Helicobacter pylori (H. pylori) is one of the most common human pathogens, affecting half of the world’s population. Approximately 20% of the infected patients develop gastric ulcers or neoplastic changes in the gastric stroma. An infection also leads to the progression of epithelial–mesenchymal transition within gastric tissue, increasing the probability of gastric cancer development. This paper aims to review the role of H. pylori and its virulence factors in epithelial–mesenchymal transition associated with malignant transformation within the gastric stroma. The reviewed factors included: CagA (cytotoxin-associated gene A) along with induction of cancer stem-cell properties and interaction with YAP (Yes-associated protein pathway), tumor necrosis factor α-inducing protein, Lpp20 lipoprotein, Afadin protein, penicillin-binding protein 1A, microRNA-29a-3p, programmed cell death protein 4, lysosomal-associated protein transmembrane 4β, cancer-associated fibroblasts, heparin-binding epidermal growth factor (HB-EGF), matrix metalloproteinase-7 (MMP-7), and cancer stem cells (CSCs). The review summarizes the most recent findings, providing insight into potential molecular targets and new treatment strategies for gastric cancer. Helicobacter pylori (H. pylori) is a helix-shaped, Gram-negative, microaerophilic, flagellated bacterium that is capable of biofilm formation and converting from spiral to coccoid form ..51].As a physiological process, EMT is observed during organogenesis, tissue development, remodeling, and wound healing ,53,54; cc-MYC [During this process, epithelial cells undergo a series of biochemical changes, which lead to the loss of polarity and migratory capacity of cells, resulting in cell shape changes (cell elongation). EMT promotes the transformation of immobile epithelial cells into motile mesenchymal cells, enhancing the metastatic properties ,70. Furtc-MYC ,77,78,79c-MYC . Moreovec-MYC ,82,83.H. pylori infection significantly affects the gastric microenvironment by induction of several inflammatory responses via infiltrating macrophages, neutrophils, regulatory T-cells, and natural killer cells [H. pylori virulence factors are considered being associated with the promotion of EMT in gastric cells, which consequently causes neoplasia and malignant transformation. This review summarizes several mechanisms associated with epithelial–mesenchymal transition, gastric tumor microenvironment, and the influence of H. pylori infection, although some described mechanisms are not only H. pylori-specific. Even though H. pylori-induced carcinogenesis is not fully understood, several mechanisms have already been deciphered.er cells ,85. Infler cells ,87,88. Ter cells ,90,91. AH. pylori, cag pathogenicity island (cagPAI) probably plays a key role in carcinogenesis [CagA oncoprotein [CagA into a cell, transforming its shape into the so-called “hummingbird phenotype” characterized by an elongated cell shape commonly observed in EMT [CagA into the cell via the T4SS induces signal transduction, with one of the most relevant mecahnisms being the nuclear factor κB (NF-κB) signaling pathway involving extracellular regulated kinases 1/2 (ERK-1/2) [H. pylori [CagA in host cells is tyrosine phosphorylated and interacts with protein tyrosine phosphatase 2 (SHP-2), also inducing the progression of the “hummingbird phenotype” [CagA enhances EMT via the stabilization of Snail protein, which is essential in carcinogenesis, mainly by the reduction of glycogen synthase kinase-3 (GSK-3) activity [CagA-positive H. pylori strains with CagA containing phosphorylation-functional EPIYA motifs present significantly higher expression of mesenchymal markers such as vimentin, Snail, and ZEB-1 and the stem cell marker CD44 [CagA-positive H. pylori strains induce a higher probability of gastric carcinogenesis and induction of EMT process [CagA-positive H. pylori infection present poor clinical outcome, and higher invasion and metastatic characteristics [Among many virulence factors of ogenesis ,93,94. Ioprotein ,96. The d in EMT ,98. InjeERK-1/2) ,100,101.ERK-1/2) . The inh. pylori . CagA inenotype” ,105. Cagactivity . CagA-poker CD44 ,109,110. process ,112,113.eristics . Besideseristics ,116.CagA-positive H. pylori strain infection [Recent research suggests that cells that undergo EMT obtain the ability to acquire cancer stem cell (CSC) properties ,122,123.nfection ,129. Hignfection ,135,136.H. pylori in the generation of cells with CSC properties, including several gastric epithelial cell lines  [CagA in the induction of CSC properties was studied on H. pylori 7.13 wild-type (WT) CagA-positive strain and its knock-out mutants—7.13CagA-negative and 7.13DCagE-negative H. pylori strains. Only the wild-type (WT) (7.13CagA-positive) strain-induced mesenchymal changes and EMT in cells; cells infected by CagA-positive strains, presented higher expression of mesenchymal markers—CD44, Snail1, vimentin, or zinc finger E-box binding homeobox 1 (ZEB1), while an expression of epithelial markers—cytokeratin 7 (CK7) or osteopontin (SSP-1) was lowered [CagA-positive H. pylori strains. The relevance of the infection by CagA-positive H. pylori strain on EMT in gastric cancer was confirmed by significantly higher expression of CD44 and mesenchymal markers in tumor samples.Bessède et al. (2014) studied the role of  MKN-74) . The rol lowered . The migCagA-positive H. pylori EMT and the induction of CSC properties [CagA-positive H. pylori infection with N-nitrosoguanidine (MNNG) stimulation causes CSC features with dysplastic lesions and mesenchymal phenotype. Amieva and Peek (2016) observed that CD44+ cells with CSC features displayed increased CagA half-life [CagA was confirmed in CD44+ gastric CSCs [H. pylori-infected (CagA-positive) gastric cancer cells exhibit CSC properties via increased expression of surface markers—CD44 and Lgr5 with Nanog, Oct4, and c-myc upregulation [CagA impairs transcription factor CDX1 expression, promoting EMT, and CSC features in gastric epithelial cells [The Wnt/β-catenin signaling pathway is involved in operties . Likewisalf-life . Accumulric CSCs . H. pylogulation ,140. Cagal cells .MYC oncogene [Yes-Associated-Protein (YAP) is an element of the Hippo tumor suppressor signaling pathway, which plays a crucial role in the maintenance of the proper size of organs, tissue homeostasis, cell proliferation, and stem cell maintenance ,143,144.oncogene ,149. Besoncogene .CagA-positive H. pylori strain induces higher YAP expression and decreases E-cadherin, N-cadherin, and Slug levels, promoting EMT.Li et al. (2018) showed a significantly higher expression of YAP and TAZ  in cancerous gastric tissues with a positive correlation between YAP, TAZ levels, and tumor size . During CagA, in the enhancement of the YAP pathway [CagA-positive H. pylori upregulates YAP1 and large tumor suppressor 2 (LATS2) expression in gastric epithelial cells [YAP1 is associated with aggressiveness and poor prognosis [YAP1/SLC35B4 axis; another mechanism involves altered IL-1β levels [Other studies have shown a potential role of GSK3/β-catenin or AMP-activated protein kinase (AMPK) pathways, which are regulated by  pathway ,153. Molal cells . Overexprognosis . Gastricβ levels ,157. TheH. pylori strains produce a high quantity of tumor necrosis factor-α (TNF-α)-inducing protein (Tipα protein), which binds to the cell surface nucleolin and induces carcinogenic alterations [H. pylori virulence [α induces mesenchymal changes of cells via EMT progression [α, as a carcinogenic factor, activates NF-κB, promoting gastric carcinogenesis by inhibiting miR-3178 expression [α release, Tipα also induces the expression of several chemokines, including chemokine (C-C motif) ligand 2 (Ccl2), chemokine (C-C motif) ligand 7(Ccl7), chemokine (C-C motif) ligand 20 (Ccl20), C-X-C motif chemokine 11 (Cxc11), chemokine (C-X-C motif) ligand 2 (Cxcl2), C-X-C motif chemokine 5 (Cxcl5), and C-X-C motif chemokine 10 (Cxcl10) [α (TNF-α), or hypoxia-inducible factor 1α (HIF1α) [α, along with CagA and VacA, plays an underlying role primarily in mucosal damage [erations ,162,163.irulence ,165. By gression ,167,168.pression ,170. Bes(Cxcl10) ,173,174. (HIF1α) ,176. In l damage ,178.H. pylori infection increases the quantity of Tipα protein in gastric cancer tissue, inducing tumor progression in H. pylori carcinogenesis [α reduces cell stiffness and phosphorylates various oncoproteins; it also enhances filopodia formation, morphological, and conformational changes within cells, and expression of vimentin via MEK-ERK phosphorylation, confirming its role in EMT progression [Suganuma et al. showed that ogenesis . Tipα regression ,180.α and nucleolin receptor (88 kDa) on the migration properties of gastric cancer cells [α could be inhibited via small interfering RNAs targeted for the nucleolin receptor, showing its role in EMT progression [H. pylori infection by inhibiting Tipα and nucleolin interaction [Researchers have suggested the role of Tiper cells ,181. Fujgression . Thus, peraction ,184.α induces the formation of filopodia and a reduction of cell stiffness, predisposing cells to higher motility [α induces higher levels of IL-1β, TNF-α, and IL-8 in SGC7901 cells [H. pylori infection promotes the Il-6/STAT3 signaling pathway in AGS cells, which is one factor promoting EMT [α did successfully eradicate H. pylori infection [Tipmotility . It has 01 cells . Furtherting EMT . Inoue enfection .H. pylori virulence factor, promoting its colonization and survival properties [H. pylori, which enhances the stimulation of cell proliferation. Lpp20 expression is increased in the acidic pH of stomach juice and during iron depletion [The Lpp20 protein plays an important role as an operties . Lpp20 iepletion ,190.α), is one of the carcinogenic factors released by H. pylori via NF-κB pathway activation [α and Lpp20 induce EMT mainly by stimulating cell proliferation, migratory properties of cells, and filopodia formation [Lpp20 , these proteins show similar effects on gastric cancer cells by stimulating EMT and gastric cancer progression.Even though Lpp20 and TipH. pylori lowered the expression of Afadin protein independently of CagA and VacA in MKN74 and NCI-N87 cell lines [H. pylori.Afadin is a cytoplasmic actin-filament-binding protein responsible for the formation and stabilization of tight and adherens junctions ,194,195.ll lines . DownregH. pylori with an affinity for penicillin binding; any modifications within PBP might induce resistance to β-lactam antibiotics  [PBP2X, PBP2B, and PBP1A [H. pylori PBPs might present unique characteristics because of several putative genes encoding PBPs.Penicillin-binding protein (PBP) is a specialized acyl serine transferase released by icillin) . These mnd PBP1A . It was PBP1A mutation-positive H. pylori (H. pylori CagA+/P+) strain and CagA+/P− in terms of clinicopathological characteristics of gastric cancer and EMT induction [H. pylori CagA+/P+ infection resulted in progressive and clinically significant EMT and gastric cancer severity. Researchers have observed decreased levels of E-cadherin and increased α-smooth muscle actin (α-SMA) levels; likewise, it was observed that PBP1A mutation is associated with decreased miR-134 levels. MiR-134 suppresses gastric carcinogenesis by targeting the KRAS gene and Golgi phosphoprotein 3 (GOLPH3) downregulation; another essential target gene of miR-134 is FoxM1 [H. pylori activity decline [Huang et al. 2019) showed significant differences between nduction . H. pylois FoxM1 ,201. The9 showed  decline . Marcus  decline .Micro-RNAs constitute a family of small noncoding RNAs, which are regulators of posttranscriptional gene expression by binding to the 3′-untranslated region of mRNA, promoting an inappropriate translation of targeted mRNA. There is an increasing interest in micro-RNA involvement in many diseases and malignancies ,206,207.H. pylori [H. pylori infection induced the overexpression of microRNA-29a-3p in gastric cancer tissue samples, which resulted in an enhanced ability of the migration of gastric epithelial cells. Furthermore, the silencing of A20 increased the EMT markers Snail, vimentin, and N-cadherin, and decreased E-cadherin levels. The authors have suggested that the overexpression of micro-RNA-29a-3p might be associated with a higher probability of EMT induction in gastric cells.Sun et al. presented the relevance of upregulation of microRNA-29a-3p and its role in the decreased expression of A20 in patients infected by . pylori . A20 is . pylori . H. pyloTWIST1 gene [Programmed cell death protein 4 (PDCD4) is a tumor suppressor responsible for the inhibition of cell growth, tumor invasion, metastasis, or induction of apoptosis . It is lST1 gene .TWIST1, and vimentin levels in gastric cancerous tissues [CagA-positive H. pylori strain infection. PDCD4 overexpression reduced EMT. MicroRNA-21 overexpression in gastric cancer modulates the expression of the tumor suppressors phosphatase and tensin homolog (PTEN) and PDCD4, altering molecular pathways associated with cell growth, invasion, migration, and apoptosis [PDCD4 is one factor responsible for the induction of carcinogenesis . Yu et a tissues . PDCD4 dpoptosis .LAPTM4B) is a proto-oncogene relevant in the progression of tumorigenesis, regulating molecular mechanisms, and cellular functioning, including proliferation, migration, and invasion [LAPTM4B upregulation has been observed in various cancers, including hepatocellular carcinoma, breast cancer, and lung adenocarcinoma [Lysosomal-associated protein transmembrane 4β  while lowering E-cadherin levels and dysregulating cell–cell junctions. Additionally, there was a noticeable increase of ZEB1, β-catenin, Snail, and Slug markers, whereas tight junction protein-1  level was decreased. LAPTM4B seems to be a factor strictly associated with EMT induction since it is responsible for regulating cells’ filopodia, increasing motility of gastric cancer cells, and a higher invasion potential [LAPTM4B results in poorer prognosis in various cancers [It has been reported that ormation . Researcotential . Overexp cancers .α in fibroblasts, which is common during CagA+VacA+ H. pylori infection [Physiologically, natural gastric stroma contains only a few fibroblasts, and those are mainly myofibroblasts. The number of fibroblasts is significantly increased in tissues affected by inflammation or neoplasm ,230,231.nfection .H. pylori infection results in the transformation of fibroblasts and myofibroblasts into CAFs [CagA+VacA+ H. pylori infection results in the overexpression of fibroblast activation protein (FAP), fibroblast surface protein (FSP) mRNA, and increased levels of the proinflammatory factors IL-6, IL-8, COX-2, and SDF-1 [α-SMA, N-cadherin, vimentin, Snail, Twist) and lowered levels of E-cadherin, antiapoptotic B-cell lymphoma 2 (Bcl-2), and proliferative marker Ki-67 protein within fibroblasts [CagA+VacA+ H. pylori infection resulted in the aberrant apoptosis process, an increased level of collagen production, and impaired fibroblast and myofibroblast differentiation into CAFs. This induced desmoplastic reactions and EMT within gastric cells, promoting carcinogenesis. The increased levels of β1-integrin and COX-2, which are crucial in invasion, metastasis, and angiogenesis, enhanced the migration properties of the infected cells. CagA+Vac+ H. pylori infection results in the upregulation of HIF-1α and tenascin-C (TNC), enhancing EMT and tumor progression, the release of proangiogenic factors, and further CAFs activation [CagA+Vac+ H. pylori infection activates gastric fibroblasts and induces the release of TGF-β [nto CAFs . CagA+Vand SDF-1 . Overexpnd SDF-1 ,248,249.roblasts . CagA+Vativation ,251,252.of TGF-β .The tumor-promoting ability of CAFs is enhanced by microRNA dysregulation . CAFs stH. pylori infection results in the upregulation of HB-EGF gene expression, which is a potential serological biomarker for gastric cancer [H. pylori g-glutamyltranspeptidase, activating PI3K, and p38 kinase-dependent signal transduction pathways [Heparin-binding epidermal growth factor (HB-EGF) is an epidermal growth factor receptor (EGFR) cognate ligand with mitogenic and chemotactic properties for fibroblasts and smooth muscle cells . H. pyloc cancer ,266,267.c cancer . HB-EGF pathways .H. pylori infection and is associated with EMT [H. pylori infection [Matrix metalloproteinase-7 (MMP-7) is an enzyme responsible for the degradation of adherence between cells in the extracellular matrix, and overexpression of MMP-7 induces a more aggressive course of gastric cancer ,272,273.with EMT ,275. Liknfection ,278,279.nfection ,280.H. pylori infection results in the increased release of MMP-7 and soluble HB-EGF, promoting EMT [H. pylori-induced inflammation and damage via M1 macrophage polarization suppression [Yin et al. showed that excessive gastrin secretion due to ting EMT . HB-EGF ting EMT . Besidesting EMT ,284,285.ting EMT . Gastricting EMT ,288. Howpression .H. pylori infection, neoplasia, or cancer, being involved in the migration properties of gastric cancer cells [Mesenchymal stem cells (MSCs) present a multipotent potential to differentiate into various cell types, while keeping a capacity for self-renewal ,291. MSCer cells ,293,294.er cells ,296. Naner cells .H. pylori gained a fibroblastic phenotype, enhancing EMT progression in gastric cancer cells [H. pylori showed a significant decrease in the levels of E-cadherin and overexpression of mesenchymal markers  and inflammatory cytokines , VEGF, epidermal growth protein (EGF), granulocyte-macrophage colony-stimulating factor (GM-CSF), and monocyte chemoattractant protein 1 (MCP-1). High concentrations of IL-15 secreted by gastric cancer MSCs contribute to tumor cell metastasis; increased IL-17 levels promote gastric cancer invasiveness [Zhang et al. showed that human umbilical cord MSCs (hucMSCs) infected by er cells . Furthersiveness ,300.SOX4 induces EMT via a release of TGF-β and contributes to the development of stem cell characteristics of gastric cancer cells [CagA-positive H. pylori infection induces EMT, and CSCs feature in gastric mucosal cells via overexpression of the Wnt/β-catenin signaling pathway [KRAS, STAT3, Rac1, Wnt, Notch, PTEN, ERK, NF-κB signaling pathways, or hypoxic microenvironment [Increased expression of er cells . Furtherer cells . CagA-po pathway . The pro pathway ,306,307. pathway . Besidesironment ,317,318.ironment .H. pylori infection and gastric carcinogenesis remains controversial. In fact, some mechanisms are yet not fully understood. H. pylori infection seems to play an important role in terms of EMT induction within gastric mucosa (The association between c mucosa .H. pylori virulence factors. Likewise, cellular components that are associated with EMT progression can be influenced by H. pylori infection (H. pylori along with its properties and virulence factors (H. pylori during the early stages of infection, which could significantly decrease the number of gastric cancer incidents.The severity and progression of gastric cancer depends on the presence of specific nfection . EMT and factors . Specifi


