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This work reports a facile wafer-level fabrication for nanogap-rich gold nanoislands for highly sensitive surface enhanced Raman scattering (SERS) by repeating solid-state thermal dewetting of thin gold film. The method provides enlarged gold nanoislands with small gap spacing, which increase the number of electromagnetic hotspots and thus enhance the extinction intensity as well as the tunability for plasmon resonance wavelength. The plasmonic nanoislands from repeated dewetting substantially increase SERS enhancement factor over one order-of-magnitude higher than those from a single-step dewetting process and they allow ultrasensitive SERS detection of a neurotransmitter with extremely low Raman activity. This simple method provides many opportunities for engineering plasmonics for ultrasensitive detection and highly efficient photon collection.

Generation of distinct cell types through asymmetric cell division (ACD) is a fundamental developmental process in multicellular organisms. Therefore, controlling when and where ACDs occur is essential for the production of new cells and tissues. The Arabidopsis ( Arabidopsis thaliana ) root has emerged as a powerful model for studying this process because its cell division patterns are highly stereotyped and easily observed. Within the ground tissue, periclinal ACDs in the endodermis generate the middle cortex (MC) post-embryonically, which serves as a hallmark of root maturation. Since the first description of MC formation, extensive research has identified the genetic and environmental cues that either promote or suppress its initiation. Over the past two decades, studies have revealed that MC formation is orchestrated by a regulatory hub centered on the SHORT-ROOT (SHR)–SCARECROW (SCR) transcriptional module and its target, CYCLIND6;1 (CYCD6;1). This core pathway is fine-tuned by multiple regulators, including transcriptional co-activators, repressors, and integrators of gibberellic acid (GA) signaling. Recent advances have uncovered new roles for transcription factors, chromatin regulators, redox enzymes, and receptor-like kinases in linking hormonal signals and positional cues to the SHR–SCR–CYCD6;1 regulatory hub. Together, these pathways ensure that MC formation occurs at the right time, place, and extent. This review summarizes advances in MC regulation, highlighting how transcriptional, hormonal, and positional networks integrate to ensure developmental plasticity in plant roots.

The genus Enterococcus is increasingly recognized for its involvement in various human infections, with several species known to be pathogenic. This study characterized Enterococcus sp. SMC-9, isolated from bile of a patient with cholangitis, and compared its characteristics with those of Enterococcus montenegrensis CoE-012-22 T , recently isolated from dried beef sausage. A comprehensive analysis, encompassing phylogenetic, genomic, and phenotypic studies, confirmed that strain SMC-9 belongs to the same species as E . montenegrensis CoE-012-22 T . However, comparative genomic analysis revealed key differences in virulence and antibiotic resistance gene profiles between the two strains. Notably, genes related to exopolysaccharide biosynthesis and the L-rhamnose biosynthesis pathway were found exclusively in strain SMC-9, suggesting their role in the strain’s colonization of the biliary tract and its involvement in cholangitis. Additionally, the tetracycline resistance gene tet(M) , which was absent in E . montenegrensis CoE-012-22 T , was identified in strain SMC-9, explaining its high tetracycline minimum inhibitory concentration (>16 μg/mL). These findings highlight the unique pathogenic traits of strain SMC-9 compared to E . montenegrensis CoE-012-22 T . Our study underscores the significant genetic and phenotypic variations that can exist among strains within the same species, highlighting the critical need for strain typing to assess their potential impact on patient outcomes and public health.

Several studies have investigated the use of invasive and non-invasive stimulation methods to enhance nerve regeneration, and varying degrees of effectiveness have been reported. However, due to the use of different parameters in these studies, a fair comparison between the effectiveness of invasive and non-invasive stimulation methods is not possible. The present study compared the effectiveness of invasive and non-invasive stimulation using similar parameters. Eighteen Sprague Dawley rats were classified into three groups: the iES group stimulated with fully implantable device, the tES group stimulated with transcutaneous electrical nerve stimulation (TENS), and the injury group (no stimulation). The iES and tES groups received stimulation for 6 weeks starting immediately after the injury. Motor function was evaluated using the sciatic functional index (SFI) every week. The SFI values increased over time in all groups; faster and superior functional recovery was observed in the iES group than in the tES group. Histological evaluation of the nerve sections and gastrocnemius muscle sections were performed every other week. The axon diameter and muscle fiber area in the iES group were larger, and the g-ratio in the iES group was closer to 0.6 than those in the tES group. To assess the cause of the difference in efficiency, a 3D rat anatomical model was used to simulate the induced electric fields in each group. A significantly higher concentration and intensity around the sciatic nerve was observed in the iES group than in the tES group. Vector field distribution showed that the field was orthogonal to the sciatic nerve spread in the tES group, whereas it was parallel in the iES group; this suggested that the tES group was less effective in nerve stimulation. The results indicated that even though rats in the TENS group showed better recovery than those in the injury group, it cannot replace direct stimulation yet because rats stimulated with the invasive method showed faster recovery and superior outcomes. This was likely attributable to the greater concentration and parallel distribution of electric field with respect to target nerve.

