Finding out the hereditary predisposition to seizures in response to specif ic external stimuli is important for understanding the causes of epileptiform conditions, developing new methods for their prevention and therapies. In the water vole, individuals with convulsive seizures are found both in natural and laboratory conditions. The data of long-lasting maintenance and breeding of water voles in vivarium conditions were analyzed in order to establish a hereditary predisposition to convulsive seizures, and the inf luence of sex and age on their development. In the vivarium, seizures are provoked by handling and are observed in 2.4 % of voles caught in the natural population with cyclic f luctuations in abundance. Seizures are observed more often in individuals caught in the phases of decline and depression of abundance than in individuals caught in the phases of rise or peak. Convulsive states are probably an element of adaptive behavior formed in the predator-prey system. In natural conditions, individuals predisposed to convulsive seizures may have a selective advantage when under increasing pressure from predators. Convulsive seizures in response to handling were noted in 29.8 % of descendants of captive-bred water voles. The proportion of such individuals increased signif icantly if one or both parents had convulsive states, which indicates the presence of a hereditary predisposition to seizures. In parent–offspring pairs, a signif icant correlation was found between the average age of onset of the f irst seizures in parents and their offspring, r = 0.42, p <0.01. The minimum age of registrationof seizures in the water vole is 39 days, the maximum is 1105 days, and the median is 274 days. Predisposition toseizures is not related to sex. Genes that control the occurrence of seizures have a pleiotropic effect on life span, sinceindividuals with seizures live longer in vivarium conditions than individuals with a normal phenotype. The watervole can serve as a suitable model object for studying the nature of convulsive states and the evolution of longevity Tonic-clonic seizures – uncontrolled muscle tension orcontraction – can occur in humans and other mammals:gray rats Rattus norvegicus , housemice Mus musculus , Mongolian gerbilsMeriones unguiculatus , deer micePeromyscus maniculatus , Syrian hamstersMesocricetus auratus , meadow volesMicrotus pennsylvanicus ,dogs Canis familiaris , cats Felis catus, and rabbits .The most common cause of seizures is an imbalance betweenexcitatory and inhibitory neurotransmitter mechanisms in thebrain . Seizures are categorized intospontaneous (provoked by unknown factors) and reflex ones .In response to tactile stimulation during handling, prolongedtonic-clonic seizures take place in bank voles Myodesglareolus , meadow voles M. pennsylvanicus, and Mongolian gerbilsM. unguiculatus .Other stimuli  are not effectiveat inducing seizures in these species. The gray rat is a popularexperimental model for research on audiogenic epilepsy. It should be noted that genetic factorspredisposing to various types of reflex epilepsy are poorlyunderstood, and genes that control somatosensory epilepsyare not known at all . Clarificationof hereditary predisposition to handling-induced seizures isimportant for understanding the causes of seizures and forimproving the prevention and treatment methods.In recent decades, rodents have been widely used in neurophysiologicalinvestigation into epilepsy owing to theirsmall size, peacefulness, and rapid reproduction in captivity. Comparative studies on mammals are neededto better understand the evolutionary and genetic factors thatexplain seizures because they should help to find commoncomponents of the pathogenesis of this neurological disorderthat are of clinical relevance to humans . To elucidate the evolutionary mechanisms underlyingthe onset of convulsive states, species in which individualswith seizures occur under natural conditions are of greatinterest. L.G. Krotova (1962) – who has investigated thefunctionof adrenal glands and carbohydrate metabolism inwater voles (Arvicola amphibius L.) living in the floodplainof the Chusova River  – noticedthat some captured animals had seizures accompanied by acoma. Owing to detailed complex research data on variouscharacteristics of population ecology, genetics, and physiologyof the water vole and to excellent advances in the methods forits breeding in captivity , the watervole can become a promising experimental model for investigationinto predisposing factors of seizures at populational,genetic, and neurological levels.In this study, data are analyzed that were obtained duringlong-term breeding of water voles at an animal facility toachieve the following aims: 1) to describe the picture of theseizures; 2) to estimate the proportion of individuals withseizures among animals caught in the wild and among animalsborn in the vivarium; 3) to determine the minimal, maximal,and average age at onset of seizures; 4) to find out whether themanifestation of convulsive seizures is associated with the sexof these animals and/or the presence of this neuropathologyin the parents; and 5) to identify a possible relation betweenthe predisposition to seizures and the life span.The study was performed on two groups of water voles kept atthe animal facility of the Institute of Systematics and Ecologyof Animals of the Siberian Branch of the Russian Academy ofSciences , under natural lighting conditionswith free access to water and feed :1) the wild group (n = 589), i. e., animals that were caughtduring 1983–2017 in the Ubinsky District of NovosibirskOblast (Russia), where comprehensive ecological andphysiological studies on a population with pronouncedhigh-amplitude fluctuations of population size (6–7-yearcycle) were being conducted . After capture, these water voleslived in the vivarium for at least 3 months . Ninety-four voles from the peak phase, 32from the decline phase, 155 from the depression phase, and308 from the increase phase were analyzed;2) the vivarium group (n = 1776), i. e., water voles (parentsand offspring) that were born in the vivarium and survivedat least 22 days after birth. This group was founded byindividuals caught in the aforementioned natural population.To preserve genetic heterogeneity, the group wasregularly replenished with wild individuals: at least onceevery 3 years.This approach allows, on the one hand, to identify populationalfactors that affect the prevalence of convulsive statesin the wild, and on the other hand, to identify genetic andontogenetic factors predisposing parents and offspring toseizures under standard maintenance conditions in a vivarium.The work on the animals of the two groups throughout thestudy period was done according to a standard scheme: 1) allwater voles were individually marked, sexed, and dates ofbirth (capture) and death were recorded; 2) until the age of70 days, weekly, the water voles born in the vivarium wereweighed and their body length was measured; the weight andbody length of the vivarium animals older than 70 days andthose of individuals caught in the natural population weremeasured once a month; 3) during the reproduction period,before forming of mating pairs , vaginal smears weretaken from females, and anogenital distance was measuredin males; 4) throughout their life span, the water voles weresubjected to experiments aimed at examining the relationbetween ethological–physiological characteristics of watervoles and their reproductive ability. The following procedureswere performed: tests of social interactions, of olfactory matechoice, and of parental behavior as well as blood samplingfrom the retroorbital sinus and tail, urine collection. Duringthe experiments, water voles were subjected to handling (atleast once a month), and some animals developed convulsiveseizures. The main diagnostic criterion of a convulsive statewas assumed to be pronounced tonic or clonic contractions ofmuscles of the trunk and/or limbs. Such cases were recorded.The animals with convulsive seizures were not culled and wereallowed to reproduce. There were no spontaneous seizureswhen the animals were in home cages and were not subjectedto the handling.Statistical analysis was performed in SPSS Statistics 15.0 and Statistica 6.1 software. Comparisons of proportions (%) between groupswere conducted by Pearson’s c2 test. Effects of categoricalvariables and continuous variables on age at onset of the firstseizure (hereafter: “age at seizure onset”) or on the life spanwere assessed via the linear mixed model, with dependent variablessubjected to the natural-logarithm (ln) transformationin order to make their distributions normal. Survival curveswere constructed by the Kaplan–Meier method. Differencesbetween the curves were evaluated by the Gehan–Wilcoxontest. The text and tables show means of parameters (X ), standarderror (± SE), and sample size (n). Statistical significancewas assumed at p < 0.05.Description of the seizuresEach seizure was convulsive (tonic and/or clonic) and occurredin the first minute after a water vole was subjected tohandling. The seizure began with a twitching of the vibrissaeand tonic tension of anterior trunk muscles, followedby their rhythmic contractions; meanwhile, the animal waslikely to arch its back and tilt its head back. At the first signsof a seizure, individuals were placed into an arena, and theirstate was examined. The convulsions spread throughout thewhole body, and in some cases, the seizure was accompaniedby gnashing of teeth and very rarely by loss of consciousness.After the seizures stopped, the animals experienced locomotoragitation: “wild running”.The proportion of individuals with convulsive seizuresamong water voles taken from the natural populationduring different phases of cyclic population dynamicsSeizures were noted in 2.4 % of the 589 water voles in thewild group; the seizures were caused by handling. There wereno sex differences in susceptibility to seizures .The proportion of individuals with seizures was affected bythe population phase at which the animals were taken fromthe population , Fig. 1. Amongthe water voles from the decline or low phase, the proportionof those susceptible to seizures was significantly higher thanthat among the water voles from the increase or peak phase.Seizures among the water voles born in the vivariumSeizures were observed in 29.8 % of the 1776 animals in thevivarium group. Susceptibility to seizures was not influencedby sex . There were no seizures during the suckling period,which ended at age 3 weeks.Age at seizure onsetThe date of onset of the first seizure was recorded only for480 animals. The minimum age at seizure onset was 39 days,and the maximum age at seizure onset was 1105 days. In theseizure group, modal age was 189 days, median age 274 days,and the mean age 324.6 ± 7.8 days. A histogram of the distributionof age at seizure onset is presented in Fig. 2.Analysis of the data using a mixed linear model, taking intoaccount the year of birth and litter identity (ID) as randomfactors, showed no effect of sex as a fixed factor  on the age of onset of the first seizure. Theinfluenceof the year of birth and lD was significant .Hereditary predisposition to convulsive statesTo clarify hereditary predisposition of water voles to handlinginducedseizures, we used animals that lived in the vivariumfor at least 39 days, which was the minimum age at seizure onset in this work. Thus, 1656 water voles from 445 litterswere analyzed. The results indicated that the susceptibilityof offspring to seizures correlates with seizure status of theirparents (Table 1). Namely, the proportion of offsprings withseizures was significantly higher if one or both parents hadseizures .Within “parent–offspring” pairs with known age at seizureonset in both parents and offspring , a significant correlation between average ages atseizure onset was found , indicatinga significant additive contribution of genes to this trait and thepossibility of selection for “age at seizure onset”.The relation between the presenceof seizures and the life spanTo clear up the relation between the presence of seizuresand the life span, the linear mixed model was used, whichincluded categorical variables (sex and the presence/absenceof seizures) and the birth year and ID as random factors. As aresult, we detected significant effects of sex  and of seizure status of an individual  on the life span. Males lived longerthan females . Thelife span of animals without seizures was significantly shorterthan that of animals with seizures , Fig. 4. A significant impact of the birthyear on the life span was noticed ,whereas such an effect of ID was statistically insignificant.Figure 5 shows survival curves of two groups of water voles(with seizures and without seizures). Significant differenceswere found between them according to the Gehan–Wilcoxontest .The effect of seizure status of parentson the life span of the offspringTo clarify the dependence of life span on the convulsive statusof parents, using a linear mixed model, we assessed the influenceof fixed factors: sex, predisposition to convulsions of themother, father and interaction of parents’ phenotypes on thelife span of offspring. The birth year and ID were included inthe model as random factors. It was found that the life spanis affected by sex , by maternalseizure status , and by an interactionof these maternal and paternal phenotypes . The offspring of parental pairs in whicheither the mother or both parents did not have seizures manifesteda significantly shorter life span . The effect of paternal seizure statuson the offspring lifespan was not statistically significant, and neither was the effect of ID. Data on the mean life span ofoffspring and seizure status of the parents is given in Table 2.These studies establish the occurrence of genetically based,tonic-clonic convulsions in water voles in response to handling.The incidence of seizures did not depend on sex.Prolonged tonic-clonic seizures in response to handling havebeen documented for the bank vole , themeadow vole , and the Mongoliangerbil . Thepattern of seizures in the water vole turned out to be similar tothat in bank voles, meadow voles, and Krushinsky–Molodkinaaudiogenic rat strain, except that in bank and meadow voles,there is no “wild running” phase observed in Krushinsky–Molodkina rats  and in water voles.According to the literature, reflex epileptiform seizurescaused by tactile or acoustic stimuli are typical only for smallrodents, which is associated with the structural and functionalfeatures of their central nervous system, due to which pathological,from a human point of view, behavioral responses tostimuli that inform about danger are carried out . Convulsive states are probablya component of adaptive behavior formed in a “predator–prey” system. Short-term clonic convulsions in response totactile or acoustic stimulation can have a frightening effect onraptors that capture prey with their claws, and rapid running,as the next phase of a convulsive seizure, allows the prey toquickly take refuge in a safe place.Individuals susceptible to seizures may have a selectiveadvantage during high predator pressure. It is known thatthe number of specialized bird predators with great mobilitychanges synchronously with the dynamics of population sizeof water voles . The results of crossingsof water voles, differing in their predisposition to seizures,indicate the hereditary transmission of this trait: the proportionof offspring with seizures significantly increases if one or,especially, both parents had seizures. Therefore, an increase inthe prevalence of individuals with seizures during the phasesof decline and depression of cyclic population dynamics may be explained by positive selection for seizure susceptibilityat the peak phase of population numbers accompanied by anincrease in predator pressure, or greater inbreeding at the lowphase, or random processes.In our study, the minimum age at seizure onset amongwater voles was 39 days, the maximum age at seizure onsetwas 1105 days, and the median age at seizure onset, 274 days.Among bank voles, median age at seizure onset is 157 days. Seizures develop at a later age in volesand gerbils than in seizure-prone laboratory rats and mice.According to our findings, in the water vole, age at seizureonset is an inherited trait that can be selected for. Identificationof the main mutation that triggers seizures and elucidationof the mechanisms underlying the observed correlationsbetween age and seizures may provide insights into thepathogenesis of seizures in other mammals. It is now knownthat 70–80 % of epilepsy cases are associated with one ormore genetic factors, with the remaining cases attributableto acquired conditions such as a tumor, stroke, or head injury.Genes that control the predisposition to convulsive statesin response to handling can be stored in populations due totheir positive pleiotropic influence on life span. Here, wedemonstrated that under vivarium conditions, individuals withseizures live longer than individuals with a normal phenotype.These results contradict mouse studies that have addressed theeffects of individual genes on epileptogenesis and life span; this discrepancy