Flow induces common and specific transcriptional changes in renal tubular epithelial cells involving the PI3K pathway.

Flow-induced shear stress affects renal epithelial cells in the nephron tubule with potential implications for differential functionalities of the individual segments. Disruptions of cellular mechanosensation or flow conditions are associated with the development and progression of various renal diseases. This study investigates the effects of flow on the transcriptome of various renal tubular epithelial cell types. We analyzed the transcriptome of induced renal epithelial cells (iREC) cultured under physiological flow (0.57 ± 0.05 dyn/cm2 ) or in static conditions for 72 h. RNA sequencing showed 861 differentially expressed genes (DEGs), with 503 up- and 358 downregulated under flow. DEGs were linked to extracellular matrix (ECM) components (e.g. Col1a1, Col4a3, Col4a4, Fn1, Smoc2), junctions (Gja1, Tubb5), channel activities (Abcc4, Aqp1), and transcription factors (Foxq1, Lgr6). Next, we performed a meta-analysis comparing our data with three published datasets that subjected epithelial cell lines from distinct segments to flow, including proximal tubule and collecting duct cells. We found that TGF-ß, p53, MAPK, and PI3K are common flow-regulated pathways. Tfrc expression and thus the capability of iron uptake is commonly upregulated under flow. Many DEGs were related to kidney diseases, such as fibrosis (e.g. Tgfb1-3 and Serpine1). To obtain further mechanistic insights we investigated the role of the PI3K pathway in flow sensing. Applying flow and inhibition of PI3K showed significantly altered expression of transcripts related to ECM remodeling, angiogenesis, and ion transport. This suggests that the PI3K pathway is a critical mediator in flow-dependent cellular processes and gene expression, potentially influencing renal development and tissue remodeling. Finally, we derived a cross-cell-line summary of common as well as segment-specific transcriptomic effects, thus providing insights into the molecular mechanisms underlying flow sensing in the nephron tubule.

Coated Blade Spray Ion Mobility Spectrometry.

Coated blade spray (CBS) is a microextraction technology with blades that serve as both the extraction device and the electrospray ionization (ESI) emitter. CBS is designed for easy and rapid extraction of analytes in complex matrices as well as ESI directly from the blade. The technology selectively enriches the components of interest on a coated metal blade. The coating consists of a selective polymer. So far, CBS has only been coupled with mass spectrometry but never with ion mobility spectrometry (IMS), where ions are separated and detected based on their ion mobility in a drift gas under the influence of an electric field, while instrumentation is compact and easy to operate so that the advantages of CBS can be particularly well exploited. Therefore, this work focuses on coupling CBS with our previously described ESI-IMS. The ion mobility spectrometer has a drift length of only 75 mm and provides a high resolving power of RP = 100. In this work, preliminary measurements of CBS-IMS are presented. In particular, the detection of benzodiazepines and ketamine in drinks and the pesticide isoproturon in water samples is shown to demonstrate the feasibility of CBS-IMS.

Performance evaluation of machine-assisted interpretation of Gram stains from positive blood cultures.

Manual microscopy of Gram stains from positive blood cultures (PBCs) is crucial for diagnosing bloodstream infections but remains labor intensive, time consuming, and subjective. This study aimed to evaluate a scan and analysis system that combines fully automated digital microscopy with deep convolutional neural networks (CNNs) to assist the interpretation of Gram stains from PBCs for routine laboratory use. The CNN was trained to classify images of Gram stains based on staining and morphology into seven different classes: background/false-positive, Gram-positive cocci in clusters (GPCCL), Gram-positive cocci in pairs (GPCP), Gram-positive cocci in chains (GPCC), rod-shaped bacilli (RSB), yeasts, and polymicrobial specimens. A total of 1,555 Gram-stained slides of PBCs were scanned, pre-classified, and reviewed by medical professionals. The results of assisted Gram stain interpretation were compared to those of manual microscopy and cultural species identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The comparison of assisted Gram stain interpretation and manual microscopy yielded positive/negative percent agreement values of 95.8%/98.0% (GPCCL), 87.6%/99.3% (GPCP/GPCC), 97.4%/97.8% (RSB), 83.3%/99.3% (yeasts), and 87.0%/98.5% (negative/false positive). The assisted Gram stain interpretation, when compared to MALDI-TOF MS species identification, also yielded similar results. During the analytical performance study, assisted interpretation showed excellent reproducibility and repeatability. Any microorganism in PBCs should be detectable at the determined limit of detection of 105 CFU/mL. Although the CNN-based interpretation of Gram stains from PBCs is not yet ready for clinical implementation, it has potential for future integration and advancement.

