the Vc layer and Lm layers were easier to observe and distinguish (Fig. 2b,c) The folds shown in the egg capsule were also investigated at higher magnification by SEM (Fig. 3a). The distance between the folds was between 15 and 30 μm, while the thickness of these folds was between 10 and 12 μm (Fig. 3b). Structural and ultrastructural features of R venosa egg capsules; (a) SEM image of the capsule; scale bar = 500 µm; (b) Surface capsule details SEM image; scale bar = 50 µm; (c) Details of the capsule wall ultrastructure (cross-section TEM image) a lamellar system with striations (arrow); scale bar = 500 nm; (d) Ultrastructural details with parallel oblique lamellar orientation (Lm) and ultra-lamellar layers (ULm); scale bar = 20 nm Details are shown in a high-resolution image (Fig. 3d) in which the size of the fringe width was 1.22 nm The image shows that the microfibrils are arranged in a highly organized parallel crystalline structure venosa egg capsules: (a) storage and loss; (b) tang δ as a function of temperature; (c) swelling degree versus immersion time; (d) tensile stress behaviour The response of the material to a tensile stress at a constant rate (0.1 mm/min) showed the typical behaviour of plastic materials with high flexibility, suggesting reversible deformations slightly greater than 50% (Fig. 4d) which corresponds to the destruction of macroscopic and microscopic integrity The value of the Young modulus estimated in the initial linear area (0.25–0.05% elongation) was 0.025 MPa indicative of a high stability of the biomaterial noticeable in the sequential analysis of the mechanical curve is that the tensile fluctuations occurred with maxima and minima The only explanation for this observation is a molecular reorganization at the superstructure level The interpretation of this phenomenon (highlighted, in detail, in Fig. 4d) consisted of a cascade of local “micro-breaks” occurring under elongation which were not attributed to the destruction of chemical structural bonds but to the destruction of bonds at the supramolecular level – in particular physical bonds Any “rupture” induced the formation of other interactions in the immediate vicinity supported by the chemical nature of the surrounding environment in this material with protein consistency This cascade of local “breaks” and reorganizations allows the material to take over the mechanical effort using it as a source of regrouping and providing flexibility to the ensemble The apparent material elasticity was given by an elongation up to 50% this fluctuation was attributed to the capacity and sensitivity of the load cell used to capture the tensile load but when these results were correlated with the DMA results it was concluded that a cascade-type mechanism occurred in this case in which the local “micro-breaks” occurring under elongation were not the result of chemical bond destruction but resulted from a destruction of bonds at the supramolecular level—in particular physical bonds—that determine the crosslinking between the protein and glucosides phases This conclusion is also supported by the fact that the tested material is a natural material and not two phases (separately) ensuring the thermodynamic stability of the assembly This thermodynamic stability was observed in the structural level by the minimum value of tensile stress This phenomenon propagated throughout the test range with corresponding elongation shifts until an energy threshold was reached which destroyed the intermolecular structuring bonds The correlation between the tensile stress and the DMA results was attributed to the contribution of the rotational movements of the absorbed water molecules to the rheological changes reported in the material and was observed when a swollen sample was tested this material showed flexibility in its dry state and the flexibility increased with increasing degree of hydration The quantitative distribution of carbon, nitrogen, oxygen, sodium, magnesium, silicon, chloride, sulfur and calcium elements was evaluated on the surface of the studied capsules using SEM with energy dispersive X-ray (EDX) microanalysis (Fig. 5). Element distribution in the capsular surface of Rapana venosa using SEM–EDX microanalysis The highest average values were identified for carbon with 51.12 wt%, nitrogen (11.01%) and oxygen with 22.83 wt% (as main components), as shown in Table 1 the following components were identified: soluble proteins (46.66 ± 1.00%) chitosan (36.94%) and other components probably consisting of insoluble compounds (15.20 ± 0.80%) The existence of chitosan in the solid sample (chitosan assay I) was confirmed; as a result, this polymer was found in the structure of the capsules with an average quantity of 11.65 ± 0.60% in the superficial layers (Table 2) The analysis of solutions obtained after deproteinization (chitosan assay II) revealed a chitosan content of 25.29 ± 0.40% λem = 515–520 nm) and red (λex = 510–550 nm; λem = 580–590 nm) The same effect was observed for granular chitosan obtained from these capsules (after deproteinization) A more careful evaluation of the evolution of the three dynamic-mechanical analysis (DMA) parameters (Fig. 4a,b) revealed that the increase in temperature was accompanied by a continuous decrease in the storage modulus while the loss modulus and the loss factor (tang δ) exhibited three and two peaks the first peak was located in the range 16–111 °C at approximately 54 °C and the second peak (111–135 °C) showed a maximum at 114 °C The first peak represented the phase separation observed in the DMA study The first peak in the tang δ curve was broad and centred at 54 °C This peak represented the peak observed in the loss curve at 45 °C which was shifted it overlapped with the peak observed at 35.5 °C in the loss modulus curve the peak at 112 °C can be attributed to the α-transition corresponding to chitosan Tg The chain mobility of chitosan also increased because of a decrease in the intermolecular interactions due to dissociation of hydrogen bonds which started the process of molecular chain scission due to the breaking of glycosidic bonds height and width depend on the moisture content Peaks related to these main elements and a similar distribution of the elements were reported in the study We believe that the formation of this type of chitosan is possible at the level of gonoduct structures at the time of capsule deposition in the marine environment our hypothesis supports the possible transformation of chitin into chitosan through chitin deacetylase enzyme activity at the gonoduct level to provide specific flexibility for capsule formation venosa egg capsules that were analysed in this study at the microstructural level showed complexity in terms of the filament layout and the microfibrillar composition; furthermore the presence of chitinous structures was confirmed antifouling and optical) could explain the reproductive efficiency and the spread of the species with remarkable adaptations of this organism capsule fragments were processed as follows: prefixation in 1 M cacodylate buffer solution with 2.7% glutaraldehyde (GA) followed by fixation in 2% osmic acid solution (in small glass tubes containing 2 mL of cold fixation medium consisting of 2% (w/v) osmic acid (OsO4) dissolved in twice-distilled water) The fixation was carried out for 1 h at a temperature of 4 °C 50% and 70% cold ethanolic solutions were used The samples were maintained in each of the ethanolic baths for 10 min the samples were placed in a mixture of Epon 812 (consisting of DDSA MNA and DMP30 as polymerizing agent) and propylene oxide at an Epon 812:propylene oxide ratio of 1:1 (v/v) The fine sections were double-stained with uranyl acetate and lead citrate and lead acetate solutions and then examined by SEM and TEM A scanning electron microscope (SEM FEI QUANTA 250 equipped with an energy dispersive spectrometer) and a transmission electron microscope (TEM-Microscope Tecnai T12) were used Optical microscopy observations were performed with an epifluorescence microscope (OPTIKA B – 350); blue filter (λex = 450–490 nm; λem = 515–520 nm); green filter (λex = 510–550 nm; λem = 590 nm) The captured images were processed with Optikam Pro 3 Software Mechanical properties were determined with an Instron 3382 Universal Testing Machine with video extensometer and the result was the average of five tests for each specimen The tests were carried out at room temperature with a crosshead speed of 50 mm/min and the samples were previously hydrated in demineralized water for 24 h All measurements were performed at room temperature and the tensile strength (TS) and ductility (percent elongation EL%) of the films were calculated using the following equations: Dynamic-mechanical analysis (DMA) tests were carried out with TRITEC 2000 B equipment (Triton Technology The dried samples were analysed in single cantilever bending mode at 1 Hz frequency The data were collected from room temperature (RT) to 180 °C at a heating rate of 5 °C/min The dry samples after washing had the following dimensions: length The obtained samples were washed to remove solid impurities (traces of sands) No surface-finishing technique was employed Fractions of different egg capsules were mixed and the measurements were performed on samples that were randomly chosen These samples were further handled in dry nitrogen as much as possible to reduce the chance of moisture reducing the glass transition temperature and Young’s modulus are parameters that relate the mechanical properties of the samples to their chemical structures The tensile strength expresses the maximum stress in a sample during tensile testing dynamic-mechanical thermal analysis (DMA) of polymeric materials is a very important technique due to its high sensitivity in detecting changes in the internal molecular mobility and in probing the phase structure and morphology of polymers of polymer blends are sensitive not only to different molecular motions but also to various transitions extent of crosslinking and the morphology of multiphase systems The swelling experiments consisted of immersing one dried sample of approximately 9.6 mg into 500 mg of water and measuring the mass of the sample at specific time points The experiment was performed in triplicate The swelling degree was calculated according to the equation: where Wd is the mass of the dried sample and Ws is the mass of the swollen sample The elemental analysis of the capsular surface was performed by the SEM–EDX (ADC1 detector at 182 points copper (II) sulfate-anhydrous powder (Merck KGaA sodium carbonate-anhydrous powder (Merck KGaA sodium potassium tartrate tetrahydrate powder (Spectrum Chemical MFG CORP) Folin-Ciocalteu phenol reagent (Merck KGaA Germany) and bovine serum albumin-lyophilized powder (Sigma Aldrich The data analysis and processing were performed according to a modified Lowry protocol65 The capsules were milled and 0.6183 g were added to alkaline extractive solutions 1:40 (w/v) for 4 h the alkaline supernatant was recovered and subjected to quantitation analyses of the soluble proteins according to the Lowry procedure Sample dilutions of 1:10 (SD1) and 1:100 (SD2) (v/v) of the alkaline supernatant and distilled water were prepared Volumes of 0.1 mL from these dilutions SD1 and SD2 were taken and mixed in Eppendorf tubes with the amounts of reagents according to the Lowry procedure The samples were measured at 750 nm using a Jasco V-630 spectrophotometer The measurements were performed in triplicate The ash content (Ash) of the egg capsules was determined using MICROTERM 1206 To determine the ash content from the egg capsule samples dried and fine-cut egg capsules was placed into previously ignited The ash content (%) of samples was determined at 800 °C in triplicate using the following equation: A chitosan colorimetric assay kit (K995-100) BioVision This is the first commercial kit available for measuring chitosan content from both solid and liquid samples The capsules were washed with distilled water and introduced into chitosan analysis solutions following the analysis procedure for solid samples (chitosan assay I—solid sample) according to the determination kit the presence of chitosan was tested after deproteinization (chitosan assay II—liquid sample) using the protocol for liquid samples (K995-100) in extraction solutions An Ostwald-type capillary viscometer was used for these measurements a VELP—Digital IR Vortex mixer and a centrifuge were used Epifluorescence microscopy analyses were used for particle observation The species Rapana venosa does not have a legally established protection status at the national or European level and therefore is not subject to the procedure for approving the derogation from the protection measures for scientific purposes considering that very small quantities of non-living parts which can be classified as animal remains without value this study did not require any authorization procedure by the competent bodies The Black Sea ecosystem changes related to the 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LSU Master's Theses 1338 https://digitalcommons.lsu.edu/gradschool_theses/1338 (2004) Download references help and useful discussion on different specific aspects of this paper Costel Coroban for proofreading the final version of this manuscript The authors thank the scuba divers from Midia Marine Terminal for helping to collect the analysed samples Department of Cellular and Molecular Biology Faculty of Applied Chemistry and Materials Science Electron Microscopy Department and Center for Research and Development of the Morphological and Genetic Studies of Malignant Pathology “Vasile Goldiș” Western University From Arad Department of Naval and Port Engineering and Management carried out the analyses (chitosan extraction mechanical properties) and wrote the paper C.C.P was in charge of the spectral analyses All authors approved the final version of the manuscript and agree to be held accountable for the content therein Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations Download citation DOI: https://doi.org/10.1038/s41598-020-71348-5 Sorry, a shareable link is not currently available for this article. 