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Vibrio spp. play a crucial role in the global recycling of the highly abundant recalcitrant biopolymer chitin in marine ecosystems through their ability to secrete chitin‐degrading enzymes to efficiently hydrolyse chitinous materials and use them as their major carbon source. In this study, the first crystal structures of a complete four‐domain chitin‐active AA10 lytic polysaccharide monooxygenase from the chitinolytic bacterium Vibrio campbellii type strain ATCC BAA‐1116 are reported. The crystal structures of apo and copper‐bound VhLPMO10A were resolved as homodimers with four distinct domains: an N‐terminal AA10 catalytic (CatD) domain connected to a GlcNAc‐binding (GbpA_2) domain, followed by a module X domain and a C‐terminal carbohydrate‐binding module (CBM73). Size‐exclusion chromatography and small‐angle X‐ray scattering analysis confirmed that VhLPMO10A exists as a monomer in solution. The active site of VhLPMO10A is located on the surface of the CatD domain, with three conserved residues (His1, His98 and Phe170) forming the copper(II)‐binding site. Metal‐binding studies using synchrotron X‐ray absorption spectroscopy and X‐ray fluorescence, together with electron paramagnetic resonance spectroscopy, gave consistently strong copper(II) signals in the protein samples, confirming that VhLPMO10A is a copper‐dependent enzyme. ITC binding data showed that VhLPMO10A could bind various divalent cations but bound most strongly to copper(II) ions, with a Kd of 0.1 ± 0.01 µM. In contrast, a Kd of 1.9 nM was estimated for copper(I) ions from redox‐potential measurements. The presence of ascorbic acid is essential for H2O2 production in the reaction catalysed by VhLPMO10A. MALDI‐TOF MS identified VhLPMO10A as a C1‐specific LPMO, generating oxidized chitooligosaccharide products with different degrees of polymerization (DP2ox–DP8ox). This new member of the chitin‐active AA10 LPMOs could serve as a powerful biocatalyst in biofuel production from chitin biomass. [ABSTRACT FROM AUTHOR]

A highly sensitive and selective off–on bis-BODIPY fluorescent sensor, BODIPY-NN, could be used to detect Cu2+ ions in aqueous solutions and to image intracellular Cu2+ ions in living cells. Upon addition of Cu2+ ions, BODIPY-NN showed a turn-on fluorescence response with 4-fold enhancement at 529 nm, indicating the inhibition of photo-induced electron transfer (PET) from the phenylenediamine–triazole unit to the BODIPY skeleton. This was triggered by oxidative C–N cyclization of phenylenediamine–triazole to in situ form a benzimidazole–triazole receptor providing an appropriate coordination environment that enhances metal ion binding efficiency. A mechanistic study revealed that Cu2+ acts as both a catalyst for oxidative C–N cyclization and a coordination center. The detection limit was determined to be 85 nM. Importantly, BODIPY-NN has been successfully utilized for fluorescence imaging of Cu2+ ions in MCF-7 cells and showed no toxicity toward MCF-7 cells. The non-cytotoxic nature and high sensitivity of BODIPY-NN pave a way for its use in in vivo studies as a promising fluorescent probe for high-performance sensing and bioimaging. [ABSTRACT FROM AUTHOR]

Homogenous line widths, characterized by the phase memory time Tm, of a nitroxide spin-labeled peptide, were measured over the entire optional range (~10 (super -10)-10(super -4) s at the spectrometer resonance frequency of 9.7 GHZ using spin-echo correlated spectroscopy (SECSY) ESR.

