Streptomyces; however, recent publications have reported terpene biosynthesis by members of the genera Actinomadura, Allokutzneria, Amycolatopsis, Kitasatosporia, Micromonospora, Nocardiopsis, Salinispora, Verrucosispora, etc. It should be noted that the use of genetically modified actinomycetes is an effective tool for studying and regulating terpenes, as well as increasing productivity of terpene biosynthesis in comparison with native producers. The review includes research articles on terpene biosynthesis by Actinomycetes between 2000 and 2022, and a patent analysis in this area shows current trends and actual research directions in this field.Terpenes and their derivatives  constitute the largest class of natural compounds, which have valuable biological activities and are promising therapeutic agents. The present review assesses the biosynthetic capabilities of actinomycetes to produce various terpene derivatives; reports the main methodological approaches to searching for new terpenes and their derivatives; identifies the most active terpene producers among actinomycetes; and describes the chemical diversity and biological properties of the obtained compounds. Among terpene derivatives isolated from actinomycetes, compounds with pronounced antifungal, antiviral, antitumor, anti-inflammatory, and other effects were determined. Actinomycete-produced terpenoids and meroterpenoids with high antimicrobial activity are of interest as a source of novel antibiotics effective against drug-resistant pathogenic bacteria. Most of the discovered terpene derivatives are produced by the genus  O-containing derivatives (terpenoids) are the largest  and structurally most diverse group of secondary metabolites derived from natural sources. Based on the number of isoprene units, terpene derivatives are classified into mono- (C10), sesqui- (C15), di- (C20), sester- (C25), tri- (C30), sesquar- (C35), and tetra- (C40) terpenes. Terpene derivatives are widely used in the food, cosmetics, and fragrance industries -1-epi-cubenol, firstly obtained by fermentation, was synthesized carbonyl and 2-[(2-chlorophenyl)amino]-2-oxo-acetyl, respectively, were identified among the metabolites of Streptomyces sp. SN194 [308 R=H309 R=CH3310 R1=R2=R3=H311 R1=R2=H, R3=CH3312 R1=H, R2=Cl, R3=H313 R1=OH, R2=R3=H314 R=H315 R=CH2OH316 R=CH(OH)2317 R=CHO318 R=COOH319320321 R=CH3322 R=H323 R1=H, R2=αOH, R3=H324 R1=H, R2=αOH, R3=CH3325 R1=H, R2=OCH3, R3=H326 R=CH3327 R=CH2OH328 R=H 329 R=CH3330 dixiamycin B331332333334335336337338339340341 R1=R2=H, R3=CH3, R4=H342 R1=R2=H, R3=CH2OH, R4=H 343 R1=R2=H, R3=R4=CH3344 R1=H, R2=OH, R3=CH3, R4=H345 R1=OH, R2=H, R3=CH3, R4=H346 KO-3988 , S. gris CB00830 , and Strp. SN194  synthesip. SN194 .308 R=H347 R= 348 R=349 R=351 R=NH2350 R=352 R=353 R=Streptomyces sp. Tü6071 produced phenalinolactones A–D (354–357), tricyclic terpene glycosides, and their derivatives 359–362, 365, and 366 [Streptomyces sp. Tü6071 with inactivated oxygenase genes , dehydrogenase genes  and putative acetyltransferase gene (plaV) yielded phenalinolactone derivatives PL HS2 (364), PL X1 (363) PL HS6 (367), and PL HS7 (368) [354) and D (357) were identified as PL IM1 (370) and PL IM2 (369), respectively [S. coelicolor M512 resulted in the formation of the non-glycosylated derivative phenalinolactone E (358) [ and 366 ,237. TheS7 (368) . Later, ectively . Heterol E (358) . 371–381), close structural analogues of phenalinolactones, were discovered by genome mining of diterpene synthases in Streptomyces sp. CB03234 and Streptomyces sp. CB03238. Tiancilactones are characterized by a highly functionalized diterpene backbone, which comprises chloroanthranilate and γ-butyrolactone moieties, and exhibit antibacterial activity [γ-lactone, namely trinulactones A (382) and B (383), were isolated from Streptomyces sp. S006 [1R2R3R4R354OH3-CO-CH3CH355OH3-CO-CHH356OH3-CO-CH2-O-CH3-CH357H3-CO-CH3CH358H3-CO-CHOH3CH359H3-CO-CH3CH360HHH3CH361H3-CO-CHH3CH362HHOH3CH363HH3CH364H3-CO-CH2-O-CH3-CH367 R=αOH368 R=O365 R=αOH366 R=O369 R370371 R1=Cl, R2=CH3, R3=OCH3372 R1=H, R2=CH3, R3=OCH3373 R1=Cl, R2=CH3, R3=OH374 R1=Cl, R2=CH3, R3=oxo375 R1=Cl, R2=H, R3=OCH3376377378379 R=H380 R=CH3381 R=H382 R=OH383Tiancilactones A–K (activity . Two newsp. S006 .R1R2R3384), its isomer 385, and fusicomycin B (386) were separated from the fermentation broth of S. violascens YIM 100212 [387) and B (388) were obtained from Streptomyces sp. KCB17JA11 [389) is new meroterpenoid derived from Streptomyces sp. CNQ-027 consisting of an unprecedented dihydronaphthalenone polyketide linked to a bicyclic diterpenoid [384 R=385 R=386 R=387388389Fusicomycin A (M 100212 . Two newCB17JA11 . Actinorerpenoid .384 R=S. platensis MA7327 and S. platensis MA7339 were shown to synthesize platensimycin (390) and platencin (391), representatives of a new class of broad-spectrum antibiotics against Gram-positive bacteria, in particular S. aureus [ent-kaurene and ent-atiserene synthases in biosynthesis of 390 and 391, representing a new biosynthetic pathway for diterpenoids [ent-copalyl diphosphate synthase involved in the biosynthesis of 390 and 391 in S. platensis CB00739, was described. PtmT2 catalyzed the cyclization of GGPP to ent-CPP, which subsequently channeled into (16R)-ent-kauran-16-ol (392) or ent-atiserene (393) by two distinct type  diterpene synthases specific for biosynthesis of 390 or 391, respectively [S. platensis SB12002 and SB12600 produced 390 and 391 with yields of 323 ± 29 mg/L and 255 ± 30 mg/L, respectively, hundreds of times greater than those of wild-type strains [S. platensis SB12600, in addition to 391, accumulated eight new congeners, platencins A2–A9 (394–402) [390 using the mixed culture of S. hygroscopicus HOK021 (NITE P-02560) and Tsukamurella pulmonis TP-B0596 was patented (JP2019149945). Exemplified by 390 and 391, a method of searching for novel natural compounds based on the analysis of biosynthetic genes was proposed (WO2015200501). Data on the biosynthesis features and biological activity of natural and synthetic analogues of platensimycin and platencin were summarized in the reviews [. aureus ,246. Furrpenoids ,248,249.ectively . The met strains ,252 (US2394–402) . A metho reviews ,255.N-containing aminobacteriohopanetriol (403), and adenosylhopane (405), as well as bacteriohopanetetrol (404) and ribosylhopane (406), were determined. Orf14 and orf18 of S. coelicolor A(3)2 responsible for the synthesis of 403 were identified [The intermediates of hopanoids biosynthesis, entified . Streptomyces sp. YIM 56130, triterpene glycoside soyasaponin I (407) [408) produced by S. argenteolus A-2 (FERM BP2065) has a unique structure consisting of a tetraterpene skeleton with 2-O-methylglucuronic acid and O-succinyl benzoate moieties [390391392393394 R1=OH, R2=H395 R1=H, R2=OH396 R1= R2=H397 R1=OH, R2=H398 R1=H, R2=OH399 R=OH400401 R=SCH3402 R=OCH3403 R=NH2404 R=OH405406407408Among the secondary metabolites of  I (407)  with a w I (407)  was obtamoieties .3903913Nocardiopsis, Amycolatopsis, Isoptericola, Saccharopolyspora, Salinispora, Kitasatosporia, Verrucosispora, etc. The compounds produced are represented mainly by sesqui- and diterpenes and their derivatives.Although most of the found actinomycete terpene derivatives are synthesized by streptomycetes, there is an increasing number of publications on terpene biosynthesis by representatives of the genera Nocardiopsis chromogenes YIM 90109, two new monocyclic germacradiene-type sesquiterpenoids germacradiene-9β,11-diol (409) and 11-hydroxy-germacradien-2-one  were identified along with the known geosmin-type sesquiterpenoid 46 [Kitasatospora setae KM-6054 [Micromonospora marina DSM 45555 [411) and (−)-germacrene A (27), respectively. The ability to produce bicyclic 2-methylisoborneol (6) and geosmin (22) was described for Nocardia cummidelens and N. fluminea [S. avermitilis carrying the genes from Saccharopolyspora erythraea NRRL2338 yielded 2-methylisoborneol (6), while Micromonospora olivasterospora KY11048 synthesized 2-methyleneornane (412) [409410411412Among the secondary metabolites of enoid 46 . The TSs KM-6054  and MicrSM 45555  catalyzefluminea . The trane (412) .4094104413 and 414) were isolated from the culture medium of Amycolatopsis alba DSM 44262 [Isoptericola chiayiensis BCRC 16888, a new sesquiterpenoid isopterchiayione (415) was registered [Amycolatopsis benzoatilytica DSM 43387 catalyzing the formation of a bicyclic sesquiterpenoid (416) was described [Verrucosispora gifhornensis YM28-088 [Verrucosispora sp. FIM06031 produced bicyclic sesquiterpenoid cyperusol C (417) and FW03104 (418) (CN101898936), respectively. Two new monocyclic sesquiterpenoids (SM 44262 . Among tescribed . VerrucoYM28-088  and VerrStreptosporangium roseum DSM 43021 and Kitasatosporia setae KM-6054 afforded tricyclic sesquiterpenoids epi-cubebol (419) [420) and B (421) [Saccharothrix espanaensis DSM 44229 [E)-β-caryophyllene (93).413414415416417418419420421Terpene synthases from ol (419)  and new  B (421) ,266, resSM 44229  was incuMicromonospora marina DSM 45555 was functionally characterized to produce micromonocyclol (422), a new diterpene alcohol with a rare 15-membered ring [Mycobacterium tuberculosis H37Rvн synthesized unique bicyclic diterpenoids, which presumably block the formation of phagolysosomes in human macrophages. The Rv3377c and Rv3378c genes proved to be responsible for synthesis of tuberculosinol (5(6),13(14)-halimadiene-15-ol, 423), 13R- (424) and 13S-isotuberculosinol (5(6),14(15)-halimadiene-13-ol, 425), and nosyberkol (426) (previously identified as edaxadiene). The analogs of Rv3377c and Rv3378c were found in the virulent strains of M. tuberculosis CDC1551 and M. bovis subsp. bovis AF2122/97, but did not occur in non-pathogenic strains [The TS from red ring . Mycobac strains ,271,272. strains .427) was firstly separated from the fermentation broth of Kitasatosporia griseola MF730-N6 (syn. Streptomyces griseolosporeus MF730-N6) in 1985 [ORF11 and ORF12 in S. lividans together with the GGDP synthase gene resulted in the formation of a new cyclic diterpene ent-clerod-3,13(16),14-triene  with a structure similar to 427 [429) to 428, accepted labdane-type diterpene diphosphates (+)-CDP (430), syn-CDP (431), (−)-ent-CDP (432), as well as halimane-type diterpene diphosphate  and catalyzed the formation of corresponding derivatives (434–437) [A bicyclic terpenoid terpentecin  .terp1 operon from Salinispora arenicola CNS-205 in E. coli led to the generation of isopimara-8,15-dien-19-ol (438). It should be noted that this terpenoid was not observed in pure cultures of S. arenicola CNS-205. Apparently, the terp1 operon was expressed under certain conditions, for example, in the presence of other marine organisms [Salinispora sp. PKU-MA00418 accepted CPP and syn-CPP and produced syn-isopimaradiene/pimaradiene analogues . Compound 439 possess a unique and previously unreported 6-6-7 ring skeleton [447) and B (448), were isolated from the culture medium of Verrucosispora gifhornenensis YM28-088 [Micromonospora haikouensis G039 [Microbispora hainanensis CSR-4 [449) and 2α-hydroxy-8(14),15-pimaradien-17,18-dioic acid (450) were identified, respectively.Heterologous expression of the biosynthetic rganisms . The terskeleton . New hydYM28-088 . Among ssis G039  and Micris CSR-4 , new ditActinomadura sp. SpB081030SC-15 [Actinomadura sp. KC 191 [451) and actinomadurol (452), rare bacterial C-19 norditerpenoids. A norditerpenoid k4610422 (453), originally discovered from a mesophilic rare actinomycete of the genus Streptosporangium, was isolated from the culture extract of a thermophilic actinomycete Actinomadura sp. AMW41E2 [422423424425426427428429430 R=αH431 R=βH432433434435436437438439440441442443444445446447448449450451452453030SC-15  and Acti. KC 191  synthesi AMW41E2 .4224234Catenulispora acidiphila DSM 44928 and Saccharopolyspora spinosa NRRL 18395 produced new di- and tricyclic catenul-14-en-6-ol (454), isocatenula-2,14-diene (455), isocatenula-2(6),14-diene (456) [457), B (458), and 2,7,18-dolabellatriene (459) [Diterpene synthases from ne (456) , and spine (459) , respectNocardia testacea NBRC 100365 and N. rhamnosiphila NBRC 108938 accepted GGPP, but not GPP, FPP, or GFPP as a substrate, which was converted by both enzymes in a tetracyclic diterpene phomopsene (460) [Allokutzneria albata DSM 44149 encoded four diterpene synthases that catalyze the formation of mono-, tri-, and tetracyclic compounds: new spiroalbatene (461), bonnadiene (462) and allokutznerene (463), and known compounds: cembrene A (164), thunbergol (464), phomopsene (460), and spiroviolene (200) [Terpene synthases isolated from ne (460) . Allokutne (200) ,288.465–482) were found in the genus Frankia [483) was isolated from the cell walls of nonpathogenic mycobacteria [454455456457458459460461462463464Hopanoid lipids  (CN101921721). Saccharomonospora sp. CNQ-490 produced saccharoquinoline (485), meroterpenoid with drimane-type sesquiterpene unit [486) and B (487), were derived from Micromonospora sp. WMMC-218, a symbiont of marine ascidians Symplegma brakenhielmi [Mycobacterium tuberculosis H37Rvн revealed that this enzyme catalyzed the formation of 1-tuberculosinyladenosine (488) and its two isomers, one of which was identified as 6N-tuberculosinyladenosine (489). Compounds 488 and 489 are specific diterpene nucleosides of pathogen of Mycobacterium tuberculosis and can serve as chemical markers of infection [Rv3377c-Rv3378c from M. tuberculosis H37Rvн in M. kansasii led to the production of 1-tuberculosinyladenosine (488) [ene unit . Two newenhielmi . Furthernfection ,296,297.ne (488) .Nocardia brasiliensis IFM 0406 (now N. terpenica) to synthesize diterpene glycoside brasilicardin A (490) was first described in 1999 [490) displays a unique structure consisting of a diterpene skeleton with L-rhamnose, N-acetylglucosamine, amino acid, and 3-hydroxybenzoate components [N. terpenica IFM0406 and identified as brasilicardins B–D (491–493) [bra0-12), responsible for the synthesis of 490, in Amycolatopsis japonicum (A. japonicum::bcaAB01) led to the formation of four brasilicardin congeners, namely BraC (492), BraD (493), BraC-agl , and BraD-agl  [S. griseus::bcaAB01 (pRHAMO) transformant containing the biosynthetic cluster of brasilicardin A and a plasmid with a biosynthetic cassette for the generation of TDP-L-rhamnose resulted in increased yields of compounds 492 (1669 mg/L), 495 (926 mg/L), and a new metabolite (496) (15 mg/L). The target 490 was obtained through a five-step chemical modification of 494 [The ability of  in 1999 . Brasilimponents . Later, 491–493) . The hetaF, 495) ,304,305.n of 494 .ato) gene cluster from Amycolatopsis tolypomycina NRRL B-24205 in S. albus led to the characterization of two unprecedented tricyclic sesterterpenoids atolypenes A (497) and B (498) [499), a new highly oxygenated unique tetracyclic 6-hydroxymeroterpenoid, was derived from Nocardiopsis sp. LGO5 [484485486487488489490 R=OCH3491 R=H492 R=OCH3493 R=H494 R=OCH3495 R=H496499497 R498 RCloning and activation of the atolypene ( B (498) . Terretosp. LGO5 .4844854Streptomyces, however, terpene biosynthesis by Allokutzneria, Amycolatopsis, Frankia, Kitasatosporia, Nocardia, Salinispora, Verrucosispora, etc., have been recently reported  contains more than 3000 strains of actinomycetes with a wide range of metabolic capabilities, which are promising for biocatalytic production of terpene derivatives [An extensive development of genome-sequencing technologies and bioinformatics tools have allowed the discovery of BCGs (including silent ones) in the genome of actinomycetes. That terpenoids and meroterpenoids are predominantly found among entified . In addirganisms ,50,310. olyspora , Nocardirdiopsis , Rhodocodococcus ,314, Salinispora , Verrucoosispora , and Actnomadura  have beeMF730-N6  and N. t IFM0406  revealed IFM0406 . Genomic IFM0406 ,310,321.ivatives ,325,326 epi-zizaene, pentalenene, cucumene, (E)-biformene synthases, and other TSs isolated from streptomycetes were characterized. In turn, genome mining of streptomycetes as producers of naphthoquinone-based meroterpenoids led to the discovery of unique prenyltransferase (PTase) and vanadium-dependent haloperoxidase enzymes (VHPO) [epi-zizaene synthase, the successful application of site-directed mutagenesis of the enzyme to control the range of the compounds produced was proved [Unlike the biosynthesis of well-studied secondary metabolites, such as polyketides and nonribosomal peptides, the prediction of terpene structures requires detailed understanding of the cyclization mechanisms and the structural characteristics of bacterial TSs ,327. In s (VHPO) ,183. Fors (VHPO) . It has s (VHPO) . By the s proved ,122 (WO2S. cinnabarinus PK209 and S. hygroscopicus HOK021 (NITE P-02560) synthesize the diterpene lobocompactol and the antibiotic platensimycin in the presence of the Gram-negative Alteromonas sp. KNS-16 [Tsukamurella pulmonis TP-B0596 (JP2019149945), respectively. The effectiveness of actinomycete terpenoids and meroterpenoids, namely pentalenolactone, albaflavenone, platensimycin, platencin, terpentecin, lavanducyanin, marinocyanins A–C, furaquinocin L, 3-dechloro-3-bromonapyradiomycin A1, napyradiomycin A1, and merochlorin A, as promising antibiotics has been proven. This is true for cyslabdan, which enhances the action (1000-fold) of the antibiotic imipenem against MRSA. In addition to high antibacterial activity, many meroterpenoids, such as napyradiomycins B1, B3, B4, A80915A, B, C, furaquinocins A and B, murayaquinone, marinocyanin A–C, and saccharoquinoline, exhibit a high cytotoxic activity against different cancer cell lines , which spread Streptomyces spores in the soil [S. coelicolor M145 spores, they synthesize albaflavenone, which may coordinate the development of the producer (quorum sensing) and/or play a role in the competitive repression of microflora (quorum suppression) in the natural environment [S. coelicolor A3(2) does not produce aminobacteriohopanetriol or produces this compound in negligible amounts. However, the triterpene generation increased sharply during the formation of an aerial mycelium and sporulation, which may be associated with structural changes in the membrane and protection against water loss [Micromonospora isolates revealed TS genes, which differ significantly from other groups of characterized bacterial TSs and may be useful as markers of the genus, while Mycobacterium tuberculosis H37Rvн produced specific diterpene nucleosides, 1- and 6N-tuberculosinyladenosines, promising for development as specific diagnostic markers of tuberculosis.Actinomycete-derived terpenoids participate in specific interactions with macroorganisms , regulate the bacterial life cycle, perform protective functions, or serve as taxonomic markers. Bacterial terpenoids are often optical isomers of plant terpenoids and may represent two chemical communication channels that do not overlap even if the same habitat is occupied by prokaryotic and eukaryotic organisms producing terpenes . Soil-smthe soil . Accordithe soil . Čihák eironment . In the ter loss . In addiDespite the significant (more than 300) number of publications on terpene biosynthesis by actinomycetes, the conducted patent analysis revealed only 26 patents in this research area . TerpenoThus, the synthesis of terpenes and terpenoids is an important pathway in the secondary metabolism of actinomycetes. The compounds produced may be promising therapeutic agents for the treatment of viral, inflammatory, cancerous, and other diseases in the future. Terpenoids and meroterpenoids synthesized by actinomycetes and possessing high antibacterial activity against drug-resistant pathogenic microorganisms may be useful for the development of new antibiotics. Further study of actinomycetes, accumulation of genetic information about this group of microorganisms, and employment of modern and development of novel tools of synthetic biology and genetic engineering will open prospects for creation of ideal “cell factories” using actinomycetes.