Improved pharmacokinetics and enhanced efficacy of the vancomycin derivative FU002 using a liposomal nanocarrier.

Antibiotic resistance still represents a global health concern which diminishes the pool of effective antibiotics. With the vancomycin derivative FU002, we recently reported a highly potent substance active against Gram-positive bacteria with the potential to overcome vancomycin resistance. However, the translation of its excellent antimicrobial activity into clinical efficiency could be hampered by its rapid elimination from the blood stream. To improve its pharmacokinetics, we encapsulated FU002 in PEGylated liposomes. For PEG-liposomal FU002, no relevant cytotoxicity on liver, kidney and red blood cells was observed. Studies in Wistar rats revealed a significantly prolonged blood circulation of the liposomal antibiotic. In microdilution assays it could be demonstrated that encapsulation does not diminish the antimicrobial activity against staphylococci and enterococci. Highlighting its great potency, liposomal FU002 exhibited a superior therapeutic efficacy when compared to the free form in a Galleria mellonella larvae infection model.

Increased tibial tuberosity torsion has the greatest predictive value in patients with patellofemoral instability compared to other commonly assessed parameters.

PURPOSE: The multifactorial nature of patellofemoral instability requires a comprehensive assessment of the affected patients. While an association between tibial tuberosity (TT) torsion and patellofemoral instability is known, its specific effect has not yet been investigated. This study investigated the effect of TT torsion on patellofemoral instability. METHODS: This retrospective cohort study compared patients who underwent surgical intervention for patellofemoral instability and asymptomatic controls. TT torsion was measured in addition to other commonly assessed risk factors for patellofemoral instability using standardised computed tomography (CT) data of the lower extremities. The diagnostic performances of the assessed parameters were evaluated using receiver operating characteristic curve analysis and odds ratios (ORs) were calculated. RESULTS: The patellofemoral instability group consisted of 79 knees, compared to 72 knees in the asymptomatic control group. Both groups differed significantly in all assessed parameters (p < 0.001), except for tibial torsion (n.s.). Among all parameters, TT torsion presented the best diagnostic performance for predicting patellar instability with an area under the curve of 0.95 (95% confidence interval [CI], 0.91-0.98; p < 0.001). A cut-off value of 17.7° yielded a 0.87 sensitivity and 0.89 specificity to predict patellar instability (OR, 55.2; 95% CI, 20.5-148.6; p < 0.001). CONCLUSION: Among the evaluated risk factors, TT torsion had the highest predictive value for patellofemoral instability. Patients with TT torsions ≥ 17.7° showed a 55-fold increased probability of patellofemoral instability. Therefore, TT torsion should be included in the assessment of patients with patellofemoral instability. LEVEL OF EVIDENCE: Level III.

How to Improve the Resolving Power of Compact Electrospray Ionization Ion Mobility Spectrometers.

Every drift tube ion mobility spectrometer (IMS) has an optimum drift voltage to reach maximum resolving power. This optimum depends, among other things, on the temporal and spatial width of the injected ion packet and the pressure within the IMS. A reduction of the spatial width of the injected ion packet leads to improved resolving power, higher peak amplitudes when operating the IMS at optimum resolving power, and thus a better signal-to-noise ratio despite the reduced number of injected ions. Hereby, the performance of electrospray ionization (ESI)-IMS can be considerably improved. By setting the ion shutter opening time to just 5 µs and slightly increasing the pressure, a high resolving power RP > 150 can be achieved with a given drift length of just 75 mm. At such high resolving power, even a mixture of the herbicides isoproturon and chlortoluron having similar ion mobility can be well separated despite short drift length.

Highly Efficient Ion Manipulator for Tandem Ion Mobility Spectrometry: Exploring a Versatile Technique by a Study of Primary Alcohols.

