Strategies for capillary electrophoresis: Method development and validation for pharmaceutical and biological applications-Updated and completely revised edition.

This review is in support of the development of selective, precise, fast, and validated capillary electrophoresis (CE) methods. It follows up a similar article from 1998, Wätzig H, Degenhardt M, Kunkel A. "Strategies for capillary electrophoresis: method development and validation for pharmaceutical and biological applications," pointing out which fundamentals are still valid and at the same time showing the enormous achievements in the last 25 years. The structures of both reviews are widely similar, in order to facilitate their simultaneous use. Focusing on pharmaceutical and biological applications, the successful use of CE is now demonstrated by more than 600 carefully selected references. Many of those are recent reviews; therefore, a significant overview about the field is provided. There are extra sections about sample pretreatment related to CE and microchip CE, and a completely revised section about method development for protein analytes and biomolecules in general. The general strategies for method development are summed up with regard to selectivity, efficiency, precision, analysis time, limit of detection, sample pretreatment requirements, and validation.

[Effects of manipulator operation task and troubleshooting task on blood pressure indexes of oceanauts].

Objective: To explore the characteristics and rules of blood pressure changes in oceanauts during simulated operation of manipulator and troubleshooting tasks with different difficulty. Methods: In July 2020, 8 deep-sea manned submersible oceanauts, 6 males and 2 females, were selected as objects. In the 1∶1 model of Jiaolong deep-sea manned submersible, the oceanauts performed manipulator operation tasks and troubleshooting tasks with different difficulties, measured the continuous blood pressure of the oceanauts, filled in the NASA Task Load Index (NASA-TLX scale) after the completion of a single mission, and the changes of systolic pressure (SBP), diastolic pressure (DBP), mean arterial pressure (MAP) and mental workload were analyzed. Results: In a single task, the SBP, DBP and MAP of the oceanauts increased first and then decreased. The blood pressure values at the third minute were significantly lower than those at the first minute (P<0.01), and those at the fifth minute were significantly higher than those at the third minute (P<0.01). When performing the same task, compared with the quiet state, SBP, DBP and MAP increased when the oceanauts performed low difficulty, high difficulty, high difficulty+2-back manipulator operation task and troubleshooting task (P<0.05). When the task difficulty was the same, the SBP and MAP of oceanauts performing manipulator operation tasks were higher than those of oceanauts performing troubleshooting tasks (P<0.05). Compared with low difficulty tasks, the scores of NASA-TLX scale for oceanauts performing high difficulty manipulator operationtasks were significantly higher (P<0.05). Compared with the low difficulty task and high difficulty task, the scale score of the high difficulty+2-back troubleshooting task was significantly higher (P<0.05). When the task difficulty was the same, the scale scores of low difficulty and high difficulty manipulator operation tasks were significantly higher than those of troubleshooting tasks (P<0.05). SBP, DBP, MAP of No. 1, No. 3, No. 4, No. 5, and No. 7 oceanauts (all of whom had 6 years of diving) were positively correlated with NASA-TLX scale scores (r>0.8, P<0.05) . Conclusion: In the process of manned deep-sea diving, when the oceanauts perform manipulator operation tasks and troubleshooting tasks, with the increase of task difficulty, the mental load of oceanauts increases, and the blood pressure index increases significantly in a short time. At the same time, improving the proficiency of operation can reduce the variation range of blood pressure indexes. Blood pressure can be used as an effective reference to evaluate the difficulty of operation and guide scientific training.

A real-world experience of atrioventricular synchronous pacing with leadless ventricular pacemakers.

