Distinct echinocandin responses of Candida albicans and Candida auris cell walls revealed by solid-state NMR

Probably not a substantial claim: may not pass S3 (load-bearing).

The balance between rigid structural components and mobile, water-accessible regions allows fungi to fine-tune cell wall permeability, aiding adaptive survival.

“Recent observations suggest that the balance between rigid structural components and mobile, water-accessible regions allows fungi to fine-tune cell wall permeability, aiding adaptive survival50,65.”

• cites [50] Dickwella Widanage, M. C. et al. Adaptative survival of Aspergillus fumigatus to echinocandins arises from cell wall remodeling beyond β−1,3-glucan synthesis inhibition. Nat. Commun. 15, 6382 (2024). sourceSupported
• cites [65] Fernando, L. D. et al. Structural adaptation of fungal cell wall in hypersaline environment. Nat. Commun. 14, 7082 (2023). sourceSupported

May combine two claims.

C. albicans cell walls share a relatively simple structure, with the rigid structural framework containing only chitin microfibrils and stiff linear β-1,3-glucan backbones, which may primarily adopt the triple helix conformation, within the inner layer.

“These findings demonstrate that C. albicans cell walls share a relatively simple structure, with the rigid structural framework containing only chitin microfibrils and stiff linear β-1,3-glucan backbones, which may primarily adopt the triple helix conformation67–69, within the inner layer.”

• cites [67] Manabe, N. & Yamaguchi, Y. 3D structural insights into β-glucans and their binding proteins. Int. J. Mol. Sci. 22, 1578 (2021). sourcePartly supported

  Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

  Experimental — accuracy not yet measured
• cites [68] Saito, H. & Yokoi, M. High-Resolution 13C NMR Study of (1->3)-beta-D-glucans in the solid state: DMSO -induced conformational change and conformational characterization by spin relaxation measurements. Bull. Chem. Soc. Jpn. 62, 392–398 (1989). sourceNot checked — no full text of the source
• cites [69] Bikmurzin, R., Marsalka, A. & Kalediene, L. Solid-state 13C nuclear magnetic resonance study of soluble and insoluble β-glucans extracted from candida lusitaniae. Moleules 28, 8066 (2023). sourcePartly supported

  Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

  Experimental — accuracy not yet measured

• AssumptionRests on an assumption: The C. albicans and C. auris strains used (SC5314, AR386, I.3) are representative of their species for cell wall architecture and echinocandin response.

Probably not a substantial claim: may not pass S3 (load-bearing).

C. albicans has a well-characterized cell wall composed of an inner layer of chitin, β-1,3-glucan, and β-1,6-glucan, and an outer layer composed of proteins modified with N- and O-linked mannans and phosphomannans.

“C. albicans has a well-characterized cell wall composed of an inner layer of chitin, β−1,3-glucan, and β−1,6-glucan, and an outer layer composed of proteins modified with N- and O-linked mannans and phosphomannans, called mannoproteins34,35.”

• cites [34] Hall, R. A. et al. The Mnn2 mannosyltransferase family modulates mannoprotein fibril length, immune recognition and virulence of candida albicans. PLoS Pathog. 9, 1003276 (2013). sourceSupported
• cites [35] Gow, N. A. R. & Lenardon, M. D. Architecture of the dynamic fungal cell wall. Nat. Rev. Microbiol. 21, 248–259 (2023). sourceNot checked — no full text of the source

Classified as probably not a substantial claim.

Echinocandins inhibit the synthesis of β-1,3-glucan, an essential polysaccharide in the fungal cell wall.

“Echinocandins, which inhibit the synthesis of β−1,3-glucan, an essential polysaccharide in the fungal cell wall, are recommended as the first-line treatment of Candida infections and have been widely used in clinical settings20,21.”

• cites [20] Chen, S. C. A., Slavin, M. A. & Sorrell, T. C. Echinocandin antifungal drugs in fungal infections. Drugs 71, 11–41 (2011). sourceNot checked — no full text of the source
• cites [21] Perlin, D. S. Current perspectives on echinocandin class drugs. Future Microbiol. 6, 441–457 (2011). sourceSupported

May combine two claims.

Mobile molecules are positioned within the soft mesh and spatially distinct from the rigid core, generally remaining loosely associated and well-hydrated.

“Positioned within the soft mesh and spatially distinct from the rigid core, these mobile molecules generally remain loosely associated and well-hydrated63,64.”

• cites [63] Latgé, J. P. & Wang, T. Modern biophysics redefines our understanding of fungal cell wall structure, complexity, and dynamics. mBio 13, e01145–22 (2022). sourceSupported
• cites [64] Fernando, L. D., Zhao, W., Gautam, I., Ankur, A. & Wang, T. Polysaccharide assemblies in fungal and plant cell walls explored by solid-state NMR. Structure 31, 1375–1385 (2023). sourceNot checked — no full text of the source

May combine two claims.

Resistance to echinocandins occurs naturally in C. auris strains and is increasing in C. albicans primarily due to mutations in two hotspot regions of β-1,3-glucan synthase Fks subunit.

“Resistance to echinocandins has been widely reported in C. auris strains, where it occurs naturally, and in C. albicans, where the incidence has been increasing primarily due to mutations in two hotspot regions of β−1,3-glucan synthase Fks subunit25,26.”

• cites [25] Perlin, D. S. Echinocandin resistance in Candida. Clin. Infect. Dis. 61, S612–S617 (2015). sourcePartly supported

  The FKS Mechanism of Resistance section (page not supplied) supports the two-hot-spot mechanism and describes resistance-conferring mutations in C. albicans. The Epidemiology section (page not supplied) says resistance remains below 3% in C. albicans and identifies C. glabrata—not C. albicans—as the species with rising resistance. Our copy does not establish that resistance occurs naturally in C. auris. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [26] Walker, L. A., Gow, N. A. R. & Munro, C. A. Fungal echinocandin resistance. Fungal Genet. Biol. 47, 117–126 (2010). sourcePartly supported

  The “FKS1 hot spot mutations confer echinocandin resistance” section reports that mutations in two FKS1 hot spots confer resistance in C. albicans. But the “Breakthrough infections” section says surveillance found no emerging trend of echinocandin resistance in Candida isolates, contrary to the claimed increasing incidence. Our copy does not address naturally occurring resistance in C. auris; its discussion of intrinsic resistance names other species. (LLM, conf=0.88)

  Experimental — accuracy not yet measured

Classified as probably not a substantial claim.

Echinocandin treatment often results only in partial inhibition of β-1,3-glucan synthesis.

“However, echinocandin treatment often results only in partial inhibition of β−1,3-glucan synthesis22–24.”

• cites [22] Rosenberg, A. et al. Antifungal tolerance is a subpopulation effect distinct from resistance and is associated with persistent candidemia. Nat. Commun. 9, 2470 (2018). sourceCould not verify

  The Results section reports some growth despite echinocandin exposure, but it does not measure or describe β-1,3-glucan synthesis. Searches for glucan and partial inhibition found no passage establishing that echinocandin treatment often inhibits its synthesis only partially. (LLM, conf=0.74)

  Experimental — accuracy not yet measured
• cites [23] Fisher, M. C. et al. Tackling the emerging threat of antifungal resistance to human health. Nat. Rev. Microbiol. 20, 557–571 (2022). sourcePartly supported

  The Introduction’s Fig. 1 caption says echinocandins inhibit 1,3-β-d-glucan synthase and that exposure can cause cell-wall stress through this inhibition. It does not say the inhibition is often only partial, so it supports the mechanism but not the claimed frequency or extent. (LLM, conf=0.84)

  Experimental — accuracy not yet measured
• cites [24] Wagner, A. S., Lumsdaine, S. W., Mangrum, M. M. & Reynolds, T. B. Caspofungin-induced β(1,3)-glucan exposure in Candida albicans is driven by increased chitin levels. mBio 14, e0007423 (2023). sourcePartly supported

  The Introduction (page not provided) says echinocandins inhibit the β(1,3)-glucan synthase enzymes Fks1 and Fks2. It does not say that inhibition is often only partial; its findings about glucan exposure during caspofungin treatment do not establish how much synthesis remains. (LLM, conf=0.78)

  Experimental — accuracy not yet measured

Classified as probably not a substantial claim.

