Carba PBP: a novel penicillin-binding protein-based lateral flow assay for rapid phenotypic detection of carbapenemase-producing Enterobacterales.

Rapid phenotypic detection assays, including Carba NP and its variants, are widely applied for clinical diagnosis of carbapenemase-producing Enterobacterales (CPE). However, these tests are based on the acidification of the pH indicator during carbapenem hydrolysis, which limits test sensitivity and speed, especially for the detection of CPE producing low-activity carbapenem (e.g., OXA-48 variants). Herein, we developed a novel rapid and sensitive CPE detection method (Carba PBP) that could measure substrate (meropenem) consumption based on penicillin-binding protein (PBP). Meropenem-specific PBP was used to develop a competitive lateral flow assay (LFA) for meropenem identification. For the detection of carbapenemase activity, meropenem concentration was optimized using a checkerboard assay. The performance of Carba PBP was evaluated and compared with that of Carba NP using a panel of 94 clinical strains characterized by whole-genome sequencing and carbapenem susceptibility test. The limit of detection of PBP-based LFA for meropenem identification was 7 ng mL-1. Using 10 ng mL-1 meropenem as the substrate, Carba PBP and Carba NP could detect 10 ng mL-1 carbapenemase within 25 min and 1,280 ng mL-1 CPE in 2 h, respectively. The sensitivity and specificity were 100% (75/75) and 100% (19/19) for Carba PBP and 85.3% (64/75) and 100% (19/19) for Carba NP, respectively. When compared with Carba NP, Carba PBP showed superior performance in detecting all the tested CPE strains (including OXA-48-like variants) within 25 min and presented two orders of magnitude higher analytical sensitivity, demonstrating potential for clinical diagnosis of CPE. IMPORTANCE This study successfully achieved the goal of carbapenemase activity detection with both high sensitivity and convenience, offering a convenient lateral flow assay for clinical diagnosis of carbapenemase-producing Enterobacterales.

Repurposing harmaline as a novel approach to reverse tmexCD1-toprJ1-mediated tigecycline resistance against klebsiella pneumoniae infections.

BACKGROUND: A novel plasmid-mediated resistance-nodulation-division (RND) efflux pump gene cluster tmexCD1-toprJ1 in Klebsiella pneumoniae tremendously threatens the use of convenient therapeutic options in the post-antibiotic era, including the "last-resort" antibiotic tigecycline. RESULTS: In this work, the natural alkaloid harmaline was found to potentiate tigecycline efficacy (4- to 32-fold) against tmexCD1-toprJ1-positive K. pneumoniae, which also thwarted the evolution of tigecycline resistance. Galleria mellonella and mouse infection models in vivo further revealed that harmaline is a promising candidate to reverse tigecycline resistance. Inspiringly, harmaline works synergistically with tigecycline by undermining tmexCD1-toprJ1-mediated multidrug resistance efflux pump function via interactions with TMexCD1-TOprJ1 active residues and dissipation of the proton motive force (PMF), and triggers a vicious cycle of disrupting cell membrane integrity and metabolic homeostasis imbalance. CONCLUSION: These results reveal the potential of harmaline as a novel tigecycline adjuvant to combat hypervirulent K. pneumoniae infections.

Aeromonas spp. from hospital sewage act as a reservoir of genes resistant to last-line antibiotics.