The healthcare industry is in dire need of rapid microbial identification techniques for treating microbial infections. Microbial infections are a major healthcare issue worldwide, as these widespread diseases often develop into deadly symptoms. While studies have shown that an early appropriate antibiotic treatment significantly reduces the mortality of an infection, this effective treatment is difficult to practice. The main obstacle to early appropriate antibiotic treatments is the long turnaround time of the routine microbial identification, which includes time-consuming sample growth. Here, we propose a microscopy-based framework that identifies the pathogen from single to few cells. Our framework obtains and exploits the morphology of the limited sample by incorporating three-dimensional quantitative phase imaging and an artificial neural network. We demonstrate the identification of 19 bacterial species that cause bloodstream infections, achieving an accuracy of 82.5% from an individual bacterial cell or cluster. This performance, comparable to that of the gold standard mass spectroscopy under a sufficient amount of sample, underpins the effectiveness of our framework in clinical applications. Furthermore, our accuracy increases with multiple measurements, reaching 99.9% with seven different measurements of cells or clusters. We believe that our framework can serve as a beneficial advisory tool for clinicians during the initial treatment of infections.Label-free rapid deep-learning-based identification of bacterial species that classifies 3D refractive index tomograms into the species.

Renin and lactate, although serving individually as indirect biomarkers of tissue perfusion, exhibit enhanced diagnostic and monitoring capabilities when utilized in combination for tissue perfusion assessment. To detect these biomarkers in human blood, we improved a screen-printed carbon electrode device by incorporating a nanocomposite film consisting of carbon nanotubes modified with gold nanostars (AuNSs). The developed nanocomposite films were used to modify the working electrodes, enhancing the surface area and the conductivity. Following morphological analysis and electrical characterization, these nanostructures were employed as the active layer atop the working electrode, facilitating the binding of receptors (renin antibody and lactate dehydrogenase) around the AuNSs. The serum samples containing different concentrations of renin (31.3–1000 pg/mL) and lactate (25–600 µM) were tested in the modified device with differential pulse voltammetry (DPV). The peak currents of the recorded curves were proportional to the concentrations of the biomarkers, with high coefficients of determination. Remarkably, the limit of detection values reached 4.84 µM, for lactate, and 0.7412 pg/mL, for renin. The proposed device had a strong agreement level with the gold-standard laboratory tests. Additionally, it was possible to detect both targets on the same device using the DPV method, offering a reliable and precise method for simultaneous biomarker detection in real-world applications.

Background. Women infected with human immunodeficiency virus (HIV) are disproportionately affected by human papillomavirus (HPV)–related anogenital disease, particularly with increased immunosuppressions. AIDS Clinical Trials Group protocol A5240 was a trial of 319 HIV-infected women in the United States, Brazil, and South Africa to determine immunogenicity and safety of the quadrivalent HPV vaccine in 3 strata based on screening CD4 count: >350 (stratum A), 201–350 (stratum B), and ≤200 cells/μL (stratum C). Methods. Safety and serostatus of HPV types 6, 11, 16, and 18 were examined. HPV serological testing was performed using competitive Luminex immunoassay (HPV-4 cLIA). HPV type-specific seroconversion analysis was done for participants who were seronegative for the given type at baseline. Results. Median age of patients was 36 years; 11% were white, 56% black, and 31% Hispanic. Median CD4 count was 310 cells/μL, and 40% had undetectable HIV-1 load. No safety issues were identified. Seroconversion proportions among women at week 28 for HPV types 6, 11,16, and 18 were 96%, 98%, 99%, and 91%, respectively, for stratum A; 100%, 98%, 98%, and 85%, respectively, for stratum B, and 84%, 92%, 93%, and 75%, respectively, for stratum C. Conclusions. The quadrivalent HPV vaccine targeted at types 6, 11, 16, and 18 was safe and immunogenic in HIV-infected women aged 13–45 years. Women with HIV RNA load > 10 000 copies/mL and/or CD4 count <200 cells/μL had lower rates of seroconversion rates. Clinical Trials Registration. NCT00604175.