AIMS: The MicraTM transcatheter pacing system (TPS) (Medtronic) is the only leadless pacemaker that promotes atrioventricular (AV) synchrony via accelerometer-based atrial sensing. Data regarding the real-world experience with this novel system are scarce. We sought to characterize patients undergoing MicraTM -AV implants, describe percentage AV synchrony achieved, and analyze the causes for suboptimal AV synchrony. METHODS: In this retrospective cohort study, electronic medical records from 56 consecutive patients undergoing MicraTM -AV implants at the Mayo Clinic sites in Minnesota, Florida, and Arizona with a minimum follow-up of 3 months were reviewed. Demographic data, comorbidities, echocardiographic data, and clinical outcomes were compared among patients with and without atrial synchronous ventricular pacing (AsVP) ≥ 70%. RESULTS: Sixty-five percent of patients achieved AsVP ≥ 70%. Patients with adequate AsVP had smaller body mass indices, a lower proportion of congestive heart failure, and prior cardiac surgery. Echocardiographic parameters and procedural characteristics were similar across the two groups. Active device troubleshooting was associated with higher AsVP. The likely reasons for low AsVP were small A4-wave amplitude, high ventricular pacing burden, and inadequate device reprogramming. Importantly, in patients with low AsVP, subjective clinical worsening was not noted during follow-up. CONCLUSION: With the increasing popularity of leadless pacemakers, it is paramount for device implanting teams to be familiar with common predictors of AV synchrony and troubleshooting with MicraTM -AV devices.

Various innovative roles for 3-Fr microcatheters in pancreaticobiliary endoscopy.

With the development of newer devices and technical innovations, pancreaticobiliary endoscopy is expanding to assume more advanced therapeutic roles. As with other devices, slimmed-down "3-Fr microcatheters" are considered to be opening new windows toward entirely new therapeutic techniques for various purposes. Our practical experience with a total of 34 consecutive patients in whom 3-Fr microcatheters were applied during pancreaticobiliary endoscopic procedures clarified the potential roles of this instrument in pancreaticobiliary endoscopy. The major benefits of 3-Fr microcatheters involve their slimness and flexibility. Applications of 3-Fr microcatheters could be categorized into three groups according to the characteristics of usage: (1) utilization as a cannulation catheter for peroral digital cholangioscopy (n = 15); (2) selective advancement through deep flexures or severely stenotic ducts (n = 11); or (3) two-devices-in-one-channel technique (n = 8). The microcatheter worked successfully for cannulation of cholangioscopy in all but one case (14/15, 93.3%). For selective advancement, the microcatheter worked for troubleshooting in 9 of 11 cases (81.8%). With the two-devices-in-one-channel technique, the microcatheter proved satisfactory in all cases (8/8, 100%). In total, the microcatheter was successfully maneuvered in 31 of 34 cases (91.1%), following the failure of procedures using conventional endoscopic techniques. In terms of adverse events, cystic duct injury was only observed in two cases (5.8%), who recovered under conservative observation, because its slimness could minimize the damage. We believe that 3-Fr microcatheters offer effective and safe salvage troubleshooting during various endoscopic pancreaticobiliary procedures that face troublesome situations with conventional strategies.

The Influence of Electrode Configuration Changes on the Sensory and Motor Response During (Re)Programming in Sacral Neuromodulation.