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Volume 10 - 2023 | https://doi.org/10.3389/fmars.2023.1122668 metamorphosis controls the population dynamics of most marine invertebrates and affects the breeding of economic shellfish Rapana venosa is an economically important species in China but artificial aquaculture has hampered its metamorphosis process Previous studies have found that juvenile oysters can effectively induce the metamorphosis of R but the specific induction mechanism is not clear we investigated the mechanism underlying the response of R venosa to juvenile oysters through the RNA-seq analysis the gene set responses to metamorphosis cues (juvenile oysters) in R and GO and KEGG enrichment analyses were further performed on these gene sets The results showed that the expression of the prototype of the class of immediate early genes exhibited lower expression in the M12 group than in the control group the expression of inhibitors of apoptosis (IAPs) was significantly increased upon exposure to juvenile oysters the Wnt signaling pathway and MAPK signaling pathway were enriched in the trend analysis These pathways may also play critical regulatory roles in the response to juvenile oysters the results show that competent larvae rapidly respond to the inducing effects of oysters via some immediate early genes which may further regulate downstream pathways such as the MAPK signaling pathway to cause subsequent changes including a decrease in HSP90 and an increase in IAPs These changes together may regulate the metamorphosis of R This study provides further evidence that juvenile oysters are the metamorphosis cues of R which may enhance our understanding of the metamorphosis mechanism in this marine invertebrate This problem has also attracted widespread concern the invasion mechanism urgently needs to be elucidated This low survival rate severely restricts the development of this industry This may partly explain why these mollusks are destroying oyster resources the specific mechanism by which oysters induce metamorphosis of R a comprehensive analysis of the metamorphosis regulation mechanism in R Transcriptomics has been widely applied to investigate the mechanisms underlying metamorphosis in invertebrates, revealing the critical pathways and genes that regulate metamorphosis and providing novel insights (Shikuma et al., 2016; He et al., 2021) to further investigate the mechanism by which juvenile oysters induce the metamorphosis of R we obtained the mRNA expression profiles of competent larvae exposed to juvenile oysters for 2 hours (M2) and 12 hours (M12) and before exposure to juvenile oysters (control Con) via sequencing on an Illumina NovaSeq 6000 platform we identified the differentially expressed mRNAs (differentially expressed genes DEGs) and performed trend analysis of gene expression based on their transcripts per million (TPM) values to further screen out the genes that were consistently regulated during exposure to juvenile oysters these genes were subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses The findings of this study will enhance understanding of the metamorphosis mechanism of R which may not only aid in the development of artificial breeding and culturing but also reveal the invasion mechanism and support the protection of wild populations of oysters and other bivalves Culturing of R. venosa larvae and sample collection were performed according to Yang et al. (2022) and the competent larvae (shell height > 1,200 μm) were used to conduct the assay Three cement pools (3.5 × 5.2 × 1.5 m) were used to culture these larvae and the assay was performed with a larval density of 0.1 ind/mL we randomly collected 100 larvae from each pool as the Con group the juvenile oysters (Crassostrea gigas) were placed evenly in pools (50 oysters on each scallop added about 25000 scallop shells to each pool) and 100 larvae were collected at 2 and 12 hours thereafter from each pool as the oyster-exposed groups (M2 and M12 During the whole process of experiment and sampling the three pools were closed with no water in or out and maintain a stable experimental conditions (23 ± 1°C A total of nine samples were obtained from the Con which were washed with PBS and stored at -80°C for RNA extraction Total RNA was extracted from the larval samples using TRIzol® Reagent (Invitrogen USA) according the manufacturer’s recommendations and 1% agarose gels were used to monitor the degradation and contamination of RNA The integrity and quantification of RNA samples were evaluated using a 2100 Bioanalyzer (Agilent Technologies and RNA samples with integrity numbers (RINs) > 8.0 were used for construction of an mRNA library The library was prepared using a TruSeq™ RNA Sample Preparation Kit (Illumina USA) and sequenced on an Illumina NovaSeq 6000 platform (Majorbio and 125-bp paired-end reads were generated and deposited in the Sequence Read Archive (ncbi.nlm.nih.gov/sra) under BioProject PRJNA855327 The raw reads were trimmed and quality-controlled via SeqPrep and Sickle with default parameters, and the clean reads were separately aligned to the reference genome of R. venosa (unpublished data) with orientation mode using HISAT2 software (Kim et al., 2015). The mapped reads of each sample were assembled by StringTie via a reference-based approach (Pertea et al., 2015) and then the paired-end clean reads were aligned to the reference genome of R venosa (unpublished data) using TopHat v2.0.12 HTSeq v0.6.1 was used to count the reads mapped to each gene Known genes and novel transcripts were identified by Reference Annotation Based Transcript (RABT) assembly method the TPM of each gene was calculated based on the length of the gene and the number of reads mapped to the gene DEG analysis was conducted among the three groups and the TPM algorithm was used to normalize the expression level of each transcript DEG analysis was performed using the DESeq2 R package with the significance thresholds set to a P-adjusted value < 0.05 and a |log2FC| ≥ 1 after filtering out the genes with TPM = 0 and normalizing the TPM by log2 transformation venosa transcriptome data was performed on the Majorbio Cloud Platform by Short Time-series Expression Miner (STEM) software To further explore the biological functions of the DEGs and these genes with significant trends GO and KEGG enrichment analyses were performed GO enrichment analysis was implemented using the GOseq package in R and KOBAS was used to analyze the statistical enrichment of genes in KEGG pathways GO terms and KEGG pathways with the significance enrichment threshold set to a P-adjusted value < 0.05 corrected by the Bonferroni method To validate the accuracy of the RNA-Seq profiling results, critical genes were selected for qRT−PCR analysis. The cDNA for qRT−PCR was synthesized using the Prime Script™ RT Reagent Kit with gDNA Eraser (TaKaRa, Japan). The primers used in the mRNA qRT−PCR assay were designed using Primer 5, and 60S ribosomal protein L28 (RL28) was selected as a housekeeping gene to normalize the data (Song et al., 2017) A SYBR PrimeScript RT−PCR Kit II (TaKaRa Japan) was used to quantify the expression levels The relative expression levels of genes were estimated by the 2−ΔΔCT method All data are presented as the means ± SEs (N = 3) Statistical significance was analyzed using SPSS v.19 with a P value < 0.05 considered to indicate significance the transcriptome data were consistent with the qRT-PCR results Table 2 Statistics of gene annotation Figure 1 Validation of the 9 random DEGs involved in different pairwise comparisons by qRT-PCR RL28 was selected to normalize the gene expression levels The data are shown as means ( ± SE) of three replicates Different superscripted letters indicate significant differences (P < 0.05) Hierarchical clustering analysis (HCA) showed that the groups exposed to oysters for 12 hours (M12) were clustered as one branch, while the group exposed to oysters for 2 hours (M2) was clustered with the Con group (Figure 2A). Additionally, principal component analysis (PCA) showed that all three groups were significantly separated from each other (Figure 2B) These results may mean that exposure to oysters has a significant effect on the mRNA profile of R venosa larvae and that the effect increases with time Figure 2 Relationships analysis between samples: Hierarchical clustering analysis (A) and principal component analysis (B) Figure 3 Statistics of differentially expressed genes (A) and Venn diagram of differentially expressed genes (B) The 189 DEGs in the Con vs. M2 comparison were regarded as the initial-response genes (I-DEGs) to oyster presence, which were significantly enriched with 2 GO terms: DNA-binding transcription factor activity and transcription regular activity (Figure 4A). There were no significantly enriched pathways (P adjust < 0.05). We further analyzed the expression of these genes enriched with the 2 GO terms. As shown in Figure 4B 12 genes were upregulated in the M2 group compared with the Con group including some nuclear receptor genes (nuclear receptor subfamily 1 and nuclear hormone receptor E75) CCAAT/enhancer-binding protein beta and transforming protein v-Fos/v-Fox Five other genes were downregulated in the M2 group deformed epidermal autoregulatory factor 1 and nuclear factor interleukin-3-regulated protein Figure 4 The functional enrichment and expression analysis of initial-response genes to oyster induction (A) GO enrichment results of 92 DEGs in Con vs (B) Heat map of the expression analysis of 17 DEGs in GO:0003700 (DNA-binding transcription factor activity) and GO:0140110 (transcription regulator activity) Figure 5 The functional enrichment and expression analysis of final-response genes to oyster induction (A) KEGG enrichment results of 597 DEGs in Con vs (B) GO enrichment results of 597 DEGs in Con vs (C) Heat map of the expression analysis of 36 DEGs in the enriched pathways: “Ribosome biogenesis in eukaryotes” “Antigen processing and presentation” and “RNA transport” Figure 6 The functional enrichment and expression analysis of late-response genes to oyster induction (A) GO enrichment results of 397 DEGs in M2 vs (B) Heat map of the expression analysis of 14 DEGs in GO:0016072 (rRNA metabolic process) and GO:0034660 (ncRNA metabolic process) autophagy - animal (18 genes with adjusted P < 0.05) and no significantly enriched GO term was identified there were also no significantly enriched GO terms or pathways we analyzed the expression of the genes in the Wnt signaling pathway and animal autophagy Figure 7 Trend analysis of genes in R (A) Different expression patterns of all genes The upper number in the box represents the serial number of profiles and the lower one is the P value GO (B) and KEGG (C) enrichment analysis of genes in profile 2 The food or prey of some animals not only provides them with the necessary nutrients for growth and development but also plays an important role in their development at a critical time. One of the most dramatic examples is when the food or prey induces animal metamorphosis (Morse et al., 1979; Rodriguez et al., 1993; Taris et al., 2010) Although representative species from diverse taxa undergo food-induced metamorphosis little is known about the processes that occur within the animals as they detect the cues produced by their food To further understand the mechanism by which juvenile oysters induce the metamorphosis of R we performed RNA-seq and analyzed changes in gene expression at different time points upon exposure to metamorphosis cues we conducted functional enrichment for these genes and identified the critical signaling pathways in the process The present results may provide new insights into the regulatory mechanisms of metamorphosis in marine invertebrates we speculate that the transcription factor AP-1 may be a critical initiator in competent larvae of R venosa in response to the metamorphosis cues of juvenile oysters that regulates other subsequent processes we also found that the MAPK signaling pathway was significantly enriched in the gene set of profile 2 in the trend analysis which may mean that in competent larvae of R MAPK signaling participates in the response to metamorphosis cues which might be considered to be a signal of metamorphosis induced by the oysters but the specific function of HSP90 needs further investigation we speculate that HSP70 may also participate in the MAPK signaling pathway and be related to the metamorphosis of R the specific effect of HSP70 on metamorphosis needs further investigation especially since no previous studies have reported the relationship between HSP70 and metamorphosis in marine invertebrates the specific function of the Wnt signaling pathway in the metamorphosis of marine invertebrates needs further investigation venosa (I-DEGs and F-DEGs) were identified We found that DNA-binding transcription factor activity and transcription regulator activity were enriched for the I-DEGs which is a prototypical immediate early gene that rapidly and transiently activates transcription in response to changes in environmental conditions exhibited lower expression in the M12 group than in the Con group while the expression of IAPs was significantly increased upon exposure to juvenile oysters the results show that competent larvae rapidly respond to the presence of oysters by expressing some immediate early genes These genes may further regulate downstream pathways such as the MAPK signaling pathway and result in subsequent changes These changes together may regulate the metamorphosis of the larvae The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/Supplementary Material The studies involving animals were reviewed and approved by the Animal Welfare Committee of the Institute of Oceanology TZ and HS conceived and designed the experiments All authors contributed to the article and approved the submitted version 42206086 & 31972814 & 32002409 & 32002374) the China Postdoctoral Science Foundation (Grant No the Young Elite Scientists Sponsorship Program by CAST (Grant No the ‘Double