Cell wall adaptation to echinocandins is characterized by increased levels and exposure of chitin following depletion of a substantial portion of β-glucan.

“These effects have been associated with cell wall adaptation to echinocandins, characterized by increased levels and exposure of chitin following the depletion of a substantial portion of β-glucan; however, our understanding of echinocandin-induced remodeling mechanism of Candida cell wall architecture remains limited24,27–29.”

• cites [24] Wagner, A. S., Lumsdaine, S. W., Mangrum, M. M. & Reynolds, T. B. Caspofungin-induced β(1,3)-glucan exposure in Candida albicans is driven by increased chitin levels. mBio 14, e0007423 (2023). sourcePartly supported

  The Introduction and Discussion (pages unavailable) describe increased chitin deposition during caspofungin treatment of Candida albicans and link it to cell-wall remodeling. They identify the exposed material as β(1,3)-glucan, not chitin. The Introduction says echinocandins inhibit β(1,3)-glucan synthesis, but does not establish depletion of a substantial portion of cell-wall β-glucan. (LLM, conf=0.88)

  Experimental — accuracy not yet measured
• cites [27] Lenardon, M. D., Sood, P., Dorfmueller, H. C., Brown, A. J. P. & Gow, N. A. R. Scalar nanostructure of the Candida albicans cell wall; a molecular, cellular and ultrastructural analysis and interpretation. Cell Surf. 6, 100047 (2020). sourceCould not verify

  The Results section describes chitin and β(1,3)-glucan in the cell wall and measures their levels in genetic mutants, not after echinocandin treatment. Searches for echinocandins, caspofungin, and compensatory chitin responses found no passage addressing increased chitin levels or exposure following drug-induced β-glucan depletion. (LLM, conf=0.72)

  Experimental — accuracy not yet measured
• cites [28] Walker, L. et al. Stimulation of chitin synthesis rescues Candida albicans from echinocandins. PLoS Pathog. 4, e1000040 (2008). sourcePartly supported

  The Discussion (page not provided) says C. albicans responds to echinocandins by increasing chitin content, protecting cells from damage caused by inhibited β(1,3)-glucan synthesis. The Results section on caspofungin sensitivity (page not provided) likewise reports increased chitin content. These passages do not establish increased chitin exposure or quantify depletion of a substantial portion of β-glucan. (LLM, conf=0.84)

  Experimental — accuracy not yet measured
• cites [29] da Silva Dantas, A. et al. Crosstalk between the calcineurin and cell wall integrity pathways prevents chitin overexpression in Candida albicans J. Cell Sci.134, jcs258889 (2021). sourcePartly supported

  The Discussion (page unavailable) reports that caspofungin-treated cells have increased wall chitin alongside decreased β-1,3-glucan, and that some chitin becomes more accessible at the wall surface. It does not establish that a substantial portion of β-glucan was depleted, so that magnitude is unsupported. (LLM, conf=0.82)

  Experimental — accuracy not yet measured

May combine two claims.

Chitin content increased following caspofungin treatment, with higher intensities observed for the signature Ch4 and Ch2 peaks.

“Conversely, the chitin content increased, with higher intensities observed for the signature Ch4 and Ch2 peaks, reflecting the induction of the classical chitin upregulation compensatory mechanism that has been identified in different fungal species in response to echinocandin treatment28,77–79.”

• cites [28] Walker, L. et al. Stimulation of chitin synthesis rescues Candida albicans from echinocandins. PLoS Pathog. 4, e1000040 (2008). sourceSupported
• cites [77] Chamilos, G., Lewis, R. E., Albert, N. & Kontoyiannis, D. P. Paradoxical effect of echinocandins across candida species in vitro: evidence for echinocandin-specific and candida species-related differences. Antimicrob. Agent Chemother. 51, 2257–2259 (2007). sourcePartly supported

  The introductory passage labeled Abstract reports a compensatory increase in cell-wall chitin synthesis in a single C. albicans isolate exhibiting paradoxical growth under caspofungin. It does not establish chitin upregulation across different fungal species, and the paper reports no Ch4 or Ch2 peak measurements. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites [78] Loiko, V. & Wagener, J. The paradoxical effect of echinocandins in Aspergillus fumigatus relies on recovery of the β-1, 3-glucan synthase Fks1. Antimicrob. Agents Chemother. 61, e01690–16 (2017). sourcePartly supported

  The Results passage discussing Fig. 6 and the Discussion support a compensatory increase in cell-wall chitin during early echinocandin exposure. They do not report Ch4 or Ch2 peaks or increased intensities for those peaks; the paper measured chitin using calcofluor-white fluorescence. It also reports less chitin in paradoxically growing hyphae than in their growth-inhibited progenitors. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites [79] Valero, C. et al. Caspofungin paradoxical effect is a tolerant “Eagle Effect” in the filamentous fungal pathogen aspergillus fumigatus. mBio 13, e0044722 (2022). sourceCould not verify

  The Observation section discusses caspofungin-induced cell-wall remodeling, but it does not report increased chitin content or Ch4 and Ch2 peak intensities. Searches for those peaks and for chitin upregulation found no passage in our copy that addresses the claimed measurements; page numbers were not available. (LLM, conf=0.73)

  Experimental — accuracy not yet measured

• AssumptionRests on an assumption: The C. albicans and C. auris strains used (SC5314, AR386, I.3) are representative of their species for cell wall architecture and echinocandin response.

May combine two claims.

Introducing the biradical AsymPolPOK into intact cells achieved a 57-fold enhancement in NMR sensitivity for carbohydrates, which reduced measurement time by 3,249-fold.

“To investigate whether caspofungin treatment affects the spatial organization of polysaccharides within the mechanical core of C. auris cell wall, we introduced the biradical AsymPolPOK83 into intact cells, achieving a 57-fold enhancement in the NMR sensitivity for carbohydrates (Fig. 5a), which reduced measurement time by 3,249-fold.”

• cites [83] Mentink-Vigier, F. et al. Computationally assisted design of polarizing agents for dynamic nuclear polarization enhanced NMR: the asymPol family. J. Am. Chem. Soc. 140, 11013–11019 (2018). sourceSupported

Classified as probably not a substantial claim. May combine two claims.

AsymPolPOK and other biradicals are small, water-soluble molecules that diffuse through porous cell walls without disrupting their structure.

“AsymPolPOK and other biradicals are small (2–3 nm), water-soluble molecules that diffuse through porous cell walls without disrupting their structure, enabling widespread use in the structural analysis of plant, fungal, algal, and bacterial cell wall materials as well as intact mammalian cells84–90.”