C5 convertase cleaves C5 in common complement pathways yielding the anaphylatoxin C5a and C5b, part of the membrane attack complex (MAC) that mitigates bacterial infections . C5 can also be cleaved “extrinsically” by other enzymes. C5a is elevated in severe COVID-19 patients (Pt). Yet, inhibiting C5 cleavage increases infections without improved survival in critically ill COVID-19 Pt . In addition, eculizumab, a C5 blocker does not prevent C5a from being generated in COVID-19 Pt . Vilobelimab (Vilo), an anti-C5a monoclonal antibody that preserves MAC, was tested in a Phase 3 multicenter, double-blind, randomized, placebo (Plc)-controlled study for its effect on survival in critically ill COVID-19 Pt .COVID-19 Pt recruited worldwide  intubated within 48 hrs before treatment, received up to 6, 800mg infusions of Vilo or Plc on top of standard-of-care. The primary endpoint was 28-Day all-cause mortality with key secondary endpoints of 60-Day mortality and safety. Blood samples were assessed by ELISA at screening, Day 8 and at hospital discharge for Vilo and C5a levels. No meningococcal vaccination or prophylactic antibiotics were used in the study.vs 41.6% for Plc  with similar 60-Day results. On Day 8 after 3 infusions, mean Vilo trough levels were 21,799.3 to 302,972.1 ng/mL . C5a was highly elevated and comparable between groups at screening: Vilo mean 130.3 ng/mL, Plc mean 123.2 ng/mL. By Day 8, C5a levels were reduced 87% for Vilo  vs an increase for Plc (mean 129.8 ng/mL). Though post Day 8 sampling was sparse, C5a levels remained elevated for Plc and low for Vilo throughout the study. Serious AEs were 58.9% for Vilo and 63.5% for Plc. Treatment-emergent AEs were 90.9% for Vilo vs 91.0% for Plc. Infection incidence per 100 Pt days was comparable in both groups.Kaplan-Meier estimates showed a 28-Day mortality rate of 31.7% for Vilo Results from this Phase 3 study show that direct C5a inhibition by vilobelimab while presumably preserving MAC, as opposed to upstream C5 blockade, results in a survival benefit for critically ill COVID-19 Pt without increasing infections.Bruce P. Burnett, PhD, InflaRx GmbH, InflaRx Pharmaceuticals, Inc.: Employee Alexander Vlaar, MD, PhD, InflaRx: Advisor/Consultant Maria Habel, PhD, InflaRx GmbH: Employee Claus Thielert, PhD, InflaRx GmbH: Employee James Dickinson, MSc, InflaRx GmbH: Employee simon Rückinger, PhD, Metronomia Clinical Research GmbH: Advisor/Consultant Robert Zerbib, MSc, InflaRx GmbH: Advisor/Consultant Dorothee Neukirchen, PhD, InflaRx GmbH: Employee Renfeng Guo, MD, inflarx: Board Member|inflarx: I am an Inventor for Vilobelimab|inflarx: Ownership Interest|inflarx: Stocks/Bonds|InflaRx GmbH, InflaRx Pharmaceuticals, Inc.: Board Member|InflaRx GmbH, InflaRx Pharmaceuticals, Inc.: Founder|InflaRx GmbH, InflaRx Pharmaceuticals, Inc.: Employee|InflaRx GmbH, InflaRx Pharmaceuticals, Inc.: Ownership Interest Niels Riedemann, MD, PhD, InflaRx GmbH, InflaRx Pharmaceuticals, Inc.: Board Member|InflaRx GmbH, InflaRx Pharmaceuticals, Inc.: Founder|InflaRx GmbH, InflaRx Pharmaceuticals, Inc.: Employee|InflaRx GmbH, InflaRx Pharmaceuticals, Inc.: Ownership Interest