Hundred’ Blue Industry Leader Team of Yantai (Recipient: Tao Zhang) Qingdao National for Marine Science and Technology (no decision to publish or preparation of the manuscript The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fmars.2023.1122668/full#supplementary-material The multi-gene family of transcription factor AP-1 CrossRef Full Text | Google Scholar regulation of metamorphosis in ascidians involves NO/cGMP signaling and HSP90 NO/cGMP signaling and HSP90 activity represses metamorphosis in the sea urchin lytechinus pictus The influence of bacteria on animal metamorphosis ERK- and JNK signalling regulate gene networks that stimulate metamorphosis and apoptosis in tail tissues of ascidian tadpoles Transcriptome and quantitative proteome analysis reveals molecular processes associated with larval metamorphosis in the polychaete pseudopolydora vexillosa PubMed Abstract | CrossRef Full Text | Google Scholar Wnt signaling in axial patterning and regeneration: lessons from planaria PubMed Abstract | CrossRef Full Text | Google Scholar Evidence for the involvement of p38 MAPK activation in barnacle larval settlement Adenosine triggers larval settlement and metamorphosis in the mussel Mytilopsis sallei through the ADK-AMPK-FoxO pathway Induction of metamorphosis decreases nitric oxide synthase gene expression in larvae of the marine mollusc Ilyanassa obsoleta (Say) Mechanisms of heat and hypoxia defense in hard clam: Insights from transcriptome analysis Google Scholar CrossRef Full Text | Google Scholar Repulsive wnt signaling inhibits axon regeneration after CNS injury Purification of a larval settlement-inducing protein complex (SIPC) of the barnacle doi: 10.1002/(SICI)1097-010X(19980501)281:1<12::AID-JEZ3>3.0.CO;2-F induces planktonic abalone larvae to settle and begin metamorphosis HSP27 and HSP70 in osmotic stress in renal vs blood cells: A comparative study (University of the Sciences in Philadelphia) Google Scholar HSP70 facilitates memory consolidation of fear conditioning through mapk pathway in the hippocampus A new puffing pattern induced by temperature shock and DNP in drosophila CrossRef Full Text | Google Scholar Settlement of benthic marine invertebrates CrossRef Full Text | Google Scholar Effects of autophagy and apoptosis on the regulation of metamorphosis and seedling of Rapana venosa CrossRef Full Text | Google Scholar Stepwise metamorphosis of the tubeworm hydroides elegans is mediated by a bacterial inducer and MAPK signaling Gene expression patterns during the early stages of chemically induced larval metamorphosis and settlement of the coral Acropora millepora Selection of housekeeping genes as internal controls for quantitative rt-pcr analysis of the veined rapa whelk (rapana venosa) Arginine biosynthesis by a bacterial symbiont enables nitric oxide production and facilitates larval settlement in the marine-sponge host Comprehensive and quantitative proteomic analysis of metamorphosis-related proteins in the veined rapa whelk Transcriptomic analysis of differentially expressed genes during larval development of rapana venosa by digital gene expression profiling Metamorphosis in warming oceans: a microbe-larva perspective Metabolomic analysis of competent larvae and juvenile veined rapa whelks (rapana venosa) Experimental induction of larval metamorphosis by a naturally-produced halogenated compound (dibromomethane) in the invasive mollusc Crepidula fornicata (L.) 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This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) *Correspondence: Tao Zhang, emhhbmd0YW9jYXNAMTI2LmNvbQ==; Hao Song, aGFvc29uZ0BxZGlvLmFjLmNu Metrics details Polyandry is a common mating strategy in animals with potential for sexual selection to continue post-copulation through sperm competition and/or cryptic female choice Few studies have investigated the influences of population density on polyandry and sperm usage and paternity distribution in successive broods of marine invertebrates The marine gastropod Rapana venosa is ideal for investigating how population density influences the frequency of polyandry and elucidating patterns of sperm usage Two different population density (12 ind/m3 and 36 ind/m3) treatments with two replications were set to observe reproductive behaviors Five microsatellite markers were used to identify the frequency of multiple paternity and determine paternal contributions to progeny arrays in 120 egg masses mean number of sires and mean egg-laying frequency were higher at high population density treatment relative to low population density treatment indicating population density is an important factor affecting polyandry The last sperm donors achieved high proportions of paternity in 74.77% of egg masses which supported the “last male sperm precedence” hypothesis high variance in reproductive success among R which might have an important influence on effective population size Though gastropod species reproduce by multi-oviposition to date few studies have investigated the paternal contribution in successive egg masses despite growing evidence that population density can have substantial effects on polyandry little is known about their effects on mating rates in gastropod venosa herd together in the wild during the reproductive season which indicates population density may be an important factor that leads to the high level of multiple paternity detected in R a combination of behavioral observation and genetic paternity analysis was applied in captivity to assess the effects of population density on polyandry and to elucidate patterns of sperm usage in R Two different population density treatments with two replications were set and mating behaviors were observed Five polymorphic microsatellite markers were used to identify the extent of female multiple mating and to address the following questions: (i) What is the relationship between population density and multiple paternity (ii) What might be the benefits obtained for females from multiple mating (iii) What are the patterns of sperm storage and usage Copulation was first observed when water temperature was above 16 °C Whelks were aggregated at corners of the tanks for high density treatment male approached female from the back of body and mounted the shell which was at about 45 degree angle to the central axis of females’ shell Following fixation of the copulatory position male everted its highly motile penis searching for the female’s copulatory bursa and retained the copulatory position for hours to several days which was likely to be a way of post-copulatory guarding to increase male’s fertilization success Both females and males were observed copulating with multiple partners No significant deviations from Hardy-Weinberg equilibrium and linkage disequilibrium between pair of loci were identified. Genetic exclusion probabilities with the five microsatellite loci were uniformly 0.999 under the one-parent-known models (Supplementary Table S1) Paternity of 21 larvae could not be unambiguously assigned (i.e which were classified as larvae with unassigned paternity and were removed from subsequent analyses To assess whether there were possible differences in a sire’s paternity contributions to different capsules in a single egg mass two to four capsules (a total of 79 capsules) were randomly selected from 36 egg masses(27 for high density treatment Multiple paternity was detected in 25 (69.44%) of the 36 egg masses Of the 54 assayed capsules from the 25 multiply sired egg masses 26 (48.15%) involved all the identified sires for the entire egg masses and 7 (12.96%) only contained offspring from the dominant sire To test for possible differences in a sire’s relative contributions to offspring in different capsules from a single egg mass a total of 108 Fisher’s exact tests were conducted for the 25 multiply sired broods Statistical significance (P < 0.05) was reached in 6 (5.56%) of these comparisons Paternity analyses of egg masses spawned by six females (F01 F32 and F35) suggested that some non-sampled males had copulated with females prior to capture demonstrating that sperm stored in the female’s reproductive tract remained viable for at least 41 days which had not mated with any males during the whole experiment period laid three egg masses sired by a non-sampled male So the genetic and behavioral observation data of female F09 and the information of non-sampled males which had copulated with females prior to capture were removed from subsequent analyses To effectively examine the effects of population density on multiple paternity the parameters of female reproductive behavior (i.e the number of sires per female) for downstream analysis were based on the genetic paternity inferences female mating frequency and the number of sires per female ranged from 2 to 10 with an average of 5.71 and from 1 to 8 with an average of 4.18 Females laid 2–10 egg masses with an average of 5.12 during the whole reproduction period female mating frequency ranged from 2 to 7 with an average of 4.67 and the number of sires per female from 1 to 5 with an average of 3.11 Females laid 2–6 egg masses with an average of 3.33 during the whole reproduction period and egg-laying frequency per female in two density treatments for R significant positive linear relationships between female egg-laying frequency and mating frequency (R2 = 0.413 P < 0.001) and between egg-laying frequency and the number of sires (R2 = 0.298 which indicated that female egg-laying frequency increased with female mating frequency and the number of sires per female The same patterns were detected between the fecundity per female and mating frequency (R2 = 0.230 P = 0.011) and between the fecundity per female and the number of sires (R2 = 0.189 These might be the reason why the average fecundity of females in high density treatment was significantly higher than that in low density treatment (t = −3.135 no relationship between female shell length and mating frequency (R = −0.089 and egg-laying frequency per female (R = −0.070 Since only 24–96 larvae were selected from each egg mass for paternity analysis it was possible that not all the paternal contributions were detected especially for males with paternity contribution less than 10% the large genetic and behavioral data set were robust to analyze the patterns of sperm usage Relative contribution of different sires within all egg masses of each female in low density treatment (a) and high density treatment (b) venosa with different population density to estimate the effects of mate encounter rates on polyandry the mating order of males could be determined based on behavioral observation so we could discuss patterns of sperm usage by comparing mating order of males and their contributions to each egg mass paternal contribution of different sires to progeny arrays across all sequential egg masses laid by individual female throughout the spawning season could be ascertained to analyze the pattern of sperm storage this study provided new insights into the frequency and mechanisms of polyandry of R as well as new information relevant to sperm storage and usage which indicated that multiple paternity could thus play an important role in determining the extent of pelagic larval duration and consequently the range of dispersal distances achieved by larvae Futher experimental design is required to investigate whether polyandous females of R venosa acquire genetic benefits and to quantify direct and indirect genetic benefits of polyandry all these earlier findings as well as the current study support the hypothesis that mate encounter rate (population density) is an important factor affecting the degree of successful female multiple mating This might be one important reason why rapa whelks herd together in the wild during the reproductive season female remating rate and sperm usage rate may not be independent because females often remate to reload sperm supply according to our behavioral observations Due to lack of the relevant knowledge about the morphological traits of male genitalia and the components of seminal fluid for R the mechanism of avoiding or reducing sperm competition is addressed in respect of male mate choice sperm removal and post copulatory guarding behavior and therefore preferred recently spawned females as mates when facing sperm competition Further studies on sperm removal mechanism and ultrastructure of female reproductive tract could improve our understanding of the dynamics of sperm competition in R It is thus possible that females have some ability to stratify sperm in the seminal receptacle or to dump sperm from the copulatory bursa venosa can selectively store and use sperm from the seminal receptacles is not known no indication that paternity is biased according to male-female genetic similarity was detected and the last males sired most of the larvae in 85 (76.57%) egg masses These results indicated that post-copulatory female choice might not be the dominant factor to determine the distribution of paternal contributions we have applied both molecular analyses and behavioral observation in elucidating the effects of population density on polyandry and patterns of sperm storage and usage in R venosa were highly polyandrous and that sperm of several males were likely to be mixed in female’s reproductive tract Reproductive stimulation might be a direct benefit obtained by a female from polyandry which further ensures a high reproductive success Population density was an important factor affecting frequency of polyandry Paternity was significantly turned in favor of the last male partner in the majority of the analyzed egg masses and fertilization could be proportional to sperm number The high variance in reproductive success among males of R venosa might have a great effect on effective population size Although this study provides the first estimates of the effect of population density on paternity and the patterns of sperm competition for R it is still unclear the genetic benefits obtained from polyandry as well as the mechanism of sperm competition and cryptic female choice Future research should investigate