OBJECTIVES: This study aimed to assess the neurophysiological basis behind troubleshooting in sacral neuromodulation (SNM). Close follow-up of SNM patients with program parameter optimization has proven to be paramount by restoring clinical efficacy and avoiding surgical revision. MATERIALS AND METHODS: A total of 34 successful SNM patients (28 overactive bladder wet, six nonobstructive urinary retention) with an implantable pulse generator were included. All possible bipolar and monopolar electrode settings were tested at sensory threshold (ST) to evaluate sensory (mapped on a perineal grid with 1 cm2 coordinates) and motor (peak-to-peak amplitude and latency of muscle action potential) responses of the pelvic floor. Pelvic floor muscle electromyography was recorded using a multiple array probe, placed intravaginally. Parametric tests were used for paired data: repeated-measures ANOVA or t-test. A nonparametric test was used for paired data: Friedman ANOVA or Wilcoxon signed rank (WSR) test; p < 0.05 was considered statistically significant. If significant, ANOVA was followed by Dunn-Bonferroni post hoc analysis. RESULTS: Monopolar configurations showed significantly lower STs-1.38 ± 0.73 V vs 1.76 ± 0.89 V (paired t-test: p < 0.0001)-and presented with significantly higher peak-to-peak amplitudes-115.67 ± 79.03 µV vs 90.77 ± 80.55 µV (WSR: p = 0.005)-than bipolar configurations. When polarity was swapped, configurations with the cathode distal to the anode showed significantly lower STs, 1.73 ± 0.91 V vs 1.85 ± 0.87 V (paired t-test: p = 0.003), and mean peak-to-peak amplitudes, 81.32 ± 72.82 µV vs 100.21 ± 90.22 µV (WSR: p = 0.0001). Cathodal changes resulted in more changes in sensory responses than anodal changes (χ2 test: p = 0.044). In cathodal changes only, peak-to-peak amplitudes were significantly higher when the distance between electrodes was maximally spread (WSR: p = 0.046). CONCLUSIONS: From a neurophysiological point of view, monopolar configurations stimulated more motor nerve fibers at lower STs, therefore providing more therapeutic efficiency. Swapping polarity or changing the position of the cathode led to different sensory and motor responses, serving as potential reprogramming options.

Solutions for Shared Resource Lab Remote Quality Control and Instrument Troubleshooting during a Pandemic.

The COVID-19 pandemic has dramatically affected shared resource lab (SRL) staff in-person availability at institutions globally. This article discusses the challenges of ensuring reliable instrument performance and quality data output while facility staff and external service provider on-site presence is severely limited. Solutions revolve around the adoption of remote monitoring and troubleshooting platforms, provision of self-service troubleshooting resources specific to facility instruments and workflows, development of an assistance contact policy, and ensuring efficiency of limited in-person staff time. These solutions employ software and hardware tools that are already in use or readily available in the SRL community, such as remote instrument access tools, video hosting and conferencing platforms, and ISAC shared resources. © 2020 International Society for Advancement of Cytometry.

Troubleshooting in thoracoscopic anatomical lung resection for lung cancer.

Video-assisted thoracoscopic surgery (VATS) anatomical lung resection (ALR) has been gaining popularity in the treatment of lung cancer in line with remarkable advances in both equipment and technique. The development and refinement of its technique have allowed thoracic surgeons to perform a wide variety of challenging and complex procedures in a minimally invasive fashion. Careful and meticulous preparation may shift in the future with the increasing sophistication and capabilities of VATS ALR. Moreover, constant awareness and a structured plan of the procedure are imperative to reducing or preventing complications. Intraoperative major complications during VATS ALR are infrequent, but can have catastrophic consequences. The decision to continue with VATS should take into consideration the surgeon's skill level and ease with the approach and the relative potential benefit against the risk to the patient. We conducted this study to investigate the possible problems during VATS ALR and identify how to solve them based on the previous literature and our institutional data sampling.

Reprogramming Sacral Neuromodulation for Sub-Optimal Outcomes: Evidence and Recommendations for Clinical Practice.

OBJECTIVES: In some patients treated for urinary or fecal incontinence with sacral neuromodulation (SNM) persistence of symptoms, a reduction in efficacy or adverse effects of stimulation can occur. In such situations, further programming of the SNM device can help resolve problems. Infrequently hardware failure is detected. This article aims to provide practical guidance to solve sub-optimal outcomes (troubleshooting) occurring in the course of SNM therapy. MATERIALS AND METHODS: A systematic literature review was performed. Collective clinical experience from an expert multidisciplinary group was used to form opinion where evidence was lacking. RESULTS: Circumstances in which reprogramming is required are described. Actions to undertake include changes of electrode configuration, stimulation amplitude, pulse frequency, and pulse width. Guidance in case of loss of efficacy and adverse effects of stimulation, developed by a group of European experts, is presented. In addition, various hardware failure scenarios and their management are described. CONCLUSIONS: Reprogramming aims to further improve patient symptoms or ensure a comfortable delivery of the therapy. Initial changes of electrode configuration and adjustment of stimulation parameters can be performed at home to avoid unnecessary hospital visits. A logical and stepwise approach to reprogramming can improve the outcome of therapy and restore patient satisfaction.