Eminium intortum (Banks & Sol.) Kuntze and E. spiculatum (Blume) Schott (Araceae). A total of 83 metabolites, including 19 phenolic acids, 46 flavonoids, 11 amino, and 7 fatty acids were identified by UHPLC-HRMS in the studied extracts for the first time. E. intortum flower and leaf extracts had the highest total phenolic and flavonoid contents . Significant radical scavenging activity (32.20 ± 1.26 and 54.34 ± 0.53 mg TE/g for DPPH and ABTS) and reducing power (88.27 ± 1.49 and 33.13 ± 0.68 mg TE/g for CUPRAC and FRAP) were observed in leaf extracts. E. intortum flowers showed the maximum anticholinesterase activity (2.72 ± 0.03 mg GALAE/g). E. spiculatum leaves and tubers exhibited the highest inhibition towards α-glucosidase (0.99 ± 0.02 ACAE/g) and tirosinase (50.73 ± 2.29 mg KAE/g), respectively. A multivariate analysis revealed that O-hydroxycinnamoylglycosyl-C-flavonoid glycosides mostly accounted for the discrimination of both species. Thus, E. intortum and E. spiculatum can be considered as potential candidates for designing functional ingredients in the pharmaceutical and nutraceutical industries.The study aimed at the metabolite profiling and evaluation of antioxidant and enzyme inhibitory properties of methanol extracts from flowers, leaves, and tubers of unexplored  The search for bioactive natural compounds derived from medicinal plants is of great importance in modern medicine . NaturalEminium (Blume) Schott is a genus of the Araceae family, which includes over 300 species that grow mostly in tropical areas − at m/z 191.018 showed fragment ions at 173.008 [M-H-H2O]−, 147.028 [M-H-CO2]−, a base peak at m/z 111.07 [M-H-CO2-2H2O]−, and was related to citric acid − at m/z 623.162 gave a base peak at m/z 299.056, corresponding to the concomitant loss of two hexoses, and was annotated as chrysoeriol 7-O-dihexoside. MS/MS spectrum of 54 [M-H]− at m/z 577.1568 showed fragment ions at m/z 431.098 [M-H-dHex]− and 413.088 [M-H-dHex-H2O]−, and a base peak at m/z 269.045 [M-H-dHex-Hex]−. The absence of the interglycosidic linkage breakdown is favored for 7-neohesperidoside. Thus, 54 was tentatively identified as apigenin 7-O-neohesperidoside -120]−, [(M-H)-90]−, and [(M-H)-30]−. C-8 isomers were characterized by the base peak 0,2 X− [(M-H)-120]− as MS/MS of orientin (30), while C-6 isomers showed the base peaks [M-H]− as homoorientin (27), isovitexin (37), and chrysoeriol 6-C-hexoside (44), together with abundant fragment 0,2 X− [(M-H)-120]− (about 60%). In addition, the fragment ion 0,3 X− [M-H-90]− was favored by C-6 isomers (20 and 22 gave indicative ions at m/z 519.113 and 503.119 [(M-H)-90]−, 489.103, and 473.109 [(M-H)-120]−, 429.082 and 413.089 [(M-H)-120-60]−, 399.072 and 383.077 [(M-H)-120-90]−, 369.061 and 353.066 [(M-H)-2 × 120]−, respectively. Based on 1,3 B− fragments at m/z 133.028 (20) and 117.033 (22), 20 and 22 were tentatively annotated as 6,8-C-diglucosides of luteolin and apigenin, respectively  and 0,3 X1 (−90) or 0,2 X1 (−120) at m/z 357.062 and 327.051, respectively  was discernable by the fragment ions at m/z 341.067 ([(M-H)-132-90]− and 311.0561 ([(M-H)-132-120]−, suggesting the presence of both O-pentosyl (X1) and C-hexosyl (X0) moieties. Additionally, the aforementioned structure was assigned on the basis of a series of fragment ions at 283.061 , 282.055 , 281.045 , and 237.055  (Among three isobars indicated as ectively . In the  X0/2CO) .29 with ([M-H]− at m/z 771.179 was acquired. In (-) ESI mode, 29 yielded prominent fragment ions at m/z 447.093 [(M-H)-2 × 162.05]−, 357.062 [(M-H)-2 × 162.05-90]−, 327.051 ([(M-H)-2 × 162.05-120]−, suggesting the presence of both O-dihexosyl and C-hexosyl moieties. The absence of the interglycosidic linkage breakdown is favored for O-sophoroside. Thus, 29 was annotated as luteolin O-sophoroside-6-C-hexosyl-orientin. Compound 32 ([M-H]− at m/z 593.151) yielded prominent ions at m/z 413.088 ([(M-H)-(162.05 + H2O)]−, 353.067 ([(M-H)-(162.05 + H2O)-60]−, 311.056 ([(M-H)-(162.05)-120]−, 293.047 [(M-H)-(162.05 + H2O)-120]−, and 281.045 Y1−/0,2 X0/CH2O indicating an O-hexosyl unit at the 2”of the primary hexose [32 was assigned to 2″-O-hexosyl-6-C-hexosyl-apigenin  and 45 ([M-H]− at m/z 799.210) shared the same fragmentation patterns yielding prominent ions at m/z 447.094 (34) and 431.098 (35), respectively, indicating a concomitant loss of hexose (162.05 Da) and sinapoyl residue  (m/z 205.049 [(sinapic acid-H)-H2O]−, 190.027 [(sinapic acid-H)-H2O-CH3]−, and 175.003 [(sinapic acid-H)-H2O-2·CH3]−. The presence of a C-hexosyl moiety on the flavonoid skeleton was deduced from the relative abundances of the ions at m/z 327.051 [M-H-120]− (79.7%) (34) and 311.056 (100%) (45) suggesting a 6-C- and an 8-C-linkage, respectively. Thus, 34 was ascribed as luteolin O-sinapoylhexosyl-6-C-hexoside, while 45 was annotated as apigenin O-sinapoylhexosyl-6-C-hexoside (Two compounds 11H10O4) . Moreovehexoside .36 and 41 were discernable by the transition [M-H]−→Y0 resulting from the indicative loss of 324.085 Da (C15H16O8). Precursor ions gave a series of fragment ions involved in the C-glycosyl flavon pattern: m/z 353.064 , 341.067 , 311.056 , and 283.061  (m/z 161.023 [(caffeic acid-H)-H2O]− and 133.028[(caffeic acid-H)-H2O-CO]−. Accordingly, 36 and 41 were assigned to apigenin O-caffeoylhexosyl-8-C-hexoside and chrysoeriol O-caffeoylhexosyl-8-C-hexoside (Two caffeoyl esters 2 X1/CO) . Caffeoyhexoside .O-hexosyl-8-C-hexoside  were deduced from the loss of feruloylhexosyl residue  and the subsequent transition of m/z 447.094/431.099/461.109→327.051/311.056/341.067, respectively, arising from the hexose cross ring cleavage  (m/z 175.039 [(ferulic acid-H)-H2O]−, 160.015 [(ferulic acid-H)-CH3]−, and [(ferulic acid-H)-H2O-CH3-CO]− point out to the feruloyl moiety  . The ionl moiety   in the fragmentation patterns of the aforementioned compounds, accompanied with the ions at m/z 163.039 [p-coumaric acid-H]−, 145.028 [(p-coumaric acid-H)-H2O]−, and 117.033 [(p-coumaric acid-H)-H2O-CO]− confirmed the presence of a coumaroyl moiety (O-deoxyhexosyl-6-C-hexoside (60 and 62) were evidenced on the base of the loss of 292 Da (C15H16O6)    .25, 27, 30, 35, 37, 42, 47, 50, 52, 55, 63, and 64 was confirmed by comparison with reference standards.The identification of flavonoids 71 and 73), a dipeptide (74), and six amino acid hexosides were tentatively annotated , a series of fragment ions resulting from neutral losses were registered at m/z 241.119 [M-H-H2O]−, 223.108 [M-H-2H2O]−, 215.140 [M-H-CO2]−, 197.1287 [M-H-H2O-CO2]−, and a base peak at m/z 128.033, corresponding to the loss of leucine [M-H-131.096]− from the glutamyl residue. Thus, 74 was related to γ-glutamyl-leucine [66, 67, 68, 69, 70, and 72 demonstrated base peaks, resulting from the loss of a hexose moiety, with the appearance of the corresponding amino acid residue. Hence, they were annotated as hexosides of glutamic acid, valine, tyrosine, leucine, phenylalanine, and tryptophan, respectively , three monounsaturated , and five polyunsaturated  free fatty acids were tentatively annotated in Eminium extracts (78–82) were dihydroxylated and three (75–77) contained three hydroxyl groups , l groups . Compounuwolffii .E. intortum and E. spiculatum methanolic extracts, we used six different methods including DPPH, ABTS cation, FRAP, CUPRAC, phosphomolybdenum, and metal chelate assays. As shown in E. intortum exhibited a much stronger antioxidant activity compared to the extracts of E. spiculatum, with values of 32.20 ± 1.26 mg TE/g for leaves, 27.87 ± 0.75 mg TE/g for flowers, and 26.90 ± 1.61 mg TE/g for tubers. The methanolic extract of the tuber of E. spiculatum also showed good antioxidant activity with a value of 33.67 ± 1.26 mg TE/g.To study the antioxidant activity of E. intortum and 86.27 ± 2.74 for E. spiculatum. The methanolic extracts of the tubers and flowers of both plants showed lower antioxidant activity compared to the leaves.E. spiculatum and E. intortum showed very significant reducing activity, with values of 41.33 ± 1.25 mg TE/g and 32.58 ± 4.02 mg TE/g, respectively. The other extracts of both plants also showed significant results of antioxidant activity in this assay. In summary, our findings suggest that the extracts are rich in phenolic compounds with radical-scavenging activity and proton-donating ability. Polyphenols have been acknowledged for their antioxidant activity, which may account for their potential ability to prevent various diseases associated with oxidative stress. This assertion is supported by several studies [E. intortum and 61.55 ± 3.97 mg EDTAE/g for E. spiculatum. It should be noted that this method is also highly specific.In the FRAP assay, a high absorbance indicates a high reducing power. The methanolic extracts of the tubers of both  studies ,28,29,30The antioxidant activity of a substance is influenced by how it interacts with radicals in the reaction medium. These interactions are facilitated by active molecules that trap the radicals, and previous studies have highlighted their importance ,32,33. TEminium genus. Alkofahi et al. [E. spiculatum extract against oxidative DNA damage using the 8-hydroxydeoxyguanosine assay, while no beneficial effect on alleviating oxidative damage was observed. Al-Ismail et al. [E. spiculatum ethanolic extract using DPPH, FRAP, and vegetable oil emulsion systems, where the extract exhibited significant properties with lower IC50 values. Additionally, Janat and Al-Thnaibat [E. spiculatum exhibited higher antioxidant activity in the phosphomolybdenum assay when compared to the aqueous extract.Limited literature exists regarding the antioxidant properties of the i et al.  conductel et al.  reportedThnaibat  reportedIn the intestinal tract, the breakdown of complex sugars into simple sugars is facilitated by two essential enzymes, namely α-amylase and α-glucosidase. The simple products, particularly glucose, are subsequently absorbed and can cause a rise in blood sugar levels. To manage diabetes, one potential therapeutic approach is to inhibit the enzymes responsible for carbohydrate hydrolysis, which can decrease postprandial blood sugar levels ,41. By iE. spiculatum also exhibited α-glucosidase inhibition, with values of 0.99 ± 0.02 ACAE/g and 0.89 ± 0.01 ACAE/g, respectively. Among E. intortum extracts, the leaf extract was only active against α-glucosidase (0.35 mmol ACAE/g). In addition, both tuber extracts showed no inhibitory effect on α-glucosidase. Our results demonstrate the potential of the studied leaf extracts to act as inhibitors of carbohydrate hydrolyzing enzymes. By delaying the absorption of dietary carbohydrates in the small intestine and reducing postprandial hyperglycemia, these inhibitors may be a valuable component in the development of antidiabetic drugs. This mechanism of action has been previously reported [reported ,46,47.Our study is the first to investigate the antidiabetic activity of the two plants, and we found that all studied extracts of the plants inhibited α-amylase activity, with values lower than 0.31 ± 0.01 ACAE/g. The inhibitory effect of phenolic compounds on carbohydrate hydrolysis enzymes has been reported previously ,49,50, aAcetylcholinesterase (AChE) and butyrylcholinesterase (BChE) are enzymes found primarily in nerve tissues and neuromuscular junctions. They are responsible for rapidly hydrolyzing acetylcholine, a neurotransmitter, into inert choline and acetate. Overexpression or excessive catalysis of AChE can cause neuronal disturbances and lead to neurological disorders. A potential strategy for neuroprotection is to inhibit AChE activity ,52,53. OE. spiculatum and E. intortum tubers exhibited strong inhibitory activity with values of 50.73 ± 2.29 KAE/g and 48.13 ± 0.24 KAE/g, respectively. Natural compounds capable of inhibiting tyrosinase activity are of great interest, with increasing demand in the fields of cosmetics and pharmaceuticals [In skin cells, tyrosinase plays a crucial role in the aging process, and inhibiting its activity is a key strategy for delaying skin aging. This enzyme catalyzes the initial two steps of melanogenesis, making it a rate-limiting factor in this process. Mutations in the tyrosinase gene or its absence can cause a reduction or cessation of pigmentation ,58. In oeuticals ,60. Numeeuticals ,63,64,65E. spiculatum. In contrast, the flower and leaves of E. intortum were grouped into Cluster III and IV, respectively  was mainly associated with antioxidant assays and AChE. The second component (PC2) contained amylase, BChE, and tyrosinase, while the third component (PC3) included glucosidase and DPPH B. Althouectively C.E. spiculatum, which contained high levels of chrysoeriol O-coumaroylhexoside-8-C-hexoside, luteolin-6,8-C-diglucoside, and apigenin 7-O-synapoylhexosyl-8-C-hexoside. The leaves and flowers of E. intortum were classified in Cluster II, which were characterized by high levels of luteolin 7-O-coumaroylhexosyl-6-C-hexoside and apigenin O-feruloylhexosyl-8-C-hexoside. Cluster III contained both tuber samples. Furthermore, we conducted a correlation analysis between the biological activities and individual components, and the results are shown in C-hexoside-8-C-pentoside was the main contributor to DPPH scavenging ability, while sinapic acid-O-dihexoside and isorhamnetin-3-O-rutinoside were positively correlated with ABTS and CUPRAC. Consistent with our findings, the compounds have been described as important antioxidants by several researchers [The classifications of the tested extracts based on their chemical components is presented in earchers ,67,68. Aearchers .Eminium species in Turkey (Eminium intortum (Banks & Sol.) Kuntze: Between Kızıltepe and Mardin, 533 m, GPS: 37°14′20″ N, 40°38′10″ E; Eminium spiculatum (Blume) Schott: Derik, Mardin, 661 m, GPS: 37°18′44″ N, 40°16′16″ E). The plant specimens were identified by one of our co-authors, Dr. Ugur Cakilcioglu, and one specimen from the plants was deposited at the Harran University herbarium. Prior to extraction, the plant materials were carefully washed with tap and distilled water to eliminate any soil and contaminants. After being air-dried for 10 days (in shade at room temperature), the flowers, leaves, and roots were powdered.In the summer of 2021, we collected For extraction, we employed the maceration method, in which 5 g of plant material was mixed with 100 mL of methanol and left to macerate at room temperature for 24 h. After maceration, the extracts were filtered through Whatman filter paper, and the solvents were evaporated using a rotary-evaporator. To preserve the extracts, we stored them at 4 °C until analysis.p-coumaric, o-coumaric, ferulic acids, saponarin, homoorientin, orientin, rutin, isovitexin, isoquercitrin, luteolin 7-O-glucoside, kaempferol 3-O-rutinoside, kaempferol 3-O-glucoside, isorhamnetin 3-O-glucoside, apigenin 7-O-glucoside, quercetin, and apigenin. Chlorogenic and caffeic acids were supplied from Phytolab .Acetonitrile (hypergrade for LC–MS), formic acid (for LC–MS) and methanol  were purchased from Merck . The reference standards used for compound identification were obtained from Extrasynthese  for We determined the total phenolic and flavonoid content of the extracts using the Folin–Ciocalteu and AlCl3 assays, respectively, according to Zengin and Aktumsek’s protocol (2014). The results of these tests were reported in terms of gallic acid equivalents (mg GAE/g dry extract) and rutin equivalents (mg RE/g dry extract) .m/z range from 100 to 1000. The mass spectrometer parameters were as follows: spray voltage 3.5 kV (+) and 2.5 kV (−); sheath gas flow rate 38; auxiliary gas flow rate 12; spare gas flow rate 0; capillary temperature 320 °C; probe heater temperature 320 °C; S-lens RF level 50; scan mode: full MS , and MS/MS . The chromatographic separation was achieved on a reversed phase column Kromasil EternityXT C18  at 40 °C. The UHPLC analyses were run with a mobile phase consisting of 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B). The run time was 33 min. The flow rate was 0.3 mL/min. The gradient elution program was used as follows: 0–1 min, 0–5% B; 1–20 min, 5–30% B; 20–25 min, 30–50% B; 25–30 min, 50–70% B; 30–33 min, 70–95%; 33–34 min 95–5% B. Equilibration time was 4 min [The UHPLC-HRMS analyses were carried out on a Q Exactive Plus mass spectrometer  equipped with a heated electrospray ionization (HESI-II) probe (ThermoScientific). The equipment was operated in negative and positive ion modes within the as 4 min . Data weas 4 min .We analyzed the extracts for a range of antioxidant and enzyme inhibitory activities, including, cupric reducing antioxidant capacity (CUPRAC), DPPH, and ABTS radical scavenging, metal chelating activity (MCA), ferric reducing antioxidant power (FRAP), phosphomolybdenum (PBD), and inhibition of amylase, tyrosinase, glucosidase, acetylcholinesterase (AChE), and butyrylcholinesterase (BChE). We employed the previously described methods to evaluate these activities . Each sap < 0.05). Then, Principal Component Analysis (PCA) and hierarchical clustered analysis (HCA) were performed to emphasize the distinct clusters in terms of their bioactivities. Furthermore, hierarchical clustered analysis (HCA) was performed to assess the (dis)similarity between samples in terms of their molecules. All used data were scale and molecules data and were log transformed before doing multivariate analysis. R v.4.2.3 statistical program was used for all analyses.All data were given as mean ± standard deviation (SD). Statistical analysis was performed by analysis of variance (ANOVA). A post hoc test (Tukey) was performed when the differences shown by data were significant  varied according to the plant parts used. Generally, the leaf extract of both species exhibited higher antioxidant effects compared to flowers and tubers. However, we obtained different results for each enzyme inhibition assay, and the leaf extracts provided good anti-amylase and anti-glucosidase actions. The extracts were rich in flavonoids. These findings provide a valuable scientific basis for evaluating the potential of the Eminium genus and suggest that the tested species could be considered as a source of natural bioactive agents for health-promoting applications. Nevertheless, additional research is necessary to elucidate the potential toxicity of both the extracts and the individual chemical constituents.In our study, we explored the detailed chemical composition and biological effects of two