points to diversityof genetic mechanisms mediating the association of such abehavioral phenotype with viability.The presence of common links in the effects of genes thatcontrol the onset of handling-induced seizures and life spanconfirms the positive effect of the mother’s convulsive statuson the life span of the offspring in water voles. The offspringof mothers with seizures were found to live longer than theoffspring of mothers with the normal phenotype, whereas theoffspring life span was not affected by seizure status of thefather. It is possible that genetic predisposition of femalesto seizures correlates with physiological parameters duringpregnancy, with lactation capacity, or with maternal behavior,which in turn determine offspring viability. We are planningon investigating this topic in the future.Thus, the water vole can serve as a suitable experimentalmodel for researching not only the nature of convulsive statesbut also the evolution of longevity. Further studies are neededregarding physiological and genetic mechanisms behind thehandling-induced seizures in the water vole as well as regardingthe relation of seizure susceptibility to life historytraits.It is shown for the first time that the water vole can have convulsiveseizures in response to handling. When water volesare kept in a vivarium, the proportion of animals susceptibleto convulsive seizures is higher among water voles caught inthe decline or depression phase of a cycling natural populationthan among water voles caught in the increase or peak phase. The susceptibility to seizures is an inherited trait correlatingwith the life span; this finding implies the possibility ofnatural and artificial selection for this trait. Our data open upopportunities for the creation of water vole strains that differin their susceptibility to handling-induced epileptiform seizures,for research on the mechanisms of epileptogenesis andlongevity.The authors declare no conflict of interest.ReferencesBronson F.H., De La Rosa J. Tonic-clonic convulsions in meadowvoles. Physiol. Behav. 1994;56(4):683-685. DOI 10.1016/0031-9384(94)90227-5.Buchhalter J.R. Animal models of inherited epilepsy. Epilepsia. 1993;34(Suppl. 3): S31-S41. DOI 10.1111/j.1528-1157.1993.tb05921.Buckmaster P.S. 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DOI10.1038/nn.3934Gülersoy E., İyigün S.S., Erol B.B. An overview of seizures and epilepsyin rabbits: etiological differences and clinical management.Nauk. Vìsn. Vet. Med. 2021;1(165):159-164. DOI: 10.33245/2310-4902-2021-165-1-159-164.Jackson R.K. Unusual laboratory rodent species: research uses, care,and associated biohazards. ILAR J. 1997;38(1):13-21. DOI 10.1093/ilar.38.1.13.Krotova L.G. Changes in the adrenal glands and carbohydrate metabolismin the water vole (Arvicola terresrris) in the spring-summerperiod. In: Issues of the intraspecific variability of mammals. Proceedingsof the Institute of Biology. Sverdlovsk, 1962;29:129-140.(in Russian)Ludvig N., Farias P.A., Ribak C.E. An analysis of various environmentaland specific sensory stimuli on the seizure activity of the Mongoliangerbil. Epilepsy Res. 1991;8(1):30-35. DOI 10.1016/0920-1211(91)90033-c.Marshall G.F., Gonzalez-Sulser A., Abbott C.M. Modelling epilepsy inthe mouse: challenges and solutions. Dis. Model. Mech. 2021;14(3):dmm047449. DOI 10.1242/dmm.047449.Muñoz L.J., Carballosa-Gautam M.M., Yanowsky K., García-AtarésN., López D.E. The genetic audiogenic seizure hamster fromSalamanca: The GASH:Sal. Epilepsy Behav. 2017;71(Pt. B):181-192. DOI 10.1016/j.yebeh.2016.03.002.Myers C.T., Mefford H.C. Advancing epilepsy genetics in the genomicera. Genome Med. 2015;7(1):91. DOI 10.1186/s13073-015-0214-7.Okudan Z.V., Özkara Ç. Reflex epilepsy: triggers and managementstrategies. Neuropsychiatr. Dis. Treat. 2018;14:327-337. DOI10.2147/NDT.S107669.Pakozdy A., Halasz P., Klang A. Epilepsy in cats: theory and practice.J. Vet. Intern. Med. 2014;28(2):255-263. DOI 10.1111/jvim.12297.Poletaeva I.I., Kostyna Z.A., Surina N.M., Fedotova I.B., Zorina Z.A.The Krushinsky–Molodkina genetic rat strain as a unique experimentalmodel of seizure states. Vavilovskii Zhurnal Genetiki i Selektsii= Vavilov Journal of Genetics and Breeding. 2017;21(4):427-434. DOI 10.18699/VJ17.261. (in Russian)Schønecker B. 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Mactra veneriformis  is a bivalve mollusk of major economic importance in China. Decreased natural yields of M. veneriformis have led to an urgent need for genomic resources. To address this problem and the currently limited knowledge of molecular evolution in this genus, we here report a high-quality chromosome-level genome assembly of M. veneriformis. Our approach yielded a 939.32 Mb assembled genome with an N50 contig length of 7,977.84 kb. Hi-C scaffolding of the genome resulted in assembly of 19 pseudochromosomes. Repetitive elements made up ∼51.79% of the genome assembly. A total of 29,315 protein-coding genes (PCGs) were predicted in M. veneriformis. Construction of a genome-level phylogenetic tree demonstrated that M. veneriformis and Ruditapes philippinarum diverged around 231 million years ago (MYA). Inter-species comparisons revealed that 493 gene families have undergone expansion and 449 have undergone contraction in the M. veneriformis genome. Chromosome-based macrosynteny analysis revealed a high degree of synteny between the 19 chromosomes of M. veneriformis and those of Patinopecten yessoensis. These results suggested that M. veneriformis has a similar karyotype to that of P. yessoensis, and that a highly conserved 19-chromosome karyotype was formed in the early differentiation stages of bivalves. In summary, the genomic resources generated in this work serve as a valuable reference for investigating the molecular mechanisms underlying biological functions in M. veneriformis and will facilitate future genetic improvement and disease treatment in this economically important species. Furthermore, the assembled genome greatly improves our understanding of early genomic evolution of the Bivalvia. Mactra veneriformis is a type of infaunal suspension-feeding bivalve that is ubiquitous and abundant in tidal flats, subtidal areas, and shallow seas along the coast of China, Korea, and Japan  broughtonii . DNA was then sheared using a sonication device to enable the construction of short-insert paired-end (PE) libraries. The short-insert (500 bp) libraries were constructed following the instructions provided with the Illumina Nextera DNA Library Prep Kit. All libraries were sequenced on an Illumina X-TEN platform . After the Illumina sequencing adaptors were removed, low-quality bases at the ends of the raw reads were trimmed. Reads were then scanned using a 4-bp sliding window for further trimming using an average quality per base threshold of 15. The resulting clean data were used in downstream analyses. For long-read sequencing, a PacBio SMRTbell library (20 kb) for PacBio Sequel was constructed using the SMRTbell Express Template Prep Kit 2.0 . Fragments smaller than 7 kb were filtered out using BluePippin . The library was bound to polymerase using the Sequel II Binding Kit 2.0 (PacBio) and loaded onto a PacBio Sequel II sequencing platform using HiFi Bundle (v2) sequencing reagent and 8M SMRT cells. M. veneriformis muscles fixed in 1% (v/v) formaldehyde were used for in situ Hi-C library construction. Nucleus extraction, permeabilization, chromatin digestion, proximity-ligation treatments, and subsequent DNA manipulation were performed as described previously  (k-mer (k = 17) depth distribution, and roughly estimated the genome size by dividing the total number of k-mers by their respective coverage.The genome size and heterozygosity of v2.2.10)  to obtaiWe integrated the assembled PacBio HiFi reads and PE Hi-C reads using Hifiasm . The asshttp://www.repeatmasker.org; accessed 2022 September 8) was used to perform de novo identification and masking of the repeat sequences in the assembly. To identify protein-coding regions, we obtained annotations via de novo prediction, homology-based searches, and transcriptome-assisted methods. We used the de novo gene prediction packages Augustus v3.2.3  genome database and aligned them to the M. veneriformis genome assembly using BLASTP  were identified in different species and compared to those in M. veneriformis to understand gene family evolution in this species.To understand the evolutionary relationship between tic tree . Gene gaM. veneriformis and P. yessoensis were separated in the early stages of bivalve evolution. P. yessoensis possessing a highly conserved 19-chromosome