[1.5 T Nuclear Magnetic Resonance Imaging System Failure Treatment and Rectification Summary].

Quench of magnetic resonance imaging system refers to the process that the superconducting condition inside the magnet is destroyed due to some reason. The large current stored in the coil is quickly converted into heat at the place where the resistance is formed, and a large amount of liquid helium in the magnet is evaporated. If it happens, it will cause huge loss to the user. We introduce the real cases of 1.5 T magnetic resonance imaging system's quench fault, maintenance treatment and management improvement, which can be used for reference by various medical institutions, so as to better strengthen the operation and maintenance management of magnetic resonance imaging system, so as to avoid the occurrence of out of tolerance fault, and do a good job in the guarantee work after the out of tolerance fault.

Resurrecting the phoenix: When an assay fails.

Understanding protein-small-molecule interactions is a critical component of rational drug-design. Structure-activity relationship (SAR)-guided medicinal chemistry is informed by the biological outcome, as assessed by biochemical activity or cellular effect, of chemical modifications on small molecules. The effectiveness of SAR is reliant on the sturdiness and durability of assay design and the quality of information garnered from assays. Lack of quality data at this step can lead to obstruction of the drug discovery pipeline with profound implications for the timelines of introducing a drug into the market. Hence, it would not be an overstatement to consider biochemical/biological assays as the backbone of drug-discovery. Enzyme assays can fail for many different reasons, with the enzyme and the substrate being the principal players. Lack of clarity can hamper progress and can lead to mounting costs and potentially losing competitive advantage. Although each assay is unique and requires a specific approach to troubleshoot the problem at hand, there are general guidelines that can be followed to maximize the chances of success. This review is a step-by-step attempt at reintroducing fundamental biochemical concepts within the context of an enzyme assay, delineating probable causes for failure and potential approaches to get an assay back up and running.

Multiphoton intravital microscopy in small animals: motion artefact challenges and technical solutions.

Since its invention 29 years ago, two-photon laser-scanning microscopy has evolved from a promising imaging technique, to an established widely available imaging modality used throughout the biomedical research community. The establishment of two-photon microscopy as the preferred method for imaging fluorescently labelled cells and structures in living animals can be attributed to the biophysical mechanism by which the generation of fluorescence is accomplished. The use of powerful lasers capable of delivering infrared light pulses within femtosecond intervals, facilitates the nonlinear excitation of fluorescent molecules only at the focal plane and determines by objective lens position. This offers numerous benefits for studies of biological samples at high spatial and temporal resolutions with limited photo-damage and superior tissue penetration. Indeed, these attributes have established two-photon microscopy as the ideal method for live-animal imaging in several areas of biology and have led to a whole new field of study dedicated to imaging biological phenomena in intact tissues and living organisms. However, despite its appealing features, two-photon intravital microscopy is inherently limited by tissue motion from heartbeat, respiratory cycles, peristalsis, muscle/vascular tone and physiological functions that change tissue geometry. Because these movements impede temporal and spatial resolution, they must be properly addressed to harness the full potential of two-photon intravital microscopy and enable accurate data analysis and interpretation. In addition, the sources and features of these motion artefacts are varied, sometimes unpredictable and unique to specific organs and multiple complex strategies have previously been devised to address them. This review will discuss these motion artefacts requirement and technical solutions for their correction and after intravital two-photon microscopy.