Pseudomonas aeruginosa.To evaluate the efficacy of ceftazidime-avibactam (CZA) and aztreonam-avibactam (AZA) against bloodstream infections (BSIs) or lower respiratory tract infections (LRTIs) – caused by extensive drug-resistant or pan drug-resistant (XDR/PDR) P. aeruginosa. Whole-genome sequencing was used to analyze the resistance determinants of each isolate. Monte Carlo simulations (MCSs) were used to evaluate the probability of target attainment (PTA) and the cumulative fraction of response (CFR) of each CZA/AZA dosing regimen via traditional infusion (TI)/optimized two-step-administration therapy (OTAT).The two-fold dilution method was used to determine the minimum inhibitory concentrations (MICs) of CZA/AZA against XDR/PDR P. aeruginosa isolates producing IMP-45, VIM-1, or VIM-2 were inhibited by AZA at a concentration of 2 to 8 mg/L. All isolates producing IND-6 plus other serine β-lactamases were high-level resistant to CZA/AZA (MICs >64 mg/L). All simulated dosing regimens of CZA/AZA against BSIs-causing XDR/PDR P. aeruginosa achieved 100% PTA when the MIC was ≤32 mg/L.We found that XDR/PDR P. aeruginosa may carry some rare MBLs . P. aeruginosa producing IMP-45, VIM-1, or VIM-2. OTAT with sufficient pharmacodynamic exposure may be an optimal treatment option for XDR/PDR P. aeruginosa with a high-level MIC of CZA/AZA.AZA has been considered as an option for the treatment of infections caused by XDR/PDR  P. aeruginosa displays resistance to various antibiotics, making treatment challenging . CZA has good safety and was regarded as a vital treatment option for P. aeruginosa infections. CZA showed good capabilities against P. aeruginosa with a sensitivity rate ranging from 76.2% to 97.8% (P. aeruginosa infections ranged from 64.3% to 90.6% (Ceftazidime/avibactam (CZA) is a novel β-lactam/β-lactamase inhibitor (BL/BLIs). And it was approved by the Chinese National Medical Products Administration (CNMPA) in 2019 for the treatment of complicated intra-abdominal infections (cIAIs), hospital-acquired pneumonia (HAP), and ventilator-associated pneumonia (VAP), and in adult patients with limited treatment options for infections caused by the following gram-negative bacteria sensitive to this product: to 97.8% . In termto 90.6% .β-lactamases  against which ATM is ineffective. Avibactam (AVI) is a β-lactamase inhibitor with a wide enzyme inhibition spectrum, including Ambler class A , class C (AmpC), and class D (OXA-48 type) β-lactamase. The combination of ATM and AVI can potentially inhibit MBL-producing bacteria (Aztreonam (ATM) was the first monobactam antibiotic to be used in clinical therapy. It was approved by the U.S. Food and Drug Administration (FDA) in 1986 for treatment of various infections caused by sensitive aerobic gram-negative bacteria. ATM was stable to hydrolysis by Ambler class B Metallo-β-lactamases (MBLs) . Howeverbacteria .P. aeruginosa in this study. Whole-genome sequencing and bioinformatic analysis were used to identify resistance genes of each isolate. Besides, we combine the population pharmacokinetic parameters (PPKs) of CZA/AZA with minimum inhibitory concentration (MIC) distribution of XDR/PDR P. aeruginosa to evaluate the efficacy of various dosing regimens. Therefore, the objectives of our work are as follows. Firstly, our work aims to compare the in vitro activity of CZA and AZA against XDR/PDR P. aeruginosa. Secondly, our team wants to evaluate the relationship between resistance genes and CZA/AZA sensitivity rates of XDR/PDR P. aeruginosa. Thirdly, we assess the efficacy of CZA and AZA for the treatment of critically ill patients with BSIs/LRTIs caused by XDR/PDR P. aeruginosa.We performed antimicrobial susceptibility testing (AST) on XDR/PDR 22.1P. aeruginosa from critically ill patients admitted to the First Medical Centre of Chinese PLA General Hospital from January 2016 to November 2021. A total of 10 P. aeruginosa strains were categorized as XDR according to CLSI criteria. Moreover, 57 P. aeruginosa strains were categorized as PDR . Ceftazidime, avibactam, and aztreonam standards were purchased from MedChemExpress. The resistance rates of XDR/PDR Pseudomonas aeruginosa to cefoperazone-sulbactam, imipenem, ciprofloxacin, piperacillin-tazobactam, ceftazidime, levofloxacin, meropenem, tobramycin, amikacin and gentamicin were 89.6%, 100%, 89.6%, 100%, 97%, 89.6%, 100%, 97.1%, 100%, 100%, respectively.We collected 67 d as PDR . All P. 2.2P. aeruginosa. A fixed concentration of AVI at 4 mg/L, 8mg/L, and 16mg/L combined with 2-fold diluted CAZ and ATM were used in ASTs. The quality control strains of our tests were E. coli ATCC 25922 and P. aeruginosa ATCC 27853. The MICs are defined as the lowest concentration of antibiotics that inhibits the growth of bacteria. The definition of MIC50 is a drug concentration that inhibits the growth of bacteria by 50%. Similarly, MIC90 is a drug concentration inhibiting 90% of bacterial growth. MIC distributions of CZA/AZA against P. aeruginosa were represented by cumulative inhibition rates (CIRs). Besides, all experiments were conducted three times following Clinical and Laboratory Standards Institute (CLSI) standards.We used the broth two-fold dilution method to determine the minimum inhibitory concentrations (MICs) of CZA/AZA against XDR/PDR 2.3P. aeruginosa were subjected to whole-genome sequencing (WGS) using Illumina MiSeq short-read sequencing . Sequenced isolates were evaluated using FASTQC, version 0.11.6, and MultiQC, version 1.6. Trimmomatic, version 0.39, removed adapters and trimmed low-quality paired end reads. Comprehensive Antibiotic Resistance Database v.1.2.0  was used to identify drug resistance genes in the strains.67 2.4fT > MIC is the best indicator for assessing BSIs. Besides, %50fT > 5 × MIC is the best indicator for assessing LRTIs. When combined with CAZ, %50fT > CT of 1 mg/L was considered the Pharmacokinetic (PK)/Pharmacodynamics (PD) target of avibactam. PK/PD targets of ATM for BSIs was %60fT > MIC. And PK/PD targets of ATM for LRTIs were %60fT > 5 × MIC. As for avibactam, %50fT > CT of 2.5 mg/L was considered appropriate for guiding dosage selection for AZA (fT > n × MIC equation was based on previous studies (Population pharmacokinetic (PPK) parameters of CZA and AZA were obtained from previously published articles . CAZ is  for AZA . Optimiz studies .2.5d, CL, t1/2) followed a log-normal distribution. All simulated dosing regimens via traditional infusion (TI)/optimized two-step-administration therapy (OTAT) were listed in MICi means each MIC value. p(MICi) means the percentage of each MIC value. A CFR ≥ 90% is adequate PD exposure for this dosing regimen.We conducted 10000-patient Monte Carlo simulations (MCSs) using Oracle Crystal Ball version.11.1.24. PK parameters  were listed in P. aeruginosa with an OXA-101 also produced OXA-850. CZA was as effective as AZA against these isolates. For isolates with an OXA gene plus IMP-45, VIM-1, or VIM-2, AZA was much more potent than CZA against these isolates, with all isolates being inhibited by a concentration of 8 mg/L. For isolates with an IND-6 plus more serine β-lactamases, all isolates produced CZA/AZA MICs of >64 mg/L. For XDR/PDR P. aeruginosa with other genotypes, the efficacy of CZA against these isolates did not differ significantly from that of AZA.The comparative MICs (mg/L) of CZA and AZA against 67 XDR/PDR 3.43.4.1The probability of target attainment (PTA) of each simulated dosing regimen for BSIs was listed in 3.4.2The PTA of each simulated dosing regimen for LRTIs was listed in 3.53.5.1via traditional infusion, the CFR was 95.63% for 4g q6h. If CZA was administered via OTAT, the CFR was 96.67% for 1.25g (0.5h) +1.25g (2h) q8h, 97.35% for 1.25g (0.5h) +1.25g (2h) q6h, 93.60% for 0.675g (0.5h) +0.675g (2h) q8h and 93.53% for 0.675g (0.5h) +0.675g (2h) q8h. If AZA was administered via traditional infusion, the CFR was 92.50% for 2.5g q6h and 96.36% for 4g q6h. If AZA was administered via OTAT, the CFR was 96.95% for 1.25g (0.5h) +1.25g (2h) q8h, 98.25% for 1.25g (0.5h) +1.25g (2h) q6h. The above dosing regimens are considered to provide adequate PD exposures for the treatment of CZA/AZA against XDR/PDR P. aeruginosa BSIs.The cumulative fraction of response (CFR) of each simulated dosing regimen for BSIs was listed in 3.5.2P. aeruginosa. CFRs were less than 90% for all simulated dosing regimens  + 1.25 g (2h)] q6h, 2.5 g  q8h, 4g  q6h, 4g  q8h).4P. aeruginosa has spread widely worldwide and the treatment of BSIs or LRTIs caused by XDR/PDR P. aeruginosa has become a tough problem . AVI is ineffective against MBL-producing isolates. However, the combination of AVI and ATM was a treatment option for MBL-producing isolates. Lee et al. also found that the combination of ATM and CZA may be a treatment option for VIM-2-producing P. aeruginosa . IND-6 is a highly divergent IND-type MBL. It was first isolated from Chryseobacterium indologenes strain 597 in Burkina Faso   . Besidesutations . Mobile fections . Our worinfusion . Besideshe blood .P. aeruginosa was confined to a small sample size and northern China. Secondly, our study only focused on partial beta-lactamases (class B β-lactamases and class D β-lactamases). Therefore, large-scale animal or clinical trials are needed in the future to confirm the efficacy of CZA/AZA against BSIs caused by XDR/PDR P. aeruginosa.Our study had several limitations that should be noted. Firstly, the collection of P. aeruginosa are intrinsic, mutational, and horizontally acquired resistomes (P. aeruginosa drug resistant. The efflux pump was also considered in the study of this article (see the whole genome sequencing results in the The main resistance mechanisms in sistomes . We founP. aeruginosa harbouring IMP-45, VIM-1, or VIM-2. Secondly, OTAT with sufficient PD exposure may be an optimal treatment option for BSI caused by XDR/PDR P. aeruginosa with a high-level MIC of CZA/AZA.In conclusion, our work has the following results. Firstly, AZA was considered as an option for the treatment of XDR/PDR The original contributions presented in the study are publicly available. This data has been deposited into the NCBI repository under accession: PRJNA967114.Ethical approval was granted by the Chinese People’s Liberation Army General Hospital.YK and JC designed and managed the project. YK performed all the experiments and wrote the manuscript. LX analyzed bioinformatics analysis. JY provided technical guidance. All authors contributed to the article and approved the submitted version.


