Mitochondria fulfil a plethora of cellular functions ranging from energy production to regulation of inflammation and cell death control. The fundamental role of mitochondria makes them a target of choice for invading pathogens, with either an intracellular or extracellular lifestyle. Indeed, the modulation of mitochondrial functions by several bacterial pathogens has been shown to be beneficial for bacterial survival inside their host. However, so far, relatively little is known about the importance of mitochondrial recycling and degradation pathways through mitophagy in the outcome (success or failure) of bacterial infection. On the one hand, mitophagy could be considered as a defensive response triggered by the host upon infection to maintain mitochondrial homeostasis. However, on the other hand, the pathogen itself may initiate the host mitophagy to escape from mitochondrial-mediated inflammation or antibacterial oxidative stress. In this review, we will discuss the diversity of various mechanisms of mitophagy in a general context, as well as what is currently known about the different bacterial pathogens that have developed strategies to manipulate the host mitophagy. In the light of their unique and intricate evolution, mitochondria developed into complex, motile, and highly dynamic organelles providing a variety of functions and signalling hubs for eukaryotic cells . BesidesMitophagy is a selective form of macroautophagy (hereafter referred to as autophagy) that selectively targets mitochondria to lysosomal degradation . The phyFor their degradation, mitochondria undergo several steps as it first need to be molecularly primed with several autophagy signals , recognised by specific adapters or receptors, which nature depends on the type of stress mitochondria encounter. This molecular targeting allows, in a second step, the recruitment of the autophagy machinery and the engulfment of targeted mitochondria inside a double membrane vacuole, called autophagosome, which subsequently fuse with lysosomes allowing its degradation . The autWhile the autophagosome formation process is common to all autophagic responses, different types of pathways control the induction of mitophagosome formation. These pathways are classified upon their dependency or not on ubiquitin and present a panel of mitophagy-promoting receptors . These rcarbonylcyanure m-chlorophenylhydrazone) [The first described and therefore most characterised mitophagy pathway is composed of the two key proteins, PINK1 (phosphatase and tensin homolog (PTEN)-induced putative kinase 1) and Parkin, which have been identified in Parkinson’s disease, the second most common neurodegenerative disease. From the molecular point of view, the PINK1/Parkin axis is known to be induced upon several mitochondrial stresses such as a drop in MMP  as triggered by uncoupling reagents such as CCCP (drazone) , oxidatidrazone) , or hypodrazone) . The mecdrazone)  but the MUL1  is another E3 ubiquitin ligase that triggers ubiquitin-dependent mitophagy and has been shown to be activated in parallel with the PINK/Parkin pathway, notably during the degradation of paternal mitochondria after oocyte fertilisation . In addiFUNDC1: FUNDC1 (FUN14 domain-containing protein 1) is a mitophagy receptor localised in the OMM that has been described as a major effector of mitophagy initiation upon hypoxia [ hypoxia . FUNDC1- hypoxia . Mechani hypoxia . In unst hypoxia  and FUND hypoxia  triggers hypoxia  Fig 2B)FUNDC1: F hypoxia .BNIP3 and BNIP3L/NIX: BNIP3 (BCL2/adenovirus E1B 19 kDa protein-interacting protein 3) and the closely related BNIP3L  were first described as OMM-resident proapoptotic proteins considering their typical Bcl2 homology 3 (BH3) domain [) domain . Indeed,) domain . This le) domain . In addi) domain . However) domain ,36. In h) domain . To fulf) domain , both BN) domain  Fig 2B)BNIP3 and) domain .Although BNIP3 and BNIP3L mitophagy pathways were first thought to be distinct of the ubiquitin-dependent PINK1/Parkin pathway, recent studies demonstrate a tight regulation between these two mechanisms. Indeed, BNIP3 also suppresses the proteolytic cleavage and inactivation of PINK1 to promote ubiquitin-dependent mitophagy . In addiFKBP8: In basal conditions, FKBP8  is anchored to the OMM where it localises and stabilises the antiapoptotic factors Bcl-2 and Bcl-Xl to the mitochondria to suppress apoptosis [poptosis . Upon nupoptosis , as wellpoptosis . More repoptosis  Fig 2B)FKBP8: Inpoptosis ,48. The BCL2-L13: Bcl2-L-13 (Bcl2-like protein 13) is an OMM-anchored protein that corresponds to the mammalian ortholog protein of the yeast Atg32 mitophagy receptor [receptor . Bcl2-L-receptor  Fig 2B)BCL2-L13:PHB2: Upon OMM rupture through proteasomal degradation of OMM proteins, IMM exposure to the cytosol can be a key signal for mitophagy to clear damaged mitochondria. In this case, PHB2 (Prohibitin 2), an IMM integral protein found as heterodimers with PHB1, is a mitophagy receptor that interacts with LC3-II when exposed to the cytosol [ cytosol . The mol cytosol . This acATAD3B: ATAD3B (ATPase Family AAA Domain Containing 3B) is a protein that regulates the stabilisation of large mtDNA-proteins complexes called nucleoids [ucleoids  and actsucleoids . ATAD3B ucleoids .In eukaryotic cells, the cardiolipin is a mitochondria-exclusive glycerophospholipid found in the IMM and promotes, when aggregated, the IMM curvature due to its typical shape defined by a double glycerophosphate backbone and four fatty acyl chains . While iP. aeruginosa is a motile gram-negative rod-shaped opportunistic extracellular pathogen and represents a major cause of life-threating nosocomial infections including hospital-acquired pneumonia or chronic obstructive pulmonary disease (COPD) [P. aeruginosa infections can eventually evolve into a chronic status through to the development of a variety of virulence effectors allowing pathological adaptation and colonisation of the epithelium into a mucoid biofilm [e (COPD) . In lung biofilm . These v biofilm .P. aeruginosa is found inside the host cell, iron extraction and transport by the pyoverdine leads to mitochondrial homeostasis disruption and subsequent host cell death [P. aeruginosa infection [P. aeruginosa as it helps the clearance of the pathogen in a similar way as xenophagy, that was already reported for that bacteria [P. aeruginosa infection, other mitophagy mechanisms are also induced by the host itself to limit inflammation. Indeed, mitophagy triggered by P. aeruginosa-induced mtDNA release attenuates the activation of the NLRC4 (NOD-like receptor family caspase recruitment domain containing 4) inflammasome [P. aeruginosa infection cluster activates a PHB2-dependent mitophagy pathway to increase P. aeruginosa clearance and limit inflammation through a negative regulation of NF-kB [P. aeruginosa since it induces a PINK1-TBK1-p62-dependent mitophagy response associated with bacterial elimination and reduction of inflammation [When ll death . As a minfection  Fig 3A)P. aerugibacteria . In addiammasome . Moreoveof NF-kB . In addiammation .M. tuberculosis is a rod-shaped intracellular pathogen harbouring one peculiar cell membrane with arabinogalactan associated to peptidoglycan and covered by a thick layer of mycolic acids [M. tuberculosis is the etiologic agent of tuberculosis, the leading cause of death from bacterial infectious disease worldwide [M. tuberculosis is a particularly adaptable pathogen that coevolved with its human host since most infections are asymptomatic (latent tuberculosis) until the host immune response is compromised. Indeed, M. tuberculosis resides in the lung alveoli where it manipulates the host immunity leading to the recruitment and aggregation of diverse types of immune cells in the infection site [M. tuberculosis difficult to eradicate. Moreover, the yearly rise of MDR strains supports the need for novel therapeutic approaches.ic acids . M. tubeorldwide . M. tubeion site . These nM. tuberculosis elimination through their direct antibacterial activity and induction of host mitophagy [M. tuberculosis infection [M. tuberculosis and M. bovis infections are known to promote M1 macrophage polarisation with a pro-inflammatory phenotype among other polarisation programs [M. tuberculosis treatment. However, whether mitophagy is directly linked to bacterial elimination is not clear. Indeed, a recent study suggests that M. bovis-induced mitophagy is rather beneficial for the bacteria as mitophagy suppresses host xenophagy for intracellular survival [M. bovis requires phosphorylated TBK1 (p-TBK1), which is needed for xenophagy of the bacteria [M. bovis to escape from degradation through xenophagy [M. tuberculosis and M. bovis-mediated mitophagy responses require further research to clarify whether mitophagy is beneficial for the bacteria or represents a defensive response from the host, or both.Recently, a study highlighted thiopeptides as emerging clinical antibiotic candidates for itophagy . This stnfection  Fig 3B)M. tubercprograms . In the programs . Hencefosurvival . The PINbacteria . The comenophagy  L. monocytogenes is a gram-positive rod-shaped facultative intracellular pathogen that causes listeriosis, a foodborne associated infection that can affect animals as well as humans [L. monocytogenes relies on its ability to cross the intestine barrier and reach the liver and spleen. In immunocompromised individuals, L. monocytogenes crosses the blood–brain barrier