A rod-shaped, obligate anaerobic, Gram-stain-positive bacterium isolated from the human blood was designated as the strain HJ-01 T . Analysis of the 16S rRNA gene sequence revealed that the strain HJ-01 T belonged to the genus Lacrimispora , and was most closely related to L. celerecrescens strains DSM 105336 and MCM B-936, with both 99.3% similarity. The average nucleotide identity values between the strain and the most closely related type strains ranged from 75.3% to 91.4%, while the values between the strain and the two non-type strains of L. celerecrescens , DSM 105336 and MCM B-936, were 98.8% to 98.9%. The digital DNA-DNA hybridization values between the strain and the most closely related type strains ranged from 19.8% to 44.5%, whereas the values between the strain and L. celerecrescens strains DSM 105336 and MCM B-936 were 89.7% to 91.6%. The phylogenomic analysis revealed that the strain formed a cluster adjacent to L. celerecrescens strains DSM 105336 and MCM B-936. The main fatty acids identified were C 16:0 and C 18:1 cis 11 DMA. The cell wall contained the meso -diaminopimelic acid-based peptidoglycan. The end products of the fermentation were acetic acid and formic acid. The strain HJ-01 T and the related Lacrimispora strains shared similar antibiotic resistance profiles, including high resistance to clindamycin (8–256 µg ml−1), linked to the cfr (C) gene located within a 3,378-bp chromosomal transposed unit. Given the chemotaxonomic, phenotypic, and phylogenetic properties, HJ-01 T (= KCTC 25933 T  = JCM 37550 T ) represent a novel species of the genus Lacrimispora , for which the name Lacrimispora sanguinis sp. nov. is proposed. Additionally, we suggest that L. celerecrescens DSM 105336 and MCM B-936 be transferred to Lacrimispora sanguinis sp. nov.

Staphylococcus aureus biofilms complicate the treatment of device-related infections. We hypothesized that combining rifampin with fluoroquinolones could eradicate biofilms even in antimicrobial-resistant S. aureus strains. We determined the synergistic interactions of these combinations in a biofilm model. Thirty methicillin-resistant S. aureus (MRSA) isolates with varying susceptibility profiles were evaluated. Minimum biofilm eradication concentrations (MBECs) were determined using the Calgary Biofilm Device, and the synergy was assessed using the fractional biofilm eradication concentration (FBEC) index. Scanning electron microscopy (SEM) was performed on one strain, and confocal laser scanning microscopy (CLSM) was conducted on four strains for visualizing and evaluating the biofilm viability. The MBEC90 for rifampin and levofloxacin were 512 mg/L and 256 mg/L, respectively, and exceeded 1024 mg/L for ciprofloxacin. Synergy was observed in 56.7% of strains for both the rifampin + ciprofloxacin and rifampin + levofloxacin combinations, with no difference between the combinations. A higher ciprofloxacin MBEC (≥16 mg/L) increased the likelihood of synergy with rifampin by 18-fold. SEM and CLSM analyses in a subset of strains confirmed the enhanced biofilm disruption with rifampin + ciprofloxacin compared to ciprofloxacin alone. Our findings suggest that rifampin combined with ciprofloxacin or levofloxacin may synergistically eradicate MRSA biofilms, offering a potential treatment option for device-related infections when alternatives are limited.

In the ongoing global fight against coronavirus disease 2019 (COVID-19), the sample preparation process for real-time reverse transcription polymerase chain reaction (rRT-PCR) faces challenges due to time-consuming steps, labor-intensive procedures, contamination risks, resource demands, and environmental implications. However, optimized strategies for sample preparation have been poorly investigated, and the combination of RNase inhibitors and Proteinase K has been rarely considered. Hence, we investigated combinations of several extraction-free protocols incorporating heat treatment, sample dilution, and Proteinase K and RNase inhibitors, and validated the effectiveness using 120 SARS-CoV-2 positive and 62 negative clinical samples. Combining sample dilution and heat treatment with Proteinase K and RNase inhibitors addition exhibited the highest sensitivity (84.26%) with a mean increase in cycle threshold (Ct) value of + 3.8. Meanwhile, combined sample dilution and heat treatment exhibited a sensitivity of 79.63%, accounting for a 38% increase compared to heat treatment alone. Our findings highlight that the incorporation of Proteinase K and RNase inhibitors with sample dilution and heat treatment contributed only marginally to the improvement without yielding statistically significant differences. Sample dilution significantly impacts SARS-CoV-2 detection, and sample conditions play a crucial role in the efficiency of extraction-free methods. Our findings may provide insights for streamlining diagnostic testing, enhancing its accessibility, cost-effectiveness, and sustainability.