Two novel reassortant highly pathogenic avian influenza viruses (H5N1) clade 2.3.4.4b.2 were identified in dead migratory birds in China in November 2021. The viruses probably evolved among wild birds through different flyways connecting Europe and Asia. Their low antigenic reaction to vaccine antiserum indicates high risks to poultry and to public health. Since the Gs/GD/96-lineage highly pathogenic avian influenza virus (HPAIV) (H5N1) was identified in 1996, H5 HPAIVs have evolved into divergent clades and caused continuous outbreaks in birds (https://wahis.woah.org/#/event-management). Human cases of H5N1/H5N6/H5N8 infection were sporadically documented , highlighting the zoonotic risk of H5 HPAIVs. Since 2021, H5 HPAIVs have caused at least 9 outbreaks in wild birds rather than poultry in mainland China . However, large outbreaks of H5N1 HPAIVs in domestic poultry were reported during 2021–2022 in Europe and the United States  . In this study, we explored the genetic origin, spread patterns, and antigenicity of H5N1 viruses identified from 2 dead migratory birds in China.Starting during 2020–2021, a new wave of HPAIV H5N1/H5N8 clade 2.3.4.4b outbreaks was reported in wild and domestic birds in Eurasia (http://www.mabsky.com).We collected oral swab specimens and lung tissues from a dead whooper swan in northern China (Inner Mongolia) on November 3, 2021, and a deceased black swan in eastern China (Zhejiang) on November 15, 2021. We performed virus isolation in 10-day-old specific pathogen-free chicken embryos  and GISAID . We reconstructed phylogenetic trees for each gene of the 3 H5N1 isolates together with reference viruses from GISAID and the National Center for Biotechnology Information (https://www.ncbi.nlm.nih.gov/genomes/FLU/Database/nph-select.cgi), using the maximum-likelihood method with a general time-reversible model plus gamma distribution in RAxML 8.2.12 (https://cme.h-its.org/exelixis/web/software/raxml) , A/whooper swan/Northern China/11.03 IMEEDSAK2-O/2021 (Ws/NC/AK2-O/2021), and A/black swan/Eastern China/11.15 ZJHZ74-Lg/2021 (Bs/EC/74-Lg/2021). We deposited whole genomes in NMDC (e/raxml)  Table. We/raxml) .https://www.who.int/publications/i/item/manual-for-the-laboratory-diagnosis-and-virological-surveillance-of-influenza) to test the reactivities of antiserum of H5 Re-11/Re-13/Re-14 vaccines against these new H5N1 isolates and H5N8 HPAIVs identified in 2020 (http://www.moa.gov.cn/govpublic).We performed hemagglutination inhibitor (HI) assays ; however, they have only been identified in wild birds in mainland China. Compared with high HI antibody titers (256) between homologous antiserum and antigens of H5 vaccines, the recent H5N1/H5N8 viruses presented low HI titers (2–16) against Re-11/Re-13 antiserum  and Bs/2021-like (Bs/EC/74-Lg/2021) reassortants  Figure 4We reconstructed the genetic reassortment history for these H5N1 viruses  clade 2.3.4.4b reassortants in migratory birds, China.Additional information for study of novel avian influenza virus (H5N1) clade 2.3.4.4b reassortants in migratory birds, China.


Integrase strand-transfer inhibitors (INSTIs) are associated with body weight gain among women living with HIV (WLH) in the Women’s Interagency HIV Study (WIHS). The role of antiretroviral therapy (ART) pharmacokinetics in INSTI-associated weight gain is unknown. We report the relationship between INSTI plasma concentrations and weight change in WLH.Model-adjusted estimates of percent body weight change by integrase inhibitor plasma concentrationModel-adjusted estimates of percent body weight change with 95% confidence band (y-axis) by integrase inhibitor plasma concentration in ng/mL (x-axis). Models were adjusted for baseline age, race/ethnicity, baseline obesity (body mass index ≥ 30 kg/m2), history of prior non-nucleoside reverse transcriptase use, and self-reported antiretroviral therapy adherence (100% vs <100%). Abbreviations: RAL=raltegravir, DTG=dolutegravir, EVG=elvitegravirData from 2006-2017 were analyzed from virologically suppressed (< 200 copies/mL) WLH in the WIHS who switched/added raltegravir (RAL), dolutegravir (DTG), or elvitegravir (EVG) to ART. Percent body weight change was calculated using weights obtained 6-12 months pre- and post- INSTI switch/add. INSTI random plasma concentrations were measured with validated LC/MS-MS assays 6-12 months post INSTI switch/add. Area under the curve (AUC) was calculated using estimates from published models and updating individual-level parameters with plasma concentrations. Linear models assessed the relationship between plasma concentrations or AUC and weight change for each INSTI separately. Models were adjusted for age, race/ethnicity, baseline obesity, history of non-nucleoside reverse transcriptase use, and self-reported ART adherence.2 (±8.8), and 57% reported 100% ART adherence. Mean percent body weight change was +2.3% (±6.0) for RAL, +1.8% (±9.0) for DTG, and +2.8% (±10.5) for EVG. AUC was able to be calculated in 167 (95%) of women. In adjusted linear models, higher RAL plasma concentration was associated with greater percent body weight change (p=0.016), Figure 1. No association of drug AUC with body weight change for any INSTI was observed.176 WLH contributed plasma drug concentrations with a mean ±SD 1.5 (±0.1) years follow-up: 43 (24%) were on RAL, 87 (49%) on DTG, and 47 (27%) on EVG. Mean age was 51 years (± 9), 64% were Non-Hispanic Black, baseline BMI was 31.6 kg/mIn WLH, random RAL plasma concentrations, but not the RAL AUC, was associated with body weight gain. In contrast, the effect of DTG and EVG plasma drug exposure on short-term body weight change appears to be limited. Mechanisms contributing to body weight gain may be INSTI-specific. In addition to further pharmacologic assessments, other mechanisms to explain INSTI-associated weight gain should be examined.Cecile D. Lahiri, MD, MS, Merck: Grant/Research Support|Theratechnologies, LLC: Advisor/Consultant|Theratechnologies, LLC: Honoraria Julie B. Dumond, PharmD, Merck: Grant/Research Support Cyra Christina Mehta, PhD, MSPH, Merck: Grant/Research Support Igho Ofotokun, MD, MSc, FIDSA, Merck: Grant/Research Support Maria L L. Alcaide, MD, Discidium Biosciences: Board Member|Gilead: Honoraria|Merk & Co: Honoraria|Senhwa Biosciences: Honoraria|Virology Education: Honoraria Jordan Lake, MD, Gilead Sciences: Grant/Research Support|Theratechnologies: Advisor/Consultant Phyllis C. Tien, MD, MSc, Merck: Grant/Research Support


Rheumatology key messageVEXAS syndrome is venulitis with severe thickening of the walls of large veins.Dear Editor, VEXAS  syndrome is an often fatal, X-linked disease characterized by haematological dysplasia overlapping with severe autoinflammatory manifestations. Acquired somatic mutations of the ubiquitin-like modifier activating enzyme (UBA1) gene impairs ubiquitylation, causing activation of the innate immune system and release of pro-inflammatory cytokines, TNF-α, IL-8, IL-6, IFN-γ and IFN-inducible protein-10 (IP-10) that culminate in severe systemic inflammation , which is independent of age, disease duration, history of thrombosis and major organ involvement, disease activity, acute phase response and corticosteroid treatment [s.d. 0.1)] but not significant with APS [0.49 mm (s.d. 0.1)] [Rheumatology online).Previous studies have shown that venous inflammation causes VWT . Varicosm s.d. 0.4 vs 0.27reatment . Additiom s.d. 0.4 vs 0.27The pathogenesis of organ inflammation in VEXAS syndrome is not entirely understood. UBA1 is essential for the initiation of ubiquitylation, a widely used post-translational protein modification mechanism that regulates many biological processes, including intracellular signalling, proteasomal/lysosomal protein degradation and antimicrobial defence . Consequkead168_Supplementary_DataClick here for additional data file.