karyotype similar to that of bilaterian ancestors. To determine whether M. veneriformis also had the conservative chromosome collinearity relationship, macrosynteny analysis was performed on M. veneriformis and P. yessoensis. A comparison of gene synteny between M. veneriformis and P. yessoensis was conducted to visualize putative homologous gene pairs or marker pairs as inferred by sequence similarity. BLASTP was used in the gene-to-gene comparison with default e-values of 1e−5. A macrosynteny conservation index was calculated as measure of conservation  and DNA transposable elements represented 2.94% and 1.27%, respectively. In contrast, long terminal repeat (LTR) elements represented only 0.65% of the assembly (Pinctada fucata) genome (Crassostrea gigas (Repetitive elements made up a large portion of the assembly . It was M. veneriformis with an average coding sequence (CDS) length of 1526.9 bp  analyzed, a total of 101,358 gene families were identified, 403 of which were present in all 20 species . Furthermore, the expanded and contracted gene families contained a total of 184 and 219 significantly enriched KEGG pathways, respectively (FDR < 0.05). The enrichment analyses suggested that the expanded gene families were involved in biological processes associated with pathogen responses and disease resistance (To compare different genomic traits between species, we performed a comparative genomic analysis of the 20 selected species using CAFE . We founsistance .P. yessoensis and T. granosa, have a highly conserved 19-chromosome karyotype (M. veneriformis was also shown to have the 19-chromosome karyotype; however, the results of the phylogenetic analyses showed that M. veneriformis and P. yessoensis were in different groups of bivalves, meaning that the 2 species separated in the early stages of bivalve evolution. To determine whether M. veneriformis also had the conservative chromosome collinearity relationship, macrosynteny analysis was performed on M. veneriformis and P. yessoensis. The results showed high karyotype conservation between M. veneriformis and P. yessoensis, with the latter possessing a highly conserved 19-chromosome karyotype similar to that of bilaterian ancestors (M. veneriformis and those of P. yessoensis. Chromosome 01 of P. yessoensis corresponded to chromosomes 07 and 19 of M. veneriformis, whereas chromosome 07 of M. veneriformis corresponded to chromosomes 01 and 16 of P. yessoensis (P. yessoensis chromosomes 01 and 16 and M. veneriformis chromosomes 07 and 19, we performed a collinearity analysis of those 4 chromosomes. The results showed that chromosome fission or fusion events may have been caused by a Robertsonian translocation (P. yessoensis chromosome 01 also corresponded to 2 chromosomes of T. granosa, supporting the results of the present study. The results also suggest that Robinson chromosome translocation may have occurred in the P. yessoensis genome. The genomic collinearity analysis showed that there were few inter-chromosomal translocations between the genomes of M. veneriformis and P. yessoensis, whereas intra-chromosomal translocations occurred more frequently. This may be the explanation for the highly conserved 19-chromosome karyotype observed among bivalves.Previous studies have shown that scallops and many other bivalves, such as ssoensis . To clarlocation . PreviouM. veneriformis using a combined strategy encompassing 3 distinct sequencing technologies: Illumina short reads, PacBio single-molecule sequencing, and Hi-C. We described features of the newly assembled genome and performed phylogenetic and gene family evolution analyses using 19 mollusk relatives. We performed chromosomal macrosynteny analysis and identified chromosomal fusion or fission events between the genomes of M. veneriformis and P. yessoensis, and determined that bivalves have a relatively low frequency of inter-chromosome translocation and a high frequency of intra-chromosome translocation. This newly assembled M. veneriformis genome constitutes an excellent resource for genomic, biological, and ecological studies of bivalve mollusks. High-quality genomic data for this species will contribute to development of molecular breeding techniques to generate bivalves with high yield, rapid growth, and disease resistant phenotypes with favorable economic traits.In this study, we assembled the first known high-quality chromosome-level genome for jkac229_Supplemental_Material_LegendsClick here for additional data file.jkac229_Supplementary_Figure_S1Click here for additional data file.jkac229_Supplementary_Figure_S2Click here for additional data file.jkac229_Supplementary_Figure_S3Click here for additional data file.jkac229_Supplementary_Figure_S4Click here for additional data file.jkac229_Supplementary_Figure_S5Click here for additional data file.jkac229_Supplementary_Figure_S6Click here for additional data file.jkac229_Supplementary_Figure_S7Click here for additional data file.jkac229_Table_S1Click here for additional data file.jkac229_Table_S2Click here for additional data file.jkac229_Table_S3Click here for additional data file.jkac229_Table_S4Click here for additional data file.jkac229_Table_S5Click here for additional data file.


Strongyloides spp. infections. We assessed the prevalence and intensity of STH species in Solomon Islands by conducting a province-wide survey using quantitative PCR (qPCR) for diagnosis, which can provide much better characterisation of STH burden than microscopy.The Kato-Katz microscopy technique is the global standard for assessment of soil-transmitted helminth (STH) burden. However, major limitations include its poor sensitivity, requirement for rapid sample processing, and inability to differentiate hookworm species nor detect Necator americanus (54.5% [range 17.5–89.4%]), followed by Ancylostoma ceylanicum (15.5% [range 2.8–45.8%]), Trichuris trichiura (9.1% [range 0–79.2%]), and Strongyloides spp. (3.2% [range 0–29.2%]). Most infections were of light intensity for N. americanus (85.7%) and T. trichiura (90.7%). Owning a household latrine was associated with a lower risk of N. americanus infection  while greater precipitation was linked to more common T. trichiura infection .We conducted a cross-sectional survey in 18 villages in Western Province to detect infections with six STH species and quantify intensity with three. We used linear mixed model regression to identify potential water, sanitation, and hygiene (WASH) and environmental risk factors for infection. We collected stool specimens from 830 village residents. Overall STH prevalence was 63.3% (range 27.5 to 91.5% across villages), led by T. trichiura and Strongyloides spp., both of which are poorly responsive to albendazole. Furthermore, One Health strategies are needed for improved A. ceylanicum and Strongyloides spp. control, WASH access and use should be improved to complement deworming programmes, and control efforts should ideally be expanded to entire communities.In this first large-scale population survey of STH in the Pacific using qPCR, we found evidence that ivermectin should be incorporated into STH control programmes because of the presence of ACTRN12618001086257.ClinicalTrials.gov Australian and New Zealand Clinical Trials Registry  Strongyloides spp. infections. It is important to assess the burden of each of these infections to design control approaches beyond the current core strategy of albendazole preventive chemotherapy. Use of qPCR for diagnosis could address these gaps because it can distinguish all relevant intestinal worm species. We completed a province-wide intestinal worm infections prevalence survey using qPCR in Western Province, Solomon Islands. Overall prevalence was extremely high (62.5%). We identified a high burden of Necator americanus (54.5%) and detected Ancylostoma ceylanicum (15.5%), Trichuris trichiura (9.1%), and Strongyloides spp. (3.2%) infections. Increased age and precipitation were associated with higher prevalence of infection, while owning a household latrine was associated with lower odds of N. americanus infection. Our findings support the need to incorporate ivermectin into deworming programmes for Strongyloides spp. and T. trichiura control as these species are poorly responsive to albendazole, implement One Health strategies to address the zoonotic A. ceylanicum and Strongyloides spp., and expand control efforts to reach entire communities. WASH access should also be improved to complement deworming programmes.Routine assessments of the burden of intestinal worm infections rely on microscopy-based diagnostic methods, such as the Kato-Katz