Common Features and Intra-Species Variation of Cutibacterium modestum Strains, and Emended Description of the Species.

Cutibacterium modestum is a new species coined in 2020 as the fifth species of genus Cutibacterium, which includes Cutibacterium acnes. The species is predicted as a minor but common member of skin microbiome and includes a group tentatively named as "Propionibacterium humerusii". The description of the species has been provided only with a single strain. To establish the characteristics of C. modestum and search for possible disease-related subtypes, we investigated the biochemical characteristics of eight live strains and performed in silico comparison of nine genomes. The common features, which included the morphology of Gram-stain positive short rods, the negativity of phenylalanine arylamidase, and several unique MALDI-TOF MS spectral peaks, were considered useful in laboratory identification. Pairwise comparisons of the genomes by in silico DNA-DNA hybridization showed similarity values of 98.1% or larger, which were far higher than the subspecies cutoff of 79-80%. The 16S rRNA gene sequences of thirteen isolates and genomes were identical. Their recA gene sequences were identical except for two strains, HM-510 (HL037PA2) and Marseille-P5998, which showed unique one-nucleotide polymorphisms. The biochemical features using API kits were slightly different among the isolates but far closer than those of the nearest other species, C. acnes and Cutibacterium namnetense. Spectra of MALDI-TOF mass spectrometry showed slight differences in the presence of m/z 10,512 (10 kD chaperonin GroS) and three other peaks, further clustering the eight isolates into three subtypes. These results indicated that these isolates did not separate to form subspecies-level clusters, but subtyping is possible by using recA gene sequences or MALDI-TOF mass spectrometry spectra. Moreover, this work has confirmed that a group "P. humerusii" is included in C. modestum.

What Do We See in Spectra?: Assignment of High-Intensity Peaks of Cutibacterium and Staphylococcus Spectra of MALDI-TOF Mass Spectrometry by Interspecies Comparative Proteogenomics.

Matrix-assisted laser-desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry is a widely used and reliable technology to identify microbial species and subspecies. The current methodology is based on spectral fingerprinting, analyzing protein peaks, most of which are yet to be characterized. In order to deepen the understanding of these peaks and to develop a more reasonable identification workflow, we applied proteogenomic approaches to assign the high-intensity peaks of MALDI-TOF spectra of two bacterial genera. First, the 3-22 kD proteomes of 5 Cutibacterium strains were profiled by UPLC-MS/MS, and the amino acid sequences were refined by referring to their genome in the public database. Then, the sequences were converted to m/z (x-axis) values based on their molecular masses. When the interspecies comparison of calculated m/z values was well-fitted to the observed peaks, the peak assignments for the five Cutibacterium species were confirmed. Second, the peak assignments for six Staphylococcus species were performed by using the above result for Cutibacterium and referring to ribosomal subunit proteins coded on the S10-spc-alpha operon (the S10-GERMS method), a previous proteomics report by Becher et al., and comprehensive genome analysis. We successfully assigned 13 out of 15 peaks for the Cutibacterium species and 11 out of 13 peaks for the Staphylococcus species. DNA-binding protein HU, the CsbD-like protein, and 50S ribosomal protein L7/L12 were observed in common. The commonality suggests they consist of high-intensity peaks in the MALDI spectra of other bacterial species. Our workflow may lead to the development of a more accurate species identification database of MALDI-TOF mass spectrometry based on genome data.

A phase III, 52-week, open-label study to evaluate the safety and efficacy of 5% sofpironium bromide (BBI-4000) gel in Japanese patients with primary axillary hyperhidrosis.