and the fetoplacental barrier leading to fatal meningitis or abortion [L. monocytogenes escapes from the endolysosomal pathway through permeabilisation of its vacuole, then it polymerises actin at one pole to acquire motility and spread to a neighbouring cell [s humans . The virabortion . At the ing cell .L. monocytogenes is also able to manipulate the host mitochondria, notably by triggering mitochondrial fragmentation mediated by its virulence effector listeriolysin O (LLO) [L. monocytogenes-induced mitochondrial fragmentation [L. monocytogenes intracellular lifecycle. Mitochondrial fission being one hallmark of mitophagy, L. monocytogenes also triggers mitophagy by a recently identified mitophagy receptor, the NOD-like receptor NLRX1 [L. monocytogenes-induced mitochondrial fragmentation. Therefore, the NLRX1-mediated mitophagy triggered by LLO is beneficial for L. monocytogenes survival in the host cell as it avoids bacteria to be killed by oxidative stress generated by damaged mitochondria [L. monocytogenes needs to induce mitochondrial fragmentation through LLO for its lifecycle, since it generates bacterial-killing mtROS release. However, L. monocytogenes could actively trigger mitophagy (through LLO) to modulate or compensate for the deleterious accumulation of mtROS caused by the presence of the bacteria. O (LLO)  Fig 4A)L. monocyentation ,78. Howeor NLRX1 . Mechanichondria . It coulY. pestis is a gram-negative facultative intracellular coccobacilli that is the causative agent of the plague inoculated by flea vectors , which are injected inside host cells through a T3SS [Y. pestis virulence also occurs through intracellular trafficking and persistence inside the host. Following phagocytosis, Y. pestis is able to target several Rab GTPases  to the phagosome to inhibit its acidification and disrupt host cell recycling, therefore leading to bacterial replication inside a nonacidic autophagosome [ew, read ). The tyh a T3SS . Howeverhagosome .Y. pestis has been identified and is linked to manipulation of host mitochondria [L. monocytogenes, Y. pestis infection leads to mitochondrial fragmentation causing an increase in mtROS release, which display antibacterial properties. However, requiring the YopH effector, Y. pestis triggers a PINK1/Parkin-dependent mitophagy response to clear dysfunctional mitochondria induced by the infection, limiting the accumulation of mtROS and allowing bacterial persistence [Y. pestis and L. monocytogenes are thus two pathogens for which mitophagy is activated by a virulence effector to clear the host cell from pathogen-induced damaged mitochondria and therefore preserve the replicative niche.More recently, another prosurvival mechanism deployed by chondria  Fig 4B)Y. pestissistence . Y. pestB. abortus is a gram-negative facultative intracellular coccobacilli that causes brucellosis, a worldwide zoonosis affecting domestic animals and humans [B. abortus virulence relies on the expression of a type 4 secretion system (T4SS) named VirB, which allows effector injection inside the host cell [B. abortus to reach its replicative niche inside the ER, called replicative Brucella-containing vacuoles (rBCVs) [B. abortus subverts the host cell autophagy initiation machinery  to help the formation of autophagic-like BCVs (aBCVs), which are necessary for bacterial egress from the host cell [d humans . Animal d humans . As an iost cell . These e (rBCVs) . Eventuaost cell .B. abortus also interacts with the host mitochondria, inducing severe mitochondrial network fragmentation during the late steps of cellular infection [B. abortus simultaneously triggers mitophagy in an iron-HIF-1α-BNIP3L-dependent manner [B. abortus exit of host cells through an alteration of aBCVs formation [B. abortus-induced mitophagy would favour the spreading of the bacteria from cell to cell, and even towards other tissues. However, as no B. abortus virulence effector has been identified yet as a driver of mitophagy induction, further research would be needed to decipher whether BNIP3L-mediated mitophagy is an active response mediated by B. abortus or not. However, in the case of B. abortus, the induction of mitophagy is linked to bacterial dissemination, and not to bacterial replication. This highlights a novel process by which mitophagy could be beneficial for the persistence of some pathogens.Recent studies showed that nfection  Fig 4C)B. abortut manner . As for ormation . This coH. pylori is a gram-negative spiral multiflagellar extracellular pathogen that colonises the epithelium of the human stomach [H. pylori infection leads to the development of several gastric diseases such as chronic gastritis, ulcers, mucosa-associated lymphoid tissue lymphoma, and gastric cancers. One of the main and most studied toxin of H. pylori is VacA (vacuolating cytotoxin A) [ stomach . H. pylotoxin A) . VacA isH. pylori-induced mitophagy might also be initiated through the translocation of STAT3  phosphorylated on the serine 727 to the mitochondria [H. pylori-mediated apoptosis and/or mitophagy eventually leads to alteration of the gastric mucosa contributing to gastric disease for the host [H. pylori survival. Indeed, VacA-dependent gastric epithelial cell death is essential for nutrient release required for H. pylori growth in the gastric environment [H. pylori thus represents a model of extracellular pathogen able to manipulate the host mitophagy for its own survival.VacA manipulates several host cell signaling pathways to induce its death either through apoptosis and/or mitophagy . Indeed,chondria . Gastricthe host , withoutironment . H. pyloV. splendidus is a widespread gram-negative rhabdoid extracellular pathogen for aquatic animals including shellfish (such as the oyster Crassostrea gigas), turbot fish , or commercially important species of sea cucumber in Chinese aquaculture (Apostichopus japonicus) [V. splendidus have been identified during A. japonicus infection. They include quorum sensing, extracellular metalloproteases, siderophores, as well as the hemolysin 4-hydroxyphenylpyruvate dioxygenase (4-hppD), which elicit a strong skin ulceration syndrome causing great economic losses in the A. japonicus aquaculture [ponicus) . Severalaculture .V. splendidus infection of A. japonicus coelomocytes, MMP alterations are observed and lead to a massive load of mtROS, which sustains mitochondrial damage and the host cell death by apoptosis [C. gigas also triggers mitophagy during V. splendidus infection, highlighting, for the first time, the existence of mitophagy in mollusks [H. pylori, V. splendidus represents a model of extracellular pathogen for which the host mitophagy is a defensive response, not sufficient, however, to completely eradicate the pathogen and resolve the infection.However, recent studies have shown that the host can engage prosurvival mitophagy-dependent programs to counteract the infection . Upon V.poptosis . In respmollusks . In oppoMitochondria are complex organelles that acquired through evolution machineries to cope with and eliminate bacterial pathogens that can infect eukaryotic host cells. These mechanisms notably include the regulation of many cellular responses such as mtROS production, regulation of apoptosis, and an important role in innate immunity. Yet several bacterial pathogens, whether they display an intracellular or extracellular lifestyle, evolved as well, being able to manipulate the biology of mitochondria to their advantage for survival purposes. One of the major means would be to drive the host cell to induce degradation of damaged mitochondria that can be deleterious for the pathogen.M. tuberculosis, which induces mitophagy to impair xenophagy. Amplification and relocalisation of molecular actors such as p-TBK1 from the xenophagy machinery to the mitophagy machinery benefit to pathogen survival and dissemination. In addition, like xenophagy, mitophagy is also described to help bacterial clearance upon P. aeruginosa , or mtROS-derived oxidative stress , which could lead to the activation of the inflammasome, the maturation of pro-inflammatory IL-1β and IL-18 cytokines, and subsequent elimination of the infection [P. aeruginosa and L. monocytogenes infection. In addition, pathogen-induced mitophagy is also related to bacterial fitness and dissemination as it helps bacterial egress and infection of neighbouring cells (in the case of L. monocytogenes and B. abortus)  . HoweverL. monocytogenes (LLO), Y. pestis (YopH), and H. pylori (VacA). Further experimental strategies should also be considered to identify putative virulence effectors from M. bovis and B. abortus, which could be required for their active manipulation of p-TBK1 and BNIP3L, respectively.When mitophagy appears to be beneficial for the pathogen survival, virulence effectors might be responsible for actively manipulating the initiation mechanisms of the host mitophagy. Several bacterial effectors have already been identified as such during cellular infection by In conclusion, even if mitochondria and bacteria could be evolutionary considered as two relatives, there is a real struggle between them to determine survival outcome during infection. Further research in the relatively recent field of pathogen-induced mitophagy would be of great interest to better understand these mechanisms at the molecular level and potentially identify new molecular actors that could pinpoint new therapeutic approaches to fight the growing threat of multidrug resistance pathogens.