• cites [84] Viger-Gravel, J. et al. Topology of pretreated wood fibers using dynamic nuclear polarization. J. Phys. Chem. C. 123, 30407–30415 (2019). sourceNot checked — no full text of the source
• cites [85] Ghosh, R., Xiao, Y., Kragelj, J. & Frederick, K. K. In-cell sensitivity-enhanced NMR of intact viable mammalian cells. J. Am. Chem. Soc. 143, 18454–18466 (2021). sourcePartly supported

  The Results and Discussion (page unavailable) show that AMUPol, a small molecule, was used to study intact viable mammalian cells while maintaining cellular organization. They do not describe AsymPolPOK, a 2–3 nm size, or diffusion through porous plant, fungal, algal, or bacterial cell walls. Instead, the Results say most AMUPol was likely excluded by the mammalian plasma membrane, so the proposed diffusion mechanism is not supported by this paper. (LLM, conf=0.87)

  Experimental — accuracy not yet measured
• cites [86] Poulhazan, A. et al. Molecular-level architecture of Chlamydomonas reinhardtii’s glycoprotein-rich cell wall. Nat. Commun. 15, 986 (2024). sourcePartly supported

  The Methods (page unavailable) describe an aqueous AMUPol biradical solution applied to extracted algal cell walls and intact microalgae. The Results (page unavailable) suggest that the polarizing radical localizes in the cell wall, but do not establish diffusion through pores or that the biradical leaves wall structure undisturbed. The copy does not give a 2–3 nm size, identify AsymPolPOK as the reagent used, or establish use across the other listed materials and cells. (LLM, conf=0.85)

  Experimental — accuracy not yet measured
• cites [87] Byeon, C. H. et al. Ultrasensitive characterization of native bacterial biofilms via dynamic nuclear polarization-enhanced solid-state NMR. Angew. Chem. 64, e202418146 (2025). sourcePartly supported

  The Results and Discussion (page unavailable) report that AsymPol-POK was dissolved in a predominantly water-based buffer and was uniformly distributed within a wet bacterial biofilm. They do not establish its 2–3 nm size, diffusion through porous cell walls, or preservation of cell-wall structure after incubation; the micrographs showing intact bacteria were taken before radical incubation. The paper also does not extend those properties to other biradicals or the full range of organisms named in the citing sentence. (LLM, conf=0.84)

  Experimental — accuracy not yet measured
• cites [88] Zhao, W., Deligey, F., Shekar, S. C., Mentink-Vigier, F. & Wang, T. Current limitations of solid-state NMR in carbohydrate and cell wall research. J. Magn. Reson. 341, 107263 (2022). sourceNot checked — no full text of the source
• cites [89] Takahashi, H. et al. Solid-state NMR on bacterial cells: selective cell wall signal enhancement and resolution improvement using dynamic nuclear polarization. J. Am. Chem. Soc. 135, 5105–5110 (2013). sourceNot checked — no full text of the source
• cites [90] Wang, T. et al. Sensitivity-enhanced solid-state NMR detection of expansin’s target in plant cell walls. Proc. Natl Acad. Sci. USA 110, 16444–16449 (2013). sourceNot checked — no full text of the source

Classified as probably not a substantial claim. May combine two claims.

A reduction in β-1,6-glucan level has been reported in the literature for C. albicans during caspofungin treatment, along with an increase in the molecular size of β-1,6-glucan and cross-linking of glucan and chitin.

“In addition to the inhibition of the biosynthesis of the primary target, β-1,3-glucan, a reduction in β-1,6-glucan level has also been reported in the literature for C. albicans during caspofungin treatment80, along with an increase in the molecular size of β-1,6-glucan and cross-linking of glucan and chitin36.”

• cites [80] Stevens, D. A., Ichinomiya, M., Koshi, Y. & Horiuchi, H. Escape of Candida from caspofungin inhibition at concentrations above the MIC (Paradoxical effect) accomplished by increased cell wall chitin;: Evidence for β-1,6-glucan synthesis inhibition by caspofungin. Antimicrob. Agents Chemother. 50, 3160–3161 (2006). sourceSupported
• cites [36] Bekirian, C. et al. β−1,6-glucan plays a central role in the structure and remodeling of the bilaminate fungal cell wall. eLife 13, RP100569 (2024). sourceNot checked — no full text of the source

May combine two claims.

Extending to a 3D hCCH TOCSY experiment with DIPSI-3 mixing allowed the resolution of all carbon sites, revealing five distinct 1,2-linked mannose forms, three glucopyranose forms from β-1,3-glucan in the cell walls, five forms of galactose or glucose derivatives, such as glucose-phosphates, alongside α- and β-glucoses in the media.

“Extending to a 3D hCCH TOCSY experiment with DIPSI-3 mixing73 allowed the resolution of all carbon sites (Supplementary Table 2), revealing five distinct 1,2-linked mannose forms (Fig. 2d), three glucopyranose forms from β-1,3-glucan (Fig. 2e) in the cell walls, five forms of galactose or glucose derivatives, such as glucose-phosphates (Fig. 2e)74, alongside α- and β-glucoses in the media (Fig. 2g”

• cites [73] Bahri, S. et al. 1H-detected characterization of carbon-carbon networks in highly flexible protonated biomolecules using MAS NMR. J. Biomol. NMR 77, 111–119 (2023). sourcePartly supported

  Split into 2 atoms — 1 confirmed, 0 partially supported, 0 disputed, 0 misattributed, 1 unverifiable. Expand the claim to see per-atom verdicts.

  Experimental — accuracy not yet measured
• cites [74] Acosta-Zaldivar, M. et al. Candida albicans’ inorganic phosphate transport and evolutionary adaptation to phosphate scarcity. PLoS Genet 20, e1011156 (2024). sourceCould not verify

  Split into 2 atoms — 0 confirmed, 0 partially supported, 0 disputed, 0 misattributed, 2 unverifiable. Expand the claim to see per-atom verdicts.

  Experimental — accuracy not yet measured

• AssumptionRests on an assumption: The C. albicans and C. auris strains used (SC5314, AR386, I.3) are representative of their species for cell wall architecture and echinocandin response.

May combine two claims.

Among these two forms, the former corresponds to type-a β-1,3-glucan observed at room-temperature, representing the bulk β-1,3-glucans in the model triple-helix configuration, exhibiting no interaction with chitin.

“Among these two forms, the former corresponds to type-a β-1,3-glucan observed at room-temperature (Fig. 2e), representing the bulk β-1,3-glucans in the model triple-helix configuration67–69, exhibiting no interaction with chitin.”

• cites [67] Manabe, N. & Yamaguchi, Y. 3D structural insights into β-glucans and their binding proteins. Int. J. Mol. Sci. 22, 1578 (2021). sourceSupported
• cites [68] Saito, H. & Yokoi, M. High-Resolution 13C NMR Study of (1->3)-beta-D-glucans in the solid state: DMSO -induced conformational change and conformational characterization by spin relaxation measurements. Bull. Chem. Soc. Jpn. 62, 392–398 (1989). sourceNot checked — no full text of the source
• cites [69] Bikmurzin, R., Marsalka, A. & Kalediene, L. Solid-state 13C nuclear magnetic resonance study of soluble and insoluble β-glucans extracted from candida lusitaniae. Moleules 28, 8066 (2023). sourcePartly supported

  The Results (page not available) suggest that β-1,3-glucans in the alkali-extracted fractions may have an ordered backbone, possibly a triple helix, but also offer a double helix as an alternative. The paper does not identify this as room-temperature type-a glucan or establish that it represents the bulk β-1,3-glucan. Its Results also describe possible β-1,3-glucan–chitin linkages, rather than establishing no interaction with chitin. (LLM, conf=0.86)

  Experimental — accuracy not yet measured

May combine two claims.

None of these changes were observed in the C. auris I.3 strain, which exhibits echinocandin resistance due to the S639F mutation in β-1,3-glucan synthase FKS1.

“None of these changes were observed in the C. auris I.3 strain, which exhibits echinocandin resistance due to the S639F mutation in β-1,3-glucan synthase FKS176.”

• cites [76] Li, J., Coste, A. T., Bachmann, D., Sanglard, D. & Lamoth, F. Assessment of the in vitro and in vivo antifungal activity of NSC319726 against Candida auris. Microbiol. Spectr. 9, e01395–21 (2021). sourcePartly supported

  The Results section identifies C. auris I.3 as an echinocandin-resistant isolate. It does not identify an S639F mutation in FKS1 or establish that mutation as the cause of resistance. The cited paper also does not describe the unspecified 'changes' whose absence the sentence attributes to I.3 (page unavailable). (LLM, conf=0.84)

  Experimental — accuracy not yet measured

• AssumptionRests on an assumption: The C. albicans and C. auris strains used (SC5314, AR386, I.3) are representative of their species for cell wall architecture and echinocandin response.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

These α-1,2-mannoses can be connected to the α-1,6-mannan backbone, as well as to β-1,2-mannose, α-1,3-mannose, and phosphate groups found in the sidechains.