A long-term study was conducted in Japanese patients with primary axillary hyperhidrosis who completed the preceding 6-week phase III, confirmatory study of 5% sofpironium bromide gel (hereinafter referred to as sofpironium) to evaluate the safety and efficacy of 52-week treatment with sofpironium. In the long-term study, 185 patients who completed the confirmatory study (94 and 91 patients in the vehicle and sofpironium groups, respectively) started to receive sofpironium (switching and extension groups, respectively), and all these patients were included in both the full analysis set (FAS) and the safety analysis set (SAF). In the FAS, there were more females than males (73.0% vs. 27.0%), and median age was 38.0 years. A total of 161 patients (86 and 75 patients in the switching and extension groups, respectively) completed the study at week 52. The proportions of patients with hyperhidrosis disease severity score of 1 or 2 and a 50% or more reduction in total gravimetric weight of sweat were 57.4% in the switching group and 58.2% in the extension group at week 52. The proportions of patients who achieved this efficacy end-point in the long-term study were similar to that (53.9%) in the sofpironium group in the confirmatory study. In the SAF, the incidences of adverse events (AEs) were 80.9% in the switching group and 83.5% in the extension group, and the incidences of adverse drug reactions were 39.4% and 45.1%, respectively. AEs that occurred in at least 20% of patients in both treatment groups were application site dermatitis (25.5% and 33.0%, respectively) and nasopharyngitis (31.9% and 23.1%, respectively). Reported AEs were generally mild, and there were no deaths. Serious AEs occurred in three patients, but none were considered related to the study drug. In this study, the efficacy of sofpironium was maintained during 52-week treatment, and no new safety risk was observed.

Electron Beam Irradiation of Lead Halide Perovskite Solar Cells: Dedoping of Organic Hole Transport Materials despite Hardness of the Perovskite Layer.

Organic-inorganic lead halide perovskite solar cells (PSCs) are highly efficient, flexible, lightweight, and even tolerant to radiation, such as protons, electron beams (EB), and γ-rays, all of which makes them plausible candidates for use in space satellites and spacecrafts. However, the mechanisms of radiation damage of each component of PSC [an organic hole transport material (HTM), a perovskite layer, and an electron transport material (ETM)] are not yet fully understood. Herein, we investigated the EB irradiation effect (100 keV, up to 2.5 × 1015 cm-2) on binary-mixed A site cations and halide perovskite (MA0.13FA0.87PbI2.61Br0.39, MA:methylammonium cation and FA:formaminidium cation), a molecular HTM of doped SpiroOMeTAD, and an inorganic ETM of mesoporous TiO2. Despite the decreased power conversion efficiency of PSCs upon EB exposure, the photoconductivities of the perovskite, HTM, and ETM layers remained intact. In contrast, significant dedoping of HTM was observed, as confirmed by steady-state conductivity, photoabsorption, and X-ray photoelectron spectroscopy measurements. Notably, this damage could be healed by exposure to short-wavelength light, leading to a partial recovery of the PSC efficiency. Our work exemplifies the robustness of perovskite against EB and the degradation mechanism of the overall PSC performance.

A phase 3, multicenter, randomized, double-blind, vehicle-controlled, parallel-group study of 5% sofpironium bromide (BBI-4000) gel in Japanese patients with primary axillary hyperhidrosis.

A phase 3 study was conducted to verify the efficacy and safety of 5% sofpironium bromide (BBI-4000) gel (hereinafter referred to as sofpironium) administrated for 6 weeks in Japanese patients with primary axillary hyperhidrosis. The primary efficacy end-point was the proportion of patients who satisfied both criteria of a Hyperhidrosis Disease Severity Score (HDSS) of 1 or 2 at the end of 6-week treatment and a 50% or more reduction in total gravimetric weight of sweat at the end of treatment relative to baseline. A total of 281 patients were randomized to receive 5% sofpironium (141 patients) or vehicle (140 patients), and all patients were included in the full analysis set (FAS). In the FAS, 70.1% of patients were female, and the median age was 35.0 years. The proportion of patients who achieved the primary efficacy end-point was 53.9% in the sofpironium group and 36.4% in the vehicle group, with a statistically significant difference of 17.5% (95% confidence interval, 6.02-28.93) between these two groups (P = 0.003). The incidence of adverse events was 44.0% in the sofpironium group and 30.7% in the vehicle group, and the incidence of adverse drug reactions was 16.3% in the sofpironium group and 5.0% in the vehicle group. Reported adverse events were generally mild or moderate in severity. In the sofpironium group, common events (incidence, ≥5%) were nasopharyngitis (14.2%) and dermatitis/erythema at the application site (8.5%/5.7%), with no serious adverse events reported. This study demonstrated the efficacy and safety of 5% sofpironium.