Candida auris (C. auris) is an emerging multi-drug resistant pathogen that is a health threat for hospitalized patients. C. auris may survive on surfaces where standard disinfectants may not achieve adequate disinfection. Continuous dry hydrogen peroxide (DHP) reduces microbial bioburden on surfaces and may reduce the risk of C. auris transmission in the clinical care setting. This study reviews the efficacy of DHP exposure, coupled with routine environmental cleaning, on reducing microbial burden of C. auris on surfaces.C. auris, baseline surface sampling of the patient room and other unit areas was performed beginning on the first day of positive culture, defined as Day 0. The DHP systems were activated and surface sampling collected to include days 1, 7, and 14. Select surfaces were sampled using polyurethane foam neutralizing buffer swabs. Individual swabs were grouped into three composites: Inner Patient Halo, Outer Patient Halo, and Mobile Shared Equipment (Table 1). Bioburden was assessed through cultures of the swabs and the identification and enumeration of resulting colony forming units (CFU). A multivariate regression model was used to evaluate the potential relationship between duration of DHP exposure and the logarithmically transformed counts of C. auris yielded by the samples.DHP-emitting systems were installed on burn intensive care and burn stepdown units. After install, for each patient newly identified with C. auris CFU were obtained from all composite swabs, with observed decrease in count over time . In a regression model including included covariates for time, room and patient proximity, there was a statistically significant relationship between C. auris bioburden and exposure to DHP =6.999, p< 0.001, adjusted R2=0.461), with duration of DHP exposure adding significantly to the relationship, p=0.Three patients were identified, resulting in collection of 36 composite swabs. C. auris colonization/infection. To fully understand the synergism between routine environmental cleaning and DHP, further research is needed.Continuous DHP, coupled with routine environmental cleaning, augments disinfection and is effective in reducing surface microbial bioburden in rooms of patients with All Authors: No reported disclosures


Psettodes flatfishes on a global scale. This study focuses on the matrilineal evolutionary path of these primitive groups, with a specific emphasis on the complete mitogenome of the Psettodes belcheri and casting light on its genetic composition, structural traits, and evolutionary chronicle. Exploring genetic variations and phylogenetic relationships uncovers the intricate evolutionary links between Psettodes species and their broader context within the Pleuronectiformes species. The complex interplay of hydrographic conditions, ocean currents, and ecological factors emerges as pivotal in shaping the evolutionary landscape of these flatfishes. Given the potential consequences for conservation, this study highlights the necessity for a holistic comprehension of marine environments and the ramifications of climate change and human interventions on flatfish species.The present research enriches our comprehension of the mitogenomic genetic features, genetic diversity, evolutionary past, and conservation prerequisites of Psettodes flatfishes is still poorly known from their range distribution in eastern Atlantic and Indo-West Pacific Oceans. The study delves into the matrilineal evolutionary pathway of these primitive flatfishes, with a specific focus on the complete mitogenome of the Psettodes belcheri species, as determined through next-generation sequencing. The mitogenome in question spans a length of 16,747 base pairs and comprises a total of 37 genes, including 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNA genes, and a control region. Notably, the mitogenome of P. belcheri exhibits a bias towards AT base pairs, with a composition of 54.15%, mirroring a similar bias observed in its close relative, Psettodes erumei, which showcases percentages of 53.07% and 53.61%. Most of the protein-coding genes commence with an ATG initiation codon, except for Cytochrome c oxidase I (COI), which initiates with a GTG codon. Additionally, four protein-coding genes commence with a TAA termination codon, while seven others exhibit incomplete termination codons. Furthermore, two protein-coding genes, namely NAD1 and NAD6, terminate with AGG and TAG stop codons, respectively. In the mitogenome of P. belcheri, the majority of transfer RNAs demonstrate the classical cloverleaf secondary structures, except for tRNA-serine, which lacks a DHU stem. Comparative analysis of conserved blocks within the control regions of two Psettodidae species unveiled that the CSB-II block extended to a length of 51 base pairs, surpassing the other blocks and encompassing highly variable sites. A comprehensive phylogenetic analysis using mitochondrial genomes (13 concatenated PCGs) categorized various Pleuronectiformes species, highlighting the basal position of the Psettodidae family and showed monophyletic clustering of Psettodes species. The approximate divergence time (35−10 MYA) between P. belcheri and P. erumei was estimated, providing insights into their separation and colonization during the early Miocene. The TimeTree analysis also estimated the divergence of two suborders, Psettodoidei and Pleuronectoidei, during the late Paleocene to early Eocene (56.87 MYA). The distribution patterns of Psettodes flatfishes were influenced by ocean currents and environmental conditions, contributing to their ecological speciation. In the face of climate change and anthropogenic activities, the conservation implications of Psettodes flatfishes are emphasized, underscoring the need for regulated harvesting and adaptive management strategies to ensure their survival in changing marine ecosystems. Overall, this study contributes to understanding the evolutionary history, genetic diversity, and conservation needs of Psettodes flatfishes globally. However, the multifaceted exploration of mitogenome and larger-scale genomic data of Psettodes flatfish will provide invaluable insights into their genetic characterization, evolutionary history, environmental adaptation, and conservation in the eastern Atlantic and Indo-West Pacific Oceans.The mitogenomic evolution of the  Psettodes is a “primitive” flatfish group within the monotypic Psettodidae family, with three recognized species: Psettodes belcheri, Psettodes bennettii, and Psettodes erumei [P. erumei, commonly referred to as the Indian halibut, ranges expansively across the Red Sea and the Indo-West Pacific Ocean, the other two species are confined to the eastern Atlantic. Intriguingly, the ranges of P. belcheri and P. bennettii exhibit partial overlap, spanning from the west Sahara to the Liberian coast.Flatfishes, also known as flounders, soles, halibuts, turbots, plaices, and tonguefishes, are classified as Pleuronectiformes. This order is split into two suborders: Psettodoidei, which includes the single-family Psettodidae, and Pleuronectoidei, which includes 13 families . Their es erumei . While PP. erumei) is heavily harvested within the tropical fishing zone as defined by the United Nations Food and Agriculture Organization (FAO) [Psettodes species as ‘Data Deficient.’ [Psettodes, display varying degrees of dextral to sinistral polymorphism [The economic significance of flatfishes has led to their frequent capture through demersal trawling in marine ecosystems . The Indon (FAO) . Despiteicient.’ . Furthericient.’ . While mmorphism . This semorphism .Psettodes has stirred argument, especially in the context of analyses involving nuclear and mitogenomic data. This genus consistently diverges from other flatfishes (Pleuronectiformes), instead aligning with other carangimorphs [Psettodes genus within the context of other carangimorphs and pleuronectoids [Psettodes taxon (P. erumei) from the Indo-Pacific region, highlighting the need for expanded inquiries involving other congeners from the Atlantic Ocean.The integration of molecular data has become integral in the exploration of global flatfish species, encompassing diverse aspects such as species identification , systemagimorphs ,24,25,26gimorphs . Later snectoids ,29. HowePsettodes’ maternal evolutionary path, this study seeks to construct the complete mitogenome of P. belcheri, elucidate its genomic attributes, and establish its phylogenetic relationships. This endeavor contributes to understanding the maternal evolution of these ancient lineages in marine environments, strengthening the mitogenomic repository of flatfishes, and expanding our scientific insight into the Psettodidae family on a global scale. While marine habitats harbor a diverse array of life forms, our understanding of speciation in marine ecosystems is comparatively limited compared with freshwater systems, necessitating urgent exploration of the mechanisms driving speciation and adaptation [P. belcheri and P. erumei) and investigate potential evolutionary scenarios within the marine environment.An integrated strategy that looks beyond superficial investigation is required for an accurate appraisal of speciation within maritime ecosystems. Advances in technology, spanning maritime engineering to genomics, are facilitating the investigation of marine speciation ,31. To eaptation . Given tP. belcheri, was acquired from the estuaries of the Kineke River  located in Kribi, Cameroon (P. belcheri and its congeners (P. erumei and P. bennettii), were obtained in .shp file format from the IUCN database   .® Genomic DNA extraction kit, manufactured by Bioneer in Daejeon, Republic of Korea, was utilized to extract genomic DNA, following the established standard protocol. The quality and quantity of the resulting genomic DNA were meticulously evaluated employing a NanoDrop spectrophotometer . To obtain the comprehensive mitogenome of P. belcheri, sequencing procedures were executed using the NovaSeq platform, provided by Illumina and accessible at Macrogen  in Daejeon, Republic of Korea. The sequencing libraries were prepared following the manufacturer’s instructions for the TruSeq Nano DNA High-Throughput Library Prep Kit . In short, 100 ng of genomic DNA underwent fragmentation using adaptive focused acoustic technology , resulting in blunt-ended dsDNA molecules with 5′-phosphorylation. After the end-repair step, DNA fragments were size selected using a bead-based method. These fragments were then modified with the addition of a single ‘A’ base and ligated with TruSeq DNA UD Indexing adapters. Subsequently, the products were purified and enriched through PCR to create the final DNA library. Library quantification was performed using qPCR, following the qPCR Quantification Protocol Guide (KAPA Library Quantification kits for Illumina Sequencing platforms), and quality assessment was carried out using Agilent Technologies 4200 TapeStation D1000 screentape . Finally, paired-end (2 × 150 bp) sequencing was performed by Macrogen using the NovaSeq platform . Over 20 million raw reads underwent processing using the Cutadapt tool  to trim adapters and remove low-quality bases, with a Phred quality score (Q score) cutoff of 20. The Geneious Prime version 2023.0.1 was used to assemble the targeted genome from the high-quality paired-end NGS reads. This