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Abstract Background Some clinically important genetic variants are not easily evaluated with next‐generation sequencing (NGS) methods due to technical challenges arising from high‐ similarity copies (e.g., PMS2, SMN1/SMN2, GBA1, HBA1/HBA2, CYP21A2), repetitive short sequences (e.g., ARX polyalanine repeats, FMR1 AGG interruptions in CGG repeats, CFTR poly‐T/TG repeats), and other complexities (e.g., MSH2 Boland inversions). Methods We customized our NGS processes to detect the technically challenging variants mentioned above with adaptations including target enrichment and bioinformatic masking of similar sequences. Adaptations were validated with samples of known genotypes. Results Our adaptations provided high‐sensitivity and high‐specificity detection for most of the variants and provided a high‐sensitivity primary assay to be followed with orthogonal disambiguation for the others. The sensitivity of the NGS adaptations was 100% for all of the technically challenging variants. Specificity was 100% for those in PMS2, GBA1, SMN1/SMN2, and HBA1/HBA2, and for the MSH2 Boland inversion; 97.8%–100% for CYP21A2 variants; and 85.7% for ARX polyalanine repeats. Conclusions NGS assays can detect technically challenging variants when chemistries and bioinformatics are jointly refined. The adaptations described support a scalable, cost‐effective path to identifying all clinically relevant variants within a single sample.

This paper covers direct sub-atmospheric pressure ionization mass spectrometry (MS). The discovery, applications, and mechanistic aspects of novel ionization processes for use in MS that are not based on the high-energy input from voltage, laser, and/or high temperature but on sublimation/evaporation within a region linking a higher to lower pressure and modulated by heat and collisions, are discussed, including how this new reality has guided a series of discoveries, instrument developments, and commercialization. A research focus, inter alia, is on how best to understand, improve, and use these novel ionization processes, which convert volatile and nonvolatile compounds from solids (sublimation) or liquids (evaporation) into gas-phase ions for analysis by MS providing reproducible, accurate, sensitive, and prompt results. Our perception on how these unprecedented versus traditional ionization processes/methods relate to each other, how they can be made to coexist on the same mass spectrometer, and an outlook on new and expanded applications (e.g., clinical, portable, fast, safe, and autonomous) is presented, and is based on ST's Opening lecture presentation at the Nordic Mass spectrometry Conference, Geilo, Norway, January 2023. Focus will be on matrix-assisted ionization (MAI) and solvent-assisted ionization (SAI) MS covering the period from 2010 to 2023; a potential paradigm shift in the making., (© 2024 John Wiley & Sons Ltd.)

Background: Atrial fibrillation (AF) has been proposed as a risk factor for exercise-induced pulmonary haemorrhage (EIPH) due to increased pressure in the left atrium., Objective: To evaluate if AF was associated with EIPH following a standardised exercise test (SET) to fatigue., Study Design: Two-arm controlled experiment., Methods: Ten untrained Standardbred mares mean (standard deviation [SD]) age 6 (2) years performed a SET on the treadmill in sinus rhythm (SR) (SET1) and 25-44 days after induction of self-sustained AF (SET2). AF was induced by tachypacing using a pacing device. Endoscopy, including tracheal wash and bronchoalveolar lavage (BAL), was performed 48-72 h before and 24 h after the two SETs. In addition, endoscopic grading of tracheal blood was performed 2 h after each SET., Results: After SET1, none of the horses showed blood in the trachea, and two horses showed erythrophagocytosis. Following SET2, two horses had grade 1 blood in the trachea and free erythrocytes and erythrophagocytosis in the BAL, while another two horses had erythrophagocytosis in the BAL. In SET2, the overall performance on the treadmill was decreased with a lower maximum velocity (SET1 10.3 ± 0.8 m/s vs. SET2 8.9 ± 0.9 m/s, p = 0.004), a higher heart rate (284 ± 21 vs. 221 ± 18 bpm, p = 0.003) and more abnormal QRS complexes (p < 0.001) compared with SET1., Conclusions: Two horses showed signs of EIPH, resulting in visible blood in the trachea, when exercising in AF compared with SR. However, a possible link between EIPH, pulmonary pressure and AF needs to be further elucidated., (© 2023 The Authors. Equine Veterinary Journal published by John Wiley & Sons Ltd on behalf of EVJ Ltd.)