In this work, we present a tandem ion mobility spectrometer (IMS) utilizing a highly efficient ion manipulator allowing to store, manipulate, and analyze ions under high electric field strengths and controlled ion-neutral reactions at ambient conditions. The arrangement of tandem drift regions and an ion manipulator in a single drift tube allows a sequence of mobility selection of precursor ions, followed by storage and analysis, mobility separation, and detection of the resulting product ions. In this article, we present a journey exploring the capabilities of the present instrument by a study of eight different primary alcohols characterized at reduced electric field strengths E/N of up to 120 Td with a water vapor concentration ranging from 40 to 540 ppb. Under these conditions, protonated alcohol monomers up to a carbon number of nine could be dissociated, resulting in 18 different fragmented product ions in total. The fragmentation patterns revealed regularities, which can be used for assignment to the chemical class and improved classification of unknown substances. Furthermore, both the time spent in high electrical field strengths and the reaction time with water vapor can be tuned precisely, allowing the fragment distribution to be influenced. Thus, further information regarding the relations of the product ions can be gathered in a standalone drift tube IMS for the first time.

Detection of Triacetone Triperoxide by High Kinetic Energy Ion Mobility Spectrometry.

High Kinetic Energy Ion Mobility Spectrometry (HiKE-IMS) is a versatile technique for the detection of gaseous target molecules that is particularly useful in complex chemical environments, while the instrumental effort is low. Operating HiKE-IMS at reduced pressures from 10 to 60 mbar results in fewer ion-neutral collisions than at ambient pressure, reducing chemical cross-sensitivities and eliminating the need for a preceding separation dimension, e.g., by gas chromatography. In addition, HiKE-IMS allows operation over a wide range of reduced electric field strengths E/N up to 120 Td, allowing separation of ions by low-field ion mobility and exploiting the field dependence of ion mobility, potentially allowing separation of ion species at high E/N despite similar low-field ion mobilities. Given these advantages, HiKE-IMS can be a useful tool for trace gas analysis such as triacetone triperoxide (TATP) detection. In this study, we employed HiKE-IMS to detect TATP. We explore the ionization of TATP and the field-dependent ion mobilities, providing a database of the ion mobilities depending on E/N. Confirming the literature results, ionization of TATP by proton transfer with H3O+ in HiKE-IMS generates fragments, but using NH4+ as the primary reactant ion leads to the TATP·NH4+ adduct. This adduct fragments at high E/N, which could provide additional information for reliable detection of TATP. Thus, operating HiKE-IMS at variable E/N in the drift region generates a unique fingerprint of TATP made of all ion species related to TATP and their ion mobilities depending on E/N, potentially reducing the rate of false positives.

Ultra-Fast Ion Mobility Spectrometer for High-Throughput Chromatography.

Fast chromatography systems especially developed for high sample throughput applications require sensitive detectors with a high repetition rate. These high throughput techniques, including various chip-based microfluidic designs, often benefit from detectors providing subsequent separation in another dimension, such as mass spectrometry or ion mobility spectrometry (IMS), giving additional information about the analytes or monitoring reaction kinetics. However, subsequent separation is required at a high repetition rate. Here, we therefore present an ultra-fast drift tube IMS operating at ambient pressure. Short drift times while maintaining high resolving power are reached by several key instrumental design features: short length of the drift tube, resistor network of the drift tube, tristate ion shutter, and improved data acquisition electronics. With these design improvements, even slow ions with a reduced mobility of just 0.94 cm2/(V s) have a drift time below 1.6 ms. Such short drift times allow for a significantly increased repetition rate of 600 Hz compared with previously reported values. To further reduce drift times and thus increase the repetition rate, helium can be used as the drift gas, which allows repetition rates of up to 2 kHz. Finally, these significant improvements enable IMS to be used as a detector following ultra-fast separation including chip-based chromatographic systems or droplet microfluidic applications requiring high repetition rates.

The dependence of reduced mobility, ion-neutral collisional cross sections, and alpha values on reduced electric field strengths in ion mobility.