assembly was accomplished by employing reference mapping with the mitogenome of a closely related species as a reference, and we employed default mapping algorithms. Mitogenome assembly was accomplished by scrutinizing the alignment of overlapping regions via MEGA X [http://mitos.bioinf.uni-leipzig.de, accessed on 15 August 2023) and MitoAnnotator  web servers [https://www.ncbi.nlm.nih.gov/orffinder/, accessed on 15 August 2023), based on the vertebrate mitochondrial genetic code. The resultant mitogenome of P. belcheri was duly submitted to the global GenBank database.The AccuPrepa MEGA X . The bou servers ,36. To fhttp://mitofish.aori.u-tokyo.ac.jp/annotation/input/, accessed on 15 August 2023). A comprehensive comparative analysis was carried out to assess the mitogenomic architecture and variations within our generated sequence in relation to two pre-existing mitogenomes from a single congener, P. erumei . The calculation of intergenic spacers, which separate adjacent genes, and overlapping regions was performed manually. To determine the nucleotide compositions of protein-coding genes (PCGs), ribosomal RNA (rRNA), transfer RNA (tRNA), and the control region (CR), we employed MEGA X [https://tandem.bu.edu/trf/trf.html, accessed on 15 August 2023) [A spherical representation of the generated mitogenome was crafted using MitoAnnotator  .P. bennettii, intra-species and inter-species distances were determined using the widely employed mitochondrial COI gene. To elucidate the matrilineal phylogenetic connections, 14 mitogenomes from 13 Pleuronectiformes species were sourced from the GenBank database (accessed on 15 August 2023) [Cyclopsetta fimbriata, was categorized within the recently established family Cyclopsettidae . Additionally, two Cynoglossidae species were included: Cynoglossus gracilis (=Cynoglossidae I) and Symphurus orientalis (=Cynoglossidae II). The mitogenome of Lates calcarifer (DQ010541), from the Centropomidae family, was incorporated as an outgroup. Concatenation of all 13 PCGs was executed using the iTaxoTools 0.1 tool to construct the dataset for phylogenetic analysis [https://itol.embl.de/login.cgi, accessed on 15 August 2023) [Genetic distances were computed using the Kimura 2-parameter (K2P) method within MEGA X. Due to the unavailability of the complete mitogenome of st 2023) ,47,48,49st 2023) ,50. The analysis . In ordeanalysis ,53,54. Eanalysis . Visualist 2023) . Additiost 2023) . This apst 2023) ,59. Aftest 2023) .P. belcheri  was characterized and assigned the GenBank accession number OR231239. The circular mitogenome of P. belcheri comprised 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), 2 ribosomal RNA genes (rRNAs), and a non-coding AT-rich control region (CR). Among these, 28 genes  were positioned on the heavy strand, while NAD6 and 8 tRNAs  were situated on the light strand  to 18,706 bp . Variability in mitogenomic lengths was largely attributed to duplications in the control regions (CRs) [P. belcheri showcased five overlapping regions, spanning a total of 14 bp. The longest overlap (7 bp) was observed between ATP synthase 8 (atp8) and ATP synthase 6 (atp6). Similarly, both P. erumei mitogenomes (FJ606835 and AP006835) displayed five overlapping regions totaling 23 bp, with the longest overlap (10 bp) between atp8 and atp6, mirroring P. belcheri. Moreover, P. belcheri exhibited 15 intergenic spacer regions spanning a total of 106 bp, with the longest (38 bp) situated between trnN and trnC. Conversely, P. erumei mitogenomes contained 11 intergenic spacer regions spanning 77–78 bp, with the longest (37–38 bp) found between trnN and trnC (P. belcheri mitogenome (54.15%), encompassing 28.09% A, 16.24% G, 29.56% C, and 26.06% T. Similar AT richness was observed in the nucleotide composition of the other two P. erumei mitogenomes, ranging from 53.07% to 53.61%  . The mitand trnC . Nucleoto 53.61% . The AT o −0.328 . This co species . This reP. belcheri mitogenome was occupied by a total of 13 protein-coding genes (PCGs), accounting for 68.27% of the whole sequence. Among these, the shortest PCG was ATP8, spanning 165 bp, while NAD5 represented the longest PCG with a length of 1839 bp. In both P. erumei mitogenomes, the total length of PCGs ranged from 11,426 bp (68.49%) to 11,427 bp (65.99%). The PCGs of P. belcheri were characterized by an AT bias of 52.73%, accompanied by AT skew and GC skew values of −0.052 and −0.323, respectively  initiation codon, except for COI, which began with a GTG  codon. A parallel pattern of initiation codons was apparent in all PCGs of the other two P. erumei mitogenomes. Among the PCGs, the conventional TAA termination codon was observed in four instances , while seven PCGs featured incomplete stop codons (T--/TA-). Additionally, two PCGs, NAD1 and NAD6, terminated with AGG and TAG stop codons, respectively. A corresponding distribution of stop codons was also noted in P. erumei mitogenomes  measuring 961 bp and a large ribosomal RNA (16S rRNA) with a length of 1702 bp. Comparatively, the total length of P. erumei’s rRNAs (2680 bp) was shorter than that of P. belcheri (P. belcheri) to 52.99% (P. erumei). Comparative analysis indicated AT skew values ranging from 0.190 (P. belcheri) to 0.227 (P. erumei), while GC skew values ranged from −0.113 (P. erumei) to −0.073 (P. belcheri) in the ribosomal RNA  to 75 bp (trnK), collectively constituting 1556 bp or 9.29% of the entire mitogenome. The combined tRNA length in both P. erumei mitogenomes was shorter (1553 bp and 1554 bp) than that of P. belcheri (P. belcheri (55.14%) and P. erumei (54.02% to 54.12%). AT skew values ranged from 0.006 (P. erumei) to 0.009 (P. belcheri), while GC skew values ranged from 0.032  to 0.036   . Most tRJ606835) .P. belcheri control region (CR) reached 1015 bp, comprising 63.65% AT content. Conversely, the complete lengths of P. erumei’s CRs varied, ranging from 968 bp to 1601 bp. The AT skew fluctuated between −0.050 (P. belcheri) and 0.074 (P. erumei), while the GC skew ranged from −0.337  to −0.241   and more than twelve copies (AP006835) of 56 bp repeats. Four conserved blocks  were detected in both P. belcheri and P. erumei mitogenomes, consistent with their presence in other teleost fishes [Psettodes species via these variable nucleotides. As demonstrated in other species, such conserved domains are integral for mitochondrial genome replication and transcription [The comprehensive length of J606835) . P. belct fishes ,69. Of tcription . Intrigucription   and S. orientalis (=Cynoglossidae II)  . The appatfishes ,83,84,85P. belcheri and P. erumei. These two species have a wide divergence time during the late Eocene to early Miocene, approximately 35 to 10 million years ago (MYA)  . Remarkago (MYA) . The othgo (MYA) . The disnditions ,87. Thusnditions .The Miocene epoch, spanning from around 23 million to 5.3 million years ago, marked a period of substantial environmental transformation on Earth . This erPsettodes flatfishes, particularly following their colonization in the eastern Atlantic and Indo-West Pacific regions by the demersal lineage, a critical integration of the maximum-likelihood time-tree computation framework and marine ecological factors becomes essential. Notably, while P. belcheri is distributed across both the South and North Atlantic Oceans, spanning the western coast of Africa from Angola to western Sahara, P. bennettii is confined solely to the North Atlantic, with a distribution encompassing Gambia, Guinea, Guinea-Bissau, Mauritania, Morocco, Senegal, and Western Sahara. The sympatric speciation of these two species may be attributed to ecological selection within the pelagic environment. Since ancient time, the hydrographic and climatic conditions in the North and South Atlantic Oceans have differed significantly due to the Coriolis effect induced by Earth’s rotation (In the context of understanding the evolutionary scenarios of rotation A. In therotation B. Converrotation C.P. belcheri and P. bennettii, to the continental shelves of western Africa. The other congener, P. erumei, likely evolved during the early Miocene and colonized into the Red Sea to the Indo-West Pacific Ocean due to the formation of the Antarctic Circumpolar Current (P. erumei between the Indian Ocean and the Atlantic Ocean. Despite the differences in hydrography, the high saline outflow from the Red Sea to the Indian Ocean results in similar characteristics of these two marine environments, allowing for similar flatfish communities to thrive. However, the Indian Ocean and Western Pacific Ocean are seismically active due to the presence of tectonic plate boundaries. Ecological speciation scenarios are frequently invoked to explain such patterns, yet distinguishing between ecological adaptation and allopatric speciation remains challenging [P. erumei from the Indian Ocean to the West Pacific Ocean may have been driven by the Equatorial Counter Current of the Indian Ocean gyre. Such ecological features greatly influence the evolution and adaptation of other flatfish species, including Psettodes, and have the potential to increase endemism in certain demersal ecosystems in marine settings. An intriguing case of rapid ecological speciation has been observed in Baltic flounder species, involving the development of a new ecological niche through a demersal spawning behavior, constituting the fastest speciation event documented for a marine vertebrate [Psettodes flatfishes, as well as ecological variables that may lead to reproductive isolation, suggest a process of ecological speciation.The Indian Ocean, characterized by the presence of the Arabian Sea and the Bay of Bengal, as well as warm and cold currents such as the Western Australian cold current, forms the Indian Ocean gyre. The divergence time of Psettodidae from other Pleuronectiformes during the late Paleocene to early Eocene suggests that these primitive flatfishes may have emerged in the eastern Atlantic Ocean. The currents and salinity levels of both the North and South Atlantic Oceans may have constrained the distribution of the two Psettodidae species,  Current D. Notablllenging . The disrtebrate . As a rePsettodes flatfishes, the co-occurrence of both East Atlantic species within a portion of their range underscores the necessity for regulated harvesting within the framework of a multispecies fishery strategy. As global climate change progresses, marked alterations in environmental conditions such as elevated water temperatures, decreased oxygen levels, and modified wind patterns impact fish reproduction and recruitment on a global scale [Marine ecosystems remain relatively unexplored on a global scale, facing substantial gaps and challenges in terms of biodiversity assessment and the generation of genetic data ,97. In aal scale . Consequal scale ,103. ConP. belcheri, shedding light on its genetic composition, structural organization, and evolutionary history. Furthermore, the genetic distances and phylogenetic relationships were explored, unveiling the evolutionary relationships among Psettodes species and their broader placement within the Pleuronectiformes order. Notably, the study identified significant genetic divergence between P. belcheri and P. bennettii in the mitochondrial COI gene, suggesting reproductive isolation despite their shared habitat. The phylogenetic relationship, divergence times, and diversification patterns were also estimated, providing insights into the emergence and distribution of Psettodes species in various oceanic regions. The interplay of hydrographic conditions, ocean currents, and ecological factors played crucial roles in shaping the evolutionary landscape of these flatfishes. Given the potential conservation implications, this study highlighted the need for a comprehensive understanding of marine ecosystems and the effects of climate change and anthropogenic activities on flatfish species. Conservation strategies and the establishment of MPAs were emphasized as essential components in safeguarding the diversity and sustainability of marine life, particularly species such as Psettodes flatfishes. In summary, the present analysis of the mitogenome and evolutionary history of Psettodes flatfishes offers valuable insights into the genetic attributes and possible ecological adaptations in East Atlantic and Indo-West Pacific Oceans.In conclusion, this study investigated the detailed analysis of the mitogenome of