Melanins are one of the great natural pigments produced by a wide variety of fungal species that promote fitness and cell survival in diverse hostile environments, including during mammalian infection. In this study, we sought to demonstrate the production of melanin in the conidia and hyphae of saprophytic fungi, including dematiaceous and hyaline fungi. We showed that a melanin‐specific monoclonal antibody (MAb) avidly labeled the cell walls of hyphae and conidia, consistent with the presence of melanin in these structures, in 14 diverse fungal species. The conidia of saprophytic fungi were treated with proteolytic enzymes, denaturant, and concentrated hot acid to yield dark particles, which were shown to be stable free radicals, consistent with their identification as melanins. Samples obtained from patients with fungal keratitis due to Fusarium falciforme, Aspergillus fumigatus, Aspergillus flavus, Curvularia lunata, Exserohilum rostratum, or Fonsecaea pedrosoi were found to be intensely labeled by the melanin‐specific MAb at the fungal hyphal cell walls. These results support the hypothesis that melanin is a common component that promotes survival under harsh conditions and facilitates fungal virulence. Increased understanding of the processes of melanization and the development of methods to interfere with pigment formation may lead to novel approaches to combat these complex pathogens that are associated with high rates of morbidity and mortality. [ABSTRACT FROM AUTHOR]

Treatment of a series of (imino)pyridine ligands bearing appended hydroxyl groups 2-((pyridin-2-ylmethylene)amino)phenol (Hpyph), 2-((pyridin-2-ylmethylene)amino)ethanol (Hpyet), and 3-((pyridin-2-ylmethylene)amino)propanol (Hpypr) with one equiv. of CuCl2·2H2O afforded the corresponding Cu(ii) complexes in low to moderate yields. The crystal structure of (μ-Cl)2[CuCl(κ2-N,N-Hpyet)]2 reveals a symmetric dinuclear structure with the bidentate N,N-coordination mode of (imino)pyridine with no Cu–OH interaction. On the other hand, the dinuclear Cu(ii) complex of the related propyl ligand Hpypr possesses a significantly different crystal structure involving nucleophilic addition of the hydroxyl group to the aldehyde group of 2-pyridinecarboxaldehyde. The Cu complex/Cu0/TEMPO/Na2CO3 (TEMPO = 2,2,6,6-tetramethylpiperidinyl-1-oxyl) catalyst system generally exhibited good activity for aerobic oxidation of benzyl alcohol to benzaldehyde in H2O at room temperature. The dinuclear Cu(ii) complex (μ-Cl)2[CuCl(κ2-N,N-Hpyet)]2 was demonstrated as an effective catalyst toward aerobic oxidation of various benzyl alcohol derivatives, cinnamyl alcohol, and 2-thiophenemethanol. [ABSTRACT FROM AUTHOR]

3,4-dihydroxyphenylacetate (DHPA) dioxygenase (DHPAO) from Pseudomonas aeruginosa (PaDHPAO) was overexpressed in Escherichia coli and purified to homogeneity. As the enzyme lost activity over time, a protocol to reactivate and conserve PaDHPAO activity has been developed. Addition of Fe(II), DTT and ascorbic acid or ROS scavenging enzymes (catalase or superoxide dismutase) was required to preserve enzyme stability. Metal content and activity analyses indicated that PaDHPAO uses Fe(II) as a metal cofactor. NMR analysis of the reaction product indicated that PaDHPAO catalyzes the 2,3-extradiol ring-cleavage of DHPA to form 5-carboxymethyl-2-hydroxymuconate semialdehyde (CHMS) which has a molar absorptivity of 32.23 mM-1cm-1 at 380 nm and pH 7.5. Steady-state kinetics under air-saturated conditions at 25°C and pH 7.5 showed a Km for DHPA of 58 ± 8 μM and a kcat of 64 s-1, indicating that the turnover of PaDHPAO is relatively fast compared to other DHPAOs. The pH-rate profile of the PaDHPAO reaction shows a bell-shaped plot that exhibits a maximum activity at pH 7.5 with two pKa values of 6.5 ± 0.1 and 8.9 ± 0.1. Study of the effect of temperature on PaDHPAO activity indicated that the enzyme activity increases as temperature increases up to 55°C. The Arrhenius plot of ln(k’cat) versus the reciprocal of the absolute temperature shows two correlations with a transition temperature at 35°C. Two activation energy values (Ea) above and below the transition temperature were calculated as 42 and 14 kJ/mol, respectively. The data imply that the rate determining steps of the PaDHPAO reaction at temperatures above and below 35°C may be different. Sequence similarity network analysis indicated that PaDHPAO belongs to the enzyme clusters that are largely unexplored. As PaDHPAO has a high turnover number compared to most of the enzymes previously reported, understanding its biochemical and biophysical properties should be useful for future applications in biotechnology. [ABSTRACT FROM AUTHOR]