Cutibacterium modestum sp. nov., isolated from meibum of human meibomian glands, and emended descriptions of Cutibacterium granulosum and Cutibacterium namnetense.

An anaerobic and aerotolerant bacterium, strain M12T, was isolated from the meibum of inflamed human meibomian glands. Cells of the strain was Gram-stain-positive, non-spore-forming and non-motile rods. Growth on trypticase soy agar plates supplemented with 5 % sheep blood was fastest at 30-37 °C under anaerobic conditions. The 16S rRNA gene sequence of the strain revealed that it belongs to the genus Cutibacterium with a 98.0 % similarity value to the closest species, Cutibacterium acnes. Genome analysis of the strain with type strains of the other Cutibacterium species resulted in digital DNA-DNA hybridization values of 32.3-22.3% and average nucleotide identity (OrthoANI) values of 86.7-73.6 %. Biochemical and physiological analyses using API rapid ID 32A and API Coryne kits revealed relatively low reactivity of the strain compared with C. acnes and Cutibacterium namnetense. The most abundant major cellular fatty acid was iso-C15 : 0. Fermentation end-products from glucose were propionate, lactate, succinate and acetate. The diagnostic diamino acid of the peptidoglycan was meso-diaminopimelic acid. Major menaquinones were MK-9(H4), MK-9(H2) and MK-9. The major peaks of the MALDI-TOF mass spectrometry spectrum were at 3493, 3712, 6986 and 7424 Da. The DNA G+C content was 59.9 mol%. Based on these findings, we propose a novel species, Cutibacterium modestum. The type strain of C. modestum is M12T (=JCM 33380T=DSM 109769T). On the basis of further genomic analysis, we also provide emended descriptions of Cutibacterium granulosum (Prévot 1938) Scholz and Kilian 2016 and Cutibacterium namnetense (Aubin et al. 2016) Nouioui et al. 2018.

Abrupt Intralesional Color Change on Dermoscopy as a New Indicator of Early Superficial Spreading Melanoma in a Japanese Woman.

Diagnosis of superficial spreading melanoma in the early stage is often difficult, even with dermoscopy. We report the case of a 37-year-old Japanese woman with superficial spreading melanoma in her left buttock. The lesion developed 20 years before becoming visible and gradually enlarged over the past few years without any symptoms. Physical examination showed a well-demarcated dark-brown macule 10 mm in diameter. Dermoscopy demonstrated a central dark area with a blue-grey structureless area, a milky-red area with irregular blue-grey dots or globules suggestive of regression structures, and multifocal black pigmentation with whitish scaly areas. An abrupt intralesional change in color from a central dark area to a peripheral light-brown area was also seen. The peripheral area showed an atypical pigment network with an obscure mesh and holes. Histopathologic examination of the lesion showed acanthosis with melanocytic proliferation and nuclear atypia, a band-like lymphocytic infiltrate, melanophages and a few nests of melanocytes just beneath the epidermis. The epidermal melanocytes were positive for S-100, Melan-A and HMB-45, but the dermal nests of melanocytes were negative for HMB-45 and positive for S-100 and Melan-A. A diagnosis of superficial spreading melanoma with a tumor thickness of 0.4 mm (pT1aN0M0, stage 1A) was established based on the clinical, dermoscopic and histopathologic findings. This case suggests that dermoscopy is useful in the diagnosis of this condition. An abrupt intralesional change of color might be a new indicator of early superficial spreading melanoma.

Enzyme linking to DNA aptamers via a zinc finger as a bridge.