Correction to: Clinical Proteomics (2022) 19:4810.1186/s12014-022-09385-7Unfortunately, in the original publication of the article, the following errors were identified after online publication of the article.The Additional file In Abstract, line 11, the text that reads as “phospho-heavy-labeled-spiketide FAIMS Stepped-CV DDA (pHASED)” should read as “hospho-heavy-labeled-spiketide FAIMS stepped-CV DDA (pHASED)”.The original article has been corrected.Additional file 2: Table S1. SBDS heavy-labeled phosphorylated peptide standards. Table S2. Common and unique phosphoproteins identifed across all four CVs based on PSM acquisition. Table S3. High confdence modifcation sites identifed in LFQ (p < 0.01). Table S4. High confdence modifcation sites identifed in pHASED (p < 0.01). Table S5. Unique and common phosphoproteins identifed in LFQ and pHASED datasets. Table S6. Phosphorylated master protein kinases identifed in LFQ dataset (p < 0.01). Table S7. Phosphorylated master protein kinases identifed in pHASED dataset (p < 0.01). Table S8. FLT3-D835 mutations associated with resistance to tyrosine kinase FLT3 inhibitors. Table S9. Kinase-Substrate analysis of LFQ dataset for resistant cells in comparison to FLT3-ITD (log2 fold change±0.5). Table S10. Kinase-Substrate analysis of pHASED data?set for resistant cells in comparison to FLT3-ITD (log2 fold change±0.5). Table S11. Canonical pathways identifed as signifcantly associated with LFQ dataset for resistant cells in comparison to FLT3-ITD. Table S12. Canonical pathways identifed as signifcantly associated with pHASED dataset for resistant cells in comparison to FLT3-ITD. Table S13. Kinase activity inferred by KSEA analysis of phosphorylation changes in pHASED dataset  for resistant cells in comparison to FLT3-ITD. Table S14. Mutation-specifc response to sorafenib. IC50 compared to FLT3-ITD. Table S15. Bliss Synergy scores for sorafenib in combination with KU-60019 at diferent doses. Table S16. Unique ATM substrates identifed with increased phosphorylation (log2≥0.5) in pHASED dataset for resistant cells in comparison to FLT3-ITD. Table S17. Vector mutations in FLT3 gene.


Correction to: Sport Sciences for Health 10.1007/s11332-022-01012-0The reference no. 4 has been incorrectly published in the original publicaiton. The complete correct reference [4] is given below.Coronavirus disease (COVID-19): the need to maintain regular physical activity while taking precautions. J Sport Health Sci 9(2):103.4. Chen P et al (2020)


Korean J Women Health Nurs 2023;29(1):66-75.https://doi.org/10.4069/kjwhn.2023.03.15This corrigendum is for correcting a reference that was mistakenly reported in the above article.4. Pereira-Kotze C, Feeley A, Doherty T, Faber M. Maternity protection entitlements for non-standard workers in low-and-middle-income countries and potential implications for breastfeeding practices: a scoping review of research since 2000. Int Breastfeed J. 2023;18(1):9. https://doi.org/10.1186/s13006-023-00542-8The corrected reference is as below. The authors apologize for any inconvenience that this may have caused.4. Brandhagen M, Lissner L, Brantsaeter AL, Meltzer HM, Häggkvist AP, Haugen M, et al. Breast-feeding in relation to weight retention up to 36 months postpartum in the Norwegian Mother and Child Cohort Study: modification by socio-economic status? Public Health Nutr. 2014;17(7):1514-1523. https://doi.org/10.1017/s1368980013001869.


Cytomegalovirus (CMV) infection is associated with significant morbidity and mortality among kidney transplant recipients (KTR). Given toxicities and costs of antivirals, there is growing interest in assays that measure CMV-specific T-cell immunity (TCI), which may predict protection against clinical infection. The Viracor® CMV TCI Panel (TCIP) uses flow cytometry and intracellular cytokine staining to measure %CMV-specific CD4+ and CD8+ T-cells; it is the only commercially available TCI assay in the US. In this investigator-initiated study, we evaluated the performance of the TCIP in predicting clinically significant CMV events. To our knowledge, this is the first prospective blinded clinical study of the TCIP.We enrolled consecutive donor (D) or recipient (R) CMV seropositive KTR, who had TCIP testing monthly until either discontinuation of valganciclovir prophylaxis or the primary outcome. We also enrolled KTR with low-level untreated DNAemia, or after completion of valganciclovir treatment, to evaluate assay predictive values for progression or relapse of CMV infection, respectively. The primary outcome was CMV DNAemia (tested at the discretion of the treating provider for symptoms or laboratory findings), prompting treatment initiation. Secondary outcomes were any DNAemia and DNAemia >1000 IU/mL. Treating providers were unaware of TCIP results.p=0.024), and the CMV protection ROC AUC was significant . The positive predictive values of both CD4+ and CD8+ T-cell positivity >0.2% for CMV protection were 96.3% (95%CI 81-99.9%) for the primary outcome, 92.6% (95%CI 75.7-99.1%) for any DNAemia, and 100% (one-sided 95%CI 87.2%) for DNAemia >1000 IU/mL.We included 46 KTR. Median age was 50.5 (range 18-74) years. 73.6% identified as men, 10.8% Black, 8.7% Hispanic; 41.3% had CMV D+/R- status. KTR with CMV events had significantly lower %CMV-specific CD8+ T-cells  (5R25AI140490) Dimitrios Farmakiotis, M.D., Astellas: Grant/Research Support|Merck: Grant/Research Support|Viracor: Advisor/Consultant|Viracor: Grant/Research Support