Intraductal papillary mucinous neoplasms (IPMNs) are characterized by mucin-producing neoplastic epithelium, and, as a result, pancreatic dilatation is observedA 72-year-old man was admitted to our hospital with a MD-IPMN. No mural nodule could be detected on computed tomography or endoscopic ultrasonography. Therefore, pancreatoscopic evaluation was attempted. First, the duodenoscope was inserted, and mucin was observed flowing from the ampulla of Vater . Then, Video 1 Diagnostic pancreatoscopy in a patient with a main-duct intraductal papillary mucinous neoplasm. The pancreatoscope is inserted. The view is obscured by mucin, but the mucin is easily aspirated. A mural nodule is then clearly observed. Forceps biopsy is successfully performed.In conclusion, this novel pancreatoscope may be useful in determining surgical margins in cases of MD-IPMN.Endoscopy_UCTN_Code_CCL_1AZ_2AB


Helicobacter pylori (H. pylori) infection is one of the major reasons for chronic PUD which, if left untreated, may also result in gastric cancer. Treatment of H. pylori is always a challenge to the treating doctor because of the poor bioavailability of the drug at the inner layers of gastric mucosa where the bacteria resides. This results in ineffective therapy and antibiotic resistance. Current treatment regimens available for gastric ulcer and H. pylori infection uses a combination of multiple antimicrobial agents, proton pump inhibitors (PPIs), H2-receptor antagonists, dual therapy, triple therapy, quadruple therapy and sequential therapy. This polypharmacy approach leads to patient noncompliance during long term therapy. Management of H. pylori induced gastric ulcer is a burning issue that necessitates alternative treatment options. Novel formulation strategies such as extended-release gastro retentive drug delivery systems (GRDDS) and nanoformulations have the potential to overcome the current bioavailability challenges. This review discusses the current status of H. pylori treatment, their limitations and the formulation strategies to overcome these shortcomings. Authors propose here an innovative strategy to improve the H. pylori eradication efficiency.Peptic ulcer disease (PUD) is one of the most prevalent gastro intestinal disorder which often leads to painful sores in the stomach lining and intestinal bleeding. Untreated   In 1989, the bacterium formerly known as Campylobacter pylori (C.pyloridis) was reclassified as H. pylori [H. pylori infection causes gastritis was established through self-experiments involving the ingestion of a bacterial broth, followed by subsequent recovery from gastritis upon eradication of H. pylori, thus verifying Koch's postulates [H. pylori. The primary routes of this infection are oral-oral and oral-fecal interactions. Dirty water can also be a source of this infection in developing countries. The ability to culture H. pylori from infected individuals' stool, dung, and saliva suggests the possibility of its transmission through these routes [H. pylori infected human stomach is provided in he world . Barry M. pylori . The prostulates . Clinicae routes . A pictoH. pylori infection lead to many disease conditions related to GI tract including gastritis, non-ulcer dyspepsia, peptic ulcer disease, antral polyp, gastric adenocarcinoma, MALT lymphoma, follicular esophagitis, gastroesophageal reflux disease, esophageal adenocarcinoma, diffuse duodenal nodular lymphoid hyperplasia, immunoproliferative small intestinal disease, and inflammatory bowel disease. Some non-GI tract associations of H. pylori infection are hepatobiliary diseases, pancreatic disease, cardiovascular disorders, endocrine and metabolic disorders, dermatological disorders, haematological disorders, neurological disorders and ophthalmic disorders [the H. pylori induced gastric ulcer. This infection is associated with symptoms like abdominal pain, loss of hunger, sickness, and lethal complications like gastrointestinal bleeding. Extra-intestinal symptoms of H. pylori include but are not limited to: short stature, immune thrombocytopenic purpura, refractory iron deficiency anaemia (IDA), vitamin B12 insufficiency, eye diseases, skin disorders, respiratory disorders, and mental health issues [H. pylori infection has been reported as a major cause for peptic ulcer disease among the various other reasons like consumption of alcohol, tobacco, cigarette, non-steroidal anti-inflammatory drugs (NSAIDs), Zollinger-Ellison syndrome and idiopathic formation [Untreated isorders . A pictoh issues ,9. Varioh issues . Peptic ormation . The untH. pylori demonstrate specialized adaptations enabling it to successfully inhabit the stomach's mucus layer, an environment known for its challenging conditions. This colonization is driven by a range of essential processes, including motility, urease production, adhesion, and various other mechanisms. H. pylori after entering into the human stomach pass to the gastric lumen. In the lumen, the urease gene cluster present in the bacterium get activated in the presence of nickel cofactor. This regulates proton-gated urea channel and allow urea entry only in acidic environment. When pH is lower than 5, there is a rapid entry of urea into the bacterium which in the presence of the urease enzyme is converted to ammonia and carbon dioxide. Ammonia further produces ammonium hydroxide in the presence of water. The ammonium hydroxide neutralizes the surrounding environment of the H. pylori and protect it from the acidic surroundings [H. pylori travel through the gastric mucosal layer  with the help of flagella [H. pylori attaches to the gastric mucosa, which primarily interact with mucin 5 (MUC5AC). The elevated production of MUC5AC can be a potential mechanism for promoting bacterial adherence [H. pylori infection lead to an increased synthesis of certain inflammatory cytokines and stop the synthesis of prostaglandin, which may contribute to inflammation and ulcer formation [H. pylori can also interfere with the normal mechanisms involved in repairing damaged stomach tissue, thereby delaying the healing of existing ulcers or promoting the development of new ones [H. pylori disrupts the balance of cell proliferation and death by affecting the genes that regulate the cell cycle leading to cell apoptosis and ultimately gastric cancer [oundings ,13. Afteflagella ,15. Upondherence . Adhesinormation . The damnew ones ,15. AlsoH. pylori is present in the extracellular space of the mucosal layer of the stomach. A low population of the bacteria reside in the intracellular vacuoles and migrate to the extracellular space to repopulate the bacteria in the mucosal layer [H. pylori.Greater than 90% of al layer . While ral layer . Howeveral layer . The abithe H. pylori cases occur in middle- and low-income nations. [H. pylori in Africa ranged from 87% in South Africa [H. pylori infection in Middle East and North African nations (MENA) was 40%. However, countries like Egypt (72–88%), Iran (82%), Israel (40–75%), Lebanon (21%), Libya (76%), Oman (68.4%), Saudi Arabia (28–70%), and Tunisia (82.7%) [H. pylori is modest in developed nations like North America (37.1%), Europe (20–40%), and Australia (16–30%). The incidence of H. pylori infection was 42.7% in females and 46.3% in men. The age-wise prevalence statistics reveal that adults are more vulnerable than children. According to reports, prevalence among minors was 32.6%, while in the age groups of 18–30 years it was 40.5% and in the age group of 31–50 years, it was 43.6% [H. pylori is the leading cause of peptic ulcer. Greater than 11% of the global population are suffering from peptic ulcer induced by prolong H. pylori infection. A study revealed 70–95% co-relation between gastric ulcer and H. pylori infection worldwide [Most of nations. . The infh Africa  to 91% ih Africa . The inc (82.7%)  had highralia 16–%. The inH. pylori infection is always a challenge for the treating doctor because of the ineffectiveness of the available treatment options. Currently approved treatment for combating H. pylori is a combination of two or more antibiotics and anti-ulcer drug in the traditional delivery mode. This strategyfail in the eradication of H. pylori infection because of the insufficient reach of antibiotics to the inner mucosal layer of the stomach lining where the bacteria reside [H. pylori has the ability to reside extracellularly as well as inside intracellular vacuoles, a formulation that can deliver the drug over a long period locally as well as systemically will be able to eradicate the organism from the system [H. pylori, and novel strategies to eradicate H. pylori infection.Treatment of a reside . This ofa reside ,23,37. H22 -antagonists and proton pump inhibitors (PPIs) are generally the preferred choice for the H. pylori eradication. A detailed list of currently used drugs for treating H. pylori infection according to their therapeutic classes is mentioned in H. pylori [2 antagonist reduces acid secretion but does not affect H. pylori bacteria [H. pylori. The adverse effects of PPIs can be separated into two categories: acid inhibition-related and non-acid inhibition-related. The majority of acid inhibition-related side effects  occur during long-term use of PPI, but non-acid inhibition-related adverse effects occur in both long-term and short-term PPI users [A combination of antibiotics and gastric acid-suppressing medications such as H. pylori . Differe. pylori . H2 antabacteria . In contPI users . In addiPI users .Table 1A2.1H. pylori treatment strategy. For the first 10–14 days of therapy, patients with an H. pylori infection are given a combination of two antibiotics along with a PPI. The antibiotic of choice in the first-line treatment are amoxicillin, clarithromycin, and metronidazole. Clarithromycin based therapy is safe and effective, but it is important that before this therapy, the patient should not have previous exposure to clarithromycin and other macrolide class of drugs. It is possible that previous exposure to antibiotics can lead to drug resistance and failure of the therapy. Lee et al. reported that clarithromycin-based triple therapy had a 90% efficacy rate, but it has currently reduced to 70–80% efficacy rate [H. pylori infection; but, the availability of bismuth and associated complications is