“These α-1,2-mannoses can be connected to the α-1,6-mannan backbone, as well as to β-1,2-mannose, α-1,3-mannose, and phosphate groups found in the sidechains27,35.”

• cites [27] Lenardon, M. D., Sood, P., Dorfmueller, H. C., Brown, A. J. P. & Gow, N. A. R. Scalar nanostructure of the Candida albicans cell wall; a molecular, cellular and ultrastructural analysis and interpretation. Cell Surf. 6, 100047 (2020). sourcePartly supported

  The Introduction (page unavailable) describes an α(1,6)-mannose backbone with α- or β-mannoside side chains and says mannans can carry β(1,2)-phosphomannosides. The Results section on N-mannan fibrils (page unavailable) also places side branches on the α(1,6) backbone. Neither passage specifies α(1,2)-mannose connections to β(1,2)-mannose, α(1,3)-mannose, or phosphate groups as the claim does. (LLM, conf=0.83)

  Experimental — accuracy not yet measured
• cites [35] Gow, N. A. R. & Lenardon, M. D. Architecture of the dynamic fungal cell wall. Nat. Rev. Microbiol. 21, 248–259 (2023). sourceNot checked — no full text of the source

May combine two claims.

The mannan components exhibited flexible motion, whether as long N-mannan fibrils extending through the outer layer or as shorter O-linked mannans associated with cell wall proteins in the inner region.

“The mannan components exhibited flexible motion (Fig. 3e), whether as long N-mannan fibrils extending through the outer layer or as shorter O-linked mannans associated with cell wall proteins in the inner region27.”

• cites [27] Lenardon, M. D., Sood, P., Dorfmueller, H. C., Brown, A. J. P. & Gow, N. A. R. Scalar nanostructure of the Candida albicans cell wall; a molecular, cellular and ultrastructural analysis and interpretation. Cell Surf. 6, 100047 (2020). sourcePartly supported

  The Results section identifies long N-mannan fibrils in the outer cell-wall layer, while the Introduction describes O-mannan as a short chain. The Discussion, under “Mannosylated cell wall proteins,” places mannosylated proteins in the inner layer and depicts their stalks with O-linked mannans. These passages support the structural description, but they do not establish flexible motion of either mannan component; page numbers are unavailable in our copy. (LLM, conf=0.86)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

Reduced water permeability is a common response previously observed in several Aspergillus species under stress.

“Both C. albicans and C. auris AR386 exhibit reduced water permeability (Fig. 4a), a common response previously also observed in several Aspergillus species under stress50,53.”

• cites [50] Dickwella Widanage, M. C. et al. Adaptative survival of Aspergillus fumigatus to echinocandins arises from cell wall remodeling beyond β−1,3-glucan synthesis inhibition. Nat. Commun. 15, 6382 (2024). sourcePartly supported

  The Results section, “Reduced water retention and polymer dynamics” (page unavailable), reports reduced water accessibility in A. fumigatus cell walls after caspofungin treatment and suggests this should reduce cell-wall permeability. It does not establish reduced water permeability as a common response across several Aspergillus species or under stress generally, and permeability was inferred rather than directly measured. (LLM, conf=0.87)

  Experimental — accuracy not yet measured
• cites [53] Fernando, L. D. et al. Structural adapataion of fungal cell wall in hypersaline environment. Nat. Commun. 14, 7082 (2023). sourcePartly supported

  The Introduction and Discussion report limited water permeability in A. sydowii, including under high-salt conditions. The Results section on other halophilic Aspergillus species reports shared cell-wall composition in A. atacamensis and A. destruens, but does not establish reduced water permeability in those species. The claim that this response was observed in several Aspergillus species is therefore broader than this source supports. (LLM, conf=0.84)

  Experimental — accuracy not yet measured

May combine two claims.

This rigidification may be a result of chitin relocation from the inner to the outer layer, which also enhanced chitin-protein association.

“This may be a result of chitin relocation from the inner to the outer layer, which also enhanced chitin-protein association, as recently reported (Fig. 8)29.”

• cites [29] da Silva Dantas, A. et al. Crosstalk between the calcineurin and cell wall integrity pathways prevents chitin overexpression in Candida albicans J. Cell Sci.134, jcs258889 (2021). sourcePartly supported

  The Results and Discussion (Figs 2 and 8; page unavailable) report increased chitin exposure in the outer wall after caspofungin treatment and suggest that chitin distributed through the wall may increase rigidity. They do not report that this relocation enhanced chitin–protein association; the Discussion instead mentions chitin–glucan cross-linking as a possible influence on rigidity. (LLM, conf=0.86)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

Compared to PDSD, CORD provides higher efficiency due to the use of active recoupling pulses.

“Compared to proton-driven spin diffusion (PDSD), CORD provides higher efficiency due to the use of active recoupling pulses113.”

• cites [113] Hou, G., Yan, S., Trebosc, J., Amoureux, J. P. & Polenova, T. Broadband homonuclear correlation spectroscopy driven by combined R2nv sequences under fast magic angle spinning for NMR structural analysis of organic and biological solids. J. Magn. Reson. 232, 18–30 (2013). sourcePartly supported

  The Introduction says conventional PDSD deteriorates as MAS frequency rises and does not work above about 25–30 kHz. The Results and Discussion describe CORD’s applied recoupling pulses and its broadband polarization transfer at fast MAS. This supports the comparison under fast-MAS conditions, but not the sentence’s unqualified claim that CORD is more efficient than PDSD generally. (LLM, conf=0.85)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Certain Candida species can transition from yeast forms to filamentous structures, producing true hyphae in species like C. albicans, C. dubliniensis, and C. tropicalis, or pseudohyphae in species such as C. auris and C. parapsilosis.

“Certain Candida species can transition from yeast forms to filamentous structures, producing true hyphae in species like C. albicans, C. dubliniensis, and C. tropicalis, or pseudohyphae in species such as C. auris and C. parapsilosis96–98.”

• cites [96] Lackey, E., Vipulanandan, G., Childers, D. S. & Kadosh, D. Comparative evolution of morphological regulatory functions in Candida species. Eukaryot. Cell 12, 1356–1368 (2013). sourceNot checked — no full text of the source
• cites [97] Ruiz, G. B., Ross, Z. K., Gow, N. A. R. & Lorenz, A. Pseudohyphal growth of the emerging pathogen Candida auris is triggered by genotoxic stress through the S phase checkpoint. mSphere 5, e00151–20 (2020). sourcePartly supported

  The Results section on genes associated with hyphal growth identifies C. albicans, C. tropicalis, and C. dubliniensis as capable of forming true hyphae. The Results section on C. auris reports pseudohyphae under genotoxic stress. The paper says C. parapsilosis cannot form true hyphae, but does not say it forms pseudohyphae; it also notes that C. auris filamentation varies by strain. (LLM, conf=0.86)

  Experimental — accuracy not yet measured
• cites [98] Louvet, M. et al. Ume6-dependent pathways of morphogenesis and biofilm formation in Candida auris. Microbiol. Spectr. 12, e0153124 (2024). sourcePartly supported

  The Introduction and Discussion (page numbers unavailable) describe C. auris transitioning to pseudohyphae and refer to true hyphae of C. albicans. The copy does not substantiate the claimed morphology of C. dubliniensis, C. tropicalis, or C. parapsilosis. (LLM, conf=0.87)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

Such structural rearrangements have been observed in the xylan of the secondary cell walls across various plant species, where folding onto the flat surfaces of cellulose microfibrils induces a conformational shift from a three-fold helical structure to a flat, two-fold conformation.

“Such structural rearrangements have been observed in the xylan of the secondary cell walls across various plant species, where folding onto the flat surfaces of cellulose microfibrils induces a conformational shift from a three-fold helical structure to a flat, two-fold conformation92–94.”