A series of benzimidazole–triazole ligands (NN′) having a pendent triazole arm with different triazole substituents including CH2Ph (3a), cyclo-C6H11 (3b), and CH2SiMe3 (3c) were obtained in moderate yields from Cu-catalyzed oxidative C–N cyclization of the respective amine-triazole compounds N,N′-bis((1-R-1,2,3-triazol-4-yl)methyl)benzene-1,2-diamine (2a–2c). Treatment of CuCl2 with one equiv. of the benzimidazole–triazole ligands afforded the corresponding CuII complexes with the empirical formula of Cu(NN′)Cl2 (4a–4c). Crystal structures of 4b and 4c reveal mononuclear and dinuclear CuII complexes, respectively. Despite the differences in triazole substituents and their solid state structures, ESR spectra indicate the same molecular structures in CH3CN solution whereas CV data suggest similar redox potentials for 4a–4c. Catalytic activities for aerobic oxidation of benzyl alcohol to benzaldehyde follow this trend: 4c ＞ 4a ＞ 4b. In addition, the catalytic system 4c/TEMPO/Cu0/NMI (TEMPO = 2,2,6,6-tetramethyl-1-piperidinyloxyl, NMI = N-methylimidazole) exhibited high activities for oxidation of activated alcohols (i.e., benzyl alcohol derivatives and allylic alcohol) in CH3CN at room temperature. [ABSTRACT FROM AUTHOR]

Zinc oxide (ZnO) nanoparticles have recently been identified as a promising candidate for advanced nanophotonics applications and quantum technologies. This work reports the formation of luminescent point defects and describes their photophysical properties. In particular, it is shown using correlative photoluminescence, cathodoluminescence, electron paramagnetic resonance (EPR), and X‐ray absorption near‐edge spectroscopy that green luminescence at 2.48 eV and an EPR line at g = 2.00 belong to a surface oxygen vacancy ( V o , s +) center, while a second green emission at 2.28 eV is associated with zinc vacancy (VZn) centers. It is established that radiative point defects can be excited in the visible that exhibits nanosecond lifetimes using both above bandgap and sub‐bandgap 405 and 532 nm excitation. This work provides important knowledge towards employment of point defects in ZnO in nanophotonics technologies. [ABSTRACT FROM AUTHOR]

Melanins are synthesized by organisms of all biological kingdoms and comprise a heterogeneous class of natural pigments. Certain of these polymers have been implicated in the pathogenesis of several important human fungal pathogens. This study investigated whether the fungal skin pathogen Malassezia furfur produces melanin or melanin-like compounds. A melanin-binding monoclonal antibody (MAb) labelled in vitro cultivated yeast cells of M. furfur. In addition, melanization of Malassezia yeasts and hyphae was detected by anti-melanin MAb in scrapings from patients with pityriasis versicolor. Treatment of Malassezia yeasts with proteolytic enzymes, denaturant and concentrated hot acid yielded dark particles and electron spin resonance spectroscopy revealed that these particles contained a stable free radical compound, consistent with their identification as melanins. Malassezia yeasts required phenolic compounds, such as L-DOPA, in order to synthesize melanin. L-DOPA also triggered hyphal formation in vitro when combined with kojic acid, a tyrosinase inhibitor, in a dose-dependent manner. In this respect, L-DOPA is thought to be an essential substance that is linked to both melanization and yeast-mycelial transformation in M. furfur. In summary, M. furfur can produce melanin or melanin-like compounds in vitro and in vivo, and the DOPA melanin pathway is involved in cell wall melanization. [ABSTRACT FROM AUTHOR]