a cause of concern [The patient's history of antibiotic use should be gathered before deciding on an acy rate . Bismuthacy rate ,58. Quad concern .2.2H. pylori bacteria while levofloxacin, tinidazole, and rabeprazole depicted 90% elimination for H. pylori bacteria [A second-line therapy is administered if the first treatment fails. In this therapy levofloxacin, amoxicillin and PPI triple regimen is used. It is reported that 10 days of treatment had resulted in 84% cure in patients . Bismuthbacteria .2.3H. pylori infection. Third-line treatment is used when the other two therapies have failed, or resistance has developed to the other therapy. Third-line therapy is rifabutin-based therapy. The treatment regimen is rifabutin, levofloxacin, amoxicillin, and PPI. The eradication rate of this therapy ranges from 75 to 90% [H. pylori [H. pylori with gyrA mutations as well as imparting a higher efficacy than levofloxacin-based treatment as a third-line therapy [It is the rescue therapy for 5 to 90% . Rifabut. pylori . Sitaflo therapy .Suzuki et al. reported that Rifaximin, in comparison with rifabutin, is less absorbed into the bloodstream and hence nearly free of side effects and possess a better bioavailability in the gastrointestinal tract . Tursi e3H. pylori is 100% effective and after the completion of treatment, the chance of reinfection is still a problem [The major drawback of the available treatment is antibiotic resistance. This is mainly due to the ineffective reach of these antibiotics to the organism since it resides in the inner layer of the intestinal mucosa. The bacteria are also resistant to the highly acidic environment of the stomach. Penicillin-allergic patients cannot be administered amoxicillin-based treatment regimen. Another drawback is the use of polypharmacy, where many pills are to be taken together, which leads to a lot of inconvenience and omission/missing of dosage by patients. Usage of multiple antibiotics can affect the gut microbiome. It also adds to the cost of treatment . The una problem .44.1H. pylori infection is antibiotic resistance, which arises due to the irrational use of antibiotics and insufficient bioavailability [H. pylori are amoxicillin (AMX), metronidazole (MET), clarithromycin (CLR), tetracycline (TET), and levofloxacin (LEV). Earlier researchers have studied the resistance of these antibiotics. The primary resistance rates to H. pylori for AMX, CLR, MET, LEV, and TET were reported to be 15.0%, 34.1–55.2%, 69.4–71.3%, 18.4–27.9%, and 17.9%, respectively. The secondary resistance rates reported for these antibiotics were 9.5%, 74.9%, 61.5%, 45.7%, and 23.5%, respectively [H. pylori and leads to resistance. To solve this problem, a combination of antibiotics and PPI are used [The primary cause of the treatment failure against lability . Antibioectively . The stuare used . Increasare used .4.2H. pylori. This is because the patient must take more than ten different medications. According to one survey, patients who took less than 60% of medication had only a 30% efficacy rate and also resulted in antibiotic resistance [H. pylori eradication failure. Age, gender, illness type of treatment, length of treatment, and bismuth dosage had no influence on the eradication rate [H. pylori, patients often use antibiotics and acid-suppressing drugs for a period of 1–2 weeks. Patients may have trouble sticking to their treatment plan because of unpleasant taste, side effects and the complexity of their medication. Nonadherence to H. pylori medication may result in treatment failure, recurrent infection, and antibiotic resistance [Patient noncompliance is the major challenge in treating sistance ,80. The ion rate . In ordesistance .4.3H. pylori treatment. In the fixed-effects model, the meta-analysis revealed that the relative risk of unsuccessful eradication in CYP2C19 extensive metabolism compared with intermediate metabolism and poor metabolism was 1.21  and 1.57 , respectively. The cure rate of the eradication regimens involving omeprazole and lansoprazole in combination varied amongst CYP2C19 genotypes, in contrast to the regimens combining rabeprazole and esomeprazole (p < 0.05) [PPI is metabolized by cytochrome P450, which is made of different isoenzymes. The isoenzyme used in the metabolism of PPI is CYP2C19, which decreases the potency of PPI. To overcome this problem, it is recommended to increase the dose of PPI, ,84. Orme < 0.05) .4.4Many drugs degrade in gastric fluid due to hydrolysis and enzymatic action. Another issue plaguing this class of drugs is the ionization in the acidic GI environment which prevents the permeability . Erah et4.5H. pylori create a neutral environment surrounding it in the mucosal layer via its adaptive mechanisms and thereby, protects it from gastric acid [H. pylori [H. pylori bacteria can form biofilm, including the clinical, slandered, and mouse adapted strains [H. pylori changes its phenotype during biofilm development, which reduces its susceptibility to antibiotics. However, extracellular protease therapy can somewhat improve this resistance [H. pylori has the ability to survive in both aerobic and anaerobic conditions and also live both extracellularly and intracellularly. It can stay in the intracellular vacuoles and can repopulate after coming in the extracellular space when the antibiotic concentration is less [Bacteria change their morphology, which protects them from eradication. In general, bacteria shrink and transform into tiny spherical entities with modified gene expression, and molecular synthesis. ric acid . The bac. pylori . Biofilm. pylori . It is c. pylori . Studies strains . Hathrou strains . It is asistance ,95. Addi is less . This is4.6H. pylori quickly adapts to the gastric environment. It produces ammonia and HCO3 via urease and α-carbonic anhydrase, to produce ammonia, making a protective natural layer outside the bacteria. A higher local pH makes it easier for bacteria to pass through stomach fluid [H. pylori against harsh circumstances and acid exposure, thereby improving H. pylori motility and help to accumulate at a higher density on the stomach epithelium, resulting in a larger inflammatory response [Gastric niche protects the bacteria from the gastric juice and allows the colonization of other bacteria. ch fluid . Studiesresponse .4.7H. pylori therapy. The high density of bacteria can increase the mucosal inflammatory cytokine responses, thus decreasing the immune response. Eradication of H. pylori can decrease the inflammation in gastric mucosa and reduce the bacterial load [H. pylori bacterial load and ulcer curing rates in patients with active duodenal ulcers. Few studies showed that in the groups with mild, moderate, and marked H. pylori infection, the success rates of H. pylori treatment were 88.9%, 94.3%, and 69.7%, and that of ulcer healing were 88.0%, 94.3%, and 63.6% respectively. The statistical analysis showed that both the eradication rate and ulcer curing rate had a strong association with pretreatment H. pylori density  [H. pylori virulence factor). Result shows that both in the antrum and corpus, the bacterial burden in patients with cagA positive strain was higher than in those without it (p < 0.01). A statistically significant link existed between the intake of proton pump inhibitors and cagA [Bacterial load is also the reason for increased acute mucosal damage and for failure in ial load ,103. Liactively) . Belda eand cagA .5H. pylori and gastric ulcer ineffective. Many antibiotics show low in vivo efficacy due to the failure of the drug to reach the target site of action. H. pylori resides in the inner layer of stomach lining, where the bioavailability of a drug is low, and the plasma concentration drops based on the half-life (t1/2) of the drug [H. pylori infection. Various advantages of these novel formulations over the traditional formulations for the treatment of H. pylori infection are represented in The above discussed challenges make the treatment of the drug . To mainthe drug ,108. Nanthe drug . In the 5.15.1.1H. pylori bacteria due to its localized action, which will directly deliver antibiotics in the gastric mucosa for longer periods. According to Javadzadeh et al. the floating system is suitable for medications that act locally on the stomach's gastric mucosa, such as metronidazole. Using this system, antimicrobial drugs can be applied topically to cure H. pylori infection, which may be helpful to prevent the adverse effects of traditional triple therapy [H. pylori growth effectively in vitro and in vivo [in-vitro investigation supported the proposed system's strong antibacterial efficacy. The hollow micro balloons loaded with amoxicillin appeared to be a good strategy for localized release of amoxicillin for the successful management of H. pylori-induced ulcers [H. pylori infection. The optimized formulation showed better pharmacokinetics in comparison to marketed formulation and showed release of drug in controlled manner for more than 24 h [H. pylori bacteria showed 71% entrapment efficiency of the drug, and 82% of the microsphere floated for more than 12 h (11). Patel et al. and Emara et al., reported that the floating tablet of clarithromycin and amoxicillin has improved the prolonged release of the drug to the intestinal mucosa  or the disruption of cellular membranes, genetic components, or proteins [H. pylori [H. pylori infection.The Nobel laureate Richard P. Feyman first introduced nanotechnology in his lecture ‘There's Plenty of Room at the Bottom’ in 1959 . Nanotecproteins . It is a. pylori . Further5.2.1H. pylori were effective by inducing inflammation, oxidative stress, apoptosis, and biofilm inhibition [Tribulus terrestris showed bactericidal effect on the H. pylori bacteria [H. pylori resistant to the antibiotic metronidazole. This ZnO-PEI NPs caused excessive formation of ROS within bacteria, resulting in cell membrane damage, RNA degradation, and consequent bacterial death. This demonstrates that antibiotics may be safely employed with ZnO-PEI NP at concentrations below those at which they cause damage to human cells [Toxicodendron vernicifuum. It exhibited antibacterial action against H. pylori by inducing ROS, cell death, oxidative stress, and cytoplasmic DNA damage [H. pylori infection in combination with antibiotics [H. pylori at concentrations ranging from 60 to 100 ng/ml. Bismuth nanoparticles have been seen to interact with the Krebs cycle, nucleotide metabolism, and amino acid metabolism, causing bacteria to produce metabolites such as uracil, glutamate, acetate, glycine, formic acid, valine, and glycine [Metallic nanoparticles are fabricated from pure metal particles such as zinc, gold, and silver, which have their own distinct surface charges and hydrophobicity . This fahibition . Gopinatbacteria . Chakraban cells . SaravanA damage . ZnO NPsA damage . Bismuthibiotics . Nazari  glycine . Fig. 4 5.2.2H. pylori. Polyacrylic acid, poly lactic-co-glycolic acid (PLGA), alginate, albumin, gliadin, chitosan, and cellulose are the most extensively utilized polymers to make polymeric nanoparticles for fighting H. pylori due to their pH sensitivity, mucosal adhesiveness, and biocompatibility [H. pylori. The MIC value of the NPs was 0.003 μg/ml to 0.1 μg/ml [H. pylori medication delivery systems [H. pylori therapy [H. pylori bacteria to gastric mucosal cells [H. pylori than the standard triple treatment [Polymeric NPs are biodegradable organic NPs with nano-spherical or nano-capsular forms . Due to tibility . PLGA NPtibility . Lutfipo.1 μg/ml . One mor systems . Harsha  therapy . Umamaheal cells . Lectin-reatment . A picto5.2.3H. pylori. The stomach's acidic environment makes it challenging to eradicate H. pylori due to the insufficient reach of antibiotics in the affected region. To get over this problem, scientists have developed a urea-mediated targeted drug delivery system consisting of pH-sensitive urea-modified UCCs- 2-PLGA NPs. These NPs have been found to have greater anti-H. pylori action, lesser cytotoxicity, more specificity, and improved pH sensitivity. It is reported by Chang et al. that amoxicillin-loaded genipin cross-linked fucose-chitosan will interface with H. pylori and show efficient and precisely controlled release [H. pylori in-vivo at a dosage of 6 mg/L on C57BL/6 mice, which resulted in 40% inhibition [H. pylori. Adding ligands or targeting molecules designed to interact with bacteria to the surface of the NPs enable selective targeting of the drug carrier NPs to the target areas [H. pylori treatment  and clarithromycin for in vitro H. pylori eradication. The outcomes showed that NLCs made interaction with the bacterial call wall by sticking to its outer cell wall without harming the human cells and then disrupting the membrane, resulting in cytoplasmic leakage.Lipid nanoparticles have been widely employed as medication carriers and chemical agent delivery in biological applications. The lipid NPs have a solid lipid core surrounded by a matrix of soluble lipophilic molecules, and their sizes range from 10 to 1000 nm. Surfactants and emulsifiers were used to maintain the exterior's structural integrity . Liposomal cells . Recent . pylori ,194. LipH. pylori, which might be owing to the Hesp component causing harm to the microbial lipid layer, increasing the permeability of the antibiotic to the bacterial cell [H. pylori and a reduction of drug side effects on normal cells. The mechanism of lipid-based NPs (LPNs) for the eradication of H. pylori is shown in H. pylori infection by removing both biofilm and mucous layers blockage. Chitosan nanoparticles (CS NPs) were used in LPNs as the exterior, mixed lipid layers comprising rhamnolipids (RHL) as the center, and DSPE-PEG2000 was added to the surface of LPNs to enhance hydrophilicity. The H. pylori biofilm was totally eradicated by the formulated LPNs. PEGylated LPNs quickly pass through mucus and efficiently remove the H. pylori biofilm that was embedded in the mucus layer without reacting with mucins H. pylori as well as MGC803 biotic cells is demonstrated in H. pylori can be eliminated, even at small concentrations, using nanostructured lipid transporters that are not loaded with medicines. Cultures of H. pylori with nanostructured lipid transporters (1.25%) were examined using microscopic techniques, which revealed that after 12 h the membranes of most of the bacteria had been broken. This demonstrated that the H. pylori barrier can be destroyed by the nanostructured lipid transporters. The capacity of nanostructured lipid transporters to be kill H. pylori while having no effect on intestinal flora has been shown Furthermore, their findings revealed that NLCs had a synergistic impact on ial cell . Further5.3H. pylori [H. pylori infection. They observed a dramatic increase in the H. pylori induced ulcer development in the P-gp-expressed rats compared to the P-gp inhibited ones [H. pylori infection. The results revealed that the P-gp inhibitors effectively eradicated H. pylori infection in 20 out of 33 patients treated with p-gp inhibitors [H. pylori patients to understand the effect of P-gp on treatment failure in H. pylori infection. In comparison with the group that were H. pylori-negative, the resistant group had greater relative antral P-gp expression (p < 0.0361), and the majority of individuals showed resistance towards clarithromycin (72%), metronidazole (63.6%), or both (54.5%) [H. pylori infection. The result showed that P-gp expression differed significantly between H. pylori-positive and H. pylori-negative individuals (P < 0.028). In antral specimens, the TT genotype showed substantially less P-gp activation than the CC genotype for the MDR1 C3435T polymorphism (p < 0.041). P-gp activation was also substantially different in the homozygous TT genotype with H. pylori infection in comparison to H. pylori-negative individuals (p < 0.029) [H. pylori. The mechanism of P-gp inhibitor to stop the drug efflux shown in P-glycoprotein (P-gp) is a transmembrane glycoprotein (170 kDa) that comes under a broad superfamily of ATP-binding cassette (ABC) transporters ,198. P-g. pylori ,199. Damted ones . Babic ehibitors . Omar et (54.5%) . Omar et< 0.029) . All the6H. pylori infection and associated gastric ulcer is always a concern for the treating doctor. The current therapeutic regimens are not very effective due to antibiotic resistance and patient noncompliance. Development of a new formulation is urgently required to overcome the challenges of the current modalities. As discussed in this review, strategies like GRDDS, p-gp inhibitors, and nano-formulations have proved to be useful in vitro as well as in a few animal models. After the comprehensive review of the various approaches to overcome the challenges, authors recommend here a strategy for effective eradication of H. pylori infection and the associated gastric ulcer. We recommend a nano-formulation of antibiotics incorporated as a GRDDS system for eradication of H. pylori infection. The treatment regimen should include a P-gp inhibitor and a PPI along with the GRDDS system containing the antibiotic. The proposed novel formulation is expected to significantly improve the treatment efficacy as it can allow precise target specific drug binding. The formulation is expected to provides better stability for antibiotics from the acidic environment and assures elevated local drug concentration. By design, GRDDS extends the time duration of the drug availability in the stomach. This extension in contact time facilitates sustained interactions with bacterial colonies, substantially enhancing the probability of successful eradication. The advantages of employing gastroretentive formulations encompass elevated bioavailability of medications, reduced dosing frequency, and potentially improved patient compliance in the treatment regimen. As indicated in the literature, the mucoadhesive approach appears to offer better convenience and efficacy in patients compared to other GRDDS in the context of H. pylori infection treatment. The mucoadhesive system exhibits the capability to adhere to the mucosal surface, enabling direct and targeted contact with the bacteria. This direct interaction holds the potential of significantly enhancing the efficacy of H. pylori infection treatment. The nano-formulation owing to its diminutive scale, increased surface area, and efficient compactness offer an additional layer of advancement by the encapsulation of antibiotics thereby providing prolonged release and increased bioavailability and minimized side effects. The extended-release property of nano-formation enhances the antibiotic's effectiveness and could potentially reduce the frequency of dosing and improves patient compliance. Leveraging polymeric nanoparticles within the framework of GRDDS presents a potent strategy against H. pylori infection. Both floating and mucoadhesive GRDDS systems demonstrate compatibility with nano-formulations, showcasing their suitability for such therapeutic endeavour. Phytochemicals and natural compounds, when used as P-gp inhibitors in conjunction with standard antibiotics, prevent their efflux and can improve the treatment outcomes, offering a more comprehensive therapeutic approach. Integrating nanoparticles into a GRDDS alongside a P-gp inhibitor and PPI holds the potential of novel approach for eradication of H. pylori infections. Concurrently, the intrinsic H. pylori eradication property of the PPI contributes to the formula's effectiveness. The inclusion of a P-gp inhibitor not only eradicates biofilm formations but also impedes drug efflux mechanisms, further boosting the treatment strategy. This multifaceted approach has the capacity to redefine the landscape of H. pylori infection management. Thus, a complete eradication of H. pylori could be achieved with this innovative approach by assuring sufficient drug concentrations locally and systemically for an extended period of time, which would also prevent the development of resistance to antibiotics.Treatment of 7As previously mentioned, novel formulations offer numerous advantages; however, the formulation development process is not without its challenges. These include the need to select suitable antibiotics and excipients based on compatibility, ensuring drug stability in gastric juices, optimizing the drug-polymer ratio to prolong stomach retention, determining the appropriate drug release kinetics, addressing cost and treatment accessibility, and successfully translating the formulation into clinical applications.H. pylori bacteria. These bacteria are anaerobic in nature, demanding specialized growth conditions and specific media for cultivation. Additionally, identifying a suitable animal model that accurately replicates the conditions of the human stomach for H. pylori infection poses its own set of considerable challenges. Handling H. pylori poses a big challenge as the bacteria has a high contamination rate. Antibiotic formulations may be less effective since H. pylori has shown increased resistance to these drugs by genetic modifications. Additional difficulties arise when attempting to assess the formulation's effectiveness within the context of combination therapy. Reproducibility of the experimental results in an animal model is also challenging.Conducting in-vivo evaluations of the formulation presents another significant challenge due to the limited availability of facilities for culturing 10.13039/501100001411Indian Council of Medical Research, New Delhi, India. File no. 3/2/2/16/2022-NCD-III.The work is funded by Ashutosh Gupta: Writing – review & editing, Writing – original draft, Visualization, Methodology, Funding acquisition, Formal analysis, Data curation, Conceptualization. Shiran Shetty: Writing – review & editing, Supervision, Formal analysis, Data curation, Conceptualization. Srinivas Mutalik: Writing – review & editing, Supervision, Investigation, Formal analysis, Data curation, Conceptualization. Raghu Chandrashekar H: Writing – review & editing, Supervision, Investigation, Conceptualization. Nandakumar K: Writing – review & editing, Supervision, Methodology, Data curation, Conceptualization. Elizabeth Mary Mathew: Conceptualization, Writing – review & editing. Abhishek Jha: Writing – original draft, Data curation, Conceptualization. Brahmeshwar Mishra: Writing – review & editing, Supervision, Conceptualization. Siddheesh Rajpurohit: Writing – review & editing. Gundawar Ravi: Writing – review & editing. Moumita Saha: Writing – review & editing. Sudheer Moorkoth: Writing – review & editing, Visualization, Supervision, Resources, Investigation, Data curation, Conceptualization.The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.