Escherichia coli isolates, four (13.79%) Klebsiella pneumoniae isolates, and one (3.45%) Enterobacter cloacae isolate. The analysis based on the nucleotide sequences of the ESBL resistance genes showed that all cefotaximase-Munichs (CTX-Ms) were CTX-M-15 and that all sulfhydryl variables (SHVs) were SHV-11: 41.67% CTX-M-15-producing E. coli, 16.67% CTX-M-15+SHV-11-producing E. coli, 8.33% CTX-M-15-producing K. pneumoniae, 25% CTX-M-15+SHV-11-producing K. pneumoniae, and 8.33% CTX-M-15-produced E. cloacae. This study shows for the first time the presence of multiresistant ESBL-producing enterobacteria in fruit bats in Makokou.In Gabon, terrestrial mammals of protected areas have been identified as a possible source of antibiotic-resistant bacteria. Some studies on antibiotic resistance in bats have already been carried out. The main goal of our study was to detect extended-spectrum beta-lactamases (ESBLs) that are produced by enterobacteria from bats in the Makokou region in Gabon. Sixty-eight fecal samples were obtained from 68 bats caught in the forests located 1 km from the little town of Makokou. After culture and isolation, 66 Gram-negative bacterial colonies were obtained. The double-disk diffusion test confirmed the presence of ESBLs in six (20.69%)  Bats are an important, widespread, and abundant taxon of mammals with over 1300 described species . Bats haThe existence of antibiotic multi-resistant (AMR) enterobacteria that have been recurrently documented in wildlife in recent years provide important insights into the potential role of wildlife as reservoirs of resistant bacteria. AMR enterobacteria are also pertinent markers to understand the dynamics of zoonosis and the complex transmission routes among wildlife and humans . Many waPteropus poliocephalus), McDougall et al. detected the presence of resistance to several antibiotic families in 5.3% of wild flying foxes from different Australian colonies , CH17 (2) CTX-M-15 [MK559065], CH18 (3) CTX-M-15 [MK559057], CH38 (2) CTX-M-15 [MK559063], CH41 (1) CTX-M-15 [MK559058], CH41 (2) CTX-M-15 [MK559064], CH42 (1) CTX-M-15 [MK559059], CH42 (2) CTX-M-15 [MK559060], CH43 (1) CTX-M-15 [MK559066], CH71 (1) CTX-M-15 [MK559061], CH82 (1) CTX-M-15 [MK559062], CH38 (2) SHV-11 [MK590053], CH42 (2) SHV-11 [MK590054] and CH43 (1) SHV-11 [MK590055].Epomops franqueti and 22 Megaloglossus woermanni were caught; a total of 68 bats of the Pteropodidae family were collected. Of the 68 bacterial strains identified, 66 were Gram-negative bacteria (GNB). Among the GNB, 29 (42.65%) were enterobacteria, which consisted of 11 (37.93%) Escherichia coli, five (17.24%) Klepsiella pneumoniae, three (10.34%) Enterobacter aerogenes, two (5.88%) Enterobacter cloacae, two (6.89%) Serratia plymuthica, one Citrobacter freundii (3.45%), one (3.45%) Enterobacter hormaechei, one (3.45%) Ewingella americana, one (3.45%) Morganella morganii, one (3.45%) Pantoea sp., and one (3.45%) Proteus vulgaris  K. pneumoniae isolates, and one (3.45%) E. cloacae isolate  ESBL-producing enterobacteria. Among beta-lactams, resistance was observed by using amoxicillin (100%), ticarcillin (100%), cefotaxime (100%), ceftazidime (100%), cefpodoxime (100%), aztreonam (100%), ticarcillin/clavulanic acid (90.90%), piperacillin (90.90%), cephalexin (100%), cefepime (81.81%), amoxicillin/clavulanic acid (54.54%), cefoxitin (54.54%), ertapenem (36.36%), piperacillin/tazobactam (54.54%), and imipenem (0.00%) .As for other antibiotics tested on these positive ESBL strains, resistance was observed as follows: Erythromycin (100%), streptomycin (100%), ciprofloxacin (90.90%), kanamycin (81.81%), tetracycline (81.81%), trimethoprim/sulfamethoxazole (72.72%), gentamycin (63.63%), nalidixic acid (63.63%), tobramycin (63.63%), colistin (54.54%), ofloxacin (54.54%), levofloxacin (45.45%), netilmicin (36.36%), amikacin (27.27%), nitrofurantoin (18.18%), and chloramphenicol (0%) .E. franqueti bats species and four ESBL-producing Enterobacteriae from M. woermanni bats species were identified. These bats belong to the family of Pteropodidae -producing E. coli, 9.09% ESBL (CTX-M-15)-producing K. pneumoniae, 27.27% ESBL -producing K. pneumoniae and 9.09% ESBL (CTX-M-15)-producing E. cloacae _SHV-11, CH43 (2)_SHV-11, CH42 (2)_SHV-11 and CH38 (2)_SHV-11) clustered with human bacterial strains from Tunisia  and E. cloacae and K. pneumoniae (9.09%) or was associated with SHV-11 in K. pneumoniae (27.27%). The prevalence of ESBL-producing enterobacteria (37.93%) was higher than that obtained in E. coli sampled in bats and other wild animals in Nigeria [K. pneumoniae was found in a gorilla in its natural habitat in the Dzanga Ndoki National Park [Tadarida teniotis (bats) in Portugal [E. coli, followed by K. pneumoniae [blaCTX-M beta-lactam resistance genes are the most common in the human and veterinary strains [blaCTX-M-15 is the most prevalent ESBL gene in human samples worldwide [blaCTX-M-15 and blaSHV-11 genes are recognized as plasmid-mediated resistance genes [The beta-lactam resistance phenotypes obtained in our study are also comparable to those obtained in a study on bats in Algeria , which s Nigeria ,15 and t Nigeria , in whiceumoniae .27%. Thenal Park ,46 and iPortugal . Enterobeumoniae . However strains ,49,50. borldwide  and is porldwide . In addice genes . All of ce genes ,52,53. blaCTX-M-15 genes had already been identified in ESBL-producing Enterobacteriaceae from patients in the Albert Schweitzer Hospital in Lambaréné and in poultry [blaCTX-M-15 and blaSHV-11 genes is probably linked to antibiotics used by humans, since our phylogenetic analyses show that all the sequences obtained in bats that carry CTX-M-15 or SHV-11 clustered with one human bacterial strain from Turkey and Tunisia, respectively. This suggests that the prevalence of antibiotic resistance in wild animals depends on the antibiotics consumed by human populations at each site and the density of the human population in contact with fauna [In Gabon, the  poultry ,32. The th fauna .E. coli isolated from wildlife may be acquired from the food [E. coli has already been described as an important source of infection in mammals [E. coli infections can result from food contamination [Tetrapleura tetraptera, the fruit of which is widely consumed in Gabon) and lemon. Hence, it is possible that some fruits partly eaten by humans and contaminated have been ingested by bats. Fruit bats are potential vectors and reservoirs of pathogenic bacteria such as E. coli, which carry acquired resistance [In our samples, the highest prevalence of resistance was obtained in resistance to aminoglycosides , fluoroquinolones  and tetracycline, which are the most common antibiotics consumed by the Gabonese people ,54. The the food ,57,58,59the food  and watethe food ; resistathe food ,52. Furt mammals . Foodbormination ,63. Secomination ,64. Mostmination ,66,67,68mination ,70. Thesmination . Severalmination ,72,73. Tsistance ,59. The sistance ,75. Batssistance . In thisThe phylogeny of CTX-M-15 was constructed using v. 1.8.1 in BioEdit v. 7.0.9.0 software. These analyses were performed with a multiple alignment matrix of obtained partial CTX-M-15 sequences and the GenBank reference sequences of human, poultry and swine from Africa (Nigeria), Asia , Europe , the Middle East (Iran), and South America (Brazil). Enterobacteriaceae carrying CTX-M-15 isolated from fruit bats of Makokou in Gabon (in pink color).K. pneumoniae from bats. Enterobacteriaceae carrying SHV-11 isolated from fruit bats of Makokou in Gabon (in pink color).The phylogeny of SHV-11 was constructed using v. 1.8.1 in BioEdit v. 7.0.9.0 software. These analyses were performed with a multiple alignment matrix of obtained partial SHV-11 sequences and the GenBank reference sequences of human, water and cattle from Europe , Asia , the Middle East (Iran), and Africa (Tunisia and Egypt). All sequences of SHV-11 were from This study showed for the first time the presence of multiresistant ESBL-producing enterobacteria in Makokou fruit bats in Gabon . The source of the contamination has not been clearly determined, but the presence of this type of resistance in bats suggests that these wild mammals could spread ESBL-producing Enterobacteriacea over long distances or across the urban landscape.Our results reinforce the need to monitor antimicrobial resistance in wild animals, in protected or unprotected areas, in order to assess environmental responses to anthropogenic pressures.