BACKGROUND: Aeromonas species are opportunistic pathogens distributed widely in the ecosystem. They are known to be capable of acquiring antibiotic resistance genes, including those encoding proteins against last-line antibiotics, such as the tmexCD-toprJ, mcr and carbapenemase genes. We investigated the genomic and phenotypic characteristics of tmexCD-toprJ-positive Aeromonas strains collected from human, animals, and water samples, particularly those from hospital wastewater in China. METHODS: Samples were collected from living animals, meat, water and human. Aeromonas strains in these samples were isolated in selective media. Antimicrobial resistance profiles of all Aeromonas strains were tested by the broth microdilution method. The presence of tmexCD-toprJ was verified by polymerase chain reaction (PCR). All tmexCD-toprJ-positive (n = 36) and selected tmexCD-toprJ-negative (n = 18) Aeromonas strains were subjected to whole genome sequencing. Carriage of antimicrobial resistance genes, the genetic environment of tmexCD-toprJ and genetic diversity of tmexCD-toprJ-positive Aeromonas strains were determined by bioinformatics analysis. Phylogenetic tree of the Aeromonas strains was built by using the Harvest Suite. FINDINGS: Among the 636 Aeromonas strains isolated from different sources, 36 were positive for tmexCD-toprJ, with the highest prevalence of tmexCD-toprJ being found in fishes (8.8%, 95 CI% 3.6-17.2%), followed by hospital wastewater (6.5%, 95 CI% 4.3-9.3%), river water (2.0%, 0.1-10.9) and duck (1.2%, 95 CI% 3.6-17.2%). All tmexCD-toprJ-positive Aeromonas strains carried multiple antimicrobial resistance genes and exhibited resistance to different classes of antibiotics. Co-existence of tmexCD-toprJ, mcr and blaKPC-2 were identified in 21 strains. The tmexCD-toprJ-positive Aeromonas strains were genetically diverse and found to belong to four different species that could be clustered into three major lineages. The tmexCD-toprJ gene clusters were predominantly located in the chromosome (35/36) of Aeromonas spp., with only one strain carrying the plasmid-borne tmexCD-toprJ cluster. The tmexCD-toprJ genes were associated with seven different types of genetic environments, each of which carried distinct types of mobile elements that may be responsible for mediating transmission of this gene cluster.

Femtosecond Laser Inscribed Excessively Tilted Fiber Grating for Humidity Sensing.

We propose a humidity sensor using an excessively tilted fiber grating (Ex-TFG) coated with agarose fabricated using femtosecond laser processing. The processed grating showcases remarkable differentiation between TE and TM modes, achieving an exceptionally narrow bandwidth of approximately 1.5 nm and an impressive modulation depth of up to 15 dB for both modes. We exposed the agarose-coated TFG sensor to various relative humidity levels and monitored the resonance wavelength to test its humidity sensing capability. Our findings demonstrated that the sensor exhibited a rapid response time (2-4 s) and showed a high response sensitivity (18.5 pm/%RH) between the humidity changes and the resonant wavelength shifts. The high sensitivity, linearity, repeatability, low hysteresis, and excellent long-term stability of the TFG humidity sensor, as demonstrated in our experimental results, make it an attractive option for environmental monitoring or biomedical diagnosis.

Analysis of cuproptosis-related lncRNA signature for predicting prognosis and tumor immune microenvironment in pancreatic cancer.

Pancreatic cancer (PC) is a highly malignant digestive tract tumor, with a dismal 5-year survival rate. Recently, cuproptosis was found to be copper-dependent cell death. This work aims to establish a cuproptosis-related lncRNA signature which could predict the prognosis of PC patients and help clinical decision-making. Firstly, cuproptosis-related lncRNAs were identified in the TCGA-PAAD database. Next, a cuproptosis-related lncRNA signature based on five lncRNAs was established. Besides, the ICGC cohort and our samples from 30 PC patients served as external validation groups to verify the predictive power of the risk signature. Then, the expression of CASC8 was verified in PC samples, scRNA-seq dataset CRA001160, and PC cell lines. The correlation between CASC8 and cuproptosis-related genes was validated by Real-Time PCR. Additionally, the roles of CASC8 in PC progression and immune microenvironment characterization were explored by loss-of-function assay. As showed in the results, the prognosis of patients with higher risk scores was prominently worse than that with lower risk scores. Real-Time PCR and single cell analysis suggested that CASC8 was highly expressed in pancreatic cancer and related to cuproptosis. Additionally, gene inhibition of CASC8 impacted the proliferation, apoptosis and migration of PC cells. Furthermore, CASC8 was demonstrated to impact the expression of CD274 and several chemokines, and serve as a key indicator in tumor immune microenvironment characterization. In conclusion, the cuproptosis-related lncRNA signature could provide valuable indications for the prognosis of PC patients, and CASC8 was a candidate biomarker for not only predicting the progression of PC patients but also their antitumor immune responses.

Terahertz asymmetric metallic hole arrays with polarization-independent quasi-bound states in the continuum for membrane sensing.