Iron oxide/carbon nanocatalysts were successfully synthesized by the calcination of ferrocenium at high temperatures ranging from 500 to 900 °C. Then the synthesized nanocomposites were characterized by XRD (X-Ray Diffraction), TEM (Transmission Electron Microscopy), VSM (Vibrating-Sample Magnetometry), BET (Brunauer-Emmett-Teller surface area measurements), TGA (Thermogravimetric Analysis), XPS (X-Ray Photoelectron Spectroscopy), EPR (Electron Paramagnetic Resonance), and CHN elemental analysis. The prepared nanocatalysts were applied for the decomposition of methylene blue as a model in wastewater treatment. It was unexpected to discover that the prepared nanocatalysts were highly active for the reaction with methylene blue in the dark even though no excess of hydrogen peroxide was added. The nanocatalyst calcined at 800 °C exhibited the rod shape with the best catalytic activity. The nanocatalysts could be reused for 12 times without the significant loss of the catalytic activity. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Hexanuclear Cu(II) complexes supported by tris(triazolyl) ligands were synthesized. • New synthetic protocol of Cu(II) clusters from aerobic oxidation of Cu(I) substrate with tris(triazolyl)methanol was shown. • Magnetic studies of these complexes reveal a strong overall antiferromagnetic behavior. Abstract Treatment of tris(4-benzyl-1,2,3-triazolyl)methanol (Htbtm) with CuIBr in refluxed CH 2 Cl 2 under aerobic conditions afforded the hexanuclear CuII complexes with the formula Cu 6 (µ 3 -O⋯H⋯O-µ 3)(µ-tbtm-к3- O , N 3, N 3′) 3 (µ-Br) 6. The crystal structure reveals two Cu 3 (µ-Br) 3 (µ 3 -O) units linked by three deprotonated tbtm ligands and a proton. Magnetic studies of the hexanuclear CuII cluster show a strong overall antiferromagnetic behavior with a dominant coupling interaction between CuII⋯CuII ions of different Cu 3 (µBr) 3 (µ 3 -O) units. [ABSTRACT FROM AUTHOR]

The article reports on the use of electron spin resonance to examine long-range distances in water-soluble bis-peptide molecular rods. Designed shapes using restricted building blocks were used for the rapid synthesis of water-soluble nanoscale molecules. The building blocks were coupled through pairs of bonds to create spiro-ladder oligomers.

Melanins are synthesized by organisms of all biological kingdoms and comprise a heterogeneous class of natural pigments. Certain of these polymers have been implicated in the pathogenesis of several important human fungal pathogens. This study investigated whether the fungal skin pathogen Malassezia furfur produces melanin or melanin-like compounds. A melanin-binding monoclonal antibody (MAb) labelled in vitro cultivated yeast cells of M. furfur. In addition, melanization of Malassezia yeasts and hyphae was detected by anti-melanin MAb in scrapings from patients with pityriasis versicolor. Treatment of Malassezia yeasts with proteolytic enzymes, denaturant and concentrated hot acid yielded dark particles and electron spin resonance spectroscopy revealed that these particles contained a stable free radical compound, consistent with their identification as melanins. Malassezia yeasts required phenolic compounds, such as L-DOPA, in order to synthesize melanin. L-DOPA also triggered hyphal formation in vitro when combined with kojic acid, a tyrosinase inhibitor, in a dose-dependent manner. In this respect, L-DOPA is thought to be an essential substance that is linked to both melanization and yeast-mycelial transformation in M. furfur. In summary, M. furfur can produce melanin or melanin-like compounds in vitro and in vivo, and the DOPA melanin pathway is involved in cell wall melanization. [ABSTRACT FROM AUTHOR]