Scientific Reports 10.1038/s41598-019-41524-3, published online 01 April 2019Correction to: The Acknowledgements sectionin this Article is incomplete.“We are grateful to Dr. D.W. Stamer and his collaborators from Duke University Eye Center  for their gift of human primary TM cells. The present work was supported by Ministerio de Economia y Competitividad and Instituto de Salud Carlos III of Spain FIS PI14/00141 (X.G.), FIS P17/00296 (X.G.), RETICs Oftared RD12/0034/0003 (X.G.), RD16/0008/0014 (X.G.), RD12/0034/0015 (M.C.), RD16/0008/0023 (M.C.), Generalitat de Catalunya 2014SGR1165 (X.G.), and National Eye Institute - NIH, EY-11906 (T.B.).”should read:“We are grateful to Dr. D.W. Stamer and his collaborators from Duke University Eye Center  for their gift of human primary TM cells. The present work was supported by European Union, Fondo Europeo de Desarrollo Regional (FEDER), Ministerio de Economia y Competitividad and Instituto de Salud Carlos III of Spain FIS PI14/00141 (X.G.), FIS P17/00296 (X.G.), RETICs Oftared RD12/0034/0003 (X.G.), RD16/0008/0014 (X.G.), RD12/0034/0015 (M.C.), RD16/0008/0023 (M.C.), Generalitat de Catalunya 2014SGR1165 (X.G.), and National Eye Institute - NIH, EY-11906 (T.B.).”


In: Imsanguan W., Bupachat S., Wanchaithanawong V., Luangjina S., Thawtheong S., Nedsuwan S., et al. Contact tracing for tuberculosis, Thailand. Bull World Health Organ. 2020 Mar 1;98(3):212–18 On page 215, Table 2, should read as follows:


M-SAL yielded 29.7% ND and average concentrations of 2.26 ± 1.15 (SD) log10 PFU/L (somatic) and 0.59 ± 0.82 (SD) log10 PFU/L (F+ ). DMF performance was inferior to D-HFUF-SAL and M-SAL methods , indicating this procedure is unsuitable for 1 L surface water sample volumes. This study represents an important step toward the use of a coliphage method for recreational water quality criteria purposes.Coliphages are alternative fecal indicators that may be suitable surrogates for viral pathogens, but majority of standard detection methods utilize insufficient volumes for routine detection in environmental waters. We compared three somatic and F+ coliphage methods based on a paired measurement from 1 L samples collected from the Great Lakes (n = 74). Methods include: 1) dead-end hollow fiber ultrafilter with single agar layer (D-HFUF-SAL); 2) modified SAL (M-SAL); and 3) direct membrane filtration (DMF) technique. Overall, D-HFUF-SAL outperformed other methods as it yielded the lowest frequency of non-detects  and the highest average concentrations of recovered coliphage for positive samples (2.51 ± 1.02  log Enteric viruses are the leading cause of recreational waterborne disease outbreaks , but detE. coli or enterococci) are present at 1–2 orders of magnitude higher concentrations (Current coliphage standard culture-based methods include single-agar layer (SAL) plaque assay , double-trations . A simplIn this study, we evaluate the performance of three somatic and F+ coliphage methods with 1 L sample volumes, including dead-end hollow fiber ultrafiltration  with SALSurface waters originated from Lake Michigan  (n = 37) and nearby Trail Creek (n = 37). Samples were collected during the 2015 Great Lakes beach season at a frequency of five samples per week. The sample collection procedures are detailed elsewhere . The D-HAll data were log10 transformed and expressed as plaque forming unit (PFU) per liter (L) for positive samples only as ND samples were not included in the concentration data. Statistical analyses were performed using SigmaPlot version 13.0 . One-way analysis of variance (ANOVA) followed by Holm-Sidak multiple comparisons or Kruskal-Wallis ANOVA on ranks (followed by Tukey tests) were applied to somatic and F+ coliphage datasets (from both sites), while Wilcoxon signed rank tests were used for overall comparisons between the coliphage types or sites.10 PFU/L) were comparable to M-SAL , but concentrations obtained by DMF (0.30 ± 0.44 [SD] log10 PFU/L) (p < 0.001) lower than either D-HFUF-SAL or M-SAL methods. The DMF method did not yield any F+ coliphage results from Lake Michigan samples, but they were detected using the other two methods (10 PFU/L) (p < 0.001) for positive samples than either M-SAL or DMF methods, but there was no statistically significant difference in F+ coliphage concentrations obtained by the M-SAL and DMF methods (p = 0.899).Method performance metrics are summarized in 0 PFU/L)  were sig methods  with D-H0 PFU/L) . D-HFUF-10 PFU/L and 3.25 ± 0.44 (SD) log10 PFU/L, respectively (p = 0.252) (p < 0.001) of somatic coliphage for positive samples were observed with the DMF method (2.75 ± 0.49 [SD] log10 PFU/L) (p = 0.659) average concentrations of F+ coliphage in positive samples were found in Trail Creek waters using the D-HFUF-SAL method (1.25 ± 0.68 [SD] log10 PFU/L) and M-SAL (1.22 ± 0.85 [SD] log10 PFU/L), but concentrations obtained by the DMF method were significantly lower (p < 0.001) (p < 0.001), and concentrations of both coliphage types were typically higher in the Trail Creek samples compared to the Lake Michigan samples (p < 0.001).Somatic coliphage were consistently detected in Trail Creek water samples irrespective of the method . D-HFUF-= 0.252) . Signifi0 PFU/L) . Compara< 0.001) . OverallRegarding frequency of NDs, D-HFUF-SAL exhibited the lowest ND range overall, with 0%–2.7% (somatic) and 5.4%–35% (F+) . M-SAL r2, CaCl2, MgSO4, tryptone, glucose, Tween 80, Antifoam Y-30, sodium hexametaphosphate) but we’ve excluded common laboratory disposables such as serological pipettes, pipette tips, gloves and similar. Regarding the processing time, we have assumed that sample is being processed by a single analyst familiar with the routine water quality assessment (e.g. membrane filtration and /or defined substrate technology for enumeration of E. coli and enterococci) and possessing basic knowledge of microbiological culture techniques. The cost per sample for D-HFUF-SAL and DMF was comparable, while M-SAL was approximately ten times higher, reflecting the increase in supplies and reagents needed to process 1 L sample volumes  to 25% (F+) . M-SAL mSupp Data