In one of these symbioses, the symbiont, Richelia euintracellularis, has penetrated the cell envelope of the host, Hemiaulus hauckii, and lives inside the host cytoplasm. How the partners interact, including how the symbiont sustains high rates of N2 fixation, is unstudied. Since R. euintracellularis has evaded isolation, heterologous expression of genes in model laboratory organisms was performed to identify the function of proteins from the endosymbiont. Gene complementation of a cyanobacterial invertase mutant and expression of the protein in Escherichia coli showed that R. euintracellularis HH01 possesses a neutral invertase that splits sucrose producing glucose and fructose. Several solute-binding proteins (SBPs) of ABC transporters encoded in the genome of R. euintracellularis HH01 were expressed in E. coli, and their substrates were characterized. The selected SBPs directly linked the host as the source of several substrates, e.g. sugars , amino acids , and a polyamine (spermidine), to support the cyanobacterial symbiont. Finally, transcripts of genes encoding the invertase and SBPs were consistently detected in wild populations of H. hauckii collected from multiple stations and depths in the western tropical North Atlantic. Our results support the idea that the diatom host provides the endosymbiotic cyanobacterium with organic carbon to fuel N2 fixation. This knowledge is key to understanding the physiology of the globally significant H. hauckii–R. euintracellularis symbiosis.A few genera of diatoms are widespread and thrive in low-nutrient waters of the open ocean due to their close association with N In the association between the diatom Hemiaulus hauckii (host) and the N2-fixing, heterocyst-forming cyanobacterium Richelia euintracellularis (endosymbiont), the cyanobacterium is uncultivable. Here, we used heterologous expression of genes from the endosymbiont to identify the function of proteins involved in the utilization of organic carbon from the host. The importance of these proteins was also confirmed by estimating gene expression in environmental samples. Our results show that the metabolisms of the symbiotic partners are integrated allowing the host to sustain the physiology of the endosymbiont for an important ecological role.Diatom diazotroph associations (DDAs) between diatoms and N2-fixing bacteria (diazotrophs). Diatoms are diverse photosynthetic unicellular microalgae that typically thrive in coastal zones with high nutrients and contribute ∼20% of the Earth's photosynthesis  are stable mutualistic partnerships between a few genera of diatoms and Nynthesis . The diaea floor . DespiteRichelia, which are filamentous, terminal heterocyst-forming cyanobacteria  of its host diatom (Hemiaulus hauckii) and resides in the cytoplasm of the diatom , reducing in particular its N assimilatory pathways to only N2 fixation. Additionally ReuHH01 lacks several central metabolic enzymes, such as glutamine:2-oxoglutarate amidotransferase (GOGAT) and the proteins for sucrose biosynthesis, and has limited transporters compared to free-living cyanobacteria , in order to identify gene function. Here, we studied properties of several SBPs from ABC transporters of ReuHH01 putatively involved in the uptake of organic carbon and additionally the properties of an invertase from this organism. We then showed gene expression for the genes that encode some components of these ABC transporters and invertase in wild symbiotic populations. Knowledge of these proteins is key to understanding the physiology of the globally significant H. hauckii–R. euintracellularis symbiosis.Since ReuHH01 has evaded isolation, we used the heterologous expression of ReuHH01 genes of interest in laboratory amenable organisms, including Anabaena mediates the uptake of glucosides and some monosaccharides such as glucose and fructose . We focused on the analysis of RintHH_17430 to address the substrate specificity of the transporter. RintHH_17430 (gene ID: 2580948285) was chemically synthesized and cloned in E. coli expression vector pET28b(+)  Fig. . Because(+) Fig. , which d(+) Fig. , we clon(+) Fig. . The mat(+) Fig. , which p(+) Fig. .µm. Increasing sucrose concentration to 1 mm increased quenching only to ∼35% . The extent of RintHH_17430 sucrose-induced fluorescence quenching is similar or somewhat stronger than that described for other periplasmic binding proteins and their substrates  and 60% identity (272/456 amino acid residues) to Anabaena Alr1521 (InvA)  whereas it grew well in nitrate-containing (BG11) medium; in contrast, the invB mutant complemented with RintHH_3860 could grow fixing N2 as well as in nitrate-containing medium  and a growth rate of the RintHH_3860-complemented Anabaena strain (0.342 day−1) similar to the growth rate of wild-type Anabaena (0.366 day−1)  encodes a protein that shows homology to invertases: 74% identity (347/467 amino acid residues) to vA) Fig. . Because80946898 nditions , we purster Fig.  and clonl et al. . The Rinter Fig. . Growth E. coli expression vector, pET28b(+) ::RintHH_3860] compared with the activity in extracts of an isogenic strain lacking RintHH_3860. Cell-free extracts of E. coli bearing RintHH_3860 supplemented with sucrose showed equimolar production of glucose and fructose, indicating that both monosaccharides resulted from sucrose hydrolysis, whereas no production was observed in the extracts of the control strain  Fig. . Becauseto 36 mm , 28. It to 15 mm , 29. ThuAnabaena N-I transporter for neutral/hydrophobic amino acids and N-II transporter for acidic and neutral amino acids , RintHH_12770 (gene ID: 2580947807), and RintHH_7180 (gene ID: 2580947237), encode proteins that contain a signal peptidase II type of signal peptide and Trp residues . RintHH_11820 is 75% identical to Anabaena NatB (289/386 amino acid residues), and RintHH_12770 is 63% identical to Anabaena NatF (226/361 amino acid residues). Fluorescence quenching of RintHH_11820 was tested by the addition of the following compounds: L-Pro, L-Ala, L-Phe, Gly, L-Leu, glucose, L-Asp, and L-Arg. Added at ∼116 µm, substantial quenching was produced by the neutral amino acids  but much less quenching was produced by the sugar or by the acidic (L-Asp) or basic (L-Arg) amino acids, and ∼17 µm L-Phe was especially effective denoting high affinity . Specific oligonucleotides and assays were designed to detect gene transcripts similar to ReuHH01 invB, glsR, natF, and potD; and secA, encoding protein translocase subunit SecA, was used for normalization  to stations 23 and 25 but was sampled 3 weeks earlier, while stations 2 and 5 were sampled 3 days apart but more distantly located (557 km) , and silicate (Si) was variable (1.04–7.03 µmol L−1) and extremely high, especially at station 23 (e.g. 25.0 µmol L−1)  Fig. . In the natF were detected at particularly significant levels compared to the gene chosen as reference, secA  carbon compounds, which are needed to fuel N2 fixation in the heterocysts , a type of transporter involved in the uptake of sugars including sucrose  but to lack GOGAT, raising the question of the origin of glutamate that is used to incorporate ammonium resulting from Nfixation . It was fixation  encoded fixation . Howeverfixation  raises t highest . DissolvR. euintracellularis HH01 also encodes an ABC uptake transporter similar to the N-I system of Anabaena, which transports neutral/hydrophobic amino acids  and assessed at the Phyre2 portal for protein modeling, prediction, and analysis (http://www.sbg.bio.ic.ac.uk/∼phyre2/html/page.cgi?id=index). Amino acid composition of a protein was analyzed at https://web.expasy.org/protparam/. Signal peptides were searched at http://www.cbs.dtu.dk/services/SignalP/.The draft genome sequence of  updated ; the preE. coli after standard IPTG induction and purified by affinity chromatography using StrepTrap HP column (prepacked with StrepTactin Sepharose) or nickel columns for proteins carrying Strep-tag II or 6xHis fusions, respectively. See The genes were synthesized by Integrated DNA Technologies, Inc. (IDT) and provided in a plasmid from which the selected DNA fragment was transferred by standard recombinant DNA methods to the plasmid shown in the corresponding supplementary figures Figs. . OligodeAnabaena invB mutant described in López-Igual et al.  using a peristaltic pump. Filters were placed in sterile tubes amended with RLT buffer (Qiagen RNAeasy Plant Mini Kit) and stored at −80°C until extraction. Total RNA was extracted and reverse transcribed (RT) and used in quantitative polymerase chain (RT-qPCR) reactions using previously described methods, commercially available kits  and TaqMAN chemistry (Applied Biosystems; R. euintracellularis HH01 (Figs. glsR, invB, natF, potD, and secA are described in the An expedition to the WTNA was conducted in May 23 to June 23, 2010. Four stations (stations: ormation , 32. Seaormation . Water w01 Figs.  and S11.pgad194_Supplementary_DataClick here for additional data file.