QTRL.caas-4A.1 and QTRL.caas-4A.2), TRA (QTRA.caas-4A and QTRA.caas-4D), and NRT (QNRT.caas-5B and QNRT.caas-5D) were identified and each explaining 5.94%–9.47%, 6.85%–7.10%, and 5.91%–10.16% phenotypic variances, respectively. Among these, QTRL.caas-4A.1 and QTRA.caas-4A overlapped with previous reports, while QTRL.caas-4A.2, QTRA.caas-4D, QNRT.caas-5B, and QNRT.caas-5D were novel. The favorable alleles of QTRL.caas-4A.1, QTRA.caas-4A, and QTRA.caas-5B were contributed by Doumai, whereas the favorable alleles of QTRL.caas-4A.2, QTRA.caas-4D, and QTRA.caas-5D originated from Shi 4185. Additionally, two competitive allele-specific PCR (KASP) markers, Kasp_4A_RL (QTRA.caas-4A) and Kasp_5D_RT (QNRT.caas-5D), were developed and validated in 165 wheat accessions. This study provides new loci and available KASP markers, accelerating wheat breeding for higher yields.Identifying loci for root system architecture (RSA) traits and developing available markers are crucial for wheat breeding. In this study, RSA-related traits, including total root length (TRL), total root area (TRA), and number of root tips (NRT), were evaluated in the Doumai/Shi4185 recombinant inbred line (RIL) population under hydroponics. In addition, both the RILs and parents were genotyped using the wheat 90K single-nucleotide polymorphism (SNP) array. In total, two quantitative trait loci (QTLs) each for TRL ( Wheat production is influenced seriously by abiotic stresses. Breeding higher-yielding and more stable accessions under abiotic stress is a crucial objective in modern wheat breeding . Root syPrevious breeding programs predominantly focused on aboveground traits such as disease resistance, grain quality, and harvest index, while the application of RSA traits has been limited due to the complexity of phenotypic evaluation . PreviouIn this study, we conducted linkage mapping for RSA traits using the wheat 90K assays in a biparental recombinant inbred line (RIL) population derived from the Doumai/Shi 4185 cross. The primary objective of this study is to uncover the genetic basis of these traits and develop available KASP markers for improving wheat RSA.2:6 RILs derived from the Doumai/Shi 4185 were used for evaluating RSA-related traits. A hydroponic experiment was conducted with three replicates in a greenhouse. There were 20 seeds from each line that were surface sterilized with10% H2O2 for 20 min. Subsequently, the cleaned seeds were placed in Petri dishes with moist filter paper. When the coleoptiles reached about 2 cm in length, wheat seedlings were transferred to plastic trays (53 × 27 cm) containing Hoagland’s nutrient solution. Plastic trays were kept in 25°C with a 16-h light and 8-h darkness cycle. After 3 weeks, roots were evaluated for RSA-related traits .Three RSA-related traits, namely, total root length (TRL), total root area (TRA), and number of root tips (NRT), were assessed using the WinRHIZO root analysis system (LA6400XL). The roots were arranged systematically in a dish and scanned using the Expression 11000XL. Subsequently, the images were imported into WinRHIZO software and analyzed using a fixed threshold parameter of 40. Each accession was scored on five plants for RSA traits, and means of three replicates were obtained. Basic statistical analyses and frequency distributions were conducted using SAS v9.3  by CapitalBio Corporation. SNPs with missing data >20% or minor allele frequency (MAF) <0.5 were filtered for further analysis. The filtered SNPs were then analyzed using the BIN function of IciMapping v4.2  and grouThe inclusive composite interval mapping (ICIM) method using IciMapping v4.1  was applQTRL.caas-4A.1, QTRA.caas-4A, and QNRT.caas-5D) were converted to KASPs (http://www.polymarker.info/) (SNPs flanking QTL with higher PVE (to KASPs  and desir.info/) Table S2r.info/) .https://wheat.pw.usda.gov/GG3/). Genes, excluding hypothetical proteins, transposon proteins, and retrotransposon proteins with SNPs in the coding region, were considered as candidate genes. Quantitative real-time PCR (qRT-PCR) was conducted to test expression differences of the candidate genes between Doumai and Shi 4185. The roots were sampled for RNA extraction after phenotyping. RNA was extracted using the TRIzol method, and the cDNA was synthesized with the HiScript II 1st Strand cDNA Synthesis Kit. Primers were designed using the Primer Premier V5.0. PCR was conducted with a mixture of 20 μl, including 2 μl cDNA, 10 μl ChamQ Universal SYBR qPCR Master Mix, and 0.4 μl of each primer. qRT-PCR was conducted in the ABI StepOnePlus Real-Time PCR System with Tower, and the gene expression level was analyzed by the 2–ΔΔCT method. All assays were performed in two biological replicates and three technical replicates. TaActin1 was used as the internal control to normalize the expression levels of different samples.To identify candidate genes involved in the QTL for RSA-related traits detected in the Doumai/Shi 4185 RIL population, the genes located in the LD block region around the peak SNP  of each QTL were extracted from the wheat IWGSC v1.1 annotation (2 (range: 6.9 cm2–15.2 cm2), and 328.2 root tips (range: 94.7–772.0). The standard deviation and coefficient of variation for TRL, TRA, and NRT were 18.1 cm (24.2%), 1.7 cm2 (16.4%), and 126.4 (38.5%), respectively. A significant correlation was observed between TRL, TRA, and NRT, with the correlation coefficient of 0.603 (P < 0.05) between TRL and TRA, 0.371 (P < 0.05) between TRL and NRT, and 0.312 (P < 0.05) between TRA and NRT.All three RSA-related traits exhibited continuous and significantly wide variation across the 262 RILs (QTRL.caas-4A.1 (BobWhite_c20306_147-Tdurum_contig54973_1510) and QTRL.caas-4A.2 (BS00059454_51-Kukri_c19883_816), respectively. These QTLs explained 5.94% (additive effect: 5.17) and 9.45% (additive effect: −7.12) of the total phenotypic variances  and QTRL.caas-4D (Ex_c6665_1067-wsnp_J-D_rep_c51623_35119179), respectively, explaining 7.10% (additive effect: 0.40) and 6.85% (additive effect: −0.39) of the total phenotypic variance. The favorable allele of QTRA.caas-4A TRA was contributed by Doumai, while the favorable allele of QTRA.caas-4D was originated from Shi4185, respectively. Two QTLs for NRT were detected on chromosomes 5B and 5D and named as QNRT.caas-5B (wsnp_Ex_rep_c67320_65870601-Tdurum_contig10191_996) and QNRT.caas-5D (IAAV6218-Kukri_c46526_103), respectively, explaining 5.91% (additive effect: 26.7) and 10.16% (additive effect: 39.1) of the total phenotypic variances. The favorable alleles of QNRT.caas-5B and QNRT.caas-5D were contributed by Doumai and Shi 4185, respectively  with higher phenotypic effects were used to develop KASP markers. Although attempts were made to develop a KASP marker for QTRA.caas-4A, it was unable to effectively distinguish between the two parental genotypes in the RIL population. Therefore, the marker did not yield conclusive results. Consequently, two KASP markers, Kasp_4A_RL  and Kasp_5D_RT , were successfully developed based on the tightly linked SNP markers  exhibited higher TRL compared to the unfavorable allele  at the P = 0.05 level  showed higher NRT than unfavorable allele  at the P = 0.05 level  encoded an ethylene-regulated nuclear protein (ERT2)-like protein; TraesCS4A01G436700 (QTRL.caas-4A.1) encoded a calcium-binding protein kinase (CDPK)-related kinase; TraesCS4A01G456300 (QTRL.caas-4A.1) encoded a cellulose synthase-like protein; TraesCS4A01G469400 (QTRL.caas-4A.2) encoded an F-box protein; and TraesCS4D01G092000 (QTRA.caas-4D) encoded an ABC transporter, whereas both the TraesCS5D01G378400 and TraesCS5D01G380200 for QNRT.caas-5D encoded E3 ubiquitin-protein ligase. The expressions of the seven candidate genes in Doumai and Shi 4185 were detected using qRT-PCR. Of these, TraesCS4A01G469400, TraesCS4D01G092000, and TraesCS5D01G380200 showed no significant differences between the parents, whereas TraesCS4A01G296500, TraesCS4A01G436700, TraesCS4A01G456300, and TraesCS5D01G378400 showed more than 1.4–3.9-fold higher expression in Doumai compared to Shi4185 , TRA (QTRA.caas-4A and QTRA.caas-4D), and NRT (QTRA.caas-5B and QTRA.caas-5D) TRA. Each of these QTLs accounted for 5.94%–9.47%, 6.85%–7.10%, and 5.91%–10.16% of the phenotypic variance, respectively.Optimization of crop root systems has long been proposed. However, genetic improvement of crop roots has been rarely attempted . A compr9 RIL lines and have identified 19 QTLs mainly distributed on chromosomes 1A, 2B, 2D, 3A, 3B, 3D, 5A, and 5D. Of these, the 5D loci (415.8 Mb–457.6 Mb) overlapped with theQNRT.caas-5D (449.4 Mb–454.1 Mb) identified in this study. IWB21309, 17.01 Mb) and 5B  for RSA traits were also different with the loci identified in this study . Furthermore, MQTL4A.1 (651.78 Mb–705.73 Mb) and MQTL4A.6 (600.04 Mb–691.14 Mb) significantly associated with root length, root surface, and root tips overlapped with QTRL.caas-4A.1 (702.3 Mb–721.3 Mb) and QTRA.caas-4A (594.2 Mb–602.9 Mb). Rht2/Rht10 19.18 Mb, SVP3-4D/BM1-4D 469.46 Mb, Vrn2-4D/ZCCT1-4D 509.43 Mb), 5B , and 5D   are different from the reported wheat genes associated with plant height and vernalization. Therefore, they represent novel genes associated with RSA-related traits. Above all, compared with the previous results and meta-analysis, QTRL.caas-4A.2, QTRL.caas-4D, QNRT.caas-5B, and QNRT.caas-5D were novel.0.00 Mb) . Based oQTRL.caas-4A.1 (702.3 Mb–721.3 Mb) co-located influencing QTL clusters related to thousand kernel weight   co-located with regions affecting QTL clusters related to yield, including SN, KNS, TKW, FLW, and KL   . Additio65.6 Mb) . These rTraesCS4A01G436700 of QTRL.caas-4A.1 encodes the CDPK-related kinase. In plants, CDPKs play a critical role in various signaling pathways and are pivotal in root growth and development -like protein. Ethylene, a simple small molecule, plays a vital role in signal transduction, cell differentiation, division, and elongation (TraesCS5D01G380200, linked to QNRT.caas-5D, encodes an E3 ubiquitin-protein ligase, which plays essential roles in plant development (Four genes involved in the biological metabolism of plant hormones, cellulose, and the ubiquitin pathway were identified as high confidence candidate genes. Among these, elopment . Silencil length . TRA Anotivities . TraesCSongation . Ethylenongation . Additioelopment  and is celopment .Kasp_4A_RL and Kasp_5D_RT were successfully developed based on tightly linked SNP markers, proving to be valuable tools for MAS in breeding programs. Additionally, accessions with more favorable alleles, superior RSA traits and appropriate agronomic traits, such as Yumai 18, Jinhe 9123, Yumai 34, Bainong 3217, Lumai 6, Jinmai 61, Lumai 5, Lumai 23, and Yumai 57 are recommended as parental lines for improvement of RSA traits.Although conventional breeding has contributed to the enhancement of root system, the selection process is time-consuming and less efficient due to the challenges in field measurements of RSA traits . KASP ofIn this study, we identified six QTLs associated with wheat RSA traits and successfully developed two KASP markers that can be utilized in wheat breeding programs aimed at achieving higher and more stable yields. This research serves as a foundational step towards gene cloning and enhancement of wheat root systems.All datasets generated for this study are included in the article or YJ: Data curation, Funding acquisition, Writing – original draft. YW: Data curation, Software, Validation, Writing – original draft, Writing – review & editing. JL: Software, Validation, Writing – review & editing. FW: Formal Analysis, Resources, Software, Investigation, Writing – review & editing. XQ: Investigation, Project administration, Writing – review & editing. PL: Funding acquisition, Supervision, Validation, Writing – original draft.


PrEP with T/C can prevent COVID-19 hospitalization and death in IC patients (pts) up to 6 months after injection. However, in the USA, authorization of T/C PrEP was paused in Jan 2023 due to loss of in vitro activity of T/C against dominant circulating SARS-CoV-2 variants, although loss of clinical efficacy is unclear. We investigated in vivo mechanisms of viral breakthrough in hospitalized IC pts with vs without prior T/C exposure.We analyzed remnant clinical SARS-CoV-2 PCR-positive swabs and sera from IC pts hospitalized at UPMC. SARS-CoV-2 variants and mutants were determined by whole genome sequencing and anti-RBD IgG levels by an enzyme immunoassay.Table 1). Median age was 67 yrs; 49% were male. IC conditions included organ transplant (23%) and hematologic cancer (32%) (Table 2). In IC patients with sequenced swabs, 21% received T/C (Table 3). Variant frequency mirrored national trends (Table 3). BA.5, XBB.1, and BF.7 were less common in T/C vs non-T/C pts . BA.2 and BQ.1 were more common in T/C vs non-T/C pts . The R346T and K444T/R/N mutations were more common in T/C vs non-T/C pts: 54% vs 41% and 37% vs 22% (Table 3). Anti-RBD IgG titers from 56% pts at the time of infection were higher in T/C vs non-T/C pts . COVID-19 mortality was numerically lower in T/C vs non-T/C pts . Mortality differences were consistent across variant epochs (Table 1).From Mar 28, 2022, to Mar 3, 2023, 72% (174/243) of swabs were successfully sequenced from 170 pts : Advisor/Consultant|LabSimply: Advisor/Consultant|Merck: Advisor/Consultant|Shionogi: Advisor/Consultant|Shionogi: Honoraria John W. Mellors, MD, AstraZeneca: Grant/Research Support|Gilead Sciences: Grant/Research Support


The identification and characterization of plasma proteins in drug resistant and drug sensitive in HIV-1 infected/AIDS patients were carried out using the SWATH-MS protocol. In total, 204 proteins were identified and quantified, 57 proteinswere differentially expressed, out of which 25 proteins were down regulated and 32 proteins were up regulated in drug resistant patients. Six proteins such as complement C4-A, immunoglobulin heavy variable 1-2, carboxylic ester hydrolase, fibulin-1,immunoglobulin lambda constant7, secreted phosphoprotein 24 were differentially expressed in individuals with drug resistant HIV as compared to individuals with drug sensitive HIV. Gene ontology of 57 differentially expressed proteins was analysedand documented. The analysis of HIV-1 infected human plasma is an attractive medium with which differentially expressed proteins could be identified in certain disease states. Proteomic techniques are widely used globally to identify differentiallyexpressed plasma proteins in response to HIV-1 infection. Sequential window acquisition of all theoretical mass spectra (SWATH-MS) is a novel technique conceptualized with the protein library and the individual protein was identified andquantified for biomarker discovery . TherefoAmmonium formate, formic acid, dithiothreitol (DTT) and iodoacetamide (IAA) were procured from Sigma, USA. Modified trypsin (sequencing grade) was procured from Promega, USA. Polysulfoethyl SCX cartridge  withcartridge holder was procured from Sciex, USA. Acetonitrile, Liquid phase-chromatography-mass spectroscopy grade water, was procured from Biosolve . Analytical grade chemicals were used for this study.The study focused on patients administered six first-line ART such as ZLE, ZLN, TLE, TLN, SLE and SLN patients and they were enrolled in the anti-retroviral therapy centre, Sarojini Naidu Medical College, Agra, India from December 2009 to November2016 as per the treatment guidelines provided by NACO, Govt. of India and the ethical guidelines formulated by the ICMR, Government of India. The age of the six patients varied from 32 years to 43 years. A patient information leaflet was used for datacollection  hsa04610: Complement and coagulation cascades - Homo sapiens (human)[2] hsa05322: Systemic lupus erythematosus - Homo sapiens (human)[3] hsa05133: Pertussis - Homo sapiens (human)[4] hsa05150: Staphylococcus aureus infection - Homo sapiens (human)[1] H00080: Systemic lupus erythematosus[2] H00102: Classic complement pathway component defects[3] H01649: SchizophreniaThe six-drug respondent and drug resistant patients were in first-line ART such as ZLE, ZLN, SLN, TLE, and TLN during their course of treatment in this period. The drug interactions with these proteins were analysed (www.dgidb.org). A totalThe serial number of the down regulated and up regulated proteins in the heat map is represented in the Gene symbol: 1.C4A 2. IGHM 3. APOA4 4.HBB, 5. SERPING1 6. PZP 7. IGHG2 8.CD5L 9.HPR 10. HBA1 11. JCHAIN 12. APOL1 13. IGHV1-2 14. IGHV3-9 15.IGHV5-51 16. C4B 17. BCHE 18. LPA 19. IGLV6-57 20. SERPINA1 21. IGKV2-24 22. IGLV3-10 23. FAT-1 24.BIN3 25. PROC 26. FN1 27. IGHA1 28. GSN 29. AFM 30. TTR 31. IGLV3-21 32. KRT1 33. IGLV3-19 34. IGKV3-15 35. C4BPB 36. FBLN1 37. IGKV1-5 38. F5 39. IGKV3-11 40.IGLV7-46 41. VNN1 42. IGLC7 43. IGHA2 44. IGHV3-13 45. IGHV1OR15-1 46. SPP2 47. CDKL3 48. IGLV10-54 49. CDC42BPA 50. SP5 51. BEX5 52. PEPD 53. IGHV6-1 54. IGHV1-45 55. CDH5 56. 3 57. SERPINA 10. The heat map of differentially expressed proteins was createdby the online tool (http://www.heatmapper.ca/expression).The gene list for a category of the molecular function of the differential expressed proteins are presented in the bar diagram. The number of genes involved in the different functional role is presented in serial number in the bar diagram1. Binding (GO:0005488), 2. Catalytic activity (GO:0003824), 3. Molecular function regulator (GO:0098772), 4. Molecular transducer activity (GO:0060089), 5. Transcription regulator activity (GO:0140110), 6. Transporter activity (GO: 0005215).The molecular function of the differential expressed proteins was created by the online tool PANTHER- gene analyst ( http://www.pantherdb.org ).The red colour dots represent the down-regulated proteins and black colour dots represent the up-regulated proteins . The volcano plot was created by Graph Pad Prism 8, USA( https://www.graphpad.com /scientific-software/prism/).The details description of the protein-protein interaction network nodes represents proteins: Splice isoforms or post translational modifications are collapsed i.e. Each node represents all the proteins produced by a single protein-coding gene locus.The edges represent protein-protein associations: Network status: number of nodes: 35, number of edges:80, average node degree: 4.57, average local clustering coefficient:0.453, expected number of edges: 7, PPI enrichment p-value:< 1.0e-16. Theprotein-protein interaction network was created by using the online tool STRING Version 11.0 (https://string-db.org).The differential expression and molecular function of the Serotransferrin and Apolipoprotein A1 was identified in HIV-1 patients treated with first line ART . Now, thWe are report that proteins identified in drug resistant and drug sensitive HIV-1 infected patients who are taking first line ART over 6 years by SWATH-MS. Plasma proteins such as complement C4-A, immunoglobulin heavy variable 1-2, carboxylicester hydrolase, fibulin-1, immunoglobulin lambda constant7, secreted phosphor protein 24 were differentially expressed in individual drug resistant HIV-1 subtype C patients. The role of these proteins is reported using gene ontology analysis. Thisstudy would be helpful for further implementation in the search for biomarker discovery in HIV-1/AIDS patients taking first line ART.The project "Characterization of drug resistant HIV-1 mutants of Agra region, India by genomic and proteomic approaches" funded for senior research fellowship of Sushanta Kumar Barik (File No. 80/990/2015-ECD-I). M. M. Alam, JALMA, ICMR and Rakesh KumarMishra, ART centre, S.N Medical College are acknowledged for providing technical help in the collection of blood samples.