Correction to: BMC Public Health (2020) 20:1616https://doi.org/10.1186/s12889-020-09721-2It was highlighted that in the original article  the lettCorrect Table 1footnotesa Mann-Whitney testb χ2 testc Mortality rate per 1000 person-yearCOPD Chronic Obstructive Pulmonary Disease, CKD Chronic Kidney Disease, ICU Intensive Care Unit, IMSS Mexican Institute of Social Services, ISSSTE Institute of Social Security and Services for State Workers, SS Mexican Ministry of HealthCorrect equation in Results section2 = 31.83, df = 12, p < 0.001)(χThe original article has been updated.


O.decamerasp. nov. and O.joshiisp. nov., of the ant genus Ooceraea are described from India. These species differ from other known congeners on the basis of number of antennal segments. An illustrated key to the known species based on the worker caste is also provided.Two new species,  Ooceraea Roger, 1862 has been challenging, since its inception based on the type species O.fragosa. The taxonomic ambiguity has led to its uncertain placements in different subfamilies: in Myrmicinae  and no constrictions between abdominal segments IV, V and VI. Ooceraea can be distinguished from the closely allied Syscia Roger, 1861 on the basis of abdominal segment III relatively narrow in dorsal view and similar in size to the preceding abdominal segment II (petiole); in lateral view, abdominal tergite IV not folding over sternite and the anterior portion of the sternite visible; hind basitarsi not dilating distally, circular in cross-section and metabasitarsal glands absent  is probably native to the Asian continent, and has been introduced to Southeast Asia, the Pacific islands, Madagascar and the Caribbean islands  and Ooceraeabesucheti  with nine- and eleven-segmented antennae respectively ;SL Scape length: maximum length of antennal scape excluding basal condylar bulb;MW Mesosomal width: maximum width of promesonotum in dorsal view;ML Mesosomal length: maximum diagonal length of mesosoma in lateral view, measured from posterodorsal border of pronotal flange to posterior basal angle of metapleuron;PL Petiolar length: maximum length of petiole in lateral view;PH Petiolar height: maximum height of petiole in lateral view (including subpetiolar process);PW Petiolar width: maximum width of petiole in dorsal view;PPL Postpetiolar length: maximum length of postpetiole in lateral view;PPH Postpetiolar height: maximum height of postpetiole in lateral view;PPW Postpetiolar width: maximum width of postpetiole in dorsal view;CI Cephalic index: HW/HL × 100;SI Scape index: SL/HW × 100;PI1 Petiolar index 1: PL/PH × 100;PI2 Petiolar index 2: PW/PL × 100;PPI1 Postpetiolar index 1: PPL/PPH × 100;PPI2 Postpetiolar index 2: PPW/PPL × 100;WI Waist index: PPW/PW × 100.DepositoriesPUACPunjabi University Patiala Ant Collection at Department of Zoology and Environmental Sciences, Punjabi University, Patiala, Punjab, India;MCZCMuseum of Comparative Zoology, Harvard University, Cambridge, Massachusetts, United States.Taxon classificationAnimaliaHymenopteraFormicidae9DCFFE46-DF69-52AC-8558-AA33512A3640http://zoobank.org/182F8A89-653B-4604-8337-F7A5F258080B9.5627°N, 77.2348°E, 780 m.India, Kerala, Periyar Tiger Reserve Holotype worker and one paratype worker, both India, Kerala, Periyar Tiger Reserve 9.5627°N, 77.2348°E, 780 m, leaf litter, Winkler, 21 January 2017, Tarun Dhadwal leg. [PUAC].HL 0.57; HW 0.56; SL 0.34; MW 0.39; ML 0.68; PL 0.29; PH 0.34; PW 0.27; PPL 0.32; PPH 0.34; PPW 0.30; CI 98; SI, 61; PI1 85; PI2 93; PPI1 94; PPI2 88; WI 111. Paratype: HL 0.57; HW 0.56; SL 0.33; MW 0.39; ML 0.68; PL 0.29; PH 0.33; PW 0.26; PPL 0.32; PPH 0.34; PPW 0.30; CI 98; SI, 59; PI1 88; PI2 89; PPI1 94; PPI2 88; WI 111.Holotype: Head in full-face view, almost as long as broad, with lateral margin weakly convex and converging anteriorly, with posterior margin concave medially and posterior lateral corners rounded. Anterior clypeal margin reduced and slightly concave in the middle. Eyes present, small in size, with two ommatidia, parafrontal ridge prominently produced. Mandibles edentate, sub-triangular. Antenna 10-segmented; scape short and clavate, reaching almost mid-length of the head; apical funicular segment fusiform. Frontal lobes reduced. Antennal sockets fully exposed from above.Mesosoma in lateral view weakly convex; promesonotal suture and metanotal groove absent. Pronotum in dorsal view anteriorly marginate. Propodeum in dorsal view with posterior margin concave; propodeal declivity in lateral view slightly concave, with lateral margin slightly marginate; propodeal lobe reduced. Petiolar node in dorsal view as long as broad, rounded anteriorly, in lateral view hemiglobular; subpetiolar process well-developed, with sickle-shaped anteroventral apex. Postpetiole in dorsal view subtrapezoidal, with anterior margin transverse and posterior margin convex, in lateral view with anteroventral corner angulate. Gastral segment I  large, occupying the most part of gaster, in lateral view with dorsal margin weakly and roundly convex.Sculpture. Head foveolate-reticulate; mesosoma, petiole and postpetiole foveolate-reticulate; gaster foveolate, with foveae smaller than those of head and mesosoma.Pilosity and Pubescence. Body covered with erect or sub-erect hairs; sides of head and legs covered with shorter hairs; scape and funicular segments covered with short decumbent or subdecumbent hairs.Body coloration. Head and gaster light brown; mesosoma, petiole and postpetiole darker than the head; legs yellowish brown.Queen. Unknown.Male. Unknown.Ooceraeajoshii sp. nov. and O.decamera sp. nov. (described below) are distinctly separated from the other valid congeners by having 10-segmented antennae. Furthermore, the two new species are well distinguished from each other by a combination of the following characters: head shape ; presence of ommatidia (present in O.joshii sp. nov. and absent in O.decamera sp. nov.); propodeal lobes (reduced versus roundly produced); petiolar node in lateral view (hemiglobular versus rectangular); subpetiolar process ; pilosity (head and body comparatively more pilose in O.joshii sp. nov.); and sculpturation .The type series was found in leaf litter samples collected from the Medaganam region of the Periyar Tiger Reserve situated at an elevation of 780 meters. The region is composed of an undisturbed tropical moist evergreen forest with low light penetration, with a mean average daytime temperature of 30 °C.Known only from the type locality.The species has been named in honor of Professor Amitabh Joshi, a distinguished evolutionary biologist based at Jawaharlal Nehru Centre for Advanced Scientific Research (JNCASR), Bengaluru, India.Taxon classificationAnimaliaHymenopteraFormicidaeEFC710C7-7193-5101-BA40-0E0AD282BD0Chttp://zoobank.org/D8C9E609-7416-4081-A54A-83CAA01CEAD610.02308°N, 77.833333°E, 250–350 m alt.India: Madras, Alagarkovil, 21 km. N Madurai, Holotype worker, India, Madras, Alagarkovil, 21 km N Madurai, 10.02308°N, 77.833333°E, 250–350 m alt.; 2 November 1972; Besuchet Lobt Mussard leg. (Specimen number/barcode: MCZ-ENT00649398) [MCZC].HL 0.62; HW 0.46; SL 0.26; MW 0.38; ML 0.78; PL 0.26; PH 0.42; PW 0.30; PPL 0.34; PPH 0.41; PPW 0.40; CI 74; SI 57; PI1 62; PI2 93; PPI1 81; PPI2 118; WI 133.CI 74), with lateral sides weakly convex, with posterior margin concave medially, with occipital lobes/corners angulate. Anterior clypeal margin slightly projecting forward. Eyes absent. Parafrontal ridge prominent and elevated. Mandibles edentate but weakly serrate. Antennae with 10 segments; scape short, clavate, slightly surpassing the mid-length of head. Frontal lobes reduced. Antennal sockets fully exposed from above.Head in full-face view rectangular, distinctly longer than broad  large occupying the most part of gaster, in lateral view with dorsal margin almost straight, base of cinctus of first gastral tergite cross-ribbed.Sculpture. Head, mesosoma, petiole and postpetiole shallowly foveolate-reticulate; mandibles and dorsal surface of gaster sparsely foveolate, foveae somewhat smaller as compared to those present on head, mesosoma, petiole, and postpetiole.Pilosity and pubescence. Whole body covered with pale yellow erect and sub-erect hairs; appressed pubescence abundant on antennae and legs.Body coloration. Mandibles, antennae, legs, subpetiolar process and gaster light brown; head, mesosoma and gaster dark brown.Queen. Unknown.Male. Unknown.The two species significantly differ from each other on the basis of dimensions of head capsule and shape of subpetiolar process.Unknown.Known only from the type locality. The place has been transformed into agricultural land and is prone to anthropogenic activities. Thus, this reinforces the concept that important biodiversity components, which are already rare, are imperiled due to local extinctions.decamera refers to the ten-segmented antennal count.The species epithet


