Potential value of a rapid syndromic multiplex PCR for the diagnosis of native and prosthetic joint infections: a real-world evidence study.

Introduction: The BIOFIRE Joint Infection (JI) Panel is a diagnostic tool that uses multiplex-PCR testing to detect microorganisms in synovial fluid specimens from patients suspected of having septic arthritis (SA) on native joints or prosthetic joint infections (PJIs). Methods: A study was conducted across 34 clinical sites in 19 European and Middle Eastern countries from March 2021 to June 2022 to assess the effectiveness of the BIOFIRE JI Panel. Results: A total of 1527 samples were collected from patients suspected of SA or PJI, with an overall agreement of 88.4 % and 85 % respectively between the JI Panel and synovial fluid cultures (SFCs). The JI Panel detected more positive samples and microorganisms than SFC, with a notable difference on Staphylococcus aureus, Streptococcus species, Enterococcus faecalis, Kingella kingae, Neisseria gonorrhoeae, and anaerobic bacteria. The study found that the BIOFIRE JI Panel has a high utility in the real-world clinical setting for suspected SA and PJI, providing diagnostic results in approximately 1 h. The user experience was positive, implying a potential benefit of rapidity of results' turnover in optimising patient management strategies. Conclusion: The study suggests that the BIOFIRE JI Panel could potentially optimise patient management and antimicrobial therapy, thus highlighting its importance in the clinical setting.

Work Intensity of Postoperative Care Following Implantation of Presbyopia-Correcting versus Monofocal Intraocular Lenses.

Purpose: To compare the work intensity of postoperative care following implantation of presbyopia-correcting intraocular lenses (IOLs) to that of standard monofocal IOLs. Patients and Methods: This open-label, multicenter, comparative study retrospectively reviewed the case records of cataract surgery patients who underwent bilateral implantation of either presbyopia-correcting IOLs (presby-IOL group; N=177) or standard monofocal IOLs (monofocal group; N=177). Outcome measures included the total time the patient spent in the office, number of visits, mean duration of visits, and the number of procedures and diagnostic tests during the first postoperative year. Outcome measures were compared between the first 90 days and days 91-365 after surgery. Results: Mean (±SD) time spent in the office during the first postoperative year was 5:50 ± 3:35 hours (H:MM) over 6.6 ± 2.9 visits in the presby-IOL group, compared to 3:38 ± 1:36 hours over 4.9 ± 1.6 visits in the monofocal IOL group (p <0.001). During the first 90 days, a presby-IOL patient spent 40 minutes longer in the office than a monofocal IOL patient (3:39 ± 1:38 hours vs 2.59 ± 1:13 hours) (p <0.001). During days 91-365, time in the office was 1:32 hours longer (p <0.001), and the mean visit duration was 8 minutes longer for the presby-IOL patients (p=0.002) than those with monofocal IOL. In addition, the presby-IOL patients underwent more procedures and diagnostic tests (p ≤0.001) as compared with standard monofocal cataract surgery patients. Conclusion: Patients implanted with presbyopia-correcting IOLs require significantly more clinic time, diagnostic testing, and procedures postoperatively as compared with standard monofocal cataract surgery patients.

Triphenylphosphine oxide promoting visible-light-driven C-C coupling via desulfurization.

Triphenylphosphine oxide (TPPO) and triphenylphosphine (TPP) can form a complex in solution, promoting visible light absorption to trigger electron transfer within the complex and generate radicals. Subsequent radical reactions with thiols enable desulfurization to produce carbon radicals that react with aryl alkenes to yield new C-C bonds. Since ambient oxygen can easily oxidize TPP to TPPO, the reported method requires no explicit addition of a photocatalyst. This work highlights the promise of using TPPO as a catalytic photo-redox mediator in organic synthesis.

Fluorine-thiol displacement probes for acetaminophen's hepatotoxicity.