• cites [92] Simmons, T. J. et al. Folding of xylan onto cellulose fibrils in plant cell walls revealed by solid-state NMR. Nat. Commun. 7, 13902 (2016). sourcePartly supported

  Split into 2 atoms — 0 confirmed, 2 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

  Experimental — accuracy not yet measured
• cites [93] Grantham, N. J. et al. An even pattern of xylan substitution is critical for interaction with cellulose in plant cell walls. Nat. Plants 3, 859−865 (2017). sourceNot checked — no full text of the source
• cites [94] Kirui, A. et al. Carbohydrate-aromatic interface and molecular architecture of lignocellulose. Nat. Commun. 13, 538 (2022). sourcePartly supported

  Split into 2 atoms — 1 confirmed, 1 partially supported, 0 disputed, 0 misattributed, 0 unverifiable. Expand the claim to see per-atom verdicts.

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

At the slower MAS rate of 13.5 kHz used in this study, CORD's performance is comparable to that of DARR, while offering the advantage of lower radiofrequency power requirements during the recoupling period.

“At the slower MAS rate of 13.5 kHz used in this study, its performance is comparable to that of Dipolar-Assisted Rotational Resonance (DARR), while offering the advantage of lower radiofrequency power requirements during the recoupling period.”

• cites [113] Hou, G., Yan, S., Trebosc, J., Amoureux, J. P. & Polenova, T. Broadband homonuclear correlation spectroscopy driven by combined R2nv sequences under fast magic angle spinning for NMR structural analysis of organic and biological solids. J. Magn. Reson. 232, 18–30 (2013). sourceCould not verify

  The Introduction identifies DARR as a conventional method used at MAS rates of 30 kHz or lower, while the Methods report CORD experiments at 40 kHz. Neither those passages nor the searched Results passages compare CORD with DARR at 13.5 kHz or establish a lower recoupling-period RF power requirement for CORD. Searched for DARR, 13.5 kHz, and RF-power comparisons. (LLM, conf=0.76)

  Experimental — accuracy not yet measured

Probably not a substantial claim: may not pass S3 (load-bearing).

CORD efficiently drives homonuclear 13C-13C polarization transfer under fast MAS conditions and high magnetic fields.

“CORD efficiently drives homonuclear 13C-13C polarization transfer under fast MAS conditions and high magnetic fields113.”

• cites [113] Hou, G., Yan, S., Trebosc, J., Amoureux, J. P. & Polenova, T. Broadband homonuclear correlation spectroscopy driven by combined R2nv sequences under fast magic angle spinning for NMR structural analysis of organic and biological solids. J. Magn. Reson. 232, 18–30 (2013). sourceSupported

May combine two claims.

In both C. albicans and C. auris, caspofungin rigidifies β-1,6-glucans and mannan sidechains.

“In both species, caspofungin rigidifies β−1,6-glucans and mannan sidechains and reduces water permeability during β−1,3-glucan depletion;”

• Also stated in: § Caspofungin treatment remodels polysaccharide composition in the Candida cell walls · hedged

  “These findings suggested that some mannans and β-1,6-glucans transitioned from the mobile fraction to the rigid fraction as a result of caspofungin-induced reorganization of the cell wall.”

• AssumptionRests on an assumption: The C. albicans and C. auris strains used (SC5314, AR386, I.3) are representative of their species for cell wall architecture and echinocandin response.

May combine two claims.

The mobile fraction revealed a variety of molecules with highly overlapping signals, except for a few additional galactose and glucose signals in C. auris, which were more abundant than those in C. albicans.

“The mobile fraction revealed a variety of molecules with highly overlapping signals, except for a few additional galactose and glucose signals in C. auris, which were more abundant than those in C. albicans (Fig. 2b).”

• AssumptionRests on an assumption: The C. albicans and C. auris strains used (SC5314, AR386, I.3) are representative of their species for cell wall architecture and echinocandin response.

May combine two claims.

C. albicans and C. auris exhibited highly similar NMR signatures, with β-1,3-glucan dominating the rigid fraction when detected through dipolar-coupling-based hCH spectra, consistent with 13C-detection results.

“These species exhibited highly similar NMR signatures, with β-1,3-glucan dominating the rigid fraction when detected through dipolar-coupling-based hCH spectra (Fig. 2a), which is consistent with 13C-detection results (Fig. 1d).”

• AssumptionRests on an assumption: The C. albicans and C. auris strains used (SC5314, AR386, I.3) are representative of their species for cell wall architecture and echinocandin response.

May combine two claims.

Mannan forms fibrillar structures that particularly dominate the outer layer of the cell wall, and these fibrils are dynamically fluctuating, in contrast to chitin microfibrils that exhibit partial structural order and crystallinity, as well as stiffness.

“Mannan forms fibrillar structures that particularly dominate the outer layer of the cell wall, and our findings indicate that these fibrils are dynamically fluctuating, in contrast to the chitin microfibrils that exhibit partial structural order and crystallinity, as well as stiffness.”

• AssumptionRests on an assumption: The C. albicans and C. auris strains used (SC5314, AR386, I.3) are representative of their species for cell wall architecture and echinocandin response.

May combine two claims.

Signals from α-1,2- and α-1,6-linked mannose residues in mannan fibrils and weak signals from galactose residues were observed in the mobile phase.

“Additionally, signals from α−1,2- and α−1,6-linked mannose residues (Mn1,2 and Mn1,6) in mannan fibrils and weak signals from galactose (Gal) residues were observed in the mobile phase.”

• AssumptionRests on an assumption: The C. albicans and C. auris strains used (SC5314, AR386, I.3) are representative of their species for cell wall architecture and echinocandin response.

May combine two claims.

The use of 0.015 μg/mL caspofungin allowed C. albicans SC5314 to maintain ~90% of growth, providing sufficient material for NMR analysis.

“The use of 0.015 µg/mL caspofungin allowed C. albicans SC5314 to maintain ~90% of growth, providing sufficient material for NMR analysis (Fig. 3a).”

May combine two claims.

Complete growth inhibition for the C. auris AR386 strain occurred only above 32 μg/mL caspofungin, with a similar 90% of growth retained at the 0.05 μg/mL concentration used in this study.

“Complete growth inhibition for the C. auris AR386 strain occurred only above 32 µg/mL caspofungin, with a similar 90% of growth retained at the 0.05 µg/mL concentration used in this study.”

May combine two claims.

In apo C. albicans, β-1,3-glucans were initially insensitive to microsecond timescale motions, indicated by long 1H-T1ρ relaxation times with an average of 15.3 ms, which decreased to 8.6 ms following caspofungin treatment, indicating increased dynamics for β-1,3-glucan matrix.

“In apo C. albicans, β-1,3-glucans were initially insensitive to microsecond timescale motions, indicated by long 1H-T1ρ relaxation times with an average of 15.3 ms (Fig. 4b), which decreased to 8.6 ms following caspofungin treatment, indicating increased dynamics for β-1,3-glucan matrix (Supplementary Fig. 4 and Supplementary Table 7).”

• AssumptionRests on an assumption: The C. albicans and C. auris strains used (SC5314, AR386, I.3) are representative of their species for cell wall architecture and echinocandin response.

May combine two claims.

Chitin contents increased to 44-48% in both strains, consistent with the comparable 1D spectral pattern of these two samples post-treatment.

“In contrast, chitin contents increased to 44-48% in both strains, consistent with the comparable 1D spectral pattern of these two samples post-treatment.”

• AssumptionRests on an assumption: The C. albicans and C. auris strains used (SC5314, AR386, I.3) are representative of their species for cell wall architecture and echinocandin response.

May combine two claims.

Mannan signals predominantly originated from α-1,2-linked sidechains, identified by unique carbon 1 and carbon 2 signals at 101-102 ppm and 81-82 ppm, respectively, and later confirmed via INADEQUATE and PAR spectra.

“Notably, the mannan signals predominantly originated from α-1,2-linked sidechains, identified by unique carbon 1 and carbon 2 signals at 101-102 ppm and 81-82 ppm, respectively, and later confirmed via INADEQUATE and PAR spectra (Supplementary Fig. 5a).”