Plakobranchus cf. ocellatus (Heterobranchia: Sacoglossa), a so-called ‘solar-powered’ sea slug with long-term retention of chloroplasts. The mitochondrial genome was 14,177 bp in length containing the standard set of 13 protein-coding genes, 2 rRNAs, and 22 tRNAs. The base composition of 27.3% A, 15.6% C, 18.6% G, and 38.5% T showed a strong A + T bias. The genome organization of P. cf. ocellatus is identical to the other sacoglossan mitogenomes sequenced so far, except for Ascobulla fragilis.We present the complete mitochondrial genome sequence of  Plakobranchus cf. ocellatus van Hasselt, 1824 belongs to a small group of marine, heterobranch slugs, called Sacoglossa, of which certain species have the ability to sequester chloroplasts from its food algae  in December 2012. The heads of about 60 slugs were dissected to remove the post-pharyngeal nerve ring, which were directly deep frozen in liquid nitrogen and then stored (−80 °C). From these, genomic DNA was extracted using the QIAGEN DNeasy® Blood & Tissue Kit . Voucher material is stored in absolute ethanol at the Zoological Research Museum Alexander Koenig (voucher no. ZFMK-TIS-29490).Specimens of ® DNA PCR-Free Library Preparation Kit  according to the manufacturer’s protocol and 125 bp paired-end reads were sequenced on an Illumina HiSeq 2500 platform . All read pairs were used for mitochondrial genome assembly with MITObim 1.8  as seed. After verification of correct circularity, genome annotation was done with MITOS revision 656  were prepared using the TruSeqElysia chlorotica, E. ornata, Thuridilla gracilis, and Placida sp. 1 NY-2013, except for Ascobulla fragilis.The complete mitochondrial genome (GenBank accession number: KX853083) had a length of 14,177 bp encoding for 13 protein-coding genes, 2 rRNAs, and 22 tRNAs. The base composition is 27.3% A, 15.6% C, 18.6% G, and 38.5% T. A total of 183 bp nucleotides were observed in multiple small intergenic regions, ranging from 1 to 54 bp (found between the genes coding for COX3 and trnI(gat)). The gene order was as follows: trnK(ttt), cox1, trnV(tac), rrnL, trnL1(tag), trnA(tgc), trnP(tgg), nad6, nad5, nad1, trnW(tca), trnY(gta), nad4l, cob, trnD(gtc), trnF(gaa), cox2, trnG(tcc), trnH(gtg), -trnQ(ttg), -trnL2(taa), -atp8, -trnN(gtt), trnC(gca), -atp6, -trnR(tcg), -trnE(ttc), -rrnS, -trnM(cat), -nad3, -trnS2(tga), trnS1(gct), nad4, -trnT(tgt), -cox3, trnI(gat), nad2 and is therefore identical to the other sacoglossan mitogenomes sequenced so far, i.e. P. cf. ocellatus and all other sacoglossan species with fully sequenced mitogenome were extracted, aligned with MAFFT v7.271 (G-INS-i mode) (Katoh & Standley The amino acid sequence of the 13 protein-coding genes of


Chromatographic separation of androgen derivatives was optimized, carefully separating isobaric interferents and acceptable outputs for precision and trueness achieved following injection of 0.4 pg on column (approximately 34 pmol/L). HMP derivatives of all androgens tested could be detected in low plasma volumes: male (100 µL) and post-menopausal female (200 µL), and derivatives were stable over 30 days at -20°C. In conclusion, HMP derivatization, in conjunction with LC-MS/MS, is suitable for quantitative analysis of DHT, testosterone and androstenedione in low plasma volumes, offering advantages in sensitivity over current methodologies.Liquid Chromatography tandem mass spectrometry (LC-MS/MS) is the gold-standard approach for androgen analysis in biological fluids, superseding immunoassays in selectivity, particularly at low concentrations. While LC-MS/MS is established for analysis of testosterone and androstenedione, 5α-dihydrotestosterone (DHT) presents greater analytical challenges. DHT circulates at low nanomolar concentrations in men and lower in women, ionizing inefficiently and suffering from isobaric interference from other androgens. Even using current LC-MS/MS technology, large plasma volumes (>0.5 mL) are required for detection, undesirable clinically and unsuitable for animals. This study investigated derivatization approaches using hydrazine-based reagents to enhance ionization efficiency and sensitivity of analysis of DHT by LC-MS/MS. Derivatization of DHT using 2-hydrazino-1-methylpyridine (HMP) and 2-hydrazino-4-(trifluoromethyl)-pyrimidine (HTP) were compared. A method was validated using an UHPLC interfaced by electrospray with a triple quadruple mass spectrometer , analyzing human plasma  following solid-phase extraction. HMP derivatives were selected for validation affording greater sensitivity than those formed with HTP. HMP derivatives were detected by selected reaction monitoring (DHT-HMP  Twydrazone , specifiution MS .3.1.1.2m/z 449 , m/z 447 (androstenedione) and m/z 451 (DHT) (m/z 257 (C11H12N4F3) and m/z 269 (C12H12N4F3) for testosterone, androstenedione and EpiT resulting from the cleavage of the steroid A and B ring (m/z 288 (C19H30ON) and m/z 164 (C5H5N3F3) whereas, m/z 286 (C19H28ON) and m/z 164 (C5H5N3F3) were selected for DHA and DHEA respectively. Heterolytic cleavage of hydrazine N-N bond of derivative may form ions at m/z 288 and m/z 286, and m/z 164 characterizes the protonated trifluoromethyl-pyrimidine moiety. Ions obtained following collision induced dissociation of derivatives of stable-isotopically labeled testosterone, androstenedione and DHT corroborated potential fragmentation   14] andd B ring  14,46]. andd B r. andd B entation .3.1.23.1.2.1E and Z isomers which elute as double peaks for each androgen HMP derivatization forms two 18 columns with modified stationary phases, particle sizes and dimensions. SunFire® C18 , ACE® Excel PFP-C18 , ACE® UltraCore 2.5 SuperC18 , ACE® Excel super C18 , Acquity UPLC® BEH C18  and Acquity UPLC® BEH C18  were evaluated aiming to achieve baseline resolution of isomers and isotopologues within reasonable analytical times. Ultimately rapid chromatographic separation was achieved between isomeric HMP derivatives of each androgen and their endogenous isomers using a UPLC® BEH C18  column operated at 50°C, with run times of 10 min  and gradient method  and the estimated LOQs were 0.4, 0.8 and 0.4 pg  on column for DHT, testosterone and androstenedione respectively. Thus, estimated LOQs were improved ~21-fold and ~ 12.5-fold for HMP-DHT and HMP-androstenedione respectively in comparison to our LC-MS/MS method for underivatized androgens 3.23.2.113C3-T 97±9%, 13C3-A4 99±1% and 13C3-DHT 98±5%). Extraction from plasma was assessed using stable-isotope labeled standards as surrogates, showing similar results . Ion suppression of internal standards was also within acceptable limits . Establishing conditions which limited ion suppression reduced the potential for variability in quantitation brought about by impact of subtle variations in matrix composition, particularly for low abundance analytes such as DHT. Minimal differences (<0.05 mins) in retention time between 13C3 IS and endogenous steroid due to the presence of heavy isotope were recorded. The assessment of the whole process also demonstrated that the presence of matrix did not adversely affected derivatization efficiency.Recovery of analytes by extraction was maximized and matrix effect minimized to ensure the cleanest possible sample to be prepared for analysis, and thus maximising column life-span and limiting source contamination. Analytes were well retained on lipophilic polymeric adsorbent Oasis® HLB ,50. and 3.2.213C3-T 1.7, 13C3-A4 1.8, and 13C3-DHT 2.7), were comparable to those in plasma, showing differences less than 20% .Interferents close to the retention times of the peaks of interest were not observed visually. The ratios of peak areas of quantifier to qualifier mass transitions ratios in standards , but acceptable within the reference range , intra and inter assay deemed acceptable (<20% RSD for precision and trueness at the LOQ  and <15%3.2.5The HMP derivatives demonstrated acceptable stability upon storage in an auto-sampler (10°C) over 24 h, with limited degradation measured for derivatized testosterone , androstenedione  and DHT  in extracts of plasma from male and post-menopausal female subjects, respectively. Derivatives were stable upon short-term storage in the freezer  for 7 days; reduction from original response testosterone , androstenedione  and DHT  in extracts of samples from males and testosterone , androstenedione  and DHT  from post-menopausal female. Moreover, derivatives were stable upon long-term storage in the freezer  for 30 days; reduction from original response testosterone , androstenedione  and DHT  in extracts of samples from males and testosterone , androstenedione  and DHT  from post-menopausal females.3.3The method was applied to samples from post-menopausal females and males and compared to typical reference ranges  1,33]. ,33. 1,334. While multi-dimensional detection in conjunction with chromatography brings anticipated benefits in selectivity In summary derivatization to form HMP derivatives of androgens, in conjunction with LC-MS/MS, is suitable for quantitative analysis of androgens in low abundance in biological fluids, offering advantages in sensitivity over analysis of underivatized DHT. The approach allows concomitant analysis of testosterone, androstenedione and DHT and this method may be extended to include further steroids with ketone functions e.g. 11-oxygenated androgens. Robust measurement was achieved across typical physiological ranges found in post-menopausal women and men offering an attractive alternative to immunoassays, which lack selectivity in the lower physiological range of concentrations and typically report only one androgenAbdullah MM Faqehi: Methodology, Formal analysis, Project administration, Resources, Validation, Writing - original draft. Scott G Denham: Methodology, Formal analysis. Gregorio Naredo: Methodology, Formal analysis, Writing - review & editing. Diego F Cobice: Methodology, Formal analysis, Writing - review & editing. Shazia Khan: Methodology, Formal analysis, Writing - review & editing. Joanna P Simpson: Methodology, Formal analysis, Writing - review & editing. Ghazali Sabil: Methodology, Formal analysis, Writing - review & editing. Rita Upreti: Investigation, Writing - review & editing. Fraser Gibb: Investigation, Writing - review & editing. Natalie ZM Homer: Methodology, Validation, Project administration, Supervision, Writing - review & editing. Ruth Andrew: Conceptualization, Methodology, Validation, Funding acquisition, Project administration, Supervision, Writing - review & editing.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.