Chemicals possessing reactive electrophiles can denature innate proteins leading to undesired toxicity, and the overdose-induced liver injury by drugs containing electrophiles has been one of the major causes of non-approval and withdraw by the US Food and Drug Administration (FDA). Elucidating the associated proteins could guide the future development of therapeutics to circumvent these drugs' toxicities, but was largely limited by the current probing tools due to the steric hindrance of chemical tags including the common "click chemistry" labels. Taking the widely used non-steroidal anti-inflammatory drug acetaminophen (APAP) as an example, we hereby designed and synthesized an APAP analogue using fluorine as a steric-free label. Cell toxicity studies indicated our analogue has similar activity to the parent drug. This analogue was applied to the mouse hepatocellular proteome together with the corresponding desthiobiotin-SH probe for subsequent fluorine-thiol displacement reactions (FTDRs). This set of probes has enabled the labeling and pull-down of hepatocellular target proteins of the APAP metabolite as validated by Western blotting. Our preliminary validation results supported the interaction of APAP with the thioredoxin protein, which is an important redox protein for normal liver function. These results demonstrated that our probes confer minimal steric perturbation and mimic the compounds of interest, allowing for global profiling of interacting proteins. The fluorine-thiol displacement probing system could emerge as a powerful tool to enable the investigation of drug-protein interactions in complex biological environments.

A multicentre evaluation and expert recommendations of use of the newly developed BioFire Joint Infection polymerase chain reaction panel.

Septic arthritis is a serious condition with significant morbidity and mortality, routinely diagnosed using culture. The FDA has recently approved the rapid molecular BioFire® Joint Infection Panel (BJIP) for synovial fluid. We aimed to evaluate the BJIP compared to culture and its potential use in patient management. A multicentre retrospective evaluation of BJIP was conducted in the UK and Ireland. Positive percent agreement (PPA) and negative percent agreement (NPA) were calculated between the BJIP and routine culture. A multidisciplinary team (MDT) discussion addressing the optimal or potential case use of the assay practice was facilitated. Three hundred ninety-nine surplus synovial fluid samples (~ 70% from native joints) from eight centres were processed using BJIP in addition to routine culture. An increased yield of positive results was detected using BJIP compared to routine culture (98 vs 83), giving an overall PPA of 91.6% and overall NPA of 93% for the BJIP compared to culture results. The BJIP detected resistant markers and additional organisms that could influence antibiotic choices including Neisseria gonorrhoeae and Kingella kingae. The MDT agreed that the assay could be used, in addition to standard methods, in adult and children patients with specialist advice use based on local needs. Rapid results from BJIP were assessed as having potential clinical impact on patient management. Organisms not included in the panel may be clinically significant and may limit the value of this test for PJI.

Unprotected peptide macrocyclization and stapling via a fluorine-thiol displacement reaction.

We report the discovery of a facile peptide macrocyclization and stapling strategy based on a fluorine thiol displacement reaction (FTDR), which renders a class of peptide analogues with enhanced stability, affinity, cellular uptake, and inhibition of cancer cells. This approach enabled selective modification of the orthogonal fluoroacetamide side chains in unprotected peptides in the presence of intrinsic cysteines. The identified benzenedimethanethiol linker greatly promoted the alpha helicity of a variety of peptide substrates, as corroborated by molecular dynamics simulations. The cellular uptake of benzenedimethanethiol stapled peptides appeared to be universally enhanced compared to the classic ring-closing metathesis (RCM) stapled peptides. Pilot mechanism studies suggested that the uptake of FTDR-stapled peptides may involve multiple endocytosis pathways in a distinct pattern in comparison to peptides stapled by RCM. Consistent with the improved cell permeability, the FTDR-stapled lead Axin and p53 peptide analogues demonstrated enhanced inhibition of cancer cells over the RCM-stapled analogues and the unstapled peptides.

Wavefront-guided myopic laser in situ keratomileusis with a high-resolution Hartmann-Shack aberrometer and a new nomogram.