technique. However major limitations include its poor sensitivity, requirement for rapid sample processing, and inability to differentiate individual hookworm species and detect  Ascaris lumbricoides, Trichuris trichiura, Strongyloides stercoralis and the hookworms Necator americanus, Ancylostoma duodenale and Ancylostoma ceylanicum represent the most frequently occurring of all neglected tropical diseases (NTDs). STHs infect an estimated 895 million people worldwide ). A. lumbricoides infection was rare, at 0.08% (range 0–16.7%). Relative to those who were infected, most infections were of light intensity for A. lumbricoides (66.7%), T. trichiura (90.7%), and N. americanus (85.7%). Relative to the entire sample, the prevalence of moderate-to-heavy intensity infections were as follows: A. lumbricoides (0.2%), T. trichiura (1.9%), and N. americanus (7.6%). Regarding the geographic distribution of infections for each species, we observed that the prevalence of N. americanus infections was above 20% in 94.4% of villages. A total of 33.3% of villages had T. trichiura prevalence of above 20%, 27.8% of villages for A. ceylanicum, and 5.6% of villages for Strongyloides spp. , and there was a pattern of increasing risk of infection with age relative to young children aged 1–5 years . For T. trichiura, there was no detectable difference in infection prevalence by sex but we observed increasing prevalence by age within children  and adolescents , compared young children  and adults  than young children. There was no evidence of differences in prevalence by sex or age for A. ceylanicum. For infection intensity, there was a pattern of increasing N. americanus egg counts with age within in adolescents  and older adults , compared to young children (Table E in T. trichiura infection intensity by age or sex. Demographic differences in A. lumbricoides infections were not examined due to few positive cases (n = 6).In the analysis of risk factors Tables  and 5, mdren see . There wN. americanus than those who reported not owning a toilet . Participants from villages with less acidic soil were 3 times more likely to have an infection than those from villages with more acidic soil . Co-infection with A. ceylanicum  and T. trichiura  were associated with a higher odds of N. americanus infection.As shown in A. ceylanicum infection than those whose water was from an unimproved source . Co-infection with N. americanus  and Strongyloides spp.  were associated with a higher prevalence of A. ceylanicum infection.Participants whose main drinking water source was from an improved source had higher odds of T. trichiura infection . Greater annual precipitation was associated with marginally higher odds of infection , as was co-infection with N. americanus .Participants who reported always wearing shoes outside were significantly less likely to have Strongyloides spp. infection .We did not detect any statistically significant associations between WASH variables and nfection . ParticiStrongyloides spp. and individual hookworm species. Previous large-scale STH epidemiological surveys to use qPCR as a stand-alone diagnostic tool include three conducted in Timor-Leste in the context of trials [T. trichiura, Strongyloides spp. and the zoonotic A. ceylanicum were also abundant. Most infections were of light intensity for N. americanus, T. trichiura, and A. lumbricoides.In this study, to our knowledge, we completed the first STH prevalence survey in a low- or middle-income country of the Pacific region that used qPCR, enabling the assessment of f trials –15 and of trials . We founf trials –21. Consf trials ,18, the Solomon Islands follows WHO recommendations for STH control wherein school-based albendazole preventive chemotherapy is provided, so far only within the capital city, Honiara. Guidelines specify treatment to groups at highest risk of morbidity, including pre-school and school-aged children, in communities with prevalence above 20%, with the aim of reducing the prevalence of moderate-to-heavy intensity infections to less than 2% . Our finA. ceylanicum is a zoonoses and is a predominant hookworm of domestic dogs and cats in the Asia Pacific region [Strongyloides spp., for which WHO recently introduced control targets to be attained by 2030 [Strongyloides spp. [T. trichiura [T. trichiura [S. stercoralis [However, our findings also show that the drug-based treatment approach alone is unlikely to control STHs, for several reasons. c region . Strongy by 2030 , is also by 2030 ,36. Zoon by 2030 . Anotherdes spp.  and T. trichiura . The co-richiura  and iverrcoralis . STH conrcoralis .N. americanus, T. trichiura, and Strongyloides spp., and greater N. americanus infection intensity, increased with age. Although school-based treatment programs will benefit children, the considerable adult reservoir in the population suggests that treatment should ideally be expanded to entire communities, perhaps integrating with mass drug administration programmes targeting other NTDs.It is important to note that risk of infection for N. americanus infections, probably because of reduced exposure to contaminated faecal matter [T. trichiura infection, likely due to moist soil conditions favourable to survival and development of STH eggs and larvae [N. americanus and Strongyloides spp. consistent with in-vitro findings on the optimal pH being 6.0 [Overall, only few WASH and environmental factors emerged as significant predictors of STH infection, possibly due to the presence of homogenously poor WASH access/conditions and limited variation in the environmental data across a small geographical area. We found that owning a household latrine was associated with a lower prevalence of l matter . Higher d larvae . Precipid larvae ,34. We feing 6.0 . Given seing 6.0 , the useT. trichiura infection. While shoe-wearing can confer protection against species that enter by percutaneous penetration, T. trichiura transmission occurs via the faecal-oral route. More likely, increased shoe-wearing behaviour might reflect higher socioeconomic status, which may be a proxy of an unmeasured variable that protects against T. trichiura exposure. Surprisingly, participants whose drinking water was from an improved source had increased odds of infection with A. ceylanicum. This might be due to increased exposure to A. ceylanicum larvae near improved water sources deposited by infected dogs in communities. We did not collect data on STH burden in dogs or contamination of the environment with dog faeces, a key gap for future research to address.An unexpected finding was that individuals who reported always wearing shoes outside had reduced odds of Limitations of this study should be considered. The WHO recommended thresholds for defining infection intensity were created using infection intensity measurements derived from the Kato-Katz technique. While there have been studies, including this one, deriving epg from Ct values ,10, addiStrongyloides spp. and individual hookworm species. This enabled us to identify opportunities to strengthen STH control in Solomon Islands, particularly incorporating ivermectin preventive chemotherapy into deworming programmes, adopting a One Health framework, and expanding STH control to entire communities. Improving WASH use and access could complement deworming programmes by protecting against exposure pathways.In conclusion, by using qPCR in a province-wide epidemiological survey in Solomon Islands, we were able to comprehensively assess the burden of all STH species including S1 TextT. trichiura and N. americanus infections. Table F in S1 Text. Results of model building steps for N. americanus model. Table G in S1 Text. Results of model building steps for A. ceylanicum model. Table H in S1 Text. Results of model building steps for T. trichiura model. Table I in S1 Text. Results of model building steps for Strongyloides spp. model Table J in S1 Text. Results of model building steps for hookworm (undifferentiated) model Table K in S1 Text. Results of model building steps for STH (undifferentiated) model.Table A in S1 Text. Sequences of primers and probes used for quantitative polymerase chain reaction. Table B in S1 Text. WASH characteristics of study population (N = 619). Table C in S1 Text. Environmental characteristics of study population (N = 830). Table D in S1 Text. Unadjusted STH prevalence by species, stratified by sex and age group. Table E in S1 Text. Mean eggs per gram (epg) of stool and incidence rate ratios (IRRs) by sex and age group for (DOCX)Click here for additional data file.S1 STROBE Checklist(DOCX)Click here for additional data file.