Lactococcus lactis (L. lactis)/pNZ8150-phosphatidylglycerophosphate synthetase A (pgsA)-HAsd, in which pgsA was used as an anchor protein, and investigated the immunogenicity of HAsd in a mouse model by oral administration without the use of a mucosal adjuvant. Compared with L. lactis/pNZ8150-pgsA, mice were orally vaccinated with L. lactis/pNZ8150-pgsA-HAsd and then produced strong humoral and mucosal immune responses. Importantly, L. lactis/pNZ8150-pgsA-HAsd provided cross-protection against H5N1, H3N2 and H1N1 virus infections. Our data support the hypothesis that HAsd presented on the surface of L. lactis can provide cross-protective immunity against divergent influenza A viruses. Taken together, these findings suggest that L. lactis/pNZ8150-pgsA-HAsd can be considered an alternative approach to developing a novel universal vaccine during an influenza A pandemic.Most of the current approaches to influenza vaccine design focus on antibodies against influenza (HA). However, these influenza vaccines typically provide strain-specific protection against mostly homologous subtypes. There is an urgent need to develop a universal vaccine that confers cross-protection against influenza viruses. Of note, the HA stalk domain (HAsd) is a promising target for such an influenza vaccine. In this study, we generated recombinant Abbreviations: HA, HAsd, HA stalk domain; L. lactis, Lactococcus lactis; SDS-PAGE, sodium dodecyl sulfate-polyacrylamide gel electrophoresis; IFA, immunofluorescence assay; PBS, phosphate-buffered saline; pgsA, phosphatidylglycerophosphate synthetase A; SPF, specific pathogen-free; CFU, colony-forming unit; BSL-3, biosafety level-3 laboratory; TCID50, 50% tissue culture infective dose; ELISA, enzyme-linked immunosorbent assay; OD, optical density; LTB, liable enterotoxin B subunit; CTB, cholera toxin B subunit. Influenza A viruses cause a highly infectious respiratory disease that remains a public health problem worldwide . Humans Lactococcus lactis (L. lactis) has been engineered to express heterologous proteins [L. lactis expressing HA, HA1, neuraminidase or nucleoprotein of avian influenza H5N1 virus is a safe and effective vaccine candidate against H5N1 virus infection in mouse and chicken models with the use of a mucosal adjuvant or enteric capsule [L. lactis in the absence of a mucosal adjuvant can provide cross-protective immunity against divergent influenza A viruses.proteins . Our pre capsule . HoweverL. lactis, recombinant L. lactis/pNZ8150-pgsA-HAsd was constructed, in which pgsA derived from the chromosomal DNA of Bacillus subtilis [L. lactis/pNZ8150-pgsA-HAsd, which elicited robust humoral and mucosal immune responses against influenza A viruses. Most importantly, unadjuvanted L. lactis/pNZ8150-pgsA-HAsd could provide cross-protective immunity against lethal challenge with homologous and heterologous influenza A viruses. These data suggest that recombinant L. lactis/pNZ8150-pgsA-HAsd is expected to serve as an alternative approach for a truly protective universal influenza vaccine against divergent influenza A viruses.Thus, to investigate the immunogenicity of the HAsd presented on the surface of subtilis , and wasL. lactis was determined by western blot analysis. A highly specific band (approximately 100 kDa) was detected for the pgsA-HAsd fusion protein in the L. lactis/pNZ8150-pgsA-HAsd cells . In contrast, no band was detected in the L. lactis/pNZ8150-pgsA cells .The HAsd gene  of codon-optimized A/Vietnam/1203/2004(H5N1)  was clonsd cells , lane 3.L. lactis, L. lactis/pNZ8150-pgsA and L. lactis/pNZ8150-pgsA-HAsd cells were examined by immunofluorescence assay (IFA) and flow cytometric analysis. There were no positive signals in the L. lactis/pNZ8150-pgsA cells . By contrast, L. lactis/pNZ8150-pgsA-HAsd consistently exhibited specific fluorescent signals, indicating that HAsd was stably expressed on the surface of L. lactis (right side).To confirm that HAsd protein was expressed on the surface of L. lactis/pNZ8150-pgsA or L. lactis/pNZ8150-pgsA-HAsd group at the prime immunization. A highly significant increase (IgG titer ≥ 25) was observed in the L. lactis/pNZ8150-pgsA-HAsd group at the boost immunization, IgG titer induced by L. lactis/pNZ8150-pgsA-HAsd reach 28.4±0.894, whereas there were still no significant (IgG titer <25) changes in the PBS or L. lactis/pNZ8150-pgsA group.HA-specific antibody responses were measured by ELISA. As shown in L. lactis/pNZ8150-pgsA or L. lactis/pNZ8150-pgsA-HAsd group at the prime immunization. Only L. lactis/pNZ8150-pgsA- HAsd could induce an elevated level of IgA antibodies after the boost immunization, IgA titers in the intestine and upper respiratory washes were 27.8±1.30 and 27.4±1.14, respectively.Mucosal IgA antibodies were also detected in the intestine and upper respiratory washes  and c. TL. lactis/pNZ8150-pgsA-HAsd orally after the prime-boost immunization could produce an elevated level of HA-specific IgG and IgA antibody responses.Collectively, these results demonstrated that mice administered L. lactis/pNZ8150-pgsA-HAsd, all vaccinated mice were challenged with 20 µL of 104 EID50 of A/Vietnam/1203/2004(H5N1), A/Beijing/47/1992(H3N2) or A/California/04/2009(H1N1) and monitored for 14 days. As shown in L. lactis/pNZ8150-pgsA showed significant body weight loss and increased virus shedding in the lung and died within 8 days after challenge with the lethal dose of virus . In contrast, mice vaccinated orally with L. lactis/pNZ8150-pgsA-HAsd experienced only mild weight loss and recovered by 14 days . Importantly, L. lactis/pNZ8150-pgsA-HAsd could confer 100% (5/5), 80% (4/5) and 80% (4/5) protection against A/Vietnam/1203/2004(H5N1), A/Beijing/47/1992(H3N2) and A/California/04/2009(H1N1), respectively .Last, to assess the cross-protective efficacy of of virus ,b and c. 14 days , b and c 14 days , E and Fectively , h and iNde I or EcoR I sites underlined . The resulting Nde I/EcoR I fragment was cloned into pNZ8150-pgsA at the C-terminal end, in which pgsA was used as an anchor protein [L. lactis NZ9000. The positive clone of L. lactis/pNZ8150-pgsA-HAsd was screened, and the plasmid was expressed as described previously [L. lactis/pNZ8150-pgsA was used as a negative control for subsequent analysis.The HAsd (876 bp) gene of A/Vietnam/1203/2004(H5N1) that included part of HA1 (from 831 bp to 1041 bp) and complete HA2 gene (from 1041 bp to 1707 bp)  was PCR  protein , and theeviously . L. lactL. lactis surface was determined by western blot analysis as described previously [6-cell L. lactis/pNZ8150-pgsA-HAsd pellets were mixed with 60 µL of 6 × loading buffer, boiled for 10 minutes, subjected to sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a nitrocellulose membrane . The membrane was incubated with a 1:500-diluted monoclonal mouse anti-HA of A/Vietnam/1203/2004(H5N1) antibody  after blocking with 5% skim milk at room temperature for 2 h. After incubation overnight at 4°C, the membrane was incubated with affinity-purified horseradish peroxidase (HRP)-conjugated anti-mouse IgG . The membrane was reacted with West Pico Chemiluminescent Substrate  and imaged using a Molecular Imager ChemiDoc XRS System . Precision Plus Protein™ WesternC™  was used as a standard protein marker.The expression level of HAsd displayed on the recombinant eviously . BrieflyL. lactis surface was by an IFA . Briefly, 106L. lactis/pNZ8150-pgsA-HAsd cells were washed three times with sterile PBS, incubated with monoclonal mouse anti-HA antibody  at 4°C for 1 h, followed by FITC-conjugated goat anti-mouse IgG at 4°C for 30 minutes, and resuspended in 100 µL of sterile PBS. Finally, L. lactis/pNZ8150-pgsA-HAsd cells were used for the IFA. L. lactis/pNZ8150-pgsA cells were used as negative controls.The HAsd protein displayed on the L. lactis/pNZ8150-pgsA-HAsd was analyzed by flow cytometric analysis .Flow cytometry was performed very similarly to IFA, as described above. Five hundred microliters of Specific-pathogen-free (SPF) six-week-old female BALB/c mice were used  in this study and housed in cages ventilated under negative pressure with high efficiency particulate air filter.L. lactis/pNZ8150-pgsA-HAsd cells was adjusted to 1012 colony forming unit (CFU)/ml with sterile PBS. The mice (n = 39 per group) were vaccinated orally with 1012 CFU of L. lactis/pNZ8150-pgsA-HAsd at days 0, 1, 2, and 3 for prime immunization and boosted at day 17, 18, 19, and 20. PBS and L. lactis/pNZ8150-pgsA cells were used as controls.The concentration of recombinant At days 15 and 34 after the initial immunization, blood samples  and intestine and upper respiratory washes  were collected.4 EID50 of lethal dose of A/Vietnam/1203/2004(H5N1) (n = 8 per group), A/Beijing/47/1992(H3N2) (n = 8 per group)or A/California/04/2009(H1N1) virus (n = 8 per group). The mice were monitored for 14 days, and body weight loss and survival rate post challenge were calculated.Two weeks after the last immunization, all vaccinated mice (n = 24 per group) were transferred into an enhanced biosafety level-3 laboratory (BSL-3) containment facility and challenged intranasally with 20 µL containing 1050).Additionally, lungs from vaccinated mice were harvested for the detection of virus shedding at day 3 post infection (n = 3 per group). Tissue samples were homogenized and processed as described previously . The virAntibody responses of serum IgG and IgA in the intestinal washes and upper respiratory washes were determined by ELISA using recombinant HA protein (2 µg/mL) of A/Vietnam/1203/2004(H5N1)  as a coating antigen, as described previously . A 1:5,0p-value of less than 0.05 was considered statistically significant.A two-tailed Student’s t-test and one-way ANOVA were used for all statistical analyses. A Due to continuous antigen gene occurring mutation, the influenza A virus is considered to have the potential to cause the next pandemic . Most ofL. lactis may provide cross-protective immunity against divergent influenza A viruses. Generally, the HAsd, including HA2 and part of HA1, was displayed on the surface of L. lactis, and L. lactis/pNZ8150-pgsA-HAsd showed a specific binding profile that could be directly with anti-HA antibody and FITC-conjugated goat anti-mouse IgG. It was then tested by IFA and flow cytometric analysis. The binding profiles observed positively correlated with the display efficacy of HAsd, demonstrating that HAsd presented on the surface of L. lactis has reactive activity in vitro, which may contribute to immunogenicity in vivo.It has been hypothesized that the HAsd present on the surface of E. coli heat-liable enterotoxin B subunit (LTB) and cholera toxin B subunit (CTB) are potent mucosal adjuvants and promote the induction of protective immunity. However, the adverse effects of these adjuvants in clinical trials, including diarrhea, low-grade fever, and vomiting, limit their further application [L. lactis has an adjuvant function because of its inherent safety profile [L. lactis/pNZ8150-pgsA-HAsd could induce an adequate humoral response and mucosal response in the absence of a mucosal adjuvant. Moreover, the contributions of HA-specific antibodies for broad-spectrum protection against divergent influenza A viruses were emphasized.It is generally recognized that lication ,24. Furt profile , and sti profile ,27. ThusL. lactis/pNZ8150-pgsA-HAsd were protected completely (100%) (5/5) from homologous A/Vietnam/1203/2004(H5N1) virus challenge, as well as 80% (4/5) against heterologous H3N2 or H1N1 virus infection, and showed a reduced degree of virus shedding in the lung. These findings provide reliable evidence that L. lactis/pNZ8150-pgsA-HAsd is an effective vaccine candidate to induce cross-protective immunity against divergent influenza A viruses.Antigen epitope is crucial in designing vaccine. The HAsd designed in this study was codon-optimized for completely matching the antigen epitope with homologous H5N1 and 60% HAsd protein sequence similarity with heterologous H3N2 or H1N1, which provides the basis of cross-protective immunity. Furthermore, virus challenge is regarded as the gold standard for assessing the efficacy of an influenza vaccine. Therefore, mice administered orally with L. lactis/pNZ8110-pgsA-HA1, in which pgsA has been used as an anchor protein, provided immune protection against homologous H5N1 virus in the mouse model [L. lactis surface. The HAsd represents a promising research target for a novel universal influenza vaccine. Notably, the HAsd presented on the surface of L. lactis has the potential to induce cross-protective immunity against divergent influenza A viruses. Thus, this study strongly demonstrates that the discovery of L. lactis/pNZ8150-pgsA-HAsd may allow for an alternative design of influenza vaccines that would afford broad coverage.It has been demonstrated previously that adjuvanted se model . In this


