An Engineered OmpG Nanopore with Displayed Peptide Motifs for Single-Molecule Multiplex Protein Detection.

Molecular detection via nanopore, achieved by monitoring changes in ionic current arising from analyte interaction with the sensor pore, is a promising technology for multiplex sensing development. Outer Membrane Protein G (OmpG), a monomeric porin possessing seven functionalizable loops, has been reported as an effective sensing platform for selective protein detection. Using flow cytometry to screen unfavorable constructs, we identified two OmpG nanopores with unique peptide motifs displayed in either loop 3 or 6, which also exhibited distinct analyte signals in single-channel current recordings. We exploited these motif-displaying loops concurrently to facilitate single-molecule multiplex protein detection in a mixture. We additionally report a strategy to increase sensor sensitivity via avidity motif display. These sensing schemes may be expanded to more sophisticated designs utilizing additional loops to increase multiplicity and sensitivity.

Quantification of localised vascular wedge-shaped defects in glaucoma.

BACKGROUND: Vascular dysfunction plays a considerable role in glaucoma pathogenesis. Previous glaucoma case studies described localised wedge-shaped vascular defects, similar to retinal nerve fibre layer (RNFL) wedge defects. This study investigates the prevalence and quantification of this vessel loss, in relation to primary open angle glaucoma (POAG) parameters. METHODS: This study included 608 eyes (351 participants): 192 PROGRESSA study participants (342 eyes) with suspect, preperimetric or early manifest POAG, observed for vascular wedge defect presence (cohort one); an additional 114 individuals (cohort two-208 eyes) with POAG at various stages of progression for wedge characterisation; and 38 controls (56 eyes). Vascular wedge defects were observed using optical coherence tomography angiography (OCTA). Wedge parameters and vessel densities were quantified using ImageJ software. RNFL and ganglion cell layer inner plexiform layer (GCLIPL) from OCT scans, and mean deviation (Humphrey visual field 24-2) were also assessed. RESULTS: Vascular wedge defects were found in 45/342 eyes (13.2%) in cohort one, in 41/208 eyes (19.7%) in cohort two and were not found in controls. Wedge defects were mostly inferotemporal (80%), and present at all disease stages. They were associated with visual field loss in the opposite hemisphere, thinner RNFL (p < 0.001), thinner GCLIPL (p = 0.003), and focal RNFL loss corresponding with the vascular defect region. CONCLUSION: Vascular wedge defects are present at all POAG stages even before functional change and are strongly concordant with focal RNFL loss. Further research is needed to explore these defects in particular their temporal relationship with clinical measures of POAG.

Real-Time and Label-Free Measurement of Deubiquitinase Activity with a MspA Nanopore.

Covalently attaching ubiquitin (Ub) to cellular proteins as a post-translational modification can result in altered function of modified proteins. Enzymes regulating Ub as a post-translational modification, such as ligases and deubiquitinases, are challenging to characterize in part due to the low throughput of in-vitro assays. Single-molecule nanopore based assays have the advantage of detecting proteins with high specificity and resolution, and in a label-free, real-time fashion. Here we demonstrate the use of a MspA nanopore for discriminating and quantifying Ub proteins. We further applied the MspA pore to measure the Ub-chain disassembly activity of UCH37, a proteasome associated deubiquitinase. The implementation of this MspA system into nanopore arrays could enable high throughput characterizations of unknown deubiquitinases as well as drug screening against disease related enzymes.

Definition and diagnosis of cerebral palsy in genetic studies: a systematic review.

AIM: To conduct a systematic review of phenotypic definition and case ascertainment in published genetic studies of cerebral palsy (CP) to inform guidelines for the reporting of such studies. METHOD: Inclusion criteria comprised genetic studies of candidate genes, with CP as the outcome, published between 1990 and 2019 in the PubMed, Embase, and BIOSIS Citation Index databases. RESULTS: Fifty-seven studies met the inclusion criteria. We appraised how CP was defined, the quality of information on case ascertainment, and compliance with international consensus guidelines. Seven studies (12%) were poorly described, 33 studies (58%) gave incomplete information, and 17 studies (30%) were well described. Missing key information precluded determining how many studies complied with the definition by Rosenbaum et al. Only 18 out of 57 studies (32%) were compliant with the Surveillance of Cerebral Palsy in Europe (SCPE) international guidelines on defining CP. INTERPRETATION: Limited compliance with international consensus guidelines on phenotypic definition and mediocre reporting of CP case ascertainment hinders the comparison of results among genetic studies of CP (including meta-analyses), thereby limiting the quality, interpretability, and generalizability of study findings. Compliance with the SCPE guidelines is important for ongoing gene discovery efforts in CP, given the potential for misclassification of unrelated neurological conditions as CP.

How I do it: Same day discharge for transurethral resection of prostate using Olympus PlasmaButton and PlasmaLoop.

Benign prostatic hyperplasia (BPH) is one of the most common conditions affecting older men. Transurethral resection of the prostate (TURP) has widely been considered the gold standard in surgical treatment for BPH. However, this procedure remains largely an inpatient procedure. Inpatient admission ultimately adds to healthcare cost and patient morbidity. In this article, we present an alternative methodology to treat BPH using combination Olympus PlasmaButton and Olympus PlasmaLoop therapy. Preliminary results from our experience suggest improved hemostasis with adequate resection, allowing a majority of our patients to be discharged the same day of the procedure. We describe our novel technique as a safe and effective way to possibly treat BPH in an outpatient setting.