Resonances with both high-quality factor and polarization-independent characteristics are highly desirable for terahertz (THz) sensing. Here, THz sensors based on asymmetric metallic hole arrays (AMHAs) are experimentally demonstrated. Such sensors consisting of four-hole arrays support polarization-independent quasi-bound states in the continuum (BICs). The induced quasi-BIC presents a quality factor exceeding 2000, which enables enhanced sensing for thin membranes. Results show that the frequency shift is 97.5 GHz for the 25-µm thick polyimide (PI), corresponding to a sensitivity of 147.7 GHz/RIU. The sensing performance strongly relates to the enhanced field originating from sharp quasi-BICs. A maximum field enhancement of 15.88 in contrast to the incident field is achieved. When the PI thickness is large than the decay length of enhanced fields, the interaction strength of field-PI becomes weak, resulting in a saturation effect for the shift of quasi-BICs. The proposed sensor possessing polarization-independent quasi-BICs has great potential for practical sensing applications in real-time chemical and biomolecular.

Variable ratio blue/red upconversion in Yb3+-Tm3+ codoped fluorosilicate glasses.

Simultaneous blue-red emission in a fiber pumped by a single wavelength source is perceived as a great challenge because of the large energy difference of the emitted photons. This Letter reports the dependence of the blue-to-red upconversion (UC) emission ratio in Yb3+-Tm3+ codoped fluorosilicate glasses (FSGs) under the excitation of a 980-nm laser on the host glass silica content. Photoluminescence spectra and SEM-EDS are used to clarify the UC mechanism, indicating that the probability of the cross-relaxation (CR) process 1G4 + 3F2→3H6 + 3F4 is key to the dominance of the blue or red emissions. This research can provide a new platform for variable UC luminescence.

Correction: A redshifted photonic bandgap and wide-angle polarization selection in an all-hyperbolic-metamaterial one-dimensional photonic crystal.

Correction for 'A redshifted photonic bandgap and wide-angle polarization selection in an all-hyperbolic-metamaterial one-dimensional photonic crystal' by Feng Wu et al., Phys. Chem. Chem. Phys., 2023, 25, 10785-10794, https://doi.org/10.1039/D3CP00280B.

A redshifted photonic bandgap and wide-angle polarization selection in an all-hyperbolic-metamaterial one-dimensional photonic crystal.

According to the Bragg scattering theory, photonic bandgaps (PBGs) in all-dielectric one-dimensional (1-D) photonic crystals (PhCs) are polarization-insensitive. This polarization-insensitive property of PBGs poses a challenge in wide-angle high-performance polarization selection. Herein, we theoretically investigate the angle dependence of the PBGs in a novel kind of 1-D PhC called all-hyperbolic-metamaterial (all-HMM) 1-D PhC entirely composed of HMMs. As the incident angle increases, the PBGs in all-HMM 1-D PhCs exhibit the redshift property under transverse magnetic polarization while exhibiting the blueshift property under transverse electric polarization. Empowered by the polarization-sensitive property of the PBGs, wide-angle high-performance polarization selection can be theoretically achieved. Such a wide-angle polarizer would be useful in liquid crystal displays, quantum interferometers, and Q-switched lasers.

Rydberg state dynamics and fragmentation mechanism of N,N,N',N'-tetramethylmethylenediamine.

The non-adiabatic relaxation processes and the fragmentation dynamics of Rydberg-excited N,N,N',N'-tetramethylmethylenediamine (TMMDA) are investigated using femtosecond time-resolved photoelectron imaging and time-resolved mass spectroscopy. Excitation at 208 nm populates TMMDA in a charge-localized 3p state. Rapid internal conversion (IC) to 3s produces two charge-delocalized conformers with independent time constants and distinct population ratios. As the system explores the 3s potential surface, the structural evolution continues on a 1.55 ps timescale, followed by a slower (12.1 ps) relaxation to the ground state. A thorough comparison of the time-dependent mass and photoelectron spectra suggests that ionization out of the 3p state ends up with the parent ion, the vibrational energy of which is insufficient for the bond cleavage. On the contrary, by virtue of the additional energy acquired by IC from 3p, the internal energy deposited in 3s is available to break the C-N bond, leading to the fragment ion. The fragmentation is found to occur on the ion surface instead of the Rydberg surface.

Terahertz angle-independent photonic bandgap in a one-dimensional photonic crystal containing InSb-based hyperbolic metamaterials: erratum.

This erratum corrects the typing errors in Eq. (6) and the caption of Fig. 4 of the original paper, Appl. Opt.61, 7677 (2022).APOPAI0003-693510.1364/AO.470923 The correction does not affect the results of the original paper.