Marine sponges are well known as rich sources of biologically natural products. Growing evidence indicates that sponges harbor a wealth of microorganisms in their bodies, which are likely to be the true producers of bioactive secondary metabolites. In order to promote the study of natural product chemistry and explore the relationship between microorganisms and their sponge hosts, in this review, we give a comprehensive overview of the structures, sources, and activities of the 774 new marine natural products from sponge-derived microorganisms described over the last two decades from 1998 to 2017. The term “symbiosis” was first defined by the German mycologist Heinrich Anton de Bary in 1879 as “the living together of unlike organisms” . Fr  [14]. F124 16)  were iso JBIR-59  and new   were is [18–19)  [18] werTrichoderma sp. derived from the Caribbean sponge Agelas dispar, four novel sorbicillinoid polyketide derivatives (20–23) (20–23 showed no bioactivities . A . A 60–61 (86–88) . Compoun4 µg/mL) .Arthrinium arundinis ZSDS1-F3 cultured from a Phakellia fusca marine sponge , four new cytochalasins, arthriniumnins A–D (89–92), a new natural product, ketocytochalasin (93), three new 4-hydroxy-2-pyridone alkaloids, arthpyrones A–C (94–96), and a new natural product, phenethyl 5-hydroxy-4-oxohexanoate (97)  inhibitory activity with an IC50 value of 0.81 μM ..107–109)126–127)  showed superior cytotoxicity at nanomolar concentrations toward 36 human tumor cell lines . Fr. Fr166) ue (177)  was puri  was yieAspergillus insuetus (OY-207) originated from the Mediterranean sponge Psammocinia sp. , and it afforded three novel meroterpenoids insuetolides A–C (178–180) and one new drimane sesquiterpene (181)  (Arthrinium sp. associated with the marine sponge Sarcotragus muscarum (Turkey) . Th. ThAsperne (192)  72]. A . A Asper2) (193) , was proA-MB-231 . Examina194–201) . Their a202–207), and a new polyketide, scedogiine G (208)   , were pr209–210)  were isoe sponge ,76.211) and a monocyclic butoxyl derivative (212) , and the new phthalide herbaric acid (215) . However, their bioassays were disappointing . Th. ThDichone (262) . Compoun± 0.3 µM .263)  and ent-gloeosteretriol (266)  led to the isolation of two new natural products, evariquinone and isoemericellin (267–268) (267 showed antiproliferative activity toward KB and NCI-H460 cells at a concentration of 3.16 μg/mL . F. FPetros364–365) , were isth Korea . New α-p368–369)  were iso Island) . Two new370–371) , were deectively . A study F (373) . Both co 33.1 μM ,120. Two374–375)  were isoectively . Futher ve (376) , which e–96.4 µM . A new dµM . A. AS)-2,43) (434)  was fromioassays . A new dys anthracene derivative called mayamycin (497), and three new streptophenazines I–K (498–500)  afforded two unusual compounds, Chlorohydroaspyrones A–B (501–502)  was found to produce two new indole derivatives (503–504) (505–508) . No bioactivities were detected . F. FStrept520–521) . Lobophof 7.5 μM . A marinin 524) . Compounectively . From th9  172].  . Compou 526–527  were ideectively . Chemicaid (531) , which wne (533)  were detn, China . Chemicane (534) . Compouncavenger .l-seryl-l-prolyl-l-glutamyl) and cyclo-(glycyl-l-prolyl-l-glutamyl) (535–536)  was found to produce seven new drimane sesquiterpenoids (537–543), five new ophiobolin-type sesterterpenoids (544–548), and the two new pyrrolidine alkaloids (549–550)   and exhibited cytotoxicity against the L929 (mouse fibroblast) cell line with an IC50 value approximated to 30 μM, while compound 553 was the most effective inhibitor of NF-κB nuclear translocation with an IC50 value of 71 μM  (654) (Aspergillus ostianus strain TUF 01F313 isolated from a marine sponge collected at Pohnpei. Three new chlorinated compounds (647–649) showed antibacterial activity against the Ruegeria atlantica strain. Compounds 650–652 showed cytotoxic activity against mouse lymphocytic leukemia cells (L1210) with LD50 values of 2.1, 71.0, and 2.0 μg/mL, respectively  (654)  were isoectively ,219,220.655–656) . Both co657–658) . Both coectively .659)  (Aspergillus sp. (strain CNK-371) that was isolated from an unidentified sponge collected at Manele Bay, Hawaii. Compounds 660–661 showed moderate cytotoxicity against human colon carcinoma (HCT-116) with IC50 values of 13.2, 10.9, and 13.9 μM, respectively . C. CAspergin (690) . Compoun25 μg/mL . One newmL [691)  was isol0.07) mm .Penicillium sp. MWZ14-4, which was isolated from an unidentified sponge (collected from the South China Sea), yielded 10 new fungal metabolites, including three hydroisocoumarins, penicimarins A–C (692–694), three isocoumarins, penicimarins D–F (695–697), and four benzofurans, penicifurans A–D (698–701) (698) showed moderate inhibitory activity against Staphylococcus albus with an MIC value of 3.13 μM and weak activity against B. cereus [702–703) . Compounds 702–703 may show antiphotoaging activity in UVB-irradiated models [Chemical examination of the sponge-derived fungus 698–701) . Penicif. cereus . Two new702–703) , were isd models .704) , amycocyclopiazonic acid (706), and amycolactam (707) (Amycolatopsis sp. isolated from a sponge sample gathered from Micronesia. Amycolactam (707) displayed significant to moderate cytotoxicity against the gastric cancer cell line SNU638, the colon cancer cell line HCT116, A546, K562, and SK-HEP1 with IC50 values of 0.8, 2.0, 13.7, 9.6, and 8.3 μM, respectively [Penicillium adametzioides AS-53 from an unidentified marine sponge  afforded a new spiroquinazoline derivative, N-Formyllapatin A (708), two new bisthiodiketopiperazine derivatives, adametizines A–B (709–710), two new acorane sesquiterpenes, adametacorenols A–B (711–712), a new dithiodiketopiperazine derivative, peniciadametizine A (713), and a highly oxygenated new analogue, peniciadametizine B (714)  with an LD50 value of 4.8 μM and moderate antimicrobial potency against several microbes with MIC values ranging from 8 to 32 μg/ mL, respectively, whereas compound 710 only showed weak activity against S. aureus . Compound 712 showed significant selective activity against the NCI-H446 cell line (IC50 = 5.0 μM). Compounds 713 and 714 showed inhibitory activity against the pathogenic fungus Alternaria brassicae with MIC values of 4.0 and 32.0 μg/mL, respectively [Aspergillus terreus MXH-23, isolated from an unidentified sponge , yielded a new butyrolactone derivative, namely butyrolactone VIII (715)  , was isoendently . Three nam (707)  were isoectively . Chemica B (714) . Compounectively ,245,246.II (715) . Derivatactivity .716) sesquiterpenoid and two new (717–718) . Compounds 717–718 exhibited significant inhibitory activity against cyclooxygenase (COX-2) with IC50 values of 10.6 and 8.9 μM, respectively. Besides, compound 718 displayed activities against intestinal virus EV71 with IC50 values of 30.1 μM [Micromonospora sp. NPS2077 isolated from an unidentified marine sponge  led to the isolation of a novel β-hydroxyl-δ-lactone compound, neomacquarimicin (719) (Bacillus subtilis and Escherichia coli [720) . No bioactivities were detected [721)  [Verrucosispora sp. FIM06054 from a marine sponge sample (the East China Sea) afforded a new compound, FW054-1 (722)   xanthone 30.1 μM . The exain (719) , which ehia coli . One newli [720) , was isodetected . One newed [721)  was from, China) . Chemica-1 (722) . Compoun 6.88 μM .Aspergillus sp. OUCMDZ-1583 from an unidentified sponge XD10410  yielded 18 new compounds named aspergones A–Q (723–739) and 6-O-demethylmonocerin (740)  [Penicillium chrysogenum SYP-F-2720 from an unidentified sponge  afforded a novel benzoic acid (741)  afforded four new sesquiterpene lactones (742–745) , a new cyclohexapeptide, similanamide (747), and a new pyripyropene derivative, named pyripyropene T (748) (747 exhibited weak activity against the MCF-7 (breast adenocarcinoma), NCI-H460 , and A373 (melanoma) cell lines, and neither of them showed antibacterial activity [Fractionation of the marine fungus in (740) . Compoun37.3 μM) . Chemicaid (741) . When adc effect . Study o742–745) . Compoun193.3 μM . The inv T (748) . Only coactivity .Corynespora cassiicola XS-200900I7, isolated from an unidentified sponge XS-2009001 , yielded 12 new chromone derivatives, corynechromones A–L (749–760), two new naphthalenones, corynenones A–B (761–762), and one new depsidone, corynesidone E (763) (764–765)  [766–767)  led to the isolation of three new meroterpenoids, named austalides S–U (768–770)  . The bio764–765) , were me20.0 μM) . Two new766–767) , were meectively . Chemica768–770) . Compounof 90 µM .Streptomyces albus PVA94-07 strain derived from an unidentified sponge afforded two new deferoxamine analogues, compounds 771–772  afforded two new polyketides, aspergchromones A–B (773–774)  . Compounrculosis .As what we described above, temperate and tropical sea areas have become the main regions with sponge-derived microorganisms related to natural product chemistry. The statistical data shows that all the sponge hosts associated with microbes were distributed in the classes of Calcarea, Demospongiae, and unidentified sponges. The class Demospongiae sponges accounts for absolute majority at 77%, the class Calcarea and others sponges account for 1% and 22%, respectively. In the class Demospongiae, the order Haplosclerida provides the most sponge species accounting for 26.75% of the totality, followed by the orders Suberitida, Axinellida, Dictyoceratida, Tetractinellida, and Poecilosclerida etc. see .Aspergillus and Penicillium, followed by Trichoderma, Acremonium, Arthrinium, and Talaromyces. The genera Aspergillus and Penicillium obtained from many different sponge species take a percentage of 25% of the total microbes reported, the majority of which displayed wonderful chemistry diversity. Actinomycetes and bacteria account for 16% and 11% of sponge-derived microbes, respectively, which offered many novel and unique compounds. The actinomycetes studied were mainly derived from the genera Streptomyces, and the bacteria were mainly derived from the genera Pseudomonas, Pseudoalteromonas, and Bacillus  [50 value of 8.8 μM against H1N1 virus  [687) showed potent cell growth inhibitory activity against the murine P388 cell line with an ED50 value of 57 ng/mL  [Natural products isolated from sponge-derived microorganisms have interesting pharmaceutical activities such as cytotoxicity, antioxidant, antifungal, antiviral, and antibacterial activities. Some novel compounds showed significant cytotoxic activities in the nM levels, such as two new indolocarbazole alkaloids  and line14.8 μM) . Truncat46.5 μM) . Dankast8 ng/mL) . As we r8 ng/mL) . Another8 ng/mL) , antipla8 ng/mL) ,134, neu8 ng/mL) ,178, andA total of 774 new compounds from sponge-derived microorganisms, covering the last two decades from 1998 to 2017, were reviewed. These new compounds presented abundant chemical diversity except for the well-known types such as terpenes, alkaloids, peptides, aromatics, meroterpenoids, macrolides, polyketides, steroids, and so on.The total amount of new compounds from sponge-derived microorganisms has increased rapidly and has not yet reached a climax, especially in the last five years see . Among tWe should make more efforts to study the pharmacodynamic relationships and pharmacological effects of promising compounds, and conduct clinical trials to complete the drug-like evaluation of the compound. The study on the chemical constituents of sponges and their co-existing microorganisms will promote the study of the relationship between sponges and their co-existing microorganisms, develop and utilize medicinal resources, and systematically study the diversity of biodiversity and ecosystems. Given the structural differences and biodiversity of compounds derived from sponge symbiotic microorganisms, we believe that there are more resources waiting to be mined.