In apoptotic pathway, the interaction of Cytochrome c (Cytc) with cardiolipin in vivo is a key process to induce peroxidase activity of Cytc and trigger the release of Cytc in the inner mitochondria into cytosol. The peroxidase active form of Cytc occurs due to local conformational changes that support the opening of the heme crevice and the loss of an axial ligand between Met80 and heme Fe. Structural adjustments at the Ω-loop segments of Cytc are required for such process. To study the role of the distal Ω-loop segments comprising residues 71–85 in human Cytc (hCytc), we investigated a cysteine mutation at Pro76, one of the highly conserved residues in this loop. The effect of P76C mutant was explored by the combination of experimental characterizations and molecular dynamics (MD) simulations. The peroxidase activity of the P76C mutant was found to be significantly increased by ∼13 folds relative to the wild type. Experimental data on global denaturation, alkaline transition, heme bleaching, and spin-labeling Electron Spin Resonance were in good agreement with the enhancement of peroxidase activity. The MD results of hCytc in the hexacoordinate form suggest the important changes in P76C mutant occurred due to the unfolding at the central Ω-loop (residues 40–57), and the weakening of H-bond between Tyr67 and Met80. Whereas the experimental data implied that the P76C mutant tend to be in equilibrium between the pentacoordinate and hexacoordinate forms, the MD and experimental information are complementary and were used to support the mechanisms of peroxidase active form of hCytc. [Display omitted] • Native structure of human Cytochrome c does not support peroxidase activity. • Activating the peroxidase activity of Cytochrome c is a crucial step on the initiation of the apoptotic pathway. • Mutation in the distal Ω-loop at P76C position increases peroxidase activity of human Cytochrome c. • Molecular dynamics revealed the conformational changes of P76C mutant support the increase of peroxidase activity. [ABSTRACT FROM AUTHOR]

Zeolitic imidazolate framework-67 (ZIF-67) was synthesized in situ in the hydrogel of chitosan-grafted poly(acrylic acid) (chitosan- g -PAA) to activate peroxymonosulfate (PMS) and degrade tetracycline (TC). The catalytic performance of the composite hydrogel for TC degradation was evaluated under different conditions. The results showed rapid degradation, with enhanced degradation efficiency as the catalyst dosage, PMS dosage, and temperature increased. TC was degraded entirely within 30 min for catalyst and PMS dosages of 1 and 1 g per L, respectively. The composite hydrogel was effective across a broad pH range. A scavenging study and electron paramagnetic resonance experiments indicated that SO 4 ˙ - , HO˙, O 2 ˙ - and 1 O 2 were involved in the degradation process. The antibacterial test against E. coli showed that the products of the TC degradation were nontoxic. Additionally, the composite hydrogel was evaluated in the presence of anions and in real water samples. The reusability study showed that the composite hydrogel could be recovered through filtration and effectively used for five consecutive cycles. Moreover, the composite hydrogel could degrade 82% ciprofloxacin and 86% norfloxacin, while it could completely degrade rhodamine B, reactive red 141, and methylene blue dyes within 30 min., Competing Interests: There are no conflicts of interest to declare., (This journal is © The Royal Society of Chemistry.)

Carbon dioxide (CO 2 ) photoreduction is an intriguing approach that converts CO 2 into high-value substances with the assistance of a photocatalyst. Key to effective photoreduction is to promote the interaction of photo-induced holes and a sacrificial reagent (SCR), separating the holes from photoelectrons and enhancing the rate of the subsequent product generation. Methanol, ethanol, isopropanol, and water SCRs were tested for their ability to assist a copper-doped sodium dititanate nanosheets/graphene oxide heterostructure (CTGN) in CO 2 photoreduction. The CTGN photocatalyst was suspended in a CO 2 -saturated aqueous solution with the assigned SCR while illuminated by a mercury lamp. Product samples from the gas and liquid phases were analyzed for targeted product compositions. Methanol SCR exhibited the best performance in facilitating CO 2 photoreduction, producing ethanol as the main product at a total carbon consumption (TCC) of 6544 μmol g cat -1 . The remarkable performance of methanol is attributed to the high diffusivity and excellent stability of the hydroxymethyl radical that developed during the photoreduction. The kinetics studies revealed the first and second order for the CO 2 depletion and product generation rates, respectively, for the alcohol SCRs., Competing Interests: The authors declare no financial or personal interest that affects professional judgment regarding the validity and analysis of this research., (This journal is © The Royal Society of Chemistry.)