The application of a hollow trephine in femoral retrograde intramedullary nailing technique.

PURPOSE: The purpose of this study was to describe and evaluate the use of a specially designed hollow trephine to create the entry point through the femoral condyle during retrograde interlocking intramedullary nailing for femoral fractures. METHODS: From June 2019 to December 2021, we treated 11 patients (5 men, 6 women; mean age, 64 years; age range 40-77 years) with mid-distal femoral fractures by retrograde intramedullary femoral nailing using a self-designed hollow trephine for femoral condyle reaming and cancellous bone harvesting. The mode of all the nails is static. Patients were followed up at 1, 4, 8, and 12 weeks and for at least 6 months after surgery. The healing process and heterotopic ossification were evaluated by imaging. Partial weight bearing was permitted during the recovery period and complete weight bearing was permitted after clinical healing of the fracture displayed by X-ray. RESULTS: The operation was successful in all patients. Over mean follow-up of 9.3 months (range, 6.0-12.0 months), all patients achieved clinical healing within three months. There were no complications such as knee joint infection, heterotopic ossification, knee joint adhesion and wedge effect. CONCLUSION: The use of the hollow trephine during femoral retrograde intramedullary nailing helps avoid postoperative complications such as heterotopic ossification, knee joint adhesions, and wedge effect. It also facilitates bone graft harvesting.

ADAMTS12 promotes migration and epithelial-mesenchymal transition and predicts poor prognosis for pancreatic cancer.

BACKGROUND: ADAMTS (a disintegrin and metalloproteinase with thrombospondin-like motifs) family, a group of extracellular multifunctional enzymes, has been proven to play a pivotal role in the tumor. In pancreatic cancer, the role and mechanism of this family remain unclear. The present study aimed to figure out the hub gene of ADAMTSs and explore the exact roles in the prognosis and biological functions in pancreatic ductal adenocarcinoma (PDAC). METHODS: We used several databases to analyze the ADAMTS family and then screen out the hub genes. The expression of ADAMTS12 in 106 pairs of PDAC tumors and adjacent normal tissues was examined by immunohistochemistry, and its correlations with clinical parameters were further analyzed. The impacts of ADAMTS12 on the migration of PDAC cells were predicted by gene set enrichment analysis and confirmed by transwell assays. The potential impacts of ADAMTS12 on the epithelial-mesenchymal transition (EMT) were identified by database analysis and experimental proof of real-time quantitative polymerase chain reaction (qPCR) and Western blotting. RESULTS: Our study found that ADAMTS12 was a crucial gene in PDAC, and it was highly expressed in tumor tissues when compared to that in the adjacent tissues. ADATMS12 had predictive value of a poor prognosis for PDAC. The elevation of ADAMTS12 was parallel to the progression of PDAC. Inhibition of ADAMTS12 suppressed the migration of PDAC cells and interfered with the process of EMT. CONCLUSIONS: ADAMTS12 is a crucial member of ADAMTSs in PDAC and a predictor of poor prognosis. Additionally, based on its impacts on migration and metastasis in PDAC and the relationship with EMT, ADAMTS12 plays a role of an oncogene in PDAC and may be a promising target for treatment.

MC-PRPA-HLFIA Cascade Detection System for Point-of-Care Testing Pan-Drug-Resistant Genes in Urinary Tract Infection Samples.

Recently, urinary tract infection (UTI) triggered by bacteria carrying pan-drug-resistant genes, including carbapenem resistance gene blaNDM and blaKPC, colistin resistance gene mcr-1, and tet(X) for tigecycline resistance, have been reported, posing a serious challenge to the treatment of clinical UTI. Therefore, point-of-care (POC) detection of these genes in UTI samples without the need for pre-culturing is urgently needed. Based on PEG 200-enhanced recombinase polymerase amplification (RPA) and a refined Chelex-100 lysis method with HRP-catalyzed lateral flow immunoassay (LFIA), we developed an MCL-PRPA-HLFIA cascade assay system for detecting these genes in UTI samples. The refined Chelex-100 lysis method extracts target DNA from UTI samples in 20 min without high-speed centrifugation or pre-incubation of urine samples. Following optimization, the cascade detection system achieved an LOD of 102 CFU/mL with satisfactory specificity and could detect these genes in both simulated and actual UTI samples. It takes less than an hour to complete the process without the use of high-speed centrifuges or other specialized equipment, such as PCR amplifiers. The MCL-PRPA-HLFIA cascade assay system provides new ideas for the construction of rapid detection methods for pan-drug-resistant genes in clinical UTI samples and provides the necessary medication guidance for UTI treatment.