Dear Editor,Several results have been reported on the efficacy of tranexamic acid (TXA) in patients with acute traumatic brain injury (TBI). As such, Lawati et al. (2020) conductRowell et al. (2020) conductIn contrast, an RCT report presented that the adjusted RRs (95% CIs) of TXA for mild-to-moderate and severe TBI death were 0.78 (0.64-0.95) and 0.99 (0.91-1.07), respectively , and a The author declares no conflict of interest.


AbstractFanjingagen. nov., with type species F.digitata Yu & Yang, sp. nov., is described from the Guizhou Province of Southwest China. Habitus photos and illustrations of male genitalia of the new species are provided.A new leafhopper genus,  Alebroides group of Empoascini leafhoppers includes 25 genera and 156 species, widely distributed in the Palaearctic Region and Old World tropics . For morphological terminology we mostly follow Taxon classificationAnimaliaHemipteraCicadellidaeYu & Yanggen. nov.04EA95A4-B242-55E4-AD37-69DA953F1F0Fhttp://zoobank.org/F1EB5835-F39D-4C1A-862C-3F55C8618661Fanjingadigitata Yu & Yang, sp. nov., here designated.Alebroides group of Empoascini, the new genus runs to Nulliata Lu, Xu & Qin in the key to genera by Nulliata in having the abdominal apodemes long and from Nulliata and other genera in having a finger-like process bearing micro-setae on the lower posterior margin of the male pygofer.Within the Relatively robust Figs . Head slMale basal abdominal sternal apodemes well developed Fig. . Male pyThe genus name is an arbitrary combination of letters.China (Guizhou).Taxon classificationAnimaliaHemipteraCicadellidaeYu & Yangsp. nov.FE13424B-E8A8-5677-B624-2DA047EAD319http://zoobank.org/F427C9A4-AEFC-472A-AFAB-B158C408A80ESize. Male 3.94–4.00mm.Whole body yellowish, anterior margin of vertex marked with two brownish patches Figs . PronotuBasal sternal abdominal apodemes extending to end of segment VI Fig. . Male py♂, China, Guizhou, Fanjing Mountain, 21-August-2017, coll. Yarong Gao (GUGC). Paratypes: 3♂, same collecting data as holotype .The new species is named from the Latin “digitalis”, meaning finger, for the finger-like process of the male pygofer.Euonymus (Celastraceae).China (Guizhou).


Correction to: Genome Biol (2019) 20:134https://doi.org/10.1186/s13059-019-1732-1Following publication of the original paper , the autIn the Additional file Additional file 1: Additional text 1. Supplementary methods. Additional text 2. Mapping SMURF-seq reads. Additional text 3. Short molecule sequencing with long-read sequencers. Additional table 3. Summary of sequencing runs. Figure S1. Distribution of length between restriction sites computed by measuring the distance between the recognition sites on the human reference genome. Figure S2. Schematic of SMURF-seq protocol. Figure S3. Sequencing of restriction enzyme digested normal diploid genome without SMURF-seq. Figure S4. Sequencing normal diploid genome using SMURF-seq. Figure S5. Sequencing normal diploid genome using SMURF-seq with 1D Rapid kit. Figure S6. Sequencing SK-BR-3 cancer genome using SMURF-seq. Figure S7. Replicate sequencing run of normal diploid genome using SMURF-seq. Figure S8. Replicate sequencing run of SK-BR-3 cancer genome using SMURF-seq. Figure S9. High-resolution CNV profile generated using SMURF-seq is highly concordant with the profile generated with Illumina WGS. Figure S10. SMURF-seq generates fragments at a faster rate than sequencing short molecules directly. Figure S11. CNV profile with reads obtained in first few minutes of sequencing. Figure S12. Multiplexed sequencing of normal diploid (barcode01) and SK-BR-3 cancer genome (barcode02) in a single sequencing run. Figure S13. Speed of nanopore sequencing as a function of read length. Figure S14. Biases correlated with GC content are reduced with LOWESS smoothing.


This letter summarizes recommendations from the interdisciplinary working group of renal tumors (IAGN) of the German Cancer Society for the systemic treatment of advanced/metastatic renal cell carcinoma in the context of the current SARS-CoV-2 pandemic During the SARS-CoV-2 pandemic, oncologists faced several new challenges  of the German cancer society towards treatment of mRCC.During the process of treatment evaluation, the assessment of treatment indication must be made on an individual basis. However, once selecting treatment, patient, and tumor-specific parameters, patient´s comorbidity and the availability of local caregiving facilities should be taken into account. In our opinion, two major scenarios according to systemic treatment have to be considered: initiation or change medication of a systemic treatment upon progressing diseases and measures taken during ongoing systemic treatment.For systemic treatment, the appropriate treatment should be selected based on patient-specific factors that take the overall picture into account when searching for treatment , as well as a surprising high rate of SARS-CoV-2 diseases among cancer patients receiving surgery  benefit was gained by any of the immune combinations for patients with a favorable risk profile (HR for OS: ipilimumab/nivolumab vs. sunitinib: 1.19 (95% CI 0.77–1.85), axitinib/pembrolizumab vs. sunitinib: 0.94 (95% CI 0.43–2.07), axitinib/avelumab vs. sunitinib: 0.812 (95% CI 0.336–1.960) ; axitinib/pembrolizumab intermediate prognosis: 0.53 (95% CI 0.35–0.82), poor prognosis: 0.43 (95% CI 0.23–0.81); axitinib/avelumab: intermediate prognosis: 0.86 (95% CI 0.615–1.202), poor prognosis: 0.57 (95% CI 0.363–0.895); Cabozantinib vs. sunitinib: 0.80 (0.53–1.21)] (Motzer et al. Although precise guidelines according to mRCC treatment reflects best efficacy and QoL data, within the SARS-CoV-2 pandemic a patient-centered treatment choice, which is adapted to the local pandemic situation is warranted (s. Table


Dear Editor,9/L in the platelet count for mortality was 0.60 (0.43-0.84). They monitored changes in the platelet count during treatments and whether maintaining a normal platelet count was related to good prognosis. A meta-analysis conducted by Lippi et al.  have presented with thrombocytopenia. Figliozzi et al. (20209 have pr) reporte 0.60 0.4-0.84. ThHowever, some reports present an inverse association. Ibañez et al. (2020) describAlthough two meta-analyses presented that thrombocytopenia reflected poor prognosis in patients with COVID-19 , the poThe author declares no conflict of interest.