We propose a novel enzyme-labeling method for DNA aptamers using enzyme-fused zinc finger proteins. We achieved thrombin detection and vascular endothelial growth factor detection using zinc finger-fused firefly luciferase.

Automatic polymerase chain reaction product detection system for food safety monitoring using zinc finger protein fused to luciferase.

An automatic polymerase chain reaction (PCR) product detection system for food safety monitoring using zinc finger (ZF) protein fused to luciferase was developed. ZF protein fused to luciferase specifically binds to target double stranded DNA sequence and has luciferase enzymatic activity. Therefore, PCR products that comprise ZF protein recognition sequence can be detected by measuring the luciferase activity of the fusion protein. We previously reported that PCR products from Legionella pneumophila and Escherichia coli (E. coli) O157 genomic DNA were detected by Zif268, a natural ZF protein, fused to luciferase. In this study, Zif268-luciferase was applied to detect the presence of Salmonella and coliforms. Moreover, an artificial zinc finger protein (B2) fused to luciferase was constructed for a Norovirus detection system. In the luciferase activity detection assay, several bound/free separation process is required. Therefore, an analyzer that automatically performed the bound/free separation process was developed to detect PCR products using the ZF-luciferase fusion protein. By means of the automatic analyzer with ZF-luciferase fusion protein, target pathogenic genomes were specifically detected in the presence of other pathogenic genomes. Moreover, we succeeded in the detection of 10 copies of E. coli BL21 without extraction of genomic DNA by the automatic analyzer and E. coli was detected with a logarithmic dependency in the range of 1.0×10 to 1.0×10(6) copies.

Detection of pathogenic bacteria by using zinc finger protein fused with firefly luciferase.

We constructed a novel bacterial genome detection system using zinc finger protein (ZF) fused with firefly luciferase (ZF-luciferase). Taking advantage of the direct recognition of double-stranded DNA (dsDNA) by ZF, we previously constructed bacterial genome detection systems that did not require dehybridization processes. To detect polymerase chain reaction (PCR) products rapidly and with a high sensitivity, we constructed two kinds of ZF-luciferase, Sp1-fused luciferase (Sp1-luciferase), and Zif268-fused luciferase (Zif268-luciferase). ZF-luciferase not only maintains luciferase activity but also shows dsDNA-binding ability and specificity. Furthermore, we succeeded in the detection of 10 copies of the genome of Legionella pneumophila and Escherichia coli O157. ZF-luciferase would be a useful tool for highly sensitive detection of pathogenic bacterial genome.

Neural activation during imitation of movements presented from four different perspectives: a functional magnetic resonance imaging study.

The present study examined the effects of the perspective of movement presented for imitation in healthy volunteers, using functional magnetic resonance imaging (fMRI) to assess the magnitude and distribution of elicited brain activity. We sought to identify the pattern of brain activity associated with the performance of finger imitation tasks under four different imitation conditions. Video presentations of a hand and forearm performing random sequential contact between different fingers and the thumb were presented for imitation, while fMRI was recorded. The four types of model for imitation were: a hand and forearm pointing away from the subject, as if the subject were looking at their own limb (first-person perspective), from both anatomical (a right hand to be imitated by the subject's own right hand) and specular (a mirror image or "left" hand to be imitated by the subject's right hand) perspectives; and a hand/forearm pointing toward the subject, as if it was the hand of another person facing the subject (third-person perspective), from both anatomical (the opposite person's right hand) and specular (the opposite person's left hand) perspectives. In addition, participants completed a motor control task. The results revealed a significant difference in the magnitude of brain activation between the first- and third-person perspective conditions, suggesting that subjects used the first-person imitation model as a substitute for internal self-representation, thus requiring less effort. The first-person perspective anatomical model activated only the right posterior insula, recruiting significantly fewer brain regions than the other model types, compared with the control condition. These findings suggest that first-person anatomical perspective models may be optimal for ease of imitation in motor learning.