Conventional treatment of jaw bone infection is often ineffective at controlling bacterial infection and enhancing bone regeneration. Biodegradable composite hydrogels comprised of carboxymethyl chitosan (CMCS) and clindamycin (CDM)-loaded mesoporous silica nanoparticles (MCM-41), possessing dual antibacterial activity and osteogenic potency, were developed in the present study. CDM was successfully loaded into both untreated and plasma-treated MCM-41 nanoparticles, denoted as (p)-MCM-41, followed by the incorporation of each of CDM-loaded (p)-MCM-41 into CMCS. The resulting CDM-loaded composite hydrogels, (p)-MCM-41-CDM-CMCS, demonstrated slow degradation rates (about 70% remaining weight after 14-day immersion), while the CDM-free composite hydrogel entirely disintegrated after 4-day immersion. The plasma treatment was found to improve drug loading capacity and slow down initial drug burst effect. The prolonged releases of CDM from both (p)-MCM-41-CDM-CMCS retained their antibacterial effect against Streptococcus sanguinis for at least 14 days in vitro. In vitro assessment of osteogenic activity showed that the CDM-incorporated composite hydrogel was cytocompatible to human mesenchymal stem cells (hMSCs) and induced hMSC mineralization via p38-dependent upregulated alkaline phosphatase activity. In conclusion, novel (p)-MCM-41-CDM-CMCS hydrogels with combined controlled release of CDM and osteogenic potency were successfully developed for the first time, suggesting their potential clinical benefit for treatment of intraoral bone infection., (© 2021 The Authors.)

A series of benzimidazole-triazole ligands (NN') having a pendent triazole arm with different triazole substituents including CH 2 Ph (3a), cyclo-C 6 H 11 (3b), and CH 2 SiMe 3 (3c) were obtained in moderate yields from Cu-catalyzed oxidative C-N cyclization of the respective amine-triazole compounds N,N'-bis((1-R-1,2,3-triazol-4-yl)methyl)benzene-1,2-diamine (2a-2c). Treatment of CuCl 2 with one equiv. of the benzimidazole-triazole ligands afforded the corresponding Cu II complexes with the empirical formula of Cu(NN')Cl 2 (4a-4c). Crystal structures of 4b and 4c reveal mononuclear and dinuclear Cu II complexes, respectively. Despite the differences in triazole substituents and their solid state structures, ESR spectra indicate the same molecular structures in CH 3 CN solution whereas CV data suggest similar redox potentials for 4a-4c. Catalytic activities for aerobic oxidation of benzyl alcohol to benzaldehyde follow this trend: 4c > 4a > 4b. In addition, the catalytic system 4c/TEMPO/Cu 0 /NMI (TEMPO = 2,2,6,6-tetramethyl-1-piperidinyloxyl, NMI = N-methylimidazole) exhibited high activities for oxidation of activated alcohols (i.e., benzyl alcohol derivatives and allylic alcohol) in CH 3 CN at room temperature.