Clonal relationship of tet(X4)-positive Escherichia coli ST761 isolates between animals and humans.

OBJECTIVES: To characterize the relationship of tet(X4)-positive isolates from different hosts and environments. METHODS: PCR and MALDI-TOF MS were used to identify the tet(X4)-positive isolates. The MICs of 13 antimicrobial agents were determined by broth microdilution. Illumina technology was used to sequence all of the isolates. One isolate was randomly selected from Escherichia coli ST761 clones for long-read sequencing to obtain plasmid sequences. Bioinformatics analysis was used to determine the phylogeny of 46 tet(X4)-positive E. coli ST761 strains. RESULTS: A total of 12 tet(X4)-positive isolates, 8 E. coli and 4 Aeromonas simiae, were obtained from six lairages of a slaughterhouse. These isolates exhibited resistance to at least three classes of antimicrobials, including tigecycline. The majority of them, seven E. coli and three A. simiae, represent separate clonal groups. Notably, the seven E. coli isolates belonged to ST761, a common ST carrying the tet(X4) gene that has been identified in 39 isolates from animals, meat, wastewater and humans from seven Chinese provinces. All 46 tet(X4)-positive E. coli ST761 strains from various sources have a close phylogenetic relationship (0-72 SNPs), with a high nucleotide sequence similarity of resistance genes and the tet(X4)-carrying IncX1-IncFIA(HI1)-IncFIB(K) hybrid plasmid, indicating a clonal relationship of tet(X4)-positive E. coli ST761 among animals, food, the environment and humans. CONCLUSIONS: The clonal relationship of tet(X4)-positive E. coli ST761 between humans and animals poses a previously underestimated threat to public health. To the best of our knowledge, this is the first description of tet(X4)-positive A. simiae.

Telithromycin resistance in Campylobacter mediated by 23S rRNA A2075G mutation and erm(B).

OBJECTIVES: Recently, epidemiological research has shown an unusually high prevalence of telithromycin-resistant Campylobacter. This study was designed to investigate the potential resistance mechanism of telithromycin resistance in Campylobacter. METHODS: A total of 122 Campylobacter isolates of chicken origin collected in 2019 from three regions of China were tested for susceptibility to telithromycin. The potential mechanism of resistance to telithromycin in Campylobacter was revealed through WGS analysis and natural transformation. RESULTS: In this study, 51.3% (61/119) of Campylobacter coli and 100.0% (3/3) of Campylobacter jejuni were resistant to telithromycin. erm(B) or A2075G mutation in 23S rRNA (23S_A2075G) was identified in the telithromycin-resistant C. coli. Cloning of the erm(B) or 23S_A2075G into C. jejuni NCTC 11168 resulted in a 256-fold increase in the MIC of telithromycin. MLST results indicated that various STs were involved in the dissemination of 23S_A2075G and erm(B). Phylogenetic analysis showed that the C. coli isolates with 23S_A2075G and erm(B) from chickens and humans were closely related. CONCLUSIONS: 23S_A2075G and erm(B), which have been widely spread in different genotypes of C. coli isolated from animals and humans, could mediate high levels of resistance to telithromycin in C. coli. C. coli containing 23S_A2075G or erm(B) are clonally related and have the potential to spread zoonotic diseases.

Ultra-sensitive refractive index sensing enabled by a dramatic ellipsometric phase change at the band edge in a one-dimensional photonic crystal.

Surface plasmon polaritons (SPPs) and Bloch surface waves (BSWs) have been widely utilized to design sensitive refractive index sensors. However, SPP- and BSW-based refractive index sensors require additional coupling component (prism) or coupling structure (grating or fiber), which increases the difficulty to observe ultra-sensitive refractive index sensing in experiments. Herein, we realize dramatic ellipsometric phase change at the band edges in an all-dielectric one-dimensional photonic crystal for oblique incidence. By virtue of the dramatic ellipsometric phase change at the long-wavelength band edge, we design an ultra-sensitive refractive index sensor at near-infrared wavelengths. The minimal resolution of the designed sensor reaches 9.28×10-8 RIU. Compared with SPP- and BSW-based refractive index sensors, the designed ultra-sensitive refractive index sensor does not require any additional coupling component or coupling structure. Such ultra-sensitive refractive index sensor would possess applications in monitoring temperature, humidity, pressure, and concentration of biological analytes.