PURPOSE: To evaluate visual, refractive, aberrometric, and patient-reported outcomes of wavefront-guided (WFG) myopic laser in situ keratomileusis (LASIK) using a high-resolution Hartmann-Shack aberrometer (iDesign Advanced WaveScan system) with a new nomogram and to determine whether the new nomogram resolved the mild undercorrection that occurs with the manufacturer's default settings. SETTING: Three private LASIK practices. DESIGN: Prospective, open-label, noncomparative, multicenter study. METHODS: One hundred ninety eyes of 95 patients underwent bilateral WFG LASIK for the correction of myopia or myopic astigmatism. A new nomogram was used, which effectively adjusted the wavefront-measured refraction sphere up or down to equal the manifest refraction sphere. Patients were followed up for 6 months. RESULTS: Eighty-four patients completed the final follow up. At 6 months, 162 (96.4%) of 168 eyes achieved monocular uncorrected distance visual acuity of 20/20 or better. No eye lost 2 or more lines of corrected distance visual acuity. The safety and efficacy indices were 1.12 and 1.09, respectively; 164 (98%) of 168 eyes had manifest refraction spherical equivalent within ±0.50 diopters (D) of emmetropia, and 154 (92%) of 168 eyes had residual manifest refractive astigmatism of 0.50 D or less. Fewer patients experienced burning, stinging, soreness, and irritation postoperatively than preoperatively. Eighty-one (96%) of 84 patients reported improved quality of life. CONCLUSIONS: WFG myopic LASIK using a high-resolution Hartmann-Shack aberrometer and a new nomogram resolved the undercorrection with the manufacturer's default settings. The treatment was safe and effective with excellent visual and refractive outcomes, high patient satisfaction, and improved quality of life.

Site-specific antibody fragment conjugates for targeted imaging.

This work details the use of amber suppression-mediated genetic incorporation of unnatural amino acids (UAAs), specifically p-azido-l-phenylalanine (pAzF) and p-acetyl-l-phenylalanine (pAcF), to develop site-specifically labeled antibody Fab fragments. These antibody fragment conjugates represent a novel class of imaging agents with optimal stability, efficacy, and pharmacological properties, which have demonstrated promising potential for probing and understanding the in vivo bio-distributions of protein targets of interest. This chapter provides general guidelines for preparing these Fab conjugates, and details of follow-up bioassays such as single-agent based positron emission tomography (PET) imaging of immune-checkpoint protein PD-L1, and the use of GCN4-mediated switchable antibody conjugates for near-infrared fluorescent imaging of cancer-related biomarkers.

Simulation of a Spontaneous Vaginal Delivery and Neonatal Resuscitation in a Biocontainment Unit.

This article describes a large-scale scenario designed to test the capabilities of a US biocontainment unit to manage a pregnant woman infected with a high-consequence pathogen, and to care for a newborn following labor and spontaneous vaginal delivery. We created and executed a multidisciplinary functional exercise with simulation to test the ability of the Johns Hopkins Hospital biocontainment unit (BCU) to manage a pregnant patient in labor with an unknown respiratory illness and to deliver and stabilize her neonate. The BCU Exercise and Drill Committee established drill objectives and executed the exercise in partnership with the Johns Hopkins Simulation Center in accordance with Homeland Security and Exercise Program guidelines. Exercise objectives were assessed by after-action reporting and objective measurements to detect contamination, using a fluorescent marker to simulate biohazardous fluids that would be encountered in a typical labor scenario. The immediate objectives of the drill were accomplished, with stabilization of the mother and successful delivery and resuscitation of her newborn. There was no evidence of contamination when drill participants were inspected under ultraviolet light at the end of the exercise. Simulation optimizes teamwork, communication, and safety, which are integral to the multidisciplinary care of the maternal-fetal unit infected, or at risk of infection, with a high-consequence pathogen. Lessons learned from this drill regarding patient transportation, safety, and obstetric and neonatal considerations will inform future exercises and protocols and will assist other centers in preparing to care for pregnant patients under containment conditions.

Application of the Incident Command System to the Hospital Biocontainment Unit Setting.