Melanins are synthesized by organisms of all biological kingdoms and comprise a heterogeneous class of natural pigments. Certain of these polymers have been implicated in the pathogenesis of several important human fungal pathogens. This study investigated whether the fungal skin pathogen Malassezia furfur produces melanin or melanin-like compounds. A melanin-binding monoclonal antibody (MAb) labelled in vitro cultivated yeast cells of M. furfur. In addition, melanization of Malassezia yeasts and hyphae was detected by anti-melanin MAb in scrapings from patients with pityriasis versicolor. Treatment of Malassezia yeasts with proteolytic enzymes, denaturant and concentrated hot acid yielded dark particles and electron spin resonance spectroscopy revealed that these particles contained a stable free radical compound, consistent with their identification as melanins. Malassezia yeasts required phenolic compounds, such as L-DOPA, in order to synthesize melanin. L-DOPA also triggered hyphal formation in vitro when combined with kojic acid, a tyrosinase inhibitor, in a dose-dependent manner. In this respect, L-DOPA is thought to be an essential substance that is linked to both melanization and yeast-mycelial transformation in M. furfur. In summary, M. furfur can produce melanin or melanin-like compounds in vitro and in vivo, and the DOPA melanin pathway is involved in cell wall melanization.

Pulsed electron-electron double resonance (PELDOR, also known as DEER) has become a method of choice to measure distances in biomolecules. In this work we show how the performance of the method can be improved at high EPR frequencies (94 GHz) using variable dual frequency irradiation in a dual mode cavity in order to obtain enhanced resolution toward orientation selection. Dipolar evolution traces of a representative RNA duplex and an α-helical peptide were analysed in terms of possible bi-radical structures by considering the inherent ambiguity of symmetry-related solutions.

Dermatophytes are keratinophilic fungi that are the most common cause of fungal skin infections worldwide. Melanin has been isolated from several important human fungal pathogens, and the polymeric pigment is now recognized as an important virulence determinant. This study investigated whether dermatophytes, including Trichophyton rubrum, Trichophyton mentagrophytes, Epidermophyton floccosum and Microsporum gypseum, produce melanin or melanin-like compounds in vitro and during infection. Digestion of the pigmented microconidia and macroconidia of dermatophytes with proteolytic enzymes, denaturant and hot concentrated acid yielded dark particles that retained the size and shape of the original fungal cells. Electron spin resonance spectroscopy revealed that particles derived from pigmented conidia contained a stable free radical signal, consistent with the pigments being a melanin. Immunofluorescence analysis demonstrated reactivity of a melanin-binding mAb with the pigmented conidia and hyphae, as well as the isolate particles. Laccase, an enzyme involved in melanization, was detected in the dermatophytes by an agar plate assay using 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as the substrate. Skin scrapings from patients with dermatophytoses contained septate hyphae and arthrospores that were reactive with the melanin-binding mAb. These findings indicate that dermatophytes can produce melanin or melanin-like compounds in vitro and during infection. Based on what is known about the function of melanin as a virulence factor of other pathogenic fungi, this pigment may have a similar role in the pathogenesis of dermatophytic diseases.

We demonstrate the synthesis of a series of spin-labeled curved oligomers to determine their end-to-end lengths and distance distributions using electron spin resonance. We synthesize shape-persistent macromolecules from conformationally restricted, asymmetric monomers that are coupled through pairs of amide bonds to create water-soluble, spiro-ladder oligomers with well-defined three-dimensional structures. We synthesized seven different macromolecules, each containing eight monomers but differing in the sequence to create macromolecules with different curved shapes. The ends of the oligomers were labeled with nitroxide spin probes, and double electron-electron resonance (DEER) electron spin resonance (ESR) experiments were carried out to obtain quantitative information about the shapes and flexibility of the oligomers. The most probable end-to-end distance of the oligomers ranges from 23 to 36 A, a range of length that we previously accessed by assembling rod-like homo-oligomers that contain 4-8 bisamino acid monomers. The relative distances measured for the oligomers confirm that, by varying the sequence of an oligomer, we are able to control its shape. The shapes of the ESR-derived population distributions allow us to compare the degree of shape persistence and flexibility of spiro-ladder oligomers to other well-studied nanoscale molecular structures such as p-phenylethynylenes.