Light transmission mechanisms in a SMF-capillary fiber-SMF structure and its application to bi-directional liquid level measurement.

Capillary fiber (CF) has been extensively investigated in a singlemode fiber (SMF)-CF-SMF (SCS) sensing structure since multiple light guiding mechanisms can be easily excited by simply tuning the air core diameter (cladding diameter) and length of the CF. Understanding the light guiding principles in an SCS structure is essential for improved implementation of a CF based fiber sensor. In this work, light guiding principles in a relatively large air core diameter (≥ 20 µm) and long length of CF (> 1 mm) are investigated theoretically and experimentally. It is found that both multimode interference (MMI) and Anti-Resonant Reflecting Optical Waveguide (ARROW) light guiding mechanisms are excited in the SCS structure in the transmission configuration. However, MMI dips are not observed in the spectrum for the air core diameters of CF smaller than 50 µm in the experiment due to large transmission loss in small air core CFs. Further experimental results demonstrate that a CF with a bigger air core diameter shows a higher sensitivity to curvature, and the highest sensitivity of -16.15 nm/m-1 is achieved when an CF-100 was used. In addition, a SMF-CF-20-CF-30-SMF (SCCS) structure is proposed for high sensitivity bi-direction liquid level measurement for the first time, to the best of our knowledge. Two types of ARROW dips (Dip-20 and Dip-30) are simultaneously excited in transmission, hence both liquid level and liquid flow direction can be detected by tracing the dip strength changes of Dip-20 and Dip-30, respectively.

Construction of multiple anti-resonant light guidance mechanisms in a hollow-core fiber structure for simultaneous measurement of multiple parameters.

The construction of multiple light guidance mechanisms in a hollow-core fiber (HCF) structure is a popular way to realize the simultaneous measurement of multiple parameters. In this work, a partial coating method to excite multiple anti-resonant light guidance mechanisms (ARLGMs) in an HCF structure for the simultaneous measurement of multiple parameters is proposed. As an example, a double ARLGM based on a partially polyimide (PI)-coated HCF structure for the simultaneous measurement of relative humidity (RH) and temperature is demonstrated theoretically and experimentally. The dip (dip II) produced by the PI-coated HCF section shifts linearly with surrounding RH changes with a sensitivity of circa 58.6 ± 0.77 pm/%RH, while the dip (dip I) produced by the bare HCF section (with an air coating layer) is insensitive to RH changes. In addition, both types of dips have linear responses to temperature variations, with similar sensitivities of ∼ 17 pm/°C. Hence, the proposed sensor structure can be used as an RH sensor that is also capable of compensating for local temperature fluctuations. More importantly, the simultaneous measurement of multiple parameters (such as biomarkers) is possible using the proposed method provided the proper sensing materials are partially coated onto the HCF surface.

Drugging the "undruggable" microRNAs.

As a naturally occurring class of gene regulators, microRNAs (miRNAs) have attracted much attention as promising targets for therapeutic development. However, RNAs including miRNAs have long been considered undruggable, and most efforts have been devoted to using synthetic oligonucleotides to regulate miRNAs. Encouragingly, recent findings have revealed that miRNAs can also be drugged with small molecules that directly target miRNAs. In this review paper, we give a summary of recently emerged small-molecule inhibitors (SMIs) and small-molecule degraders (SMDs) for miRNAs. SMIs are small molecules that directly bind to miRNAs to inhibit their biogenesis, and SMDs are bifunctional small molecules that upon binding to miRNAs induce miRNA degradation. Strategies for discovering SMIs and developing SMDs were summarized. Applications of SMIs and SMDs in miRNA inhibition and cancer therapy were also introduced. Overall, SMIs and SMDs introduced here have high potency and specificity in miRNA inhibition. We envision that these small molecules will pave the way for developing novel therapeutics toward miRNAs that were previously considered undruggable.