the costs and benefits using fitness measures and the relationship between the number of sperm transferred to the sperm storage organ and the reproductive success of males Ethical approval was not required for this study because no endangered animals were involved venosa specimens was conducted in strict accordance with Animal Care Quality Assurance in China There were no significant differences in shell length between females and males (t = 0.799 nor were there significant differences in shell length between the two density treatments for both females (t = 1.086 Whelks were maintained for two weeks after the last observed egg capsules deposition by females 2012) spanned the natural spawning season of R Seawater in the rearing tanks was changed and moderately aerated every day and the water temperature in the tank ranged from 15.8 to 25.8 °C which closely matched environmental temperature Whelks were fed daily with sufficient live bivalves (Ruditapes philippinarum Mactra veneriformis) during the experimental period The large numbers of capsules (>80) and embryos (>10 000) in most egg masses precluded attempts to genetically assay all capsules or embryos so six egg capsules per egg mass were randomly selected and preserved at −80 °C before hatching of embryos foot muscle tissues were collected from each adult and preserved in 95% ethanol for genetic analyses Samples were then centrifuged at 3000 rpm for 2 min to pellet cellular debris Genomic DNA of adults was extracted from ethanol-fixed muscle tissue using the TIANamp marine animals DNA extraction kit (Tiangen Bio. PCR products were separated on the ABI3730 XL DNA Sequencer and sized with the standard GeneScan 500 ROX using GeneMarker 2.2 (SoftGenetics we were able to estimate the number of males contributing to the offspring of each female and then confirm the minimum mating frequency per female by consulting the behavioral data To examine effect of population density on polyandry the number of sires per female and egg-laying frequency were analyzed using a nested Linear Model with population density treatment as a fixed effect and replicate (L-I II for high density) nested within the density treatment as a random effect using SPSS 20.0 In order to infer benefits obtained by female from multiple mating relationships between egg-laying frequency per female and female mating frequency as well as the number of sires per female were tested by linear regression analysis using SPSS 20.0 relationships between fecundity per female and female mating frequency as well as the number of sires per female were also tested by using the same method Influences of population density on polyandry and patterns of sperm usage in the marine gastropod Rapana venosa Multiple mating and clutch size in invertebrate brooders versus pregnant vertebrates The evolution of polyandry: sperm competition Experimental removal of sexual selection reveals adaptations to polyandry in both sexes The evolution of polyandry: multiple mating and female fitness in insects Genetic benefits enhance the reproductive success of polyandrous females Multiple paternity in reptiles: patterns and processes High degree of multiple paternity in the viviparous Shiner Perch a fish with long-term female sperm storage Density drives polyandry and relatedness influences paternal success in the Pacific gooseneck barnacle Multiple paternity and population genetic structure in natural populations of the poeciliid fish Multiple paternity and mating group size in the European earwig Sperm competition games: raffles and roles Patterns of sperm use in the scorpionfly Panorpa germanica L Sperm competition in fish:bourgeois’ males and parasitic spawning Mechanisms of last male precedence in a moth: sperm displacement at ejaculation and storage sites Multiple paternity in Littorina obtusata (Gastropoda Littorinidae) revealed by microsatellite analyses Multiple paternity and female sperm usage along egg-case strings of the knobbed whelk Extreme female promiscuity in a non-social invertebrate species Sperm utilization in subadult and adult simultaneous hermaphrodite snails mating in the wild Mate choice by males and paternity distribution in offspring of triple-mated females in Neptunea arthritica (Gastropoda: Buccinidae) Seasonal effects on egg production and level of paternity in a natural population of a simultaneous hermaphrodite snail Mating system evolution and worker caste diversity in Pheidole ants ‘Late’ male sperm precedence in polyandrous wool-carder bees and the evolution of male resource defence in Hymenoptera Microsatellite Evidence for High Frequency of Multiple Paternity in the Marine Gastropod Rapana venosa and deposition of the egg capsules of the female purple shell Morphlogy of reproductive system of Rapana venosa (Valenciennes)(Gastropoda) 399–405 (1990) (in Chinese with English abstracts) The effects of female size on fecundity in a large marine gastropod Rapana venosa (Muricidae) Deposition of egg capsule and larval development of Rapana venosa (Gastropoda: Muricidae) from the south-eastern Black Sea Reproductive strategies in hermaphroditic gastropods: conceptual and empirical approaches The more the better–polyandry and genetic similarity are positively linked to reproductive success in a natural population of terrestrial salamanders (Salamandra salamandra) Fast versus Slow Larval Growth in an Invasive Marine Mollusc: Does Paternity Matter Geographical variation in the mating system of the dusky pipefish (Syngnathus floridae) Molecular Evidence for Multiple Paternity in a Population of the Viviparous Tule Perch Hysterocarpus traski Reconstructing paternal genotypes to infer patterns of sperm storage and sexual selection in the hawksbill turtle Influence of mating order on courtship displays and stored sperm utilization in Hermann’s tortoises (Testudo hermanni hermanni) (1998) Sperm competition and sexual selection (Academic Press Female infidelity and genetic compatibility in birds: the role of the genetically loaded raffle in understanding the function of extrapair paternity Male mate choice: why sequential choice can make its evolution difficult Female copulatory status and male mate choice in Neptunea arthritica (Gastropoda: Buccinidae) Copulation behaviour of Neptunea arthritica: baseline considerations on broodstocks as the first step for seed production technology development Cryptic female choice: criteria for establishing female sperm choice The effect of multiple paternity on the genetically effective size of a population Effective population sizes with multiple paternity Does variance in reproductive success limit effective population sizes of marine organisms in Genetics and evolution of aquatic organisms (ed Male reproductive competition in spawning aggregations of cod (Gadus morhua Effects of temperature and salinity on the development of embryos and larvae of the veined rapa whelk Rapana venosa (Valenciennes Polygynandry and sexual size dimorphism in the sea spider Ammothea hilgendorfi (Pycnogonida: Ammotheidae) a marine arthropod with brood-carrying males Isolation and characterization of 24 polymorphic microsatellite loci for the veined rapa whelk Genepop’007: a complete re-implementation of the genepop software for Windows and Linux GERUD 2.0: a computer program for the reconstruction of parental genotypes from half-sib progeny arrays with known or unknown parents COLONY: a program for parentage and sibship inference from multilocus genotype data Nonacs, P. Skew calculator. (2003) Available at: https://www.eeb.ucla.edu/Faculty/Nonacs/PI.html estimating and analysing relatedness and inbreeding coefficients Download references Shaojun Ban and Hongqian Pan for assistance in collecting and culturing R We are grateful to Brian Bowen for editing our manuscript and providing valuable advice This project was funded by the National Natural Science Foundation of China (No 100 Talents Program of the Chinese Academy of Sciences to JXL National Key Technology R&D Program in the 12th Five Year Plan of China (No.2011BAD13B01) and the NSFC-Shandong Joint Fund for Marine Ecology and Environmental Sciences (No.U1406403) Dong-Xiu Xue and Tao Zhang: These authors contributed equally to this work Key Laboratory of Marine Ecology and Environmental Sciences The authors declare no competing financial interests Download citation Sign up for the Nature Briefing newsletter — what matters in science Singapore’s second LNG bunker vessel has successfully completed its inaugural LNG loading operations through ship-to-ship (STS) transfer the bunkering operation was completed from Teekay’s LNG carrier Pan Africa on 30 May The milestone underscores the company’s “commitment to providing sustainable energy solutions that helps to reduce emissions in the shipping industry” FueLNG is driving positive change and advancing the industry’s energy transition goals in tandem with Singapore’s sustainability objectives,” the firm noted a joint venture between Keppel O&M and Shell Singapore Pte christened its newest LNG bunker vessel FueLNG Venosa at a ceremony held at Hyundai Mipo Dockyard in Ulsan Chartered from Korea Line LNG the LNG bunker vessel has a total capacity of 18,000 cbm and is expected to bring significant economies of scale the vessel is designed to facilitate safe and quick turnaround of vessels carrying out simultaneous cargo handling and bunkering operations; and is capable of bunkering different types of LNG fuel tanks FueLNG Venosa will provide gas-up and cool-down services to LNG carriers and LNG-fueled vessels after dry docking in Singapore or en route to loading operations a Bermuda-based shipping company and one of the world’s largest owners of LNG carriers merged with NYC-based investment firm Stonepeak As Seapeak, the company launched a new vision and identity, reflecting intentions to renew and grow its primary business of owning and operating LNG carriers.  The Pioneers of Offshore Engineering GustoMSC, part of NOV’s Marine and Construction business, is recognized for providing advanced design & engineering consultancy for mobile offshore units and reliable equipment. In close cooperation with our customers, we translate experience, science, and technical knowledge into realistic & innovative ideas. The performance of new and existing jack-ups, vessels […] a joint venture between Keppel Offshore and Marine (Keppel O&M) and Shell Singapore recently added a new vessel to its LNG bunkering fleet Built by Hyundai Mipo Dockyard of South Korea for shipowner Korea Line LNG FueLNG Venosa will operate under charter with FueLNG to provide bunkering services for ships that call at the Port of Singapore The vessel is designed to facilitate the safe and quick turnaround of vessels carrying out simultaneous cargo handling and bunkering operations and is capable of bunkering different types of LNG fuel tanks The 116- by 24.5-metre newbuild has a draught of 6.5 metres and a total LNG cargo capacity of 18,000 cubic metres The propulsion system delivers a speed of 11 knots In addition to being used for bunkering operations FueLNG Venosa will also provide gas-up and cool-down services to LNG carriers and LNG-fuelled vessels after drydocking in Singapore or en route to loading operations Click here for more news, features, and vessel reviews as part of this month's focus on naval architecture. Volume 8 - 2021 | https://doi.org/10.3389/fmars.2021.690282 This article is part of the Research TopicMolecular Physiology in Molluscs, Volume IIView all 26 articles Metamorphosis is a critical developmental event in mollusks and neuroendocrine system plays an essential role in this process Rapana venosa is an economically important shellfish in China venosa aquaculture is limited by metamorphosis food habit transition makes the mechanism of R To investigate the changes in the neuroendocrine system and to reveal its role in regulating the food habit transition and metamorphosis of R we cloned the cDNA sequences encoding 5-hydroxytryptamine receptor (Rv-5HTR) nitric oxide synthetase (Rv-NOS) and cholecystokinin receptor (Rv-CCKR) and investigated their expression by quantitative real-time PCR analysis and explore the spatio-temporal changes of 5-HT protein expression using Immunohistochemical (IHC) analysis The expression of the three geens was significantly increased in the early intramembrane veliger stage which indicates that the three genes are related to the development of digestive system expression of the three genes was decreased after metamorphosis while Rv-NOS and Rv-CCKR were increasingly expressed in competent larvae which may help the larvae find suitable environments and promote digestive system development for metamorphosis and the result of 5-HT IHC analysis also reflects the development of neuroendocrine system results show that CCK can effect the expression of digestive enzyme we hypothesized that CCK and CCK receptor may be critical regulatory factors of food habit transition and metamorphosis These results might provide information on the development of neuroendocrine system of R and new insight into the regulation of the food habit transition and metamorphosis of gastropods and the neuroendocrine system plays an important role in the regulation of metamorphosis there is no further study on the regulation of the neuroendocrine system on the metamorphosis of R 5-HT and CCK are likely to be the key mediators of the process of the food habit transition oyster-induced metamorphosis of R A previous study indicated that significant changes occur in the nervous system during the metamorphosis of R. venosa. The expression of the NOS and 5-HT receptor was found to decrease after metamorphosis (Song et al., 2016a) but no further study has examined their roles in the metamorphosis of R and whether the neuroendocrine system regulates metamorphosis by regulating the digestive system is unknown we aimed to examine the development of the neuroendocrine system by obtaining cDNA sequences and determining the expression profile of 5-HT receptor and observed the spatiotemporal expression characteristics of 5-HT by immunohistochemical (IHC) analysis the present