As ion mobility spectrometry (IMS) is used with mass spectrometry in more applications, increased emphasis is placed on the ion-neutral collisional cross sections (CCS) to identify unknown analytes in complex matrices. While CCS values can provide useful information about relative analyte size, several critical assumptions are inherent in the most common method of calculating CCS values, the Mason-Schamp equation. The largest source of error in the Mason-Schamp equation originates from not accounting for higher reduced electric field strengths, which are present in low-pressure instruments that require calibration. Previous corrections based on field strength have been proposed in literature, but their data used atomic ions in atomic gases, whereas most applications examine molecules measured in nitrogen. Here, we use a series of halogenated anilines measured in air and nitrogen between 6-120 Td on a first principles ion mobility instrument (HiKE-IMS). With this series of measurements, the average velocity of the ion packet is known allowing for direct calculation of reduced mobilities (K0), alpha functions, and finally, a detailed examination of CCS as a function of E/N. In the worst-case scenario, there is over a 55% difference in CCS values for molecular ions measured at high fields depending on the method used. When comparing CCS values to those in a database for unknown identification, this difference can lead to misidentification. To immediately alleviate some of the error in calibration procedures, we propose an alternative method using K0 and alpha functions that simulate first principles mobilities at higher fields.

Already current practice? A snapshot survey on doxycycline use for prevention of sexually transmitted infections in parts of the German MSM community.

PURPOSE: Recent studies point toward a potential benefit of doxycycline use for post-exposure prophylaxis (PEP) or pre-exposure prophylaxis (PrEP) to prevent sexually transmitted infections (STIs). Although prescribing doxycycline in a prophylactic intention is not generally recommended yet, we noticed an increasing number of inquiries from individuals within the LGBTQ community for doxycycline prescriptions. METHODS: We conducted an anonymous online survey to evaluate the current extent of doxycycline use for PEP or PrEP within the LGBTQ community using REDCap electronic data capture tools. Participants gained access to the online survey through a QR code on posters in the premises of our STI outpatient department and at LGBTQ community-related events in the south-western region of Germany. Additional access was provided by a direct link shared on social media profiles for men having sex with men (MSM), transgender, and queers. RESULTS: 96 of 99 responses were eligible for analysis. Twenty-two participants (23%) indicated to have already used doxycycline for PEP and six participants (6%) used doxycycline for PrEP. The majority of participants used pills left over from previous doxycycline treatment. Forty percent of indicated modes of access were without a regular prescription, e.g., by provision from acquaintances (with or without healthcare profession) or by ordering online. CONCLUSION: Our study shows that the concept of doxycycline use for prevention of STIs is already well known and applied in the LGBTQ community. Further analysis, especially modeling studies, are needed to evaluate strategies aiming to reduce doxycycline intake (PEP/PrEP versus repeated targeted therapies) and improve sexual health outcomes within the community.

The origin of isomerization of aniline revealed by high kinetic energy ion mobility spectrometry (HiKE-IMS).

Although aniline is a relatively simple small molecule, the origin of its two peaks observed in ion mobility spectrometry (IMS) has remained under debate for at least 30 years. First hypothesized as a difference in protonation site (amine vs. benzene ring), each ion mobility peak differs by one Dalton when coupled with mass spectrometry where the faster mobility peak is the molecular ion peak, and the slower mobility peak is protonated. To complicate the deconvolution of structures, some previous literature shows the peaks as unresolved and thus proposes these species exist in equilibrium. In this work, we show that when measured with high kinetic energy ion mobility spectrometry (HiKE-IMS), the two peaks observed in spectra of both aniline and all n-fluoroanilines are fully separated (chromatographic resolution from 2-7, Rp > 110) and therefore not in equilibrium. The HiKE-IMS is capable of changing ionization conditions independently of drift region conditions, and our results agree with previous literature showing that ionization source settings (including possible fragmentation at this stage) are the only influence determining the speciation of the two aniline peaks. Finally, when the drift and reactant gas are changed to nitrogen, a third peak appears at high E/N for 2-fluoroaniline and 4-fluoroaniline for the first time in reported literature. As observed by HiKE-IMS-MS, the new third peak is also protonated showing that the para-protonated aniline and resulting fragment ion, molecular ion aniline, can be fully separated in the mobility domain for the first time. The appearance of the third peak is only possible due to the increased separation of the other two peaks within the HiKE-IMS.

Complex reaction kinetics of a Mannich reaction in droplets under electrospray conditions.