High-consequence pathogens create a unique problem. To provide effective treatment for infected patients while providing safety for the community, a series of 10 high-level isolation units have been created across the country; they are known as Regional Ebola and Special Pathogen Treatment Centers (RESPTCs). The activation of a high-level isolation unit is a highly resource-intensive activity, with effects that ripple across the healthcare system. The incident command system (ICS), a standard tool for command, control, and coordination in domestic emergencies, is a command structure that may be useful in a biocontainment event. A version of this system, the hospital emergency incident command system, provides an adaptable all-hazards approach in healthcare delivery systems. Here we describe its utility in an operational response to safely care for a patient(s) infected with a high-consequence pathogen on a high-level isolation unit. The Johns Hopkins Hospital created a high-level isolation unit to manage the comprehensive and complex needs of patients with high-consequence infectious diseases, including Ebola virus disease. The unique challenges of and opportunities for providing care in this high-level isolation unit led the authors to modify the hospital incident command system model for use during activation. This system has been tested and refined during full-scale functional and tabletop exercises. Lessons learned from the after-action reviews of these exercises led to optimization of the structure and implementation of ICS on the biocontainment unit, including improved job action sheets, designation of physical location of roles, and communication approaches. Overall, the adaptation of ICS for use in the high-level isolation unit setting may be an effective approach to emergency management during an activation.

Dofetilide-Associated QT Prolongation: Total Body Weight Versus Adjusted or Ideal Body Weight for Dosing.

Dofetilide is an antiarrhythmic drug with dosing based on the Cockcroft-Gault formula using total body weight (TBW). We investigated the impact of calculating dofetilide dose using adjusted body weight (ABW) or ideal body weight (IBW) on subsequent dose reduction or discontinuation. We conducted a retrospective review of 265 patients admitted to an academic medical center for initiation of dofetilide using TBW. Dosing was recalculated using ABW or IBW. Patients who would have received a reduced dose using ABW or IBW (reduced dose group) were compared with patients whose dose would not have changed (same dose group). Manual measurement of QT intervals was performed. We found that Forty-one of 265 patients (15%) would have received a lower initial dose of dofetilide based on ABW. Patients in this reduced dose group had 2.95 times greater odds of drug discontinuations or dose reductions due to QTc prolongation (95% confidence interval, 1.47-5.90; P < 0.01) compared with the same dose group. Seventy-seven of 265 patients (29%) would have received a lower initial dose of dofetilide based on IBW. Patients in this reduced dose group had 1.78 times greater odds of drug discontinuations or dose reductions due to QTc prolongation (95% confidence interval, 0.98-3.21; P = 0.056) compared with the same dose group. These data suggest that caution should be used when dosing dofetilide using TBW, as it may lead to a greater frequency of dose reduction or discontinuation compared with dosing using ABW or IBW.

DNA-crowded enzyme complexes with enhanced activities and stabilities.

We present a robust and simple method to prepare DNA-crowded enzyme complexes by directly assembling long DNA duplexes on the enzyme surface. DNA-crowded enzyme complexes show boosted substrate turnover numbers, and increased stabilities against various storage conditions. They could be potentially scaled up for applications in biomaterials and biotechnology.

Utilizing Electronic Health Record Information to Optimize Medication Infusion Devices: A Manual Data Integration Approach.

Health information technology is increasingly utilized within healthcare delivery systems today. Two examples of this type of technology include the capture of patient-specific information within an electronic health record and intravenous medication infusion devices equipped with dose error reduction software known as drug libraries. Automatic integration of these systems, termed intravenous (IV) interoperability, should serve as the goal toward which all healthcare systems work to maximize patient safety. For institutions lacking IV interoperability, we describe a manual approach of querying the electronic health record to incorporate medication administration information with data from infusion device software to optimize drug library settings. This approach serves to maximize utilization of available information to optimize medication safety provided by drug library software.

Toxic peripheral keratopathy: a syndrome in laser refractive surgery.