Tracking Changes in the Endorsement of Injunctive Drinking Norms in Response to the COVID-19 Pandemic Using Longitudinal Alignment Analysis.

The onset of the pandemic saw shifts in messaging around the acceptability of alcohol consumption at different times and contexts. A psychometric analysis of responses to injunctive norms may reveal important differences in specific aspects of norms that were influenced by the pandemic. Study 1 used alignment analysis to evaluate measurement invariance in low- and high-risk injunctive norms across samples of Midwestern college students from 2019 to 2021. Study 2 used an alignment-within-confirmatory factor analysis (CFA) approach to replicate the solution from Study 1 in an independent longitudinal sample (N = 1,148) who responded between 2019 and 2021. For Study 1, the latent mean for high-risk norms was significantly higher in 2021, and the endorsement of four specific norms also differed. In Study 2, increases in latent means for low- and high-risk norms were observed across 2020 and 2021, and differential endorsement emerged for one high-risk norm item. Examining scale-level changes in injunctive drinking norms provides insight into how college students' perceptions changed in response to the COVID-19 pandemic.

Mapping the conformational energy landscape of Abl kinase using ClyA nanopore tweezers.

Protein kinases play central roles in cellular regulation by catalyzing the phosphorylation of target proteins. Kinases have inherent structural flexibility allowing them to switch between active and inactive states. Quantitative characterization of kinase conformational dynamics is challenging. Here, we use nanopore tweezers to assess the conformational dynamics of Abl kinase domain, which is shown to interconvert between two major conformational states where one conformation comprises three sub-states. Analysis of kinase-substrate and kinase-inhibitor interactions uncovers the functional roles of relevant states and enables the elucidation of the mechanism underlying the catalytic deficiency of an inactive Abl mutant G321V. Furthermore, we obtain the energy landscape of Abl kinase by quantifying the population and transition rates of the conformational states. These results extend the view on the dynamic nature of Abl kinase and suggest nanopore tweezers can be used as an efficient tool for other members of the human kinome.

Analyzing the Association Between Depression and High-Risk Sexual Behavior Among Adult Latina Immigrant Farm Workers in Miami-Dade County.

Latinas are often more affected by HIV due to their socio-economic and demographic profiles and are also less likely to receive proper mental health care. Latina immigrants are often even more vulnerable due to socio-economic and cultural factors that place them at higher risk. The current study seeks to examine the association between depression and risky sexual behaviors among adult Latina immigrants from a farm working community in South Miami-Dade County, (Florida, USA). Cross-sectional secondary data analysis was used for responses from a community-based participatory research (CBPR) study. Out of 234 Latina immigrants, 15% reported being depressed and 80% were reported as having engaged in risky sexual behavior. Although no association was found between depression and high-risk sexual behavior, significant secondary findings present associations between risky sexual behavior and low sexual relationship power, interpersonal violence, and relationship status. Implications for future research on depression and risky sexual behaviors among this population are discussed.

Recurrent Granuloma Gluteale Infantum Secondary to Fecal Overflow Incontinence.

Granuloma gluteale infantum is a rare pediatric dermatological disorder of uncertain etiology. Suggested causes include fluorinated corticosteroids, Candida albicans, and irritant contact dermatitis. We present the case of a 3-year-old boy with recurrent episodes of granuloma gluteale infantum which resolved with treatment of his fecal overflow incontinence. As each recurrence correlated with a relapse of overflow incontinence, in this case the cause was irritant contact dermatitis from the liquid stool. This is the first reported case of recurrent granuloma gluteale infantum.

Comparison of lesion-to-cerebellum uptake ratios and standardized uptake values in the evaluation of lung nodules with 18F-FDG PET.

OBJECTIVE: To evaluate the clinical performance of the lesion-to-cerebellum uptake ratio (LCR), a semiquantitative index for differentiating malignant from benign lung nodules with [F]fluorodeoxyglucose positron emission tomography (F-FDG PET). METHODS: Thirty-six patients (16 females, 20 males; median age, 73 years; range, 41-87 years) with 42 known or suspected malignant lung nodules underwent whole-body PET imaging after an intravenous injection of a mean dose of 543+/-69 MBq (14.7+/-1.9 mCi) of F-FDG. The standardized uptake value (SUV) and the LCR were calculated for each nodule and receiver operating characteristic (ROC) curves were analysed using the ROCKIT 0.9B software package. RESULTS: Surgical pathology and follow-up with serial computed tomography scans for at least 24 months revealed 18 malignant lung lesions and 24 benign lesions less than 3.0 cm in size. The mean LCR was 0.70+/-0.40 for malignant nodules and 0.23+/-0.12 for benign nodules (P<0.001, two-tailed test). The area under the estimated ROC curve was 0.8660 for SUV data and 0.9197 for LCR data (P=0.2408, two-tailed test). CONCLUSIONS: The LCR method appears to be a valuable semiquantitative index for the evaluation of malignancy in pulmonary nodules with F-FDG PET, which is simple to perform clinically and does not require accurate measurements of body weight or the residual activity in the syringe utilized for F-FDG injection.