• AssumptionRests on an assumption: The C. albicans and C. auris strains used (SC5314, AR386, I.3) are representative of their species for cell wall architecture and echinocandin response.

May combine two claims.

2D 13C-13C correlation spectra revealed a significant increase in the chitin content in caspofungin-treated C. albicans SC5314 and C. auris AR386, as well as a notable rise in the signals for β-1,6-glucan and mannan.

“The 2D 13C-13C correlation spectra revealed a significant increase in the chitin content in caspofungin-treated C. albicans SC5314 and C. auris AR386, as well as a notable rise in the signals for β−1,6-glucan and mannan (Fig. 3d).”

• AssumptionRests on an assumption: The C. albicans and C. auris strains used (SC5314, AR386, I.3) are representative of their species for cell wall architecture and echinocandin response.

May combine two claims.

After caspofungin treatment, chitin in C. albicans was significantly less flexible on both nanosecond and microsecond timescales, while β-1,3-glucan became more dynamic.

“After caspofungin treatment, chitin in C. albicans was significantly less flexible on both nanosecond and microsecond timescales, while β-1,3-glucan became more dynamic, indicating a better separation of these two polymers (Fig. 4b, c).”

May combine two claims.

C. albicans fully restructured its cell wall after caspofungin treatment, evidenced by a 1.5-fold increase in wall thickness, while C. auris strains showed no equivalent phenotypic responses.

“Notably, C. albicans fully restructured its cell wall, evidenced by a 1.5-fold increase in wall thickness, while C. auris strains showed no equivalent phenotypic responses (Fig. 3b).”

May combine two claims.

The addition of caspofungin led to similar or even higher β-1,6-glucan levels in the single deletion mutants compared to AR386: the β-1,6-glucan level in the mobile fraction of caspofungin-treated cell walls was 35%, 34%, and 53% for wild-type, kre6aΔ, and kre6bΔ samples, respectively.

“Interestingly, the addition of caspofungin led to similar or even higher β-1,6-glucan levels in the single deletion mutants compared to AR386: the β-1,6-glucan level in the mobile fraction of caspofungin-treated cell walls was 35%, 34%, and 53% for wild-type, kre6aΔ, and kre6bΔ samples, respectively (Fig. 7c, right).”

May combine two claims.

Both C. albicans and C. auris AR386 showed an increase in chitin content and a reduction in β-1,3-glucan after caspofungin treatment.

“Both species showed an increase in chitin content and a reduction in β-1,3-glucan (Fig. 8 and Fig. 3c), as expected for the canonical compensatory response to echinocandins.”

May combine two claims.

In microbroth dilution assay, micafungin MIC50 was 0.5 μg/mL for kre6a∆ and 0.25 μg/mL for AR386.

“These results were confirmed in microbroth dilution assay with micafungin MIC50 of 0.5 and 0.25 µg/mL for kre6a∆ and AR386, respectively (Fig. 6d).”

May combine two claims.

Attempts to generate a double KRE6a and KRE6b deletion strain in C. auris AR386 were unsuccessful; although the deletion cassette integrated into the KRE6a/b loci, both genes persisted.

“Attempts to generate a double KRE6a and KRE6b deletion strain in C. auris AR386 were unsuccessful (Supplementary Fig. 7 and Supplementary Table 8); although the deletion cassette integrated into the KRE6a/b loci, both genes persisted, suggesting that these genes are essential in C. auris.”

May combine two claims.

A very slight increase of susceptibility to micafungin and caspofungin of PADH1_KRE6a was observed in spotting assay, which was not confirmed in broth microdilution assay.

“Inversely, we observed a very slight increase of susceptibility to micafungin and caspofungin of PADH1_KRE6a in spotting assay, which was not confirmed in broth microdilution assay.”

May combine two claims.

The special form of β-1,3-glucan (Bb3) showed stronger interactions with chitin than the bulk form, as indicated by higher intensity of ChMe-Bb3 compared to ChMe-B3 and presence of cross-peaks for Ch2-Bb3 and Bb3-Ch2 without corresponding peaks for bulk β-1,3-glucan.

“However, the special form (Bb3) showed stronger interactions with chitin, as indicated by the higher intensity of ChMe-Bb3 compared to ChMe-B3, and the presence of cross-peaks for Ch2-Bb3 and Bb3-Ch2, without corresponding peaks for the bulk β-1,3-glucan.”

May combine two claims.

A slight but significant increase of FKS1 expression was observed in kre6a∆, but not in kre6b∆.

“A slight but significant increase of FKS1 expression was observed in kre6a∆, but not in kre6b∆.”

May combine two claims.

The protein and lipid components primarily reside in the mobile phase, exhibiting dynamic behavior similar to that of mannan.

“The protein and lipid components primarily reside in the mobile phase, exhibiting dynamic behavior similar to that of mannan (Supplementary Figs. 14 and 15).”

Probably not a substantial claim: may not pass S3 (load-bearing).

Expression of CHS1 was not affected in the deletion strains and their respective complemented strains.

“Expression of CHS1 was not affected in the deletion strains and their respective complemented strains.”

May combine two claims.

Results for caspofungin were difficult to interpret because of a paradoxical growth effect in all strains with none achieving 50% inhibition.

“Results were difficult to interpret for caspofungin because of a paradoxical growth effect in all strains with none of them achieving 50% inhibition.”

May combine two claims.

Specific protein and lipid signals persist through the PAR duration, indicating a partially ordered structure, but they do not show any detectable cross-peaks with carbohydrate components.

“While specific protein and lipid signals persist through the PAR duration, indicating a partially ordered structure, they do not show any detectable cross-peaks with carbohydrate components (Supplementary Figs. 5 and 6).”

May combine two claims.

Mobile domains regulate water accessibility and influence the diffusion of soluble molecules, including antifungals, while rigid domains serve as a physical barrier, reducing overall permeability.

“These mobile domains regulate water accessibility and influence the diffusion of soluble molecules, including antifungals, while rigid, often partially dehydrated structural domains serve as a physical barrier, reducing overall permeability.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Candida species contain long β-1,3-glucans and short β-1,6-glucans, with the latter potentially serving as a bridge between the former and cell wall proteins.

“Candida species contain long β−1,3-glucans and short β−1,6-glucans, with the latter potentially serving as a bridge between the former and cell wall proteins (Fig. 1a)61.”

• cites [61] Ruiz-Herrera, J., Elorza, M. V., Valentín, E. & Sentandreu, R. Molecular organization of the cell wall of Candida albicans and its relation to pathogenicity. FEMS Yeast Res. 6, 14–29 (2006). sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

β-1,6-glucan is primarily linear and contains short side chains of β-1,3-glucoside or laminaribioside residues.

“β−1,6-glucan is primarily linear and contains short side chains of β−1,3-glucoside or laminaribioside residues36,62.”

• cites [36] Bekirian, C. et al. β−1,6-glucan plays a central role in the structure and remodeling of the bilaminate fungal cell wall. eLife 13, RP100569 (2024). sourceNot checked — no full text of the source
• cites [62] Iorio, E. et al. Candida albicans cell wall comprises a branched β-d-(1→6)-glucan with β-d-(1→3)-side chains. Carbohydr. Res. 6, 1050–1061 (2008). sourceNot checked — no full text of the source

May combine two claims.

The splitting of BBr3 and BBr4 peaks likely arises from these units contributing to two distinct structural environments: one where β-1,3-glucan branches to another β-1,3-glucan, forming branched β-1,3/1,6-glucan, and the other where β-1,3-glucan branches to a linear β-1,6-glucan.

“This likely arises from these units contributing to two distinct structural environments: one where β-1,3-glucan branches to another β-1,3-glucan, forming branched β-1,3/1,6-glucan, and the other where β-1,3-glucan branches to a linear β-1,6-glucan.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Mannan components often account for 30-40% of the cell wall polysaccharides and are linked to the β-1,3-glucan-chitin network either via the β-1,6-glucan moiety or directly.