This retrospective observational case series describes a clinical syndrome in laser refractive surgery. Five eyes of 4 patients with noninflammatory peripheral corneal opacification in the period immediately after photorefractive keratectomy or laser in situ keratomileusis were identified and the charts abstracted. Peripheral stromal opacification, localized corneal thinning, irregular astigmatism leading to a decrease in corrected visual acuity, and gradual clearing were characteristic findings. Gradual clearing occurred without treatment in all eyes over a period of 2 weeks to 12 months. This syndrome has similarities to central toxic keratopathy but occurs in the peripheral cornea. We refer to the syndrome as toxic peripheral keratopathy, the cause of which is unknown.

Wavefront-guided photorefractive keratectomy in eyes with prior radial keratotomy: a multicenter study.

PURPOSE: To investigate the outcomes of wavefront-guided photorefractive keratectomy (WG PRK) using prophylactic mitomycin C (MMC) in eyes that had previously undergone radial keratotomy (RK). DESIGN: Retrospective, observational, consecutive case series. PARTICIPANTS: Thirty-two eyes of 27 patients with previous RK that underwent WG PRK with MMC. METHODS: The records were reviewed of consecutive RK patients whose eyes underwent WG PRK with MMC in 4 centers with postoperative follow-up of 6 months or longer (range, 6-21 months). Eyes were divided into myopic WG PRK and hyperopic WG PRK groups based on their preoperative spherical equivalent (SE). Preoperative best spectacle-corrected visual acuity (BSCVA) was compared with postoperative uncorrected visual acuity (UCVA) and BSCVA to ascertain efficacy and safety. Change in SE and attempted versus achieved SE were evaluated. Incidences of haze and other complications were recorded. MAIN OUTCOME MEASURES: Uncorrected visual acuity, BSCVA, SE, corneal haze, and other complications. RESULTS: In the myopic WG PRK group (n = 9), UCVA improved by 3 lines on average (P = 0.015) with UCVA of > or =20/20 in 56% and > or =20/40 in 100% of eyes; 55% were within 0.5 diopter (D), and 100% were within 1 D of attempted refraction. In the hyperopic WG PRK group (n = 23), UCVA improved for 3 lines on average (P<0.001), with UCVA of > or =20/20 in 48% and > or =20/40 in 100% of eyes; 57% were within 0.5 D and 74% were within 1 D of attempted refraction. One eye lost 2 lines of BSCVA as a result of the development of mild to moderate haze, but recovered in 4 months. No eyes lost more than 2 lines of BSCVA. Six eyes (19%; 6/32) experienced the development of haze in the postoperative course, with mild to moderate haze in 1 eye and trace haze in the other 5 eyes. No other complications were noted. CONCLUSIONS: Wavefront-guided PRK with MMC in eyes with prior RK improved the UCVA significantly and was safe over the short follow-up of this series. Although haze occurred, no eye suffered persistent visual loss of 2 or more lines. FINANCIAL DISCLOSURE(S): Proprietary or commercial disclosure may be found after the references.

Lack of progression of ectasia seven years after LASIK in a highly myopic keratoconic eye.

PURPOSE: To report a patient with clinically evident keratoconus with a preoperative spherical equivalent refraction of more than -10.00 diopters (D) who underwent successful LASIK with no evidence of progression of ectasia 7 years after surgery. METHODS: Interventional case report. RESULTS: A patient with clinically evident keratoconus in his left eye underwent bilateral LASIK. Preoperative best spectacle-corrected visual acuity (BSCVA) in the left eye was 20/25 with a manifest refraction of -8.50 -3.25 x 126. Seven years later he presented complaining of progressive loss of visual acuity in his left eye. A significant posterior subcapsular and nuclear cataract was present in the left eye. There was no evidence of ectasia on topography. Cataract surgery was performed and BSCVA retumed to 20/20. CONCLUSIONS: LASIK can be successful in some eyes with keratoconus, even when deep ablations are performed. It is not appropriate to conclude from this case that LASIK is safe in eyes with keratoconus. This case raises an important question: why do some eyes with normal preoperative topography develop ectasia after LASIK, whereas other eyes with significant keratoconus preoperatively remain stableert ke after LASIK?