Precise, efficient, and effective control of chemical reaction conditions is a viable measure for the environment-conscious time and energy resource management in modern laboratories and in industry. Parameter changes such as surface enlargement, pH, local reactant accumulation by solvent evaporation and polarization effects, etc., have been shown to greatly affect the reaction rate of a chemical reaction. In electrospray (ES) ionization - a soft ionization method often used for mass spectrometry - all these parameters change constantly and with high dynamics during the nebulization process that generates droplets as the ultimate confined µ-reaction vessels. Therefore, high acceleration factors are reported in literature for a manifold of such µ-droplet reactions. Here, the tri-molecular Mannich reaction was identified as a suitable candidate for studying thermal, electronic, and fluidic manipulation of the ES process to achieve high conversion rates with short reaction times and compare them to the batch reaction. Some of these manipulations were conducted separately to better quantify their individual contributions. Here, the keto-enol-tautomerism of the used ß-diketones, the high proton concentrations, and the longer reaction times in the µ-droplets are presumed to have the greatest impact on these enhancement factors. Experiments were performed to find ES conditions with small initial droplets and long droplet flight times where the highest reaction conversion rates are obtained. A sharp increase in the product peak was found at large distances between the mass spectrometry (MS) inlet and ES source at high voltages. Moreover, different trends were found for the two ketones studied, acetylacetone (AcAc) and 1,3-cyclohexanedione (Cyclo), by changing the temperature of the heated ES source. Finally, high conversion rates were obtained for the combination of formaldehyde (Fal) and piperidine (Pip) with AcAc and Cyclo, respectively, with over 90%. With respect to the batch reaction, this is mainly due to an increase in reaction kinetics as well as a shift in thermodynamics for the µ-droplet reaction environment.

Rebounds of sevoflurane concentration during simulated trigger-free pediatric and adult anesthesia.

BACKGROUND: In trigger-free anesthesia a volatile anesthetic concentration of 5 parts per million (ppm) should not be exceeded. According to European Malignant Hyperthermia Group (EMHG) guideline, this may be achieved by removing the vapor, changing the anesthetic breathing circuit and renewing the soda lime canister followed by flushing with O2 or air for a workstation specific time. Reduction of the fresh gas flow (FGF) or stand-by modes are known to cause rebound effects. In this study, simulated trigger-free pediatric and adult ventilation was carried out on test lungs including ventilation maneuvers commonly used in clinical practice. The goal of this study was to evaluate whether rebounds of sevoflurane develop during trigger-free anesthesia. METHODS: A Dräger® Primus® was contaminated with decreasing concentrations of sevoflurane for 120 min. Then, the machine was prepared for trigger-free anesthesia according to EMHG guideline by changing recommended parts and flushing the breathing circuits using 10 or 18 l⋅min- 1 FGF. The machine was neither switched off after preparation nor was FGF reduced. Simulated trigger-free ventilation was performed with volume-controlled ventilation (VCV) and pressure-controlled ventilation (PCV) including various ventilation maneuvers like pressure support ventilation (PSV), apnea, decreased lung compliance (DLC), recruitment maneuvers, prolonged expiration and manual ventilation (MV). A high-resolution ion mobility spectrometer with gas chromatographic pre-separation was used to measure sevoflurane in the ventilation gas mixture in a 20 s interval. RESULTS: Immediately after start of simulated anesthesia, there was an initial peak of 11-18 ppm sevoflurane in all experiments. The concentration dropped below 5 ppm after 2-3 min during adult and 4-18 min during pediatric ventilation. Other rebounds of sevoflurane > 5 ppm occurred after apnea, DLC and PSV. MV resulted in a decrease of sevoflurane < 5 ppm within 1 min. CONCLUSION: This study shows that after guideline-compliant preparation for trigger-free ventilation anesthetic machines may develop rebounds of sevoflurane > 5 ppm during typical maneuvers used in clinical practice. The changes in rate and direction of internal gas flow during different ventilation modes and maneuvers are possible explanations. Therefore, manufacturers should provide machine-specific washout protocols or emphasize the use of active charcoal filters (ACF) for trigger-free anesthesia.

Decrease of tibial tuberosity trochlear groove distance following mechanically aligned total knee arthroplasty.