High incidence of diaphragmatic myopotential oversensing by a specific implantable cardioverter defibrillator.

INTRODUCTION: Diaphragmatic myopotential oversensing (dMPO) by implantable cardioverter defibrillators (ICDs) is thought to be a rare condition that can be misdiagnosed as lead failure and lead to unnecessary lead replacement. We observed several cases of dMPO in patients with Sorin/LivaNova ICDs (MicroPort Sci.). We sought to systematically assess the incidence of dMPO in patients with Sorin/LivaNova ICDs. METHODS AND RESULTS: A predefined number of 100 consecutive patients with Sorin/LivaNova ICDs were prospectively included in the device clinic of our center. Stored arrhythmia episodes were checked for spontaneous dMPO. In addition, we performed provocation maneuvers by Valsalva. At least one episode of spontaneous or provoked dMPO was seen in 12 (12%) of the 100 patients included in the study (86% males, median age: 66 years). Nine of 89 patients (10%) with true bipolar and 3 of 11 patients (27%) with integrated bipolar sensing configuration were affected. Spontaneous dMPO was observed in 7 of 58 patients (12%) with sensitivity programmed to 0.4 mV and in 2 of 42 patients (5%) with sensitivity programmed to 0.6 mV (not significant). In three patients, dMPO could be provoked with no spontaneous episodes recorded. In two nonpacemaker-dependent patients with a CRT-D, ventricular pacing was temporarily inhibited. No antitachycardia therapy was triggered by dMPO in any patient. CONCLUSIONS: DMPO is frequent in patients with Sorin/LivaNova ICDs, especially with sensitivity programmed to 0.4 mV. It also frequently occurs with true bipolar sensing configuration. DMPO should not be misinterpreted as lead failure to avoid unnecessary lead replacement.

Why do we need training? - A "Training school on molecular methods used for foodborne parasite diagnostics in different matrices" is a example of knowledge transfer to foster research quality in EU.

Foodborne parasites with zoonotic potential are of particular concern for human health, being responsible for serious and potentially life threatening diseases. In the last decades, the development of molecular biology techniques have been successfully implemented for clinical diagnosis of FBPs in animal or human samples providing cheaper, less labor intensive, reliable and more sensitive tests. It is apparent from recent publications that unsubstantiated molecular methods for parasite detection that have undergone scant evaluation for sensitivity and specificity are becoming increasingly common. The aim of the organized Training Schools was to transfer knowledge on application, optimization and troubleshooting for methods used to extract, amplify, and sequence nucleic acids from contaminated matrices and isolated FBPs. The organized Training Schools fulfilled the trainees' expectations, whom acquired useful knowledge for their research activities.

His bundle pacing: Clinical consequences of unnecessary right ventricle backup pacing.

As a backup lead in right ventricle (RV) is often used in His-bundle pacing (HBP) implants, in sinus rhythm patients the His lead is connected to the left ventricular (LV) port of a CRT device. In current devices, the backup pacing will be delivered 100% of time due to cross-channel ventricular refractory periods. Beyond an impact on battery, unnecessary RV pacing could find excitable tissue and capture a portion of the myocardium tissue potentially reducing the benefits of physiological HBP as shown in this case report where the switch from biventricular to LV-only pacing improved acute and 2-month echocardiography parameters.

DCD and DBD lung transplantation optimized by ex vivo perfusion: What to do when the pump fails.

BACKGROUND: Ex vivo perfusion is a safe and feasible method of assessing and using high-risk donor organs. AIM: We describe a case of successfully ex vivo treated and transplanted human lung allografts. METHODS: Donor human lungs were assessed using ex vivo, our trouble shooting protocol allowed safe recovery. RESULTS: We successfully implanted our ex vivo treated organs.

Permanent His Bundle Pacing: A programming and troubleshooting guide.