“These mannan components, which often account for 30–40% of the cell wall polysaccharides, are linked to the β−1,3-glucan-chitin network either via the β−1,6-glucan moiety or directly36.”

• cites [36] Bekirian, C. et al. β−1,6-glucan plays a central role in the structure and remodeling of the bilaminate fungal cell wall. eLife 13, RP100569 (2024). sourceNot checked — no full text of the source

May combine two claims.

Both β-1,6-glucans and branched β-1,3-glucans have a branching topology and may exhibit a reduced degree of intermolecular association compared to chitin microfibrils and β-1,3-glucan triple helices.

“Both β-1,6-glucans and branched β-1,3-glucans have a branching topology and may exhibit a reduced degree of intermolecular association compared to chitin microfibrils and β-1,3-glucan triple helices.”

May combine two claims.

The spectral patterns of treated C. albicans SC5314 and C. auris AR386 were nearly identical post echinocandin treatment, indicating similar molecular structures of their rigid cell wall cores after treatment, despite their differences in wall thickness.

“The spectral patterns of treated C. albicans SC5314 and C. auris AR386 were nearly identical post echinocandin treatment, indicating similar molecular structures of their rigid cell wall cores after treatment, despite their differences in wall thickness.”

• AssumptionRests on an assumption: The C. albicans and C. auris strains used (SC5314, AR386, I.3) are representative of their species for cell wall architecture and echinocandin response.

May combine two claims.

Proteins and lipids exhibited a lower, 30-fold enhancement because, although some may contribute to cell wall structuring, the majority are predominantly located in the plasma membrane or intracellularly, making them less exposed than cell wall polysaccharides.

“Proteins and lipids exhibited a lower, 30-fold enhancement because, although some may contribute to cell wall structuring, the majority are predominantly located in the plasma membrane or intracellularly, making them less exposed than cell wall polysaccharides.”

Classified as probably not a substantial claim.

Sensitivity enhancement is achieved by transferring electron spin polarization to 1H nuclei in the biradical, followed by 1H spin diffusion across the solvent and finally to biomolecules, resulting in homogeneous hyperpolarization over distances of at least tens of nanometers.

“Sensitivity enhancement is achieved by transferring electron spin polarization to 1H nuclei in the biradical, followed by 1H spin diffusion across the solvent and finally to biomolecules, resulting in homogeneous hyperpolarization over distances of at least tens of nanometers84.”

• cites [84] Viger-Gravel, J. et al. Topology of pretreated wood fibers using dynamic nuclear polarization. J. Phys. Chem. C. 123, 30407–30415 (2019). sourceNot checked — no full text of the source

May combine two claims.

Due to its resistance, the β-1,3-glucan content in the I.3 strain remained largely unaffected, even though the drug concentration was five times higher than that applied to C. auris AR386 and 17 times higher than that used for C. albicans.

“Due to its resistance, the β-1,3-glucan content in the I.3 strain remained largely unaffected (Fig. 3e and Supplementary Fig. 2), even though the drug concentration was five times higher than that applied to C. auris AR386 and 17 times higher than that used for C. albicans.”

• AssumptionRests on an assumption: The C. albicans and C. auris strains used (SC5314, AR386, I.3) are representative of their species for cell wall architecture and echinocandin response.

May combine two claims.

Only certain α-1,2-linked sidechains of mannan, rather than the more embedded α-1,6-linked backbones, interact with chitin microfibrils and undergo rigidification.

“It is important to note that only certain α-1,2-linked sidechains of mannan, rather than the more embedded α-1,6-linked backbones, interact with chitin microfibrils and undergo rigidification.”

May combine two claims.

Despite C. albicans SC5314 and C. auris AR386 showing nearly identical spectra for their rigid polysaccharides, their polymer dynamics and water retention respond differently to caspofungin treatment.

“Despite C. albicans SC5314 and C. auris AR386 showing nearly identical spectra for their rigid polysaccharides (Fig. 3c, e), their polymer dynamics and water retention respond differently to caspofungin treatment.”

• AssumptionRests on an assumption: The C. albicans and C. auris strains used (SC5314, AR386, I.3) are representative of their species for cell wall architecture and echinocandin response.

May combine two claims.

In caspofungin-treated C. albicans, the remaining β-1,3-glucans were only loosely associated with chitin, preserving their hydration, while in C. auris, they became tightly integrated into chitin domains, leading to reduced hydration.

“In caspofungin-treated C. albicans, the remaining β-1,3-glucans were only loosely associated with chitin, preserving their hydration, while in C. auris, they became tightly integrated into chitin domains, leading to reduced hydration.”

May combine two claims. The quote only partly states this claim.

The tight association between chitin and β-1,3-glucan in the rigid core represents a conserved structural feature that remains unaffected by echinocandin treatment, which is crucial for the mechanical strength of the cell wall and fungal survival.

“Therefore, the tight association between chitin and β-1,3-glucan in the rigid core represents a conserved structural feature that remains unaffected by echinocandin treatment, which is crucial for the mechanical strength of the cell wall and fungal survival.”

• AssumptionRests on an assumption: The C. albicans and C. auris strains used (SC5314, AR386, I.3) are representative of their species for cell wall architecture and echinocandin response.

May combine two claims.

The increase of FKS1 expression in kre6a∆ suggests a compensatory effect resulting from downregulation of all β-1,6-glucan synthase genes (KRE6a and KRE6b) in kre6a∆, which was not the case in kre6b∆.

“This result suggests a compensatory effect resulting from the downregulation of all β-1,6-glucan synthase genes (KRE6a and KRE6b) in kre6a∆, which was not the case in kre6b∆ (maintaining normal level of KRE6a expression).”

May combine two claims.

The more pronounced dehydration in C. auris leads to a more robust cell wall that provides better protection against stress.

“However, C. auris shows more pronounced dehydration in its rigid core compared to C. albicans (Fig. 8), leading to a more robust cell wall that provides better protection against stress.”

May combine two claims.

The structure of the rigid core of the C. auris cell wall remained unperturbed by caspofungin treatment, even though the molecular composition of the cell wall was reorganized.

“This finding aligned with the observed similarity in polymer dynamics and hydration profiles between the apo and caspofungin-treated C. auris, collectively indicating that the structure of the rigid core remained unperturbed, even though the molecular composition of the cell wall has been reorganized.”

• AssumptionRests on an assumption: The solid-state NMR measurements on whole cells reflect the native cell wall structure and dynamics.

Probably not a substantial claim: may not pass S3 (load-bearing).

The partial restoration of KRE6b expression could be due to some loss of the distal portion of the KRE6b promoter resulting from insertion of the complementation cassette.

“This could be due to some loss of the distal portion of the KRE6b promoter resulting from the insertion of the complementation cassette.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

β-1,6-glucans have been shown recently to be highly responsive to environmental factors such as pH, salt, and presence of antifungals, and can be regulated to compensate for defects in O-/N-mannan elongation.

“The biosynthesis and structure of β-1,6-glucans is also a crucial but under-investigated component of the Candida cell wall, which have been shown recently to be highly responsive to environmental factors such as pH, salt, and the presence of antifungals, and can be regulated to compensate for defects in O-/N-mannan elongation36.”

• cites [36] Bekirian, C. et al. β−1,6-glucan plays a central role in the structure and remodeling of the bilaminate fungal cell wall. eLife 13, RP100569 (2024). sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

KRE6 has been considered essential in C. albicans, but recent success in creating a homozygous KRE6 deletion mutant under specific conditions has raised questions about the essentiality of this gene.

“Similarly, KRE6 has been considered essential in C. albicans, but the recent success in creating a homozygous KRE6 deletion mutant under specific conditions has raised questions about the essentiality of this gene82.”

• cites [82] Han, Q. et al. Blocking β-1,6-glucan synthesis by deleting KRE6 and SKN1 attenuates the virulence of Candida albicans. Mol. Microbiol. 111, 604–620 (2019). sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

Dehydration of chitin reflects its antiparallel hydrogen-bonded packing of the linear carbohydrate chains that exclude most water molecules.