Kachinia Tong & Li, 2018 and Promolotra Tong & Li, 2020 are recorded from China for the first time. Two new species, Kachinialonglingsp. nov. (♂♀) and Promolotralushuisp. nov. (♂♀) are described. Descriptions, diagnoses, photographs and keys to Kachinia and Promolotra species are provided.The genera  Oonopidae Simon, 1890 includes 1893 extant species in 115 genera worldwide, mainly distributed in tropical regions  were taken under high vacuum with a Hitachi S-4800 after critical-point drying and gold-palladium coating. All measurements were taken using an Olympus BX51 compound microscope and are in millimeters. The terminology used in the text and figures follows SYNU) in Shenyang, China (curator: Yanfeng Tong).The specimens were examined using a Leica M205C stereomicroscope. Details were studied under an Olympus BX51 compound microscope. Photos were made with a Canon EOS 750D zoom digital camera (18 megapixels) mounted on an Olympus BX51 compound microscope. Endogyne were cleared in lactic acid. Scanning electron microscope images , Myanmar.Taxon classificationAnimaliaAraneaeOonopidaeTong & Zhangsp. nov.83FC57DD-B207-51D0-B931-DD9CFD1538A0https://zoobank.org/651BE492-19F3-480A-9EB7-CF6675FCFF21Holotype ♂ (SYNU-640), China, Yunnan Prov., Baoshan City, Longling Co., Longxin Town, Xiaoheishan Nature Reserve, 16.02.2011, Z. Li & L. Wang; Paratypes: 1♀ (SYNU-641), 1♀ (SYNU-642), 4 ♀ (SYNU-643-646), same data as for the holotype.K.mahmolae and K.putao by the strongly elevated epigastric region of the male . Habitus as in Fig. Female. As in male except as noted. Habitus as in Fig. The specific name is a noun in apposition taken from the type locality.Known only from the type locality.Taxon classificationAnimaliaAraneaeOonopidaeGenusTong & Li, 2020956B765C-34F2-50EC-8981-3DD686DC17FAPromolotrashankhaung Tong & Li, 2020 from Myanmar.Oonopinae Simon, 1890. According to Molotra Ubick & Griswold, 2011 by the heavily sclerotized dorsal and ventral abdominal scuta, the long spines on legs I and II, and the embolar region.The genus belongs to the subfamily Promolotrahponkanrazi Tong & Li, 2020, P.lushui sp. nov., P.shankhaung Tong & Li, 2020.China (Yunnan), Myanmar.Taxon classificationAnimaliaAraneaeOonopidaeTong & Zhangsp. nov.0111FE05-4B7A-59C2-8DCA-D3F41A907301https://zoobank.org/E02029CB-E8DA-471D-9B13-A5D39E3E20C0Holotype ♂ (SYNU-647), China: Yunnan Prov., Lushui City, Pianma Town, 3.03.2011, Z. Li & L. Wang leg.; Paratypes: 2 ♀ (SYNU-648-649), 3 ♂ (SYNU-650-652), same data as for the holotype.The new species can be distinguished from congeners by the blunt end of the cymbiobulb . Habitus as in Fig. Female. As in male except as noted. Habitus as in Fig. The specific name is a noun in apposition taken from the type locality.Known only from the type locality.


Nirmatrelvir/ritonavir administered twice daily for 5 days (5D) (within 5 days of symptom onset) resulted in a clinically and statistically significant (6% absolute and 86% relative) risk reduction in COVID-19 related hospitalization or all cause death through Day 28 [1]. COVID-19 rebound regardless of treatment has been reported . Here, an integrated analysis of EPIC-HR/SR  virology data was conducted to evaluate clinical resistance to nirm/r and if it is associated with viral load rebound (VLR) or progression to severe COVID-19 .10 copies/mL. The amino acid (AA) sequence was compared to Wuhan-Hu-1 [7] and Mpro substitutions were called if AAFREQ ≥ 10%. Emergent substitutions (ES) were called if observed post-baseline only and called treatment emergent substitutions (TES) if the ES was ≥ 2.5-fold more frequent, and had ≥3 additional occurrences, in nirm/r than placebo (PBO). Viral RNA load rebound (VLR) was defined as VL increase at D10 or D14 by ≥ 0.5 log10 copies/mL from D5 and resulting in a VL ≥ 3.0 log10 copies/mL.Next generation sequencing analysis was performed from nasal swabs  with a viral RNA level ≥3.0 logpro ES did not differ between treatment arms. Among those with sequence data available , nirm/r Mpro TES were observed in: T98I/del(n=3), E166V (n=3), and W207L/del (n=3) and within the Mpro cleavage sites: A5328S/V(n=5) and S6799A/P/Y (n=4). None of the TES were associated with progression to severe COVID-19. VLR was observed in  with Mpro TES observed in few VLR patients (3.4% in nirm/r and 0% in PBO). Among the Mpro TES identified, E166V is an in vitro resistance mutation . Figure 1 shows the VL trajectories of the 3 patients with E166V, one patient experienced VLR at D10, all effectively controlled the virus by D14 and did not experience severe COVID-19.In EPIC-HR/SR, the primary SARS-CoV-2 variant was Delta (91%) followed by Omicron (7.4%). The percentage of patients with Min vitro resistance substitution emerged in few patients, but was not associated severe COVID-19. Additionally, these data show VLR is not a result of treatment resistance to nirm/r driven by TES or any Mpro substitutions.In EPIC-HR/SR, an Mary Lynn Baniecki, PhD, Pfizer Inc: Employee|Pfizer Inc: Stocks/Bonds Shunjie Guan, PhD in Statistics, AbbVie: Stocks/Bonds|Pfizer: Stocks/Bonds Zhenyu Wang, Ph.D., Alkermes: Stocks/Bonds|Moderna: Stocks/Bonds|Pfizer: Stocks/Bonds Yan Chen, Ph. D, Pfizer, Inc: Stocks/Bonds Weihang Bao, PhD, Pfizer, Inc.: Employee of Pfizer|Pfizer, Inc.: Stocks/Bonds Wen He, PhD, Pfizer Inc.: Stocks/Bonds Elizabeth Dushin, PhD, Pfizer Inc.: Stocks/Bonds Craig Hyde, PhD, Pfizer: Employed by Pfizer|Pfizer: Stocks/Bonds Yuao Zhu, PhD, Pfizer, Inc.: Stocks/Bonds Rhonda D. Cardin, PhD, Pfizer, Inc: Stocks/Bonds Jennifer Hammond, PhD, Pfizer: Employee|Pfizer: Stocks/Bonds Sandeep Menon, PhD, Pfizer: Stocks/Bonds Charlotte Allerton, PhD, Pfizer: Stocks/Bonds Holly Soares, PhD, Pfizer: Stocks/Bonds


Enterococcus faecalis.The whole-genome sequence of strain K-4, isolated from grass silage in Thailand, which constitutes a chromosome and two plasmids, is 2,914,933 bp long, has a GC content of 37.5%, and contains 2,734 predicted protein-coding genes. Average nucleotide identity based on BLAST+ (ANIb) and digital DNA-DNA hybridization (dDDH) values indicated that the strain K-4 was closely related to  Enterococcus comprises Gram-positive, non-spore-forming facultative anaerobic, spherical, or ovoid cells with low genomic GC content   were assembled de novo using Unicycler v0.4.8 (https://github.com/rrwick/Unicycler) (dnaA for the chromosome. The genome sequence was annotated with DFAST v1.2.6 (https://dfast.nig.ac.jp/)  and the Genome-to-Genome Distance Calculator 3.0 (http://ggdc.dsmz.de/ggdc.php)  and decoded on the Illumina MiSeq system. Overall, 1,192,876 reads with 299,930,918 bases were decoded with an average read length of 251.4 bp and spot length of 602 bp with 2 × 301-bp paired-ends reads. Low-quality bases  were trimmed, and short reads (<25 bp) were removed using Platanus trim v1.0.7 (us_trim) . Data reus_trim)   . Averagegdc.php)  using reE. faecalis ATCC 19433T  with a sequence depth of ~88. Overall, 2,734 coding regions, 60 tRNAs, 12 rRNAs, and 1 CRISPR region were annotated. The DNA sequences of two plasmids, namely, pEK4S  and pEK4L , were also assembled and compared with their partial sequences using Blast  and DRX314299 (Illumina).Strain K-4 genome sequence and annotation data were deposited in DDBJ/GenBank under BioProject number


Professional and regulatory agency guidelines recommend that fluoroquinolones not be used as first-line therapy to treat uncomplicated UTI. Increasingly, β-lactams are being used to treat UTI despite limited evidence supporting their effectiveness. The study aim was to compare the efficacy of β-lactams and other commonly prescribed antibiotics for UTI to fluoroquinolones.< 30d, co-diagnosis where antibiotics were indicated, hospital discharge < 7d, scheduled urological procedure < 7d, pregnancy. Outcome: UTI related admission or outpatient return visit within 3-30d. Demographic, co-morbidity, vitals, laboratory, and prescription data were extracted. Propensity scores were used to balance treatments and GEE models were used to estimate outcomes.A multi-centered retrospective cohort study of outpatient visits with a UTI diagnosis between 2019-2021 was developed. Inclusion criteria: In-person visit in the Emergency Department or Primary/Urgent Care with ICD-10 documented UTI diagnosis and oral antibiotic prescription filled within 0-2 d. Exclusion criteria: Prior UTI There were 73,334 visits in 130 VA medical centers. Mean (S.D.) cohort age was 66.0 (15.8) y, 22.9% were female. Antibiotics prescribed N (%): β-lactams  27,385 (37.3%), nitrofurantoin 15,509 (21.1%), trimethoprim / sulfamethoxazole (TMP/SMX) 15,453 (21.1%), and fluoroquinolones 14,997 (20.4%). The overall 30-day clinical failure rate was 10,525 (14.4%).Comparative Effectiveness of Oral Antibiotics for Outpatient UTIsCommonly prescribed antibiotics for outpatient UTI were associated with a modest increase in 30-day clinical failure relative to fluoroquinolones. Effectiveness differences were most pronounced in male patients. Clinicians should carefully differentiate between complicated and uncomplicated UTI in males and consider preferentially prescribing fluoroquinolones.Karl Madaras-Kelly, PharmD, MPH, BioMerioux: Grant/Research Support