study can provide new insight into the mechanism of gastropod metamorphosis from the perspective of the neuroendocrine and digestive systems and each developmental stage was sampled in triplicate and sextuplicate and stored at −80°C for qRT-PCR larvae at different developmental stages were washed in PBS and 7.5% MgCl2 solution was slowly added to completely anesthetize the larvae the samples were fixed at room temperature for 2–6 h using 4% paraformaldehyde solution transferred to 70% ethanol and stored at −20°C for use The larvae reached the four-spiral stage (shell height >1,250 μm competent larvae) were used for the CCK induction assays (CCK peptide: T510159 The following controls and treatments were included in the assays: (1) control group without CCK polypeptide (control and (2) treatment group with CCK polypeptide (1 mg/L) (CCK induction each tank (393 mm × 282 mm × 223 mm) contained 80 larvae We randomly collected three samples from each tank at 2 h (early stage which were used to analyze the mRNA expression of critical genes related to metamorphosis All samples were stored at -80°C until use All procedures involved in the animal collection rearing and dissection were conducted following the Guideline of Ethical Regulations of Animal Welfare of the Institute of Oceanology Chinese Academy of Sciences (IOCAS 2013.3) Our study protocols were approved by the Animal Welfare Committee of the IOCAS The 5′-RACE-ready cDNA was used as the template The 3′ ends of the three genes were amplified using 5HTR-3′-GSPn and NOS-3′-GSPn and UPM and 3′RACE-ready cDNA were used as the templates Confirmation of the full-length cDNAs by polymerase chain reaction (PCR) was conducted as follows: 94°C for 2 min; 35 cycles of 94°C for 30 s; 50–60°C for 30 s; 72°C for 1 min; and 72°C for 10 min (Eppendorf The PCR products were inserted into the pMD19-T vector (Takara Japan) and transformed into JM109 competent cells (Takara Japan) according to the manufacturer’s instructions and ten positive clones were selected to confirm the nucleotide sequences by TsingKe Biological Technology (Beijing The sequences were analyzed and assembled by DNAStar software to obtain the full-length cDNA sequence and MEGA 7.0 was used to construct phylogenetic trees with the neighbor-joining method First-strand cDNA was synthesized for qRT-PCR using reverse transcriptase (Takara) The SYBR Green real-time PCR assay (2 × SYBR Green qPCR Mix Sparkjade) was used with an Eppendorf Mastercycler® ep realplex (Eppendorf The 10-μl reaction mix volume was prepared containing 10 μl of 2 × SYBR Green qPCR Mix (Sparkjade 0.2 μl each of 10 pmol the forward and reverse primers 0.6 μl (100 ng) of cDNA and 4 μl sterile deionized water Amplifications were performed according to the manufacturer’s instructions Standard curves were generated with cDNA template dilutions of 10 qRT-PCRs were performed using the following thermocycler program: 95°C for 2 min and 40 cycles of 95°C for 15 s and 60°C for 30 s The relative gene expression was calculated using the 2–ΔΔ Ct method The secondary antibodies used for IHC analysis were Alexa 594-conjugated goat anti-rat IgG (Jackson Lab United States) and Alexa Fluor® 488-conjugated goat anti-rabbit IgG (Jackson Lab Negative control slides for all the IHC tests were included The calculated molecular mass and isoelectric point of the predicted Rv-CCKR protein are 48.86 kDa and 9.80 Figure 2. Phylogenetic trees based on Rv-5HTR (a), Rv-NOS (b), and Rv-CCKR (c) deduced amino acid sequences from Rapana venosa and other species. The trees were constructed based on the multiple sequence generated by Clustal X and aligned using the neighbor-joining method in MEGA 7.0. The genes which have been pharmacologically proven were highlighted with asterisk, which has been listed in Supplementary Table 3 and Rv-CCKR (C) mRNA expression and the heatmap (D) of three genes expression during Rapana venosa larval development (mean ± SEM Different superscript letters indicate significant differences (P < 0.05) the early intramembrane veliger stage (ev) the middle intramembrane veliger stage (mv) and the later intramembrane veliger stage (lv) the four-spiral whorl stage (competent larva and all three genes first increased in the early intramembrane veliger stage (ev) and decreased in the postlarval stage (J) (P < 0.05) hollow arrow) were clearly connected in a complex nerve network Immunoreactivity of 5-HT and α-tublin in early development stage of Rapana venosa No positive immune signals in the cleavage stage (a) blastula stage (b) and gastrulae stage (c) Two pairs of positive immune signals in late trochophore stage [(d) Immune-positive signals increased to 5 in early intra-membrane veliger stage (e) The 5-HT immune-positive signal extended to both cerebral nerve fibers in the middle intramembrane veliger stage [(f) Positive signals began to appear in the lower foot primordia of the later intramembrane veliger larvae and a strong signal appeared at the velum [(g) The immunohistochemical analysis revealed a complex neural network in the mantle in four-spiral whorl stage larvae (competent larvae white arrow] and the receptors in the base of the ciliate [(i) hollow arrow] were clearly connected in a complex nerve network To assess the digestive and neuroendocrine systems of larvae induced by CCK, the mRNA expression of carboxypeptidase, cellulase, 5-HTR and NOS was detected (Figure 5) The expression of the carboxypeptidase gene was significantly increased in both the early and later stages in competent larvae induced by oysters while cellulase was significantly decreased in the later stage (P < 0.05) The expression of 5-HT receptor increased significantly while the expression of NOS decreased significantly in both the early and later stage in competent larvae induced by oysters (P < 0.05) mRNA expression of critical genes (Carboxypeptidase 5-HT receptor and NOS) in Competent larva induced by CCK (mean ± SE (Different superscript letters indicate significant differences the mec hanism by which metamorphosis is regulated in carnivorous gastropods may be more complicated due to the accompanying food habit transition to further understand the regulatory mechanism of R the development and changes in the neuroendocrine system especially those related to the digestive system during the early developmental stage This study elucidated the mechanism of neuroendocrine system development in R venosa through an integrated approach of localization of the Rv-5HTR protein in the larva and examination of the molecular-metabolic responses of 5-HT and CCK in order to further investigate the regulatory effect of the neuroendocrine system on the food habit transition and metamorphosis of R These results may suggest that 5-HT and Rv-5HTR may not be positive regulators of metamorphosis in R the regulatory mechanism of 5-HT and Rv-5HTR requires further exploration The signaling pathways of NOS and NO have extensive functions and their regulation during the metamorphosis of R we show the changes of 5-HT and NOS during food habit transition in metamorphosis of R we found CCK is an important factor that regulates the development and function of the digestive system which may further suggest a close relationship between food habit transition and metamorphosis By exploring the changes in critical genes during the early stages of larval development the developmental process of the neuroendocrine system was initially identified; by further exploring the regulation of CCK we reveal the relationship between digestive system and neuroendocrine system This study provides new insight for studying the process of metamorphosis in carnivorous gastropods The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/Supplementary Material This research was supported by the National Natural Science Foundation of China (Grant Nos the Natural Science Foundation of Shandong Province (Grant No the China Post-doctoral Science Foundation (Grant No the Major Scientific and Technological Innovation Project of Shandong Provincial Key Research and Development Program (2019JZZY020708) the Earmarked Fund for Modern Agro-Industry Technology Research System (CARS-49) the Industry Leading Talents Project of Taishan Scholars (Recipient: TZ the “Double Hundred” Blue Industry Leader Team of Yantai (Recipient: TZ) Qingdao National Laboratory for Marine Science and Technology (no The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fmars.2021.690282/full#supplementary-material The role of serotonin in the control of pedal ciliary activity by identified neurons in 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Volume 11 - 2020 | https://doi.org/10.3389/fmicb.2020.01265 The symbiotic microbiota can stimulate modulation of immune system which also can promote immune system mature in critical developmental periods we have investigated the symbiotic microbiota in Rapana venosa at five early development stages using Illumina high-throughput sequencing Analysis of the symbiotic microbiota sequences identified that the most abundant phylum was Proteobacteria Beta diversity analysis indicated that the structure of the symbiotic microbiota dramatically shifted in early development stages The abundance of immune-related KEGG Orthologs (KOs) also increased in competent larval (J4 30-day post-hatching) and postlarval after 3 days of metamorphosis (Y5 Acid phosphatase activity decreased significantly in the Y5 stage and alkaline phosphatase activity also at a lower level in Y5 stage whereas lysozyme activities exhibited no remarkable change the activities of catalase and superoxide dismutase activities decreased dramatically during early development stages of R Dramatic changes in the symbiotic microbiota and the immune response mainly occurred in the initially hatched veliger (C1) competent larval (J4) and postlarval (Y5) stages during which the hosts might experience substantial environmental changes or changes in physiological structure and function These findings expand our understanding of the stage-specific symbiotic microbiota in R venosa and the close association between immune system and symbiotic microbiota in mollusks the specific relationship may need more researches are needed to investigated in the future The changes that occur in the symbiotic microbiota composition during the development of mollusks may reflect the development of the immune system which may play a regulatory role in metamorphosis we aimed to elucidate the structure changes of the symbiotic microbiota during early development stages of R and investigate the relationship between the development of the immune system and symbiotic microbiota This information could help to expound the correlations between symbiotic microbiota and the development of the immune system as well as further understand the mechanism of metamorphosis in R Each developmental stage had five replicates (30 larvae were used in each replicate) Larvae in each collected sample were euthanized and sterilized with 70% ethanol for 1 min and then washed three times using sterile water snap-frozen and preserved at −80°C for DNA extraction Environmental samples included five bait samples and five water samples Five hundred milliliters of food samples and water samples were filtered through 0.22 μm cellulose nitrate filter then subjected to bead beating and DNA extraction from the filter using the same method used for the other samples The samples from each development stage were homogenized and used to extract genomic DNA total protein and total RNA by using the DNA/RNA/Protein Coextraction Kit (DP423 China) according to the manufacturer’s protocol and the extracted products were used for PCR amplification analysis of immune enzyme activity and qRT-PCR the DNA and RNA concentrations were detected by a NanoDrop 1000 (Thermo Fisher Scientific and the purities were analyzed according to the ratio of the absorbance values at 260 and 280 nm (OD 260/OD 280) PCR amplification of 16S rRNA gene and sequencing were performed by Majorbio Co., Ltd. (Shanghai, China). PCR amplification was carried out using the 338F/806R primer targeting V3-V4 regions of the bacterial 16S rRNA gene (338F 5′-ACTCCTACGGGAGGCAGCAG-3′ and 806R 5′-GGACTACHVGGGTWTCTAAT-3′) (Chu et al., 2015) PCR products were purified by the AxyPrep DNA Gel Extraction Kit (Axygen Biosciences and quantified by QuantiFluorTM-ST (Promega then sequenced (2 × 250 bp) on the MiSeq PE300 platform (Illumina A unit of CAT activity was defined as the amount that can catalyze the conversion of 1 μmol H2O2 s–1 All immune enzyme activities were measured following the manufacturer’s protocols of assay kits (Nanjing Jiancheng Bioengineering Institute and 105-fold dilutions of each cDNA template and qRT-PCR was carried out with this program: 95°C for 5 s and 40 cycles of 95°C for 15 s defensin) or 58°C (tumor necrosis factor) for 30 s The 2–ΔΔCt method was used to analyzed the relative gene expression Levene’s test was used to determine the homogeneity of variances in analysis of immune-related enzyme activity and gene expression and mean differences were determined using Student’s t-test SPSS 19.0 was used in all statistical analysis and P-value < 0.05 indicated statistically significant difference Sequencing and OTU classification information Relative read abundances of different bacterial phyla (A) and genera (B) within the different communities Sequences that could not be classified into any known group are designated ‘unclassified.’ C1 four-spiral whorl stage (competent larvae); Y5 postlarval stage after 3 days of settling; F The overlapping and separate sections represent the number of shared and unique OTUs postlarval stage after 3 days of metamorphosis; F The alpha diversity of the microbiota community varied significantly in different development stages (Figure 3) The Shannon index ranged from 2.118 to 4.104 and significantly decreased with host development while the Simpson index ranged from 0.052 to 0.494 and increased with host development which suggests that the diversity of the symbiotic microbiota decreased with host development while the ACE index ranged from 142 to 940 and both the Chao and ACE index did not show significant change with the development of the host Boxplots showing the ranges of different alpha diversity indices The boxplots show significant differences in the ACE index (A) and Shannon index (D) at different culture stages Different lower-case letters for each bar indicate significant differences (P < 0.05) Adonis and ANOSIM tests of significant differences between the five culture stages Typing analysis (B) at the genus level was based on the weighted UniFrac distances A UPGMA tree (C) (based on the weighted UniFrac distances) and a genus-level relative abundance map are shown on the left and right Among the top 10 phyla, the Kruskal–Wallis H test demonstrated that the abundances of Proteobacteria, Cyanobacteria, Actinobacteria, Bacteroidetes, Firmicutes, Verrucomicrobia, Chlamydiae, Planctomycetes, TM6_Dependentiae, and Chloroflexi changed significantly between different culture stages (P-value < 0.05) (Figure 5) showed an increasing trend before the J4 stage and became dramatically depleted in the J4 stage The abundances of Verrucomicrobia and Bacteroidetes increased from the D2 stage while the abundances of Chlamydiae and Planctomycetes decreased with host development Abundances of the 10 most abundant phyla at different culture stages The relative abundance of each phylum at the five culture stages is shown The abundance at C1 was assigned a value of 1 and the relative abundances at other stages were calculated relative to this abundance Asterisks indicate significant differences between groups by PERMANOVA (P < 0.05) Two asterisks indicate extremely significant differences between the groups (P < 0.01) postlarval stage after 3 days of metamorphosis Bacterial taxa differentially represented between the three groups (F3 and Y5) as determined by LEfSe using default parameters The asterisks indicate significant differences between the groups (P < 0.05) J4: four-spiral whorl stage (competent larvae) and Y5: postlarval stage after 3 days of metamorphosis Relative abundances of predicted functions Relative abundances of immune-related KOs at KEGG level 2 in different development stages Different upper-case letters within each bar indicate significant differences (P < 0.05) The SOD and CAT activities in the larvae exhibited a decreasing trend with the development of R. venosa (Figure 8A) and reached their lowest levels at the Y5 stage (P < 0.05). The ALP and ACP activities were also at their lowest levels at the Y5 stage, while the LSZ activity did not change significantly during the early development stages (P < 0.05) (Figure 8B) Relative mRNA expression of Toll-like receptor 2 Values are expressed as the mean ± SD (n = 5) four-spiral whorl stage (competent larva); Y5 The relative expression of Toll-like receptor 2, tumor necrosis factor and defensin significantly changed during host development (Figure 8C) The relative expression of Toll-like receptor 2 peaked at the D2 stage after which the expression of this gene decreased significantly with host development The relative expression of tumor necrosis factor increased with host development and peaked at the Y5 stage which is similar to the observed trend for defensin Increased attention is being paid to the relationship between aquatic hosts and their symbiotic microbiota. Previous researches have indicated that symbiotic microbiota is important for the maturation of immune system (Mazmanian et al., 2005; Edelman et al., 2008) so there may be some changes in the symbiotic microbiota during host development Present results showed that significant shifts in the structure of symbiotic microbiota and immune response of R venosa occurred at early development stages which may reflect a substantial change in the physiological functions of the larvae the diversity of symbiotic microbiota showed a decreasing trend during the development stages while the richness indices showed no significant change which suggests that the role of the dominant population becomes increasingly prominent as the host develops NMDS and typing analysis also showed differences in the symbiotic microbiota between development stages The microbiota compositions of stages D2 and F3 are highly similar while those of J4 and Y5 belong to the same type Metamorphosis occurs in the late phase of the J4 stage so there should be a considerable difference in the symbiotic microbiota between the J4 and Y5 stages significant changes were observed earlier than expected which may suggest that a series of changes in the internal physiological functions of the host during metamorphosis precede the changes in apparent features including the further development of the digestive and immune system This shift in the symbiotic microbiota in R venosa occurs because the conditions of host (e.g. and microbial interactions) change dramatically during host development We found that the abundance of Vibrio decreased dramatically in the J4 stage in the present study which may be another indicator of changes in the immune system that occur during metamorphosis in the host All three KOs predicted by PICRUSt are related to immune function had significantly higher abundances in the J4 and Y5 stages than in the other stages indicating the involvement of many microbes in the host immune system the applicability of PICRUSt prediction in R although PICRUSt prediction in humans shows consistency across all body tissues The activities of CAT and SOD were dramatically decreased during the early development of R which might be related to the decreases in the abundances of pathogens CAT and SOD all exhibited high activities in the C1 stage which may be because larvae hatching from the oocysts first enter the water environment with an abundance of microorganisms and initial colonization by the symbiotic microbiota in larvae stimulates the immune system of the host Tumor necrosis factor exhibited a high expression level in the Y5 stage which may suggest that this gene is related to the shift in symbiotic microbiota during metamorphosis defensin also exhibited relatively high expression in the C1 stage which indicates that defensin is associated with not only the shift in symbiotic microbiota during metamorphosis but also the initial colonization in host by the symbiotic microbiota The host and symbiotic microbiota are components of an indivisible whole that interact with each other the development of immune system in host affects the structure of the symbiotic microbiota the colonization by and shift in the symbiotic microbiota stimulate the development of the immune system Our present study comprehensively examined the symbiotic microbiota and the immune response in R venosa during the early development stage and investigated the relationship between the symbiotic microbiota and the immune system of the host We found that dramatic changes in the symbiotic microbiota and the immune response mainly occurred in the initially hatched veliger (C1) during which the hosts experienced substantial environmental changes or changes in physiological structure and function These findings expand our knowledge of the stage-specific symbiotic microbiota in R venosa and the close association between symbiotic microbiota and immune system in mollusks additional studies are needed to explore the specific relationship between microbiota and immune system during the early development stages The datasets generated for this study can be found in the SRA under accession number PRJNA574861 This research was supported by the National Key Research and Development Program of China (2019YFD0900800) the National Natural Science Foundation of China (Grant No the Earmarked Fund for Modern Agro-industry Technology Research System (CARS-49) the Industry Leading Talents Project of Taishan Scholars (Recipient: TZ) the ‘Double Hundred’ Blue Industry Leader Team of Yantai (Recipient: TZ) Qingdao National Laboratory for Marine Science and Technology (No decision to publish and preparation of the manuscript for providing support for the experimental material The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fmicb.2020.01265/full#supplementary-material FIGURE S1 | Relative abundances of predicted functions KOs at KEGG level 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and Immune Responses in Early Development Stages of Rapana venosa (Valenciennes 1846) Provide Insights Into Immune System Development in Gastropods Copyright © 2020 Yang, Song, Yu, Hu, Zhou, Wang and Zhang. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) †These authors share first authorship Last week: Advanced to the 120-pound final of the Lewiston-Porter Tournament where he lost to Buffalo Saint Francis lightweight Hector Colom He became a sectional class champion for the first time last year Victor wrestles Saturday in the Section V Class A championships About me: Joined Victor's varsity as a seventh-grader Seeks his first trip to state championships .. His wrestling club is Superior Wrestling Academy .. "probably something with science," once in college .. Played JV football last fall as a running back and defensive back .. Must-see television for him is Sportscenter and Pawn Stars but it's not the same wrestling that we do here." .. People would be surprised: "That I spend so much time on school SEPT. 15, 2016 — Most people don’t want to spill oil onto beaches. But after the disastrous 1989 Exxon Valdez spill covered the remote rocky beaches of Alaska’s Prince William Sound with crude oil But Venosa wasn’t satisfied with the research attempts that came out of that spill He wanted to set up a more scientifically rigorous and controlled study of how effective bioremediation was under realistic conditions in the marine environment getting permission to spill oil into the environment on purpose is a very difficult Anderson began wondering how to improve the efficiency of oil spill cleanup and better protect Delaware’s abundant natural resources Anderson was listening to Ken Lee from Fisheries and Oceans Canada as he presented on bioremediation at the International Oil Spill Conference Lee mentioned how important—and difficult—it was to do controlled field studies on bioremediation The comment got Anderson thinking; maybe he could help make this happen in Delaware “Anything we can do to improve the aftermath of an oil spill in Delaware,” recalled Anderson The pair decided to work together to bring Venosa’s meticulous research approach to a study of oil bioremediation on Delaware’s beaches I worked on getting a permit with EPA and with the state,” said Anderson He and his collaborators also reached out to local environmental groups in Delaware and to NOAA and other agencies to build support for the research project building in as many safeguards as possible to limit any potential environmental impacts One issue the research team would have to work around was the fact that each May Delaware’s sandy shores are crawling with horseshoe crabs a prehistoric marine creature with armor and a long as they migrate north to nest in the Arctic each summer stop along these shores to nourish themselves with a feast of horseshoe crab eggs To avoid interfering with this ecological phenomenon Anderson and Venosa would have to start the experiment after horseshoe crab spawning season had passed With just a few days left before the experiment was to begin on July 1 1994 and with Venosa and his colleagues at EPA and the University of Cincinnati already on the road from Ohio to Delaware Anderson finally secured the needed permit the researchers set up the experiment very carefully they focused intensely on replication and randomization They cordoned off five separate blocks of sandy beach on Delaware Bay so that each block was parallel to the ocean yet would still be within reach of the tides they randomly assigned three oil treatment plots and one control plot after having weathered crude oil applied at the very beginning were sprayed daily at low tide with seawater and nutrients (nitrogen and phosphorus) This meant that each treatment and control was replicated five times reducing the chance that human error or natural variation would skew the results “We grew up our microorganisms on the beach in 55 gallon drums using the same seawater and microorganism [species],” recounted Venosa who served as the lead researcher for the study “We added them back onto these plots every week These [microbes] were adapted to the oil we used and to the climatic conditions at the site.” the research team strung oil containment boom along the waters surrounding the experimental plots to catch any oil runoff they lined up cages of filter-feeding oysters in the surf off of each study block as well as farther up and down the shoreline NOAA ecologist Alan Mearns helped facilitate this monitoring and multiple toxicity studies to determine the potential toxicity of the various treatments over time According to one of the study write-ups published at the 1997 International Oil Spill Conference the researchers found that: “oil was lost naturally because of both physical and chemical processes and biodegradation that degradation of oil alkanes and PAHs [polycyclic aromatic hydrocarbons] in upper intertidal sandy sediments could be enhanced with the continuous addition of dissolved nutrients that treatment with oil-degrading bacteria provided no additional benefit and that treatment neither enhanced nor reduced the toxicity of the oil.” While the team did detect a boost in how quickly oil broke down in plots sprayed with nutrients (which fed naturally occurring microbes) it was a pretty minor benefit in the big picture of oil spill cleanup And adding more microbes didn’t increase the rate of oil breakdown at all Delaware Bay’s waters are already rich with nutrients—and oil-eating microbes “It was probably a lot of runoff from fertilizer from agriculture and wastewater treatment plants,” speculated Venosa “We had a two to three times increase in the rate of biodegradation.” However for an area like Delaware Bay with high background levels of nutrients Venosa wouldn’t recommend going to the trouble and cost of using bioremediation techniques