Polycystic ovary syndrome (PCOS) and periodontal disease (PDD) share common risk factors. The bidirectional interaction between PCOS and PDD has been reported, but until now, the underlying molecular mechanisms remain unclear. Endocrine disorders including hyperandrogenism (HA) and insulin resistance (IR) in PCOS disturb the oral microbial composition and increase the abundance of periodontal pathogens. Additionally, PCOS has a detrimental effect on the periodontal supportive tissues, including gingiva, periodontal ligament, and alveolar bone. Systemic low-grade inflammation status, especially obesity, persistent immune imbalance, and oxidative stress induced by PCOS exacerbate the progression of PDD. Simultaneously, PDD might increase the risk of PCOS through disturbing the gut microbiota composition and inducing low-grade inflammation and oxidative stress. In addition, genetic or epigenetic predisposition and lower socioeconomic status are the common risk factors for both diseases. In this review, we will present the latest evidence of the bidirectional association between PCOS and PDD from epidemiological, mechanistic, and interventional studies. A deep understanding on their bidirectional association will be beneficial to provide novel strategies for the treatment of PCOS and PDD. PCOS co2vice versa. Several cross-sectional studies unanimously concluded that PCOS increases the risk of PDD  and clinical attachment loss (CAL). However, no difference were observed in oral-health-related quality of life (OHRQoL) scores between the two groups, which might reveal a poor periodontal status and weak awareness of oral health care in infertile women . PCOS ask of PDD –18. Womek of PDD . A retrok of PDD . The bidk of PDD . In addik of PDD . Althougk of PDD , strong k of PDD Table 1.3PCOS is featured with endocrine disorders, including HA, insulin resistance (IR), and estrogen reduction. Endocrine disorder aggravates the development of PDD. We summarize the potential mechanisms of endocrine disorders between PCOS and PDD in the following four aspects Figure 13.1Fusobacterium nucleatum (F.n) is one of the anaerobic bacteria in supra- and subgingival biofilms. F.n serves as a bridge to assist the interspecies coaggregation, including Porphyromonas gingivalis (P.g), Treponema denticola (T.d), and Streptococcus gordonii (S.g), and accelerates biofilm formation via outer membrane proteins. In addition, F.n breaks the epithelial barrier and engages in attacking the lymphocytes, resulting in the host immune imbalance  were observed in gingival crevicular fluid (GCF) in patients with comorbidity of PCOS and gingivitis rather than patients with gingivitis alone  in subgingival microflora in adolescent girls with PCOS   , inhibitnd P.g)  by inducnd P.g) . Therefo3.22  are observed in PCOS patients than in healthy controls , which in vitro . In addin vitro . It has n vitro . Decreasn vitro , 72. Pron vitro . Progestex vivo . In addiex vivo . In summ3.4+CD28− T cell and Th17/Treg ratio . The PCOS-induced OS has an impact on gingival inflammation. A higher level of malondialdehyde (MDA) (a lipid peroxidation product indicating OS) in serum and GCF was detected in women with PCOS, which was positively correlated with gingival inflammation , 27. In 3.7Epidemiological data showed that obesity affects 30%–70% PCOS women . Althoug3.8Porwal et al.  compared44.1PDD is characterized by chronic inflammation induced by the subgingival biofilm. Various studies have demonstrated that multiple proinflammatory cytokines and reactive oxygen species (ROS) are involved in the systemic effects of PDD on systemic diseases, such as DM, MS, and obesity . PDD inc4.2P.g, a major pathogen in periodontitis, can exacerbate IR   . IR altee versa . In addie versa , 94. The4.3Bacteroides vulgatus was identified in gut microbiota of PCOS women , primarily expressed in adipose tissues, exerts an influence of insulin sensitivity through regulating glucolipid metabolism . Both PC5.2Environmental factors have significant impact on the development of PCOS and PDD. Several studies have confirmed that women with low socioeconomic status have higher incidence of PCOS . Meanwhi6Multiple lines of evidence confirmed the bidirectional relationships between PCOS and PDD. Endocrine disorders, low-grade inflammation, immune imbalance, and OS in PCOS deteriorate the periodontal microenvironment, while PDD accelerates the development of low-grade inflammation, OS, and IR and increases the risk of PCOS. However, only a few intervention studies revealed the mechanisms underlying the causal relation between PCOS and PDD. In addition, both PCOS and PDD share common risk factors including genetic or epigenetic predisposition and low socioeconomic status. Further longitudinal studies are needed to elucidate the shared pathophysiology between PCOS and PDD.YD, JX, DW, and JT conceived the study question, and all authors participated in the study design. JX created the first draft of the manuscript. HX, PZ, ZZ, and WF made substantial contributions to drafting the article. YD revised the manuscript. All authors contributed to the article and approved the submitted version.


Bacillus pumilus, Enterobacter kobei, Klebsiella pneumonia, Pseudomonas mandelii, Pseudomonas putida, Pseudomonas avellanae, and Staphylococcus equorum), isolates had efficacy for cadmium tolerance and showed sequestration potential at varying MIC. Furthermore, K. pneumonia was observed to have the highest (900 ppm) tolerance for cadmium and the lowest (600 ppm) was shown by E. kobei. Besides, K. pneumonia showed the highest (75.2%) sequestration potential while the least (52.4%) potential was observed for P. putida. These cadmium tolerant species can be implemented in contaminated environments for detoxification and elimination of cadmium from these agricultural fields.Due to industrialization, the contamination of toxic metals in soils is currently one of the major concerns to scientists worldwide. The presence of high concentrations of heavy metals including cadmium in the environment is mainly attributed to human activities. Being a highly toxic metal, cadmium can enter plant cell transporters usually used for the uptake of essential cations, such as iron, calcium, and zinc. This study deals with the appraisement of response and tolerance shown by various bacteria in varied cadmium concentrations . The optical density (OD) of the isolates was measured to determine the minimum inhibitory concentration (MIC) of cadmium. Isolated bacteria have been identified using 16S rRNA gene sequence and Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Among the 72 isolates, 07 ( Pseudomonas putida, Bacillus pumilus, Klebsiella pneumonia, Pseudomonas mandelii, Pseudomonas avellanae, Enterobacter kobei, and Staphylococcus equorum) were found to have efficacy for cadmium tolerance and showed sequestration potential at varying MIC.07 Bacterial isolates (Klebsiella pneumonia was observed to have the highest (900 ppm) tolerance for cadmium and the lowest (600 ppm) was shown by E. kobei.Klebsiella pneumonia showed the highest (75.2%) sequestration potential while the least (52.4%) potential was observed for P. putida.All the isolated bacteria were identified using latest and robust techniques.Bacillus subtilis. In Bacillus licheniformis, teichoic and teichuronic acids are key binding sitesfor metals . Another report from the region also suggests that pollution amounts the highest threat to saffron quality . From the various sites, the replicates of the soil samples were collected in equal proportions (approx. 