PURPOSE: Anterior knee pain (AKP) is common following total knee arthroplasty. The tibial tuberosity trochlear groove distance (TTTG) influences patellofemoral joint loading in the native knee. Increased TTTG may lead to maltracking of the patella and anterior knee pain. The purpose of this study was to investigate potential changes in TTTG following total knee arthroplasty (TKA). METHODS: TTTG was measured on preoperative CT data on a consecutive series of patients scheduled to receive TKA with patient-specific instrumentation, and compared to a computer simulation of the postoperative TTTG. Preoperative TTTG was measured with a 3D planning software in 250 knees. The postoperative result was simulated and TTTG measured within the software. Three different groups were analysed: neutral (180° ± 3) (n = 50), valgus (> 190°) (n = 100), and varus (< 170°) (n = 100). RESULTS: Median preoperative to simulated postoperative TTTG decreased from 15.0 [interquartile range (IQR) 6.0] mm to 6.5 (IQR 5.0) mm for all axes combined. A significant postoperative reduction of TTTG was found in each group (p < 0.001). The mean change in TTTG did not differ significantly between the groups [- 8.8 (IQR 5.5) mm neutral, - 8.3 (IQR 7.0) mm valgus, - 7.5 (IQR 5.8) mm varus, p = 0.223]. CONCLUSION: This computer-based study suggests that mechanically aligned TKA significantly decreases TTTG distance in neutral, valgus and varus knees, assuming that the postoperative result coincides with the preoperative planning. Further study is warranted to evaluate the clinical relevance of this finding.

Excessive femoral torsion is not associated with patellofemoral pain or instability if TKA is functionally aligned and the patella denervated.

PURPOSE: Recent data suggest that individual morphologic factors should be respected to restore preoperative patellofemoral alignment and thus reduce the likelihood of anterior knee pain. The goal of this study was to investigate the effect of excessive femoral torsion (FT) on clinical outcome of TKA. METHODS: Patients who underwent TKA and complete preoperative radiographic evaluation including a long-leg radiograph and CT scan were included. 51 patients showed increased FT of > 20° and were matched for age/sex to 51 controls (FT < 20°). Thirteen patients were lost to follow-up. Thirty-eight matched pairs were compared after a 2 year follow-up clinically (Kujala and patellofemoral score for TKA) and radiographically (FT, frontal leg axis, TT-TG, patellar thickness, patellar tilt, and lateral displacement of patella). Functional alignment of TKA was performed (hybrid-technique). All patellae were denervated but no patella was resurfaced. RESULTS: There was no significant difference between clinical scores two years after surgery between patients with normal and excessive FT (n.s.). Kujala score was 64.3 ± 16.7 versus 64.8 ± 14.4 (n.s.), and patellofemoral score for TKA was 74.3 ± 21 versus 78.5 ± 20.7 (n.s.) for increased FT group and control group, respectively. There was no correlation between preoperative FT and clinical scores. Other radiographic parameters were similar between both groups. No correlations between clinical outcomes and preoperative/postoperative frontal leg axis or total leg axis correction were found (n.s.). CONCLUSION: If the leg axis deformity is corrected to a roughly neutral alignment during cemented TKA, including patellar denervation, then excessive FT was not associated with patellofemoral pain or instability. LEVEL OF EVIDENCE: Prospective comparative study, level II.

Long-term recurrence rate in anterior shoulder instability after Bankart repair based on the on- and off-track concept.