unless a spill happened right before something like the annual horseshoe crab spawning and bird migration “What we found was you don’t have to do any more nutrient addition,” said Anderson “Just keep adding ambient water and keep it aerated to get the [biodegradation] benefit but give it a little hand by keeping it wet on the beach face.” the research team considered the experiment a success that showed bioremediation to be a “polishing technique,” to be potentially used in oil spills when the local conditions were right and only after other quicker-acting cleanup methods had been applied first If an area showed high local levels of nutrients and oil-degrading microbes bioremediation likely wouldn’t be very effective “I was expecting more of a quantifiable effect in biodegradation but I didn’t realize the nutrients were going to be relatively high in the background,” reflected Venosa “I was expecting to see somewhat similar increases in the field as in the lab it’s different because your controls don’t have any nutrients so whenever you add nutrients that are in excess of what they need to grow As a result of this and subsequent studies in Canada, the EPA released guidance documents on implementing bioremediation methods in different environments, such as marine shorelines, freshwater wetlands [PDF], and salt marshes bioremediation is starting to mean more than just adding microbes or nutrients and now includes a range of other products meant to stimulate oil-degrading activity But not since 1994 on the shores of Delaware Bay has the United States seen another field experiment that has intentionally released oil into the environment to find out That summer was a unique opportunity for oil spill scientists to learn as rigorously and realistically as possible how well a certain cleanup method could work on an oil spill Subscribe now to get new articles and updates in your inbox » I’ve mentioned my penchant for growing vines up and over otherwise-dull shrubbery and in doing so not long ago I guess I got to realizing I was barely making use of a fraction of the opportunities But when she bloomed…the beauty was a duchess and I didn’t have the heart to toss her majesty in the heap With the help of a big spiral of jute twine rich-purple possibility….and come to think of it Yikes: It was climbing in the big bottlebrush buckeye that a storm took down last week Another viticella hybrid ‘Venosa Violacea,’ bears a distinctive star of white in its otherwise purple tepals (above) This one’s a little bigger-flowered than the others (4 inches wide along with more than 200 kinds of clematis (and 350 garden vines over all) Seems as if all my missed opportunities for vines up shrubs (and posts Δdocument.getElementById( "ak_js_1" ).setAttribute( "value" This site uses Akismet to reduce spam. Learn how your comment data is processed. @Brian: I planted a montana in my zone 5 garden and it’s doing great I probably planted it too close to a large tree and it took it many years (4 probably) to start flowering but it’s now putting an excellent show When it comes to clematis there is no such thing as overfeeding It’s also worth it to do a great job at planting time I usually dig a 20 x 20 ” hole and mix in tons of organic matter @Islandexile: When I combine clematis in the same area I make sure they belong to the same pruning group cut them all 1 foot above the ground and that’s it To me Christopher Lloyd’s book on clematis is still the best I don’t recall having seen 3 clematis together in his garden @ Great Dixter I no longer love the Clematis montana rubens perfect to capture a brisk winter wind and take down our beautiful cedar fence and have had to say good bye to the clematis I’m sticking to the daintier versions now A query for you experienced clematis growers Has anyone tried to grow clematis in pots on a second floor deck in SW Vermont Are there any varieties that might succeed I understand growing in pots can imitate winter temperatures of one or two zones colder your advice and experience with the clematis nevertheless sounds very wise I think they are a mess left to regrow from the tops and do a regular cutback as you suggest here too I hope that we will see you soon again by the way – our montanas regularly shed their leaves with the onset of winter but we leave pruning them until after they’ve flowered planting an ‘clematis’ that lived or got marched over in garden developments maybe now I’m ready to initiate something for an few to shine ‘Polish Spirit’ [& few friends] along with an ‘white Kousa Dogwood’ in an ‘garden frontal section nearby’ I’ll think about it for there is not much room nearly have to ‘pry them into such a selective spot’ perhap’s with an ‘small cedar wood lattice motif’ and benching spot to muse about excellence I am always prying things into tight spots here Are any varieties particularly powdery mildew reisistant If I recall correctly the varieties with Clematis texensis “blood” in them are the most apt to mildew if too thick and overgrown or growing in a place without good air circulation — particularly if there are hot Thinning the vines more in spring when pruning looking after watering to avoid stress earlier in the season and sometimes relocating a troubled vine to a more open spot are possible solutions The British Clematis Society has some info I have all of these with the exception of ‘Polish Spirit’ Sounds like you have a great assortment — I want to add more since they just bring extra enjoyment to an area A WAY TO GARDEN is the latest horticultural incarnation of me birthed in March 2008 with my own words as its primary DNA mingled with ideas shared in weekly expert interviews I have been the garden columnist for “The New York Times,” where I began my journalism career decades ago I host a public-radio podcast; I also teach online plus hold tours at my 2.3-acre Hudson Valley (NY) Zone 6A garden and always say no to chemicals and yes to great plants Design by Purr Modelling Risk is something that many in financial services treat now as commonplace The problem is when someone has to check the models and provide the relevant Corporate Governance to ensure that Regulators are satisfied that models within a company are doing the job they are supposed to This is where EQV and the Digital Twin of an Organization might be very useful tells #DisruptionBanking: “Our system is unique as it provides a unique core into the systems software of any organization Which means we are actually able to provide a more consistent approach to system architecture.“ “In risk management or trading we can use AI and we can develop customized solutions Our system has been quite successful in the banking industry in Italy and we almost cover the entire market The success is built on the interaction which we have with our clients Our strength is in the fact that our system can adapt to any organization We asked Olga about all the banks plunging literally billions of dollars into Tech so far this year? See our piece on Deutsche Bank from June “We recognize the dynamism of the industry the constant regulatory pressure and regulatory demand I was a risk manager in the past in the City of London,” Olga goes on to say “What I have seen is that organizations spend an incredible amount of money on the Tech side but what they forget is to use a more ‘strategic’ solution With a top down bottom up approach which is what we like to use “Very often when management decisions are taken to invest in Tech in a bank they do not communicate properly internally Credit and Market risk management are held separately in banks and can sometimes lead to doubling up on risk within the Bank Having a system like ours would negate this problem and improve the storage and analysis of the information whilst being easier for the regulator to understand internal controls “EQV’s Sphera has been classified as a Digital Twin of an Organization type of product The Digital Twin of an Organization is the capability of a system to be able to replicate all the processes of an organization The uniqueness of our Sphera DTO product is that we can adapt completely to an organization with this replication “Resilience is the key to our system.” For more insights from Olga please follow the link below to the video on our YouTube channel: For more details about how EQV’s innovative Sphera system can help your organization #EQV #Modelling #RiskManagement #Regulators #TechSpend #Sphera #DigitalTwin #Resilience #DTO #Replication #Communication #ArtificialIntelligence #Wrongdoing #CentralBank #Italy Author: Andy Samu and website in this browser for the next time I comment We publish press releases on our site only if the company's Authority Score on Google is high enough to justify it as news We do not respond to speculative inquiries from @gmail.com addresses Please represent a company when reaching out to us The newest addition to FueLNG’s fleet was named FueLNG Venosa at a ceremony held on 21 April at Hyundai Mipo Dockyard in Ulsan As described, the vessel is designed to facilitate safe and quick turnaround of vessels carrying out simultaneous cargo handling and bunkering operations; and is capable of bunkering different types of LNG fuel tanks. Besides bunkering operations, FueLNG Venosa will provide gas-up and cool-down services to LNG carriers and LNG-fueled vessels after dry docking in Singapore or en route to loading operations, FueLNG said. The naming of the FueLNG Venosa is said to mark another significant step in FueLNG’s efforts to promote the use of LNG as a marine fuel and contribute to a more sustainable future for the shipping industry. Aw Kah Peng, Chairman of Shell Companies in Singapore, who is the godmother of the vessel, said: “The FueLNG Venosa will support a growing number of LNG-fueled ships that call at the Singapore port, contributing to the country’s ambition of becoming a regional hub for LNG bunkering. As an industry, we must work together towards reducing emissions from shipping, and LNG is a key enabler in this transition.“ Saunak Rai, General Manager of FueLNG, noted: “We are thrilled to add the FueLNG Venosa to our fleet, which will help us cater to the growing demand for LNG bunkering services in the Asia Pacific region. The FueLNG Venosa will further enhance our capability to provide our customers with safe, reliable, and efficient LNG bunkering solutions.” 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, a SQL command or malformed data. 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. Cinema's ambiguous relationship with organised crime has been dramatically underlined after the jailing of an actor from the hit film Gomorrah for Mafia links. Giovanni Venosa played a local boss in the award-winning 2008 film, an unflinching look at the activities of the notorious Naples Mafia, the Camorra. The film was based on the book by the journalist Roberto Saviano, who has had 24-hour police protection since writing his exposé of the crime syndicate. In one scene lifted from the Saviano book, the character played by Venosa tells two younger Mafiosi: "Ve taglie a chep." In the local dialect, the phrase means: "I'll cut your heads off." On Monday afternoon, the court in Santa Maria Capua Vetere, near Caserta, outside Naples, sentenced Venosa to 13 years and 10 months after he was found guilty of racketeering and Mafia association. The prosecutor Luigi Landolfi had asked for a 24-year term. The court declared Venosa guilty of extortion in 2008 and 2009, even though he was spending time in this period at a correction facility in San Giuliano Saliceta, near Modena in the north of Italy. The court heard how he would still travel down to Castel Volturno, near Naples, to resume his criminal activities. Many shopkeepers and business owners in the area were terrorised into paying Venosa protection money, prosecutors said. Police and magistrates finally began an investigation in to the racket after a tip-off from the former mayor of Castel Volturno, Francesco Nuzzo. Gomorrah, an exposé of the criminal activities of the Casalesi and other Camorra clans, has been translated into numerous languages and sold millions of copies worldwide. The film, directed by Matteo Garrone, won the Grand Prix at the Cannes Film Festival in 2008. Mr Garrone deliberately recruited amateur actors from the Camorra-infested areas of Naples and surrounding towns to give his film the gritty feel of the book. Mafia expert Corrado De Rosa said: "Garrone wanted the Caravaggio look, the grimy realism as well as the beauty, that why he hired people who he knew had links with clans." New subscribers only. £8.99/mo. after free trial. Plan auto-renews until cancelled There were rumours in the Italian press the film-makers made undeclared payments to Camorra clans in exchange for their co-operation. Join thought-provoking conversations, follow other Independent readers and see their replies serious mafioso: Gomorrah star jailed for crime links","description":"  the joint venture between Keppel Offshore & Marine Ltd and Shell Singapore named bunker vessel FueLNG Venosa at a ceremony held 21 April at Hyundai Mipo Dockyard in Ulsan The company’s first vessel and Singapore’s first LNG bunker vessel “FueLNG Venosa will support a growing number of LNG-fuelled ships that call at the Singapore port contributing to the country’s ambition of becoming a regional hub for LNG bunkering we must work together towards reducing emissions from shipping and LNG is a key enabler in this transition.”  The state-of-the-art ship was built at Hyundai Mipo Dockyard Co and chartered from Korea Line LNG Co The vessel is designed to facilitate safe and quick turnaround of vessels carrying out simultaneous cargo handling and bunkering operations and is capable of bunkering different types of LNG fuel tanks FueLNG Venosa will provide gas-up and cool-down services to LNG carriers and LNG-fuelled vessels after drydocking in Singapore or en route to loading operations “We are thrilled to add FueLNG Venosa to our fleet which will help us cater to the growing demand for LNG bunkering services in the Asia Pacific region.”  