10 g). The sample was mixed systematically and subjected to segregation of cadmium-resistant bacteria by using the standard protocol .Serial dilution of soil samples with normal saline was performed for bacterial confinement accompanied by immediate direct plate  and pureThe bacterial isolates were identified using morphological characteristics, sequencing of the 16S rRNA gene, and Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) identification.viz. colony size, shape, elevation, color, opacity, and margins. The morphological description of the bacteria was carried out using Gram’s staining method followed by direct microscopy (100x Olympus IX 71 fluorescence microscope).To analyze, macro- as well as micro-morphological features of bacteria, incubation of media plates, were done for 1–3 days at 37°C. Following which the bacteria were characterized on the basis of colony characteristics The Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) technique was used to identify bacteria using the Ethanol-formic acid extraction process .GenElute bacterial Genomic DNA kit (Sigma) was used to extract and purify DNA according to manufacturing protocol followed by amplification of the 16S rRNA gene using universal bacterial primers forward primers [(27F) 5′AGAGTTTGCCTGCTCAG-3′] and reverse primers [(1429R) 5′-GGGTTACCTGACTGACTTT-3′; via a 0.45 μm pore size syringe filter to eliminate any bacterial contamination. To determine the MIC, various concentrations were prepared from the stock solution using the Muller Hinton Broth as per the existing protocol  of cadmium chloride (Himedia) screened protocol . Before colonies . Hence, viz., sulphuric acid and nitric acid were used followed by the determination of metal concentration in percentage by Atomic Absorption Spectrometer (AAS Perkin Elmer Germany).The sequestration potential of isolates tolerant to cadmium was carried out using standard methods , and ana3 and 9.5 × 104 CFU/g. Out of the 72 bacterial isolates, the percentage of Gram-negative was found to be 35.5%, whereas the percentage of Gram-positive was 64.5%. However, it was revealed by the taxonomical classification that the most dominant family among the isolated bacteria was Bacillaceace (43.24%) followed by other families like Enterobacteriaceae (21.62%) Planococcaceae(8.10%), Microbacteriaceae (8.10%), Pseudomonadaceae (5.40%), Staphylococceaceae (5.40%), and Aeromonadaceae (2.70%).Enumeration of the bacterial colonies obtained on the petri-plates was carried using a digital colony counter, and the colony forming units (CFUs/g) were used to assess the bacterial load. In summer seasons, bacterial load was found to be in elevated concentration as compared to winter seasons. In the successive years of study, we observed an overall increase in bacterial load during all four seasons of the year. In the three sampling sites, the CFU/g was found to vary between 8 × 10viz., P. putida, B. pumilus, Klebsiella pneumoniae, P. mandelii, P. avellanae, E. kobei and S. equorum.Out of 72 total isolates only seven bacterial species identified by MALDI-TOF MS and 16S rRNA gene sequencing were found to be tolerant to cadmium , 3 viz.,Based on the Tamura-Nei model  evolutioP. putida, B. pumilus, Klebsiella pneumonia, P. mandelii, P. avellanae, E. kobei, and S. equorum.All the cadmium-resistant bacteria isolates were evaluated and thus analyzed using a linear MALDI-TOF mass spectrometer analysis . The criterion score that was used for identification was as per the recommendations of the manufacturer. The isolates presenting a score of ≥2.000 reveal identification up to species-level, a score between 1.700 and 1.999 illustrate identification up to genus level, and a score of <1.700 revealed no identification. The MS spectra thus produced  and the K. pneumoniae showing nine spectra in the ion range at m/z 3250.957–4850.437; P. putida, six spectra in the ion range at m/z 3705.083–10444.950; B. pumilus, eight spectra in the ion range at m/z 4365.567–15387.201; P. avellanae, nine spectra in the ion range at m/z 3143.861–4263.179; P. mandelii showed seven spectra in the ion range at m/z 3129.834–13758.497; E. kobei, eight spectra in the ion range at m/z 4365.467–15397.201 and S. equorum showed five spectra in the ion range at m/z 4304.659–4794.126. MALDI-TOF-MS profile analysis revealed the presence of species-specific ion signals.Based on Both the sequencing of the 16Sr RNA gene and the MALDI-TOF MS analysis gave identical identification results of isolates hence, it can be inferred that both the techniques are reliable and can be used for the identification of bacteria.K. pneumoniae has the highest (900 ppm) and E. kobei the lowest (600 ppm) tolerance against cadmium.The tolerance of all isolates against cadmium and the subjective MIC . determiK. pneumoniae showed the highest sequestration potential of 75.2% among the cadmium tolerant species, whereas P. putida showed the lowest sequestration potential of 52.4% , which are involved in protecting these cells from toxic metal effects (K. pneumonia exhibited the highest range of tolerance (900 ppm) while E. kobei showed the lowest range (600 ppm). Several studies had shown that bacteria have the effectiveness to remove Cd from contaminated environments. Similar to our findings bacterial species like Pseudomonas aeruginosa and Rhodobacter sphaeroides have shown promising results in Cd removal in fixed temperatures as reported by Pseudoalteromonas sp., from the deep-sea that have shown efficacy for Cd removal via biosorption and intracellular uptake from the liquid media and the same mechanism can be attributed to the bacterial species like P. putida and P. mandelii of the current report . In the present investigation, K. pneumoniae showed the excellent ability for cadmium sequestration, i.e., 75.2%, and showed a tolerance level of 900 ppm. The other bacterial species tolerant to cadmium have shown promising results in tolerance and sequestration as well. The primary conclusion depicts the cadmium tolerance among different isolated bacterial species collected from the saffron soils of Kashmir valley and further highlighting their role in Cd sequestration. Besides, detoxification and elimination of the heavy metals from soils contaminated with cadmium, the cadmium-resistant property might prove to be beneficial, thereby offering possible alternatives for the process of bioremediation.Matrix-assisted laser desorption ionization time-of-flight mass spectrometry and 16S rRNA sequencing documented seven cadmium tolerant species of bacteria (The data presented in the study are deposited in GenBank repository and accession numbers are MK424244, MK422437, MK530126, MN239484, MK530124, MK426682, and MK426804. All the data has been released by the GenBank.BU: writing—original draft, writing—review and editing, data curation, and formal analysis. RN: concept design, data curation, formal analysis, writing, review, and editing. BG: writing—review and editing. PR: review and editing. All authors contributed to the article and approved the submitted version.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, or those of the publisher, the editors and the reviewers. 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.