BACKGROUND: Since its first proposal, the concept of on- and off-track lesions in anterior shoulder instability has gained clinical relevance as a tool to predict the failure rate of arthroscopic Bankart repair. Current literature only reports either short-term follow-up or long-term results of small sample sizes. The aim of this study was to provide a long-term evaluation of recurrent instability following arthroscopic Bankart repair in a large cohort using the on-track vs. off-track concept as a predictor for failure. METHODS: We retrospectively analyzed 271 patients who underwent primary arthroscopic Bankart repair for anterior shoulder instability between 1998 and 2007. All patients with a minimum follow-up of 78 months and a preoperative computed tomographic (CT) or magnetic resonance imaging (MRI) scan were included into the study. Preoperative CT and/or MRI scans were used to determine the glenoid track and width of Hill-Sachs lesion. Recurrence of instability was defined as presence of instability symptoms (dislocation, subluxation, and/or apprehension) or revision surgery (stabilization procedure) and was assessed as the primary outcome parameter. RESULTS: The glenoid track of 163 shoulders was assessed (female n = 51, male n = 112) with a mean follow-up of 124 months (99.4-145.6, standard deviation = 2.5) and a mean age of 24 years (20-34.). An off-track Hill-Sachs lesion was found in 77 cases (47%), and in 86 cases (53 %) it was on-track. The rate of recurrent instability in the off-track group was 74% (n = 57) compared with 27% (n = 23) in the on-track group (P < .001). The overall rate of revision surgery due to instability was 29% (n = 48) after a mean time of 50.9 months (±42.8) following Bankart repair. The rate of revision surgery in the off-track group was 48% (n = 37) after a mean of 53.5 months (±42.0) vs. 13% (n = 11) after 42.3 months (±46.3) in the on-track group (P < .001). CONCLUSION: This study shows that the on- and off-track concept helps to distinguish patients for whom an isolated arthroscopic Bankart repair yields long-term benefits. Because of the high rate of recurrent instability in the off-track group, an off-track lesion should be treated surgically in such a way that the off-track lesion is converted into an on-track lesion.

Split-Ring Resonator Based Sensor for the Detection of Amino Acids in Liquids.

Amino acids belong to the most important compounds for life. They are structural components of proteins and required for growth and maintenance of cells. Essential amino acids cannot be produced by the organism and must be ingested through the nutrition. Therefore, the detection of amino acids is of great interest when analyzing cell culture media and nutrition. In this work, we present a split-ring resonator as a simple but sensitive detector for amino acids. Split-ring resonators are RLC resonant circuits with a split capacitance and thus a resonance frequency that depends on the electromagnetic properties of a liquid sample at the split capacitance. Here, the split capacitance is an interdigital structure for highest sensitivity and covered with a fluidic channel for flow through experiments. First measurements with a vector network analyzer show detection limits in the range from 105 µM for glutamic acid to 1564 µM for isoleucine, depending on the electromagnetic properties of the tested amino acids. With an envelope detector for continuous recording of the resonance frequency, the split-ring resonator can be used in ion chromatography. At a flow rate of 0.5 mL/min, it reaches limits of detection of 485 µM for aspartic acid and 956 µM for lysine.

Defining estimands for efficacy assessment in single arm phase 1b or phase 2 clinical trials in oncology early development.

The addendum of the ICH E9 guideline on the statistical principles for clinical trials introduced the estimand framework. The framework is designed to strengthen the dialog between different stakeholders, to introduce greater clarity in the clinical trial objectives and to provide alignment between the estimand and statistical analysis. Estimand framework related publications thus far have mainly focused on randomized clinical trials. The intention of the Early Development Estimand Nexus (EDEN), a task force of the cross-industry Oncology Estimand Working Group (www.oncoestimand.org), is to apply it to single arms Phase 1b or Phase 2 trials designed to detect a treatment-related efficacy signal, typically measured by objective response rate. Key recommendations regarding the estimand attributes include that in a single arm early clinical trial, the treatment attribute should start when the first dose is received by the participant. Focusing on the estimation of an absolute effect, the population-level summary measure should reflect only the property used for the estimation. Another major component introduced in the ICH E9 addendum is the definition of intercurrent events and the associated possible ways to handle them. Different strategies reflect different clinical questions of interest that can be answered based on the journeys an individual subject can take during a trial. We provide detailed strategy recommendations for intercurrent events typically seen in early-stage oncology. We highlight where implicit assumptions should be made transparent as whenever follow-up is suspended, a while-on-treatment strategy is implied.

Rapid Development of Cefiderocol Resistance in Carbapenem-resistant Enterobacter cloacae During Therapy Is Associated With Heterogeneous Mutations in the Catecholate Siderophore Receptor cirA.

We report a case of resistance development toward cefiderocol in a patient with intra-abdominal and bloodstream infections caused by carbapenemase-producing Enterobacter cloacae within 21 days of cefiderocol therapy. Whole genome sequencing revealed heterogeneous mutations in the cirA gene, encoding a catecholate siderophore receptor, conferring phenotypic resistance to cefiderocol.