“Dehydration of chitin reflects its antiparallel hydrogen-bonded packing of the linear carbohydrate chains that exclude most water molecules.”

May combine two claims.

These spectral changes highlight compensatory responses to caspofungin treatment, characterized by a reduction in β-1,3-glucan levels, an increase in chitin content, and enhanced interactions between chitin microfibrils and mannan side chains.

“These spectral changes highlighted the compensatory responses to caspofungin treatment, characterized by a reduction in β-1,3-glucan levels, an increase in chitin content, and enhanced interactions between chitin microfibrils and mannan side chains.”

May combine two claims.

β-1,3-glucan adopts distinct structures when bound to chitin microfibrils compared to when partitioned into the solvent, with the former representing only a small fraction.

“The data clearly demonstrated that β-1,3-glucan adopts distinct structures when bound to chitin microfibrils compared to when partitioned into the solvent, with the former representing only a small fraction.”

Classified as probably not a substantial claim. May combine two claims.

The strains resistant to different antifungal classes may exhibit distinct structural characteristics-a hypothesis supported by the observed differences between strains AR386 and I.3 in this study.

“Moreover, it is crucial to investigate the link between cell wall structure and drug resistance in C. auris, because the strains resistant to different antifungal classes may exhibit distinct structural characteristics—a hypothesis supported by the observed differences between strains AR386 and I.3 in this study.”

• AssumptionRests on an assumption: The C. auris AR386 strain is representative of the C. auris species for cell wall structure and drug response.

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

The fungal material remaining in the MAS rotor after the experiment can be used to initiate new cultures, indicating that a substantial portion of the cells remain viable post-experiment.

“The fungal material remaining in the MAS rotor after the experiment can be used to initiate new cultures, indicating that a substantial portion of the cells remain viable post-experiment (Supplementary Fig. 17).”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

The NMR sensitivity enhancement (εon/off) was approximately 57 and the DNP signal buildup time ranged between 5.2 and 13 s.

“The NMR sensitivity enhancement (εon/off) was ~57 and the DNP signal buildup time ranged between 5.2 and 13 s.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

Open circle represent the mean of biological duplicates and error bars are s.d.

“Open circle represent the mean of biological duplicates and error bars are s.d.”

May combine two claims.

These changes likely resulted in weaker cell walls in kre6bΔ compared to kre6aΔ, which displayed reduced susceptibility to echinocandins.

“These changes likely resulted in weaker cell walls in kre6bΔ compared to kre6aΔ, which displayed reduced susceptibility to echinocandins (Fig. 6b).”

• AssumptionRests on an assumption: The C. auris AR386 strain is representative of the C. auris species for cell wall structure and drug response.

Probably not a substantial claim: may not pass S3 (load-bearing).

Resonance assignment of mobile carbohydrates was achieved using proton-detected 3D (and 2D) 1H-13C J-hCCH TOCSY with DIPSI-3 mixing.

“Resonance assignment of mobile carbohydrates were achieved using proton-detected 3D (as well as its 2D version) 1H-13C J-hCCH TOCSY126,127 with DIPSI-3 mixing128.”

• cites [126] Bax, A., Clore, G. M. & Gronenborn, A. M. 1H-1H correlation via isotropic mixing of 13C magnetization, a new three-dimensional approach for assigning 1H and 13C spectra of 13C-enriched proteins. J. Magn. Reson. 88, 425–431 (1990). sourceNot checked — no full text of the source
• cites [127] Clore, G. M., Bax, A., Driscoll, P. C., Wingfield, P. T. & Gronenborn, A. M. Assignment of the side-chain proton and carbon-13 resonances of interleukin-1.beta. using double- and triple-resonance heteronuclear three-dimensional NMR spectroscopy. Biochemistry 29, 8172–8184 (1990). sourceNot checked — no full text of the source
• cites [128] Shaka, A. J., Lee, C. J. & Pines, A. Iterative schemes for bilinear operators; application to spin decoupling. J. Mag. Reson. 77, 274–293 (1988). sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing).

The pathogenicity and immunogenicity of Candida species are closely linked to these morphological alterations.

“The pathogenicity and immunogenicity of Candida species are closely linked to these morphological alterations99.”

• cites [99] Sudbery, P., Gow, N. A. R. & Berman, J. The distinct morphogenic states of Candida albicans. Trends Microbiol 12, 317–325 (2004). sourceNot checked — no full text of the source

Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

The native hydration level of the fungal cells was fully retained throughout the sample preparation process.

“The native hydration level of the fungal cells was fully retained throughout the process.”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

1D 13C CP spectra were repeatedly acquired before and after 2D and 3D experiments to closely monitor sample integrity, and the 13C spectra are highly reproducible.

“1D 13C CP spectra were repeatedly acquired before and after 2D and 3D experiments to closely monitor sample integrity, and the 13C spectra are highly reproducible (Supplementary Fig. 16).”

Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

β-1,6-glucan appears to play critical roles in interacting with both mannan and chitin, as suggested by solid-state NMR results, and may exhibit different behaviors between C. albicans and C. auris in response to echinocandins.

“Another underexplored molecule is β-1,6-glucan36,82, which appears to play critical roles in interacting with both mannan and chitin, as suggested by solid-state NMR results, and may exhibit different behaviors between C. albicans and C. auris in response to echinocandins.”

• cites [36] Bekirian, C. et al. β−1,6-glucan plays a central role in the structure and remodeling of the bilaminate fungal cell wall. eLife 13, RP100569 (2024). sourceNot checked — no full text of the source
• cites [82] Han, Q. et al. Blocking β-1,6-glucan synthesis by deleting KRE6 and SKN1 attenuates the virulence of Candida albicans. Mol. Microbiol. 111, 604–620 (2019). sourceNot checked — no full text of the source

Classified as probably not a substantial claim.

The combined application of the 2D correlation and through-bond polarization transfer methods enables assignment of NMR resonances for various carbohydrate components.

“The combined application of these methods enables the assignment of NMR resonances for various carbohydrate components, with chemical shifts tabulated in Supplementary Table 1.”

Classified as probably not a substantial claim. May combine two claims.

The water-edited experiment's 1H-T2 filter (3.0 ms × 2 for apo C. auris and 5.0 ms × 2 for caspofungin-treated C. auris) eliminated 97% of polysaccharide signals while retaining 80% of water magnetization.

“followed by a 1H-T2 filter of 3.0 ms × 2 for the apo C. auris sample and 5.0 ms × 2 for the caspofungin-treated C. auris sample, which eliminated 97% of the polysaccharide signals while retaining 80% of the water magnetization.”

Probably not a substantial claim: may not pass S3 (load-bearing).

Bars represent the mean with s.d. of three biological replicates.

“Bars represent the mean with s.d. of three biological replicates.”

May combine two claims.

The combined methods facilitate differentiation of carbohydrate components' presence across different dynamic regimes and enhance the analysis of their polymorphic structure, composition, and physical packing within native cell walls.

“facilitates differentiation of their presence across different dynamic regimes, and enhances the analysis of their polymorphic structure, composition, and the physical packing of biomolecules within native cell walls.”

Probably not a substantial claim: may not pass S1 (asserted), S3 (load-bearing).

Alternatively, KRE6a and KRE6b may need to be present in equimolar amounts so that the overexpression of individual genes does not impact the overall biosynthesis process.

“Alternatively, KRE6a and KRE6b may need to be present in equimolar amounts so that the overexpression of individual genes does not impact the overall biosynthesis process.”

Probably not a substantial claim: may not pass S3 (load-bearing).

If the Kre6 proteins require stoichiometric levels of other ancillary proteins to form an active enzymatic complex, then overexpression may not result in a significant increase in enzymatic activities.

“If the Kre6 proteins require stoichiometric levels of other ancillary proteins to form an active enzymatic complex, then overexpression may not result in a significant increase in enzymatic activities.”