Shell and CMA CGM inked an LNG multi-year supply agreement last year Shell will supply LNG to CMA CGM’s 13,000-TEU container ships in the Port of Singapore The port of Singapore is the world’s largest bunkering hub for conventional heavy fuel and is preparing to secure its long-term pole position, expanding to handle LNG, biofuels, ammonia, hydrogen and methanol alongside traditional fuels. Readers can see a detailed report here Sign up for Riviera’s series of technical and operational webinars and conferences in 2023: By Pagine Ebraiche staff* the Italian Ministry for Cultural Heritage and Activities has granted material aid to value old evidence of Jewish presence in Italy €2.5 million is the sum the Government has given for the Jewish catacombs of Venosa evidence for the great presence in the area between the 3rd and 7th century who announced this commitment; she intervened yesterday at a meeting about the site and the projects for making it more known in the seat of UCEI Bibliographical Centre and organized by the Italian Jewish Heritage Foundation together with the association Daniela Di Castro As said by the President of the Foundation Dario Disegni Venosa represents the greatest effort in terms of projects and resources being also marked by the activation of a scholarship which allowed to create a presentation video recently broadcast by Rai (Italian national television) For a year now there has been a very positive dialogue with the local Superintendence and Administration That of today is a new step in an intense work in progress” underlined Disegni “Venosa community feels the responsibility of the role the history has given us Here you will always find an open door” said then mayor Iovanni It was her who walked minister Alberto Bonisoli through the visit at the catacombs in the near archaeological Roman complex on March 12th so that he stated: “Venosa is an exceptional site And tourism-wise speaking it has all the possibilities “to be another Pompei” “the minister called me to give me the news of the allocation” UCEI council member and president of the association Daniela di Castro born with the aim of treasuring Jewish culture in the name of one of its greatest and never forgotten protagonists “Southern Italy – Coen affirmed – is a mine of testimonies to be discovered and made known” The same satisfaction was also expressed by Sabrina Mutino who intervened in the name of the Superintendence for Archaeological Heritage of Basilicata and noted “the special consideration” paid to the catacombs of Venosa explaining that the public funding “will be of use mainly for their preservation” took then the floor speaking about the “initiatives for Jewish Venosa 2020” The start of this path has brought to a collaboration the benefits of which were seen also at the first exhibition held at the Museum of Italian Judaism and the Shoah in Ferrara “The catacombs deserve an ever more international interest – for the meaning of the history they witness for the symbols and the charm they evoke” Funaro said What do we know today about the Jewish presence in Venosa in those ages The reconstruction comes thanks to professor Giancarlo Lacerenza from CNR (the Italian National Research Council) who traced the environmental and occupational context with a regard for research prospects who presented new techniques of survey and fruition “Most of the data we need to reconstruct what was Jewish Venosa are already there A detailed study of historical research is fundamental virtual reconstruction too has had and will have a key role It is a huge and necessary work” acknowledged Lacerenza some thoughts of former Superintendent Maria Luisa Nava with whom a dialogue started some years ago and who expressed her own satisfaction for the level and the results this communication among different bodies has achieved Seleziona il modo in cui vuoi essere contattato da Unione delle Comunità Ebraiche Italiane: Usiamo Mailchimp come piattaforma per la realizzazione e l'invio dei nostri notiziari. Con l'iscrizione le tue informazioni saranno trasferite a Mailchimp per l'elaborazione. Scopri di più sulla privacy di Mailchimp 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 a joint venture between Shell and Seatrium has deployed LNG bunker ship FueLNG Venosa to perform the gassing up and cooling down and LNG bunkering of Seatrium’s floating living lab the first of its kind offshore floating testbed in Singapore The 18,000 cubic meter FueLNG Venosa, also known as Singapore’s second LNG bunker vessel, completed the gassing up and cooling down and bunkering operation of the floating living lab at Seatrium’s Benoi yard marking a significant milestone in its commissioning FueLNG said in a social media update on February 26 shall play a pivotal role in the trial of many new technologies paving the way for a more sustainable and innovative power production Harbercraft electrification and low carbon fueling for vessels,” FueLNG stated Southeast Asia’s first floating and stacked energy storage system (ESS) was deployed at Seatrium’s floating living lab to commence operations by the first quarter of 2024 the stacked ESS is a key component of an integrated floating energy solution that could help to overcome Singapore’s land constraints with a deployment footprint of up to 40% less than land-based ESS specifically facilitating the maritime and offshore industry knowledge and reach we can optimally meet your businesses media needs and requirements in three cohesive ways: Network with the right people Discover the world's largest classical music catalogue with Presto Music If you are a library, university or other organisation that would be interested in an institutional subscription to Gramophone please click here for further information Address: 480 New North Rd, KingslandPhone: (09) 815 6644Facebook: KingslandMMXVCost: Dishes $14.90 to $27.50; desserts $12.90 No doubt Dave Perillo wanted "venosa" rather than "KingslandMMXV" for his Facebook moniker, but it was already taken by a young woman called Terry who lives in Milan but was born in the south of Italy. Perillo named his place after his father's hometown in Basilicata (in the boot's pronounced high arch), a little-travelled part of the country that Dave calls "the wild back of beyond". The family was part of the big influx of Italians after the war: Dave's Dad married a Kiwi girl and luckily for Dave, who was born here, she liked Italian food, though he reckons he got his birthright entitlement of lamb chops and mashed spud. Now, here he is, squeezing between the tables and checking diners are happy at his tiny Kingsland restaurant, a room so small that when they need more wine from the back room, the waitress goes out the front door and down the side to fetch it. It's small enough to be buzzing any time it's open but he's happy to take bookings ("I'm not going to turn business away," he says; it's a policy that more than a few up-themselves Auckland eateries would do well to emulate). Perillo was the co-founder of the Corner Store at the top of Mt Eden Rd but a home-style Italian place is something he says he's been thinking and talking about for 30 years. If that's true, it's been worth the wait. Venosa is the kind of place you remember stumbling into in some back street in Italy and being certain you'd never find that magic again, much less on the other side of the world. The food is unpretentious though far from unrefined; simple though far from plain. In a city where it sometimes seems every place is straining for effect, Venosa makes everything look effortless. The tucker, which Perillo says is "the stuff I grew up eating", is prepared by chefs far too young to be as good as they are: Harry McAlister, who did his time at Cibo, and Rowan Maissin. I kept sneaking glances at the kitchen, expecting them to be joined by a burly Italian for whom they were kitchenhands, but no one else ever showed. The menu, which plainly changes regularly, listed nothing to surprise, but there was plenty to delight in what came out of the kitchen. Double-crumbed fried eggplant avoided, probably by being finished in the oven, the oily horrors that dish can become. Topped with rocket pesto and half-melted buffalo mozzarella, it was a sensation. With some reluctance, I had passed on the bruschetta, because the Professor said she wanted to save room for dessert and we turned our attention to the mains. More po-faced reviewers than I save five stars for Noma and the French Laundry, but I ask myself what a place is trying to do and whether it could be done better: as a homely, neighbourhood Italian joint with sensational food, this is certainly a five-star experience. 'The team have nailed it – it’s a strategy that everyone can understand.' Newlyweds Umberto Di Venosa and Susan McMonagle met while organising people's Camino holidays Umberto Di Venosa and Susan McMonagle got married recently in France: Photo: David Conachy They only got married seven weeks ago in France, and Susan McMonagle still finds it fun to call Umberto Di Venosa her husband. She made a speech at their wedding in which she discussed the Frenchman's strengths, because he is not only physically fit. "He is strong in his values around family too," she says. "Umberto is also great fun, and is not afraid to do something silly in public to make me laugh." Umberto (36), grew up in Drancy in Paris as the younger of Ciriaco and the late Fulvia's two sons. His Italian dad still lives there and he is close to him, but his non-smoking, teetotal mum sadly passed away aged 50 from lung cancer. He was talented at rugby and went on to study business and marketing at a sports university in Paris. He had a friend, Fabien, in Dublin so he came here to improve his English one summer and loved it so much he returned after completing college in 2003. Umberto and Susan (33), met when she got a job at his company in 2010. She's the youngest of Mary and Stephen's three children, and says her parents have always been extremely loving and supportive. Susan studied applied languages at DCU, followed by a diploma in legal studies at DIT, and after working with Dublin Tourism she got a job in customer service with One Foot Abroad, eventually becoming assistant operations manager. "Umberto was one of my three bosses and I probably had less contact with him than some of the others with the role I was in," she says. "There was nothing romantic between us as both of us were in other relationships. If you had told me at that point that we would be married a number of years later, I would have laughed in your face." "Susan is very beautiful and nice, and I saw how determined and hard-working she was," says Umberto. "I love her smile, and she also liked other languages and cultures and is very open-minded." Susan thought Umberto was handsome and hard-working and she also found him very funny. At the time she was doing a part-time master's in international relations, and they only told their colleagues that they were dating when Susan resigned to take up an internship with the United Nations' refugee agency, UNHCR. "It's a small company and we were afraid it would make things awkward, but we made a big announcement at the Christmas party and nobody cared," she laughs. She and Umberto waited a while before he introduced her to his children. He proposed while they were visiting Chateau de Fenelon in Dordogne, and they got married in Saint-Jean-d'Angely in August. Umberto's brother Lorenzo was best man and Eva and Theo were flower girl and page boy. They had 115 French, Irish and Italian guests, and held a big barbecue for them at Umberto's family home the next day. They went to San Sebastian on honeymoon, and have now returned to normal life. Susan's internship at UNHCR turned into a full-time job and she's also training to be a barrister at King's Inns. With the added pressure of studying, Umberto is good at keeping her calm as she likes everything to be done well. "For example, we were camping recently, and she took out wipes to clean the inside of the tent, even though we were going for a walk and would be mucky coming back," he laughs. www.followthecamino.com www.onefootabroad.com Join the Irish Independent WhatsApp channel BURSASGXHomeBy KUALA LUMPUR (Feb 21): Myra by Oriental Interest Bhd has launched Begonia comprising 91 units of double-storey link homes according to a press statement on Tuesday (Feb 21) The project is the second phase of the 70.6-acre Myra Gardens township in Kundang.  The RM62.77 million Begonia offers two layouts Venosa features four bedrooms and three bathrooms sized from 1,854 to 2,121 sq ft and a land size from 20ft by 70ft to 25ft by 70ft the intermediate units come with a backyard patio of 10ft.  have four bedrooms and three bathrooms with a built-up of 2,157 sq ft and a land size of 20ft by 70ft These units have a starting price of RM808,000 The Grandis units will be within a 100m radius of the township’s clubhouse with its backlanes directly connecting to the township’s central playground.  Myra’s brand and culture chief Akil Kalimullah said: “Myra Gardens strives to provide a harmonious balance between the fast-paced city and a tranquil retreat for our residents is our humble effort to offer a contemporary lifestyle while also providing a peaceful haven after a long day We are committed to creating homes that align with the needs and desires of our homebuyers.” The release added that Begonia is a gated and guarded development with a two-tier security system and is the closest residential project to the facilities offered in the township.  Facilities in Myra Gardens include a clubhouse Nearby amenities in the township and its surrounding area include AEON Rawang The township is also within close proximity to the recreational forest of Bukit Lagong Myra Gardens is near the Rawang and Sungai Buloh areas with easy access to major highways such as the Kuala Lumpur-Kuala Selangor Expressway (LATAR) Guthrie Corridor Expressway and North-South Expressway.  Myra Gardens will have a total of 587 landed residential units and a gross development value of RM440 million across six phases Please enable JS and disable any ad blocker