Permanent His Bundle Pacing (HBP) has recently gained popularity. However, implanting physicians and those who perform the device checks must invest in additional education in order to accurately program these devices, identify changes in morphology and perform troubleshooting to help achieve the best outcomes for the patients. This paper reviews key aspects of HBP and provides the educational tools for successful HBP follow-up and troubleshooting.

[Versa HD Volume-modulated Accelerator High Voltage Circuit Fault Maintenance].

This paper introduces a fast repair methods of Versa HD volume-modulated accelerator's high voltage circuit fault:the key points test method. To identify five key points:①Enter maintenance mode to check for AFC deviation; ② The magnetron waveform MI and PFN waveform PFN V are detected on the maintenance terminal board; ③ Detect thyratron waveform; ④ Check the supply voltage of thyratron; ⑤ HT PSU 600 V DC test 600 V normal or not. The waveform or voltage is also measured to efficiently narrow the fault range to find out the cause of the fault, quickly remove the fault.

An indissoluble knot: An unexpected troubleshooting during Micra implantation.

Micra is a transcatheter leadless pacing system delivered percutaneously through femoral vein access into the right ventricle. We are going to describe an initially standard procedure, with an unexpected troubleshooting at the end: the impossibility to release the device and retract the delivery due to a knot in the tether.

Serous-lined, stapled pouch wall plication: initial results of a simple and quick novel continence mechanism in troubleshooting secondary to continent cutaneous urinary diversion.

OBJECTIVES: To report a novel and straightforward technique of a secondary continent outlet for continent cutaneous urinary diversion (CCUD) reservoirs without the need for further bowel resection, reducing operating time and length of hospitalization. PATIENTS AND METHODS: From 2015 to 2017, six patients with unreconstructable, incontinent outlets (out of a total pool of 595 patients with CCUD) have undergone the technique described in the present paper at our department. The technique relies on the Mitrofanoff principle, using a stapled full-thickness pouch wall plication, which creates a flap-valve continence mechanism. RESULTS: All patients enjoyed full continence with ease of clean intermittent catheterization (CIC) in the postoperative period and on follow-up to a mean (range) of 12.4 (7-18) months. No major complications were encountered in any patient and the average capacity of the reservoirs was not compromised by the procedure (540 mL preoperatively vs 500 mL in further follow-up). CONCLUSION: In revisional surgery for secondary CCUD incontinence, especially if the patient has already lost a significant amount of bowel or has previously undergone radiation therapy, the technique described here represents a safe and effective alternative to restore continence.

Section detachment in immunohistochemistry: causes, troubleshooting, and problem-solving.

Section detachment in immunohistochemistry (IHC) is a common phenomenon, increasing times and costs of diagnosis and research. However, it has poorly been investigated. The aim of this study was to identify the causes of section detachment, with the purpose of defining a quality assured laboratory procedure to minimize detachment frequency. We screened 3349 IHC sections from formalin-fixed paraffin-embedded tissue, identifying 177 cases with section detachment (5.3% of the sample). Detachment regarded mainly samples of surgical breast tissue and IHC procedures in which heat pretreatment was used. Focusing on pre-analytical factors, we investigated seven main critical issues: (1) section aging; (2) section thickness; (3) slide contamination; (4) slide aging; (5) slide brand; (6) "human" influence; and (7) sample size and fixation. Each of these issues was individually investigated to establish their influence on detachment. Targeted experiments were performed by varying section age, thickness, cleanliness, slide brand and age, and sample size and fixation. Finally, to investigate operator-dependent causes, sections were cut by different operators blinded to aim. The most important factors influencing section detachment were demonstrated to be: section thickness, slide aging, slide brand, "human" influence, and size and fixation of samples. The pre-analytical phase, including all the aforementioned issues, should be standardized within a quality assurance program. By adopting these recommendations, we obtained a 34% drop in section detachment. Although section detachment remains difficult to eradicate completely, many other influences can be addressed and corrected in any laboratory leading to an increase in efficiency and cost saving.