Background: Paranasal sinus cysts (PSC) are a common cause of equine secondary sinusitis. The outcome and associated complications have not been frequently reported., Objectives: To review the associated clinical signs, associated morbidities and outcomes of horses treated for PSC., Study Design: Retrospective multicentre case series., Methods: Retrospective analysis of case records and telephone follow up survey., Results: Subjects were 37 horses 1-24 years old that were presented with nasal discharge (n = 31), facial swelling (n = 25) and epiphora (n = 19). Radiography and computed tomography allowed identification of the cyst-induced changes including concomitant tissue destruction (n = 31), leading among other things to local nerve damage causing headshaking (n = 6) and unilateral blindness (n = 1). Radiographic changes to adjacent dental apices were present in 10 horses. Horses over 10 years old showed more of the named associated problems. Post-operative complications included surgical site infection (SSI) (n = 11), nasofrontal suture periostitis (n = 6) and sequestration (n = 1) following removal of the PSC via osteotomy. The long-term response to treatment was available for 28 cases with 22 horses (78.6%) fully cured, 4 (14.3%) partially cured and 2 (7.1%) not responding to treatment. In 7 horses (18.9%) there was recurrence of the cyst post-operatively., Main Limitations: Due to the study being a multicentre retrospective case series with collection of data over an extended period, there may be inconsistency in data recording and absence of reporting of some findings., Conclusions: Overall, the diagnosis and treatment of sinus cysts is relatively straightforward and carries a good prognosis. In long-standing cases complications secondary to the expansive growth of cysts will dramatically affect the prognosis for full recovery due to pressure-induced changes to facial bones, cheek teeth and nerves. These secondary complications mainly occurring in older horses may be due to a combination of a relatively longer period of affection and the inflexibility of older horses' bones. Cyst recurrence following treatment can occur in up to 19% of cases., (© 2018 EVJ Ltd.)

Background: The objective of this study was to assess the impact of perioperative transfusion on the prognosis of patients who underwent complete (R0) resection of oral squamous cell carcinoma and reconstruction by microvascular flaps., Methods: By following an inclusion and exclusion protocol, 223 patients were included in the study who underwent R0 resection of oral squamous cell carcinoma and reconstruction by microvascular flaps at a single center. Clinical and pathologic factors as well as transfusion data were retrieved from a prospective database and analyzed retrospectively. Survival data were assessed using the method of Kaplan and Meier. For multivariate analysis the accelerated failure time model (Weibull distribution) was chosen., Results: The overall survival rate was 71% at 1 year, 67% at 3 years, and 55% at 5 years. In univariate analysis, age (P = .003), tumor size (P = .005), lymph node status (P = .008), tumor differentiation (P = .008), transfusion (P = .006), American Society of Anesthesiologists (ASA) class (P = .001), and mandibular reconstruction (P = .045) were associated significantly with overall survival. Multivariate analysis identified only age, histopathologic differentiation, and ASA class as independent risk factors (P < .001, P = .04, and P = .049, respectively). Age was identified as the strongest independent predictor for overall survival (hazards ratio for each 13-year increase in age, 1.97; 95% confidence interval, 1.36-2.85)., Conclusions: Transfusion of >4 U of blood did not appear to influence overall survival in patients who underwent primary surgery for oral squamous cell carcinoma. Because age and ASA class evolved as the strongest predictors of shortened overall survival, associated comorbidities may require more attention, particularly in elderly or socially deprived patients., ((c) 2009 American Cancer Society)