We used data from people initially free of clinical cardiovascular disease to evaluate the association between metabolic syndrome (MS) and incident preclinical heart failure (pHF).STANISLAS was a familial, single-center, longitudinal prospective cohort study composed of 1,006 families from Nancy, France . Age- and sex-adjusted logistic regression and inverse probability weighting models were used to evaluate the association between MS and pHF, which was defined by diastolic dysfunction, atrial enlargement, ventricular hypertrophy, or elevated natriuretic peptides. Among 944 people who were adults at the first and final visit, those with baseline MS were more likely to be older (63 vs. 61 vs. 59 years of age) and male (73% vs. 55% vs. 45%) compared to people who developed incident MS and people who had no baseline MS, respectively. Furthermore, compared to people without baseline MS, the risk of pHF was numerically larger among people with baseline MS  and people who developed incident MS . Concerning the metabolic determinants of MS, the risk of pHF was most elevated in people with baseline hypertension  and elevated waist circumference .Overall, HF is an important public health concern given the high risk of mortality when patients with MS or elevated fasting glucose become established with the disease. Early aggressive lifestyle modification and medical intervention among patients free of cardiovascular disease with an obese-hypertensive phenotype may be warranted to prevent HF development. There is a high prevalence of metabolic syndrome (MS) among patients with preclinical heart failure (HF) . AlthougSTANISLAS was designed as a familial, single-center, prospective longitudinal cohort study composed of 1,006 families from Nancy, France, who were recruited between 1993 and 1995. The families were healthy and free of any acute and/or chronic illnesses at recruitment. From 2011 to 2016, STANISLAS participants were revisited for a fourth examination V4) at our department as previously described ), reduced HDL-C , hypertension , and elevated fasting glucose  [ mmol/L ip value comparing the three groups' associations with preclinical HF was computed in a multinomial weighted regression model. All analyses were conducted with SAS (v9.0).For the primary analysis, we evaluated the association between MS and preclinical HF among individuals who were adults at the V1 examination. We utilized age- and sex-adjusted logistic regression and inverse probability weighting models. That is, among patients at V4, the inverse probability of treatment  was computed with a propensity score that was based on a logistic model adjusted for age and sex. The inverse probability of treatment was then balanced across the three groups  to minimize between-group clinical differences and ensure exchangeability. The association between the three groups and preclinical HF was then evaluated utilizing a generalized logistic regression model. Sensitivity analyses removing the NTproBNP criterion from the preclinical HF definition were also conducted. A global Biological samples were obtained by venipuncture at the Nancy Hospital Center's Centre d'Investigation Clinique. Using sample-handling protocols, serum and plasma samples were obtained. All samples were then preserved at temperatures ranging from −80°C to −196°C in a central biobank. NTproBNP was then quantified utilizing an electrochemiluminescence immunoassay .The median follow-up time was 17 years (1993–2016). Among the 1,705 participants with V4 examinations between 2011 and 2016, only 944 individuals were adults at the V1 examination , male (73% vs. 55% vs. 45%), have elevated waist circumference (110 vs. 99 vs. 87 cm), hypertension (79% vs. 53% vs. 16%), and have elevated fasting glucose  patients had baseline MS and 263 (28%) patients developed MS during follow-up. At the V1 examination, 5% of study participants had a BMI >30 kg/mp = 0.05) and patients who had MS at baseline  patients with complete echocardiographic and biomarker data developed preclinical HF  and among patients who had MS at baseline . Adjusting for BMI in addition to age and sex attenuated the significant association of baseline MS and preclinical HF , but a numerically increased risk was still observed.In the sensitivity analysis, 354 of the 872 (40.6%) patients developed preclinical HF with respect to the definition that omitted NTproBNP. Consistent results for the risk of preclinical HF were seen in this sensitivity analysis. Compared to patients without baseline MS, the risk of preclinical HF was numerically increased both among patients who developed incident MS  appeared to be the metabolic determinants most associated with preclinical HF development.p trend HR <0.0001) [Our analysis extends on prior work from cohorts with older patients with greater degrees of cardiovascular comorbidities. A prior analysis of 51,451 patients in the Women's Health Initiative, Multi-Ethnic Study of Atherosclerosis (MESA), and CHS suggested a dose-response relationship between BMI and the risk of HF with preserved ejection fraction (<0.0001) . In the <0.0001) . The comn = 408; 46.8%) was smaller than the population of adult patients with echocardiographic data alone (n = 872). Nevertheless, the association between baseline MS and preclinical HF was consistent in sensitivity analyses where NTproBNP was removed from the preclinical HF definition. Finally, there was no further follow-up available beyond V4 to enable analyses evaluating the association between MS and clinically confirmed acute HF.Our study had some limitations. First, our analysis did not assess the impact of diet and physical activity on modifying the relationship between MS and preclinical HF. As a result, the impact of diet and physical activity on HF development warrants further evaluation. Second, prior analyses have identified a poor association between biomarkers  and structural heart disease; as a result, the use of natriuretic peptides in our definition of preclinical HF may have limited the analysis. Nevertheless, when we removed NTproBNP from the definition, the association between baseline MS and preclinical HF was still observed . Third, Given the high risk of mortality when patients with MS or elevated fasting glucose become established with HF , HF prevOur study demonstrated that after 17 years of follow-up, patients with baseline MS, especially those with obese-hypertensive phenotypes, are at increased risk of developing preclinical HF. As a result, the primary prevention of preclinical HF should be an important public health focus given the high risk of mortality when patients with metabolic aberrations are diagnosed with clinically confirmed HF. Evaluating the preventive efficacy of early aggressive lifestyle modification and medical intervention among patients free of cardiovascular disease with obese-hypertensive phenotypes may be warranted.All STANISLAS participants consented to having their data utilized for subsequent secondary analyses. As a result, no separate ethics approval was needed for the present study. The local Ethics Committee in Nancy, France (Comité de Protection des Personnes Est III-Nancy-France) approved the initial research protocol .A.S. reports receiving support from the Fonds de Recherche Du Quebec Sante (FRQS) Junior 1 clinician scientist award, Takeda, Roche Diagnostics, BMS-Pfizer and is funded by the Alberta Innovates Health Solution Clinician Scientist Fellowship. A.R. reports receiving funding from McGill University's Graduate Excellence Award in Medicine & Ethelwyn and Ernie Nyman Fellowship. P.R. reports receiving consulting fees from Novartis, NovoNordisk, Relypsa, AstraZeneca, Grünenthal, Idorsia, Stealth Peptides, Fresenius, and Vifor; lecture fees from Bayer and CVRx; and he is the cofounder of CardioRenal. F.Z. reports consulting fees for serving on the board of Boston Scientific; consulting fees from Novartis, Takeda, AstraZeneca, Boehringer Ingelheim, GE Healthcare, Relypsa, Servier, Boston Scientific, Bayer, Johnson & Johnson, and Resmed; and speaking fees from Pfizer and AstraZeneca. There are no other conflicts to report.The STANISLAS study is sponsored by Nancy CHRU. This work is supported by the French Ministry of Health “Programme Hospitalier de Recherche Clinique Inter regional 2013,” by the Contrat de Plan Etat-Lorraine and FEDER Lorraine, and a public grant overseen by the French National Research Agency (ANR) as part of the second “Investissements d'Avenir” program FIGHT-HF (reference: ANR-15-RHU-0004), and by the French PIA project “Lorraine Université d'Excellence,” reference ANR-15-IDEX-04-LUE. It is also supported by the European Fibro-Targets Project (Grant agreement No. SP7#602904), European HOMAGE project , the MEDIA project , FOCUS-MR (reference: ANR-15-CE14-0032-01), ERA-CVD EXPERT (reference: ANR-16-ECVD-0002-02), and the Fondation de Recherche en Hypertension Artérielle.The study was conceptualized by A.S., and F.Z. A.S. and J.P.F. contributed to the study methodology. A.S. and J.P.F. conducted the study investigation and contributed to the writing  of the manuscript. A.S., A.R., J.P.F., J.L.M., E.B., N.G., P.R., and F.Z. contributed to the writing (review and editing) of the manuscript. F.Z. supervised the study. We attest that all authors have reviewed and approved the article prior to its submission.The data that support the findings of this study are not publicly available because of their containing information that could compromise the privacy of research participants but are available from the corresponding author (F.Z.) upon reasonable request.