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Improving mycotoxin enzyme immunoassay performance by optimizing formaldehyde-mediated hapten–protein conjugation : an a-cyclopiazonic acid case study
(2026) Hart, Leon; Rehagel, Christina; Schale, Daniela; Plötz, Madeleine; Usleber, Ewald
Enzyme immunoassays (EIAs) for small molecules often suffer from limited sensitivity or high non-specific background. This study presents a reproducible strategy to improve EIA performance by optimizing hapten–protein conjugation conditions. Using an immunoassay for the mycotoxin α-cyclopiazonic acid (α-CPA) as a case study, key parameters of the formaldehyde-mediated (Mannich) condensation reaction for synthesizing a hapten–bovine serum albumin conjugate were systematically investigated. Variation of reaction temperature (8–37°C) and time (0.25–72 h) showed that short reaction times at low temperature markedly improved assay performance. Under optimized conditions (8°C, 15 min), the detection limit improved from >100 ng/mL to 2.4 ± 0.2 ng/mL, with a strong reduction in non-specific background. The results demonstrate that targeted optimization of hapten–protein conjugation can significantly enhance EIA sensitivity and specificity, which helps to reduce the number of laboratory animals for antibody production. The approach is readily transferable to other indole-containing mycotoxins and was successfully applied to determine α-CPA in fungal agar plugs from mold-ripened cheeses. This study highlights an important aspect of conjugation conditions during immunoassay optimization.
Behavioral and autonomic manifestations of suspected focal impaired awareness seizures supported by interictal electroencephalographic findings in three dogs : a case series
(2026) Czerwik, Adriana; Olszewska, Agnieszka; Farke, Daniela; Schmidt, Martin Jürgen
Focal epileptic seizures with sensory impairment can be difficult to recognize and are often mistaken for behavioral disorders. Electroencephalography (EEG) can provide important supportive evidence for their epileptic origin when interpreted in conjunction with clinical presentation. This article describes three cases of canine patients: a 2-year-old Border Terrier, a 1-year-old Poodle mix, and a 4-year-old mixed-breed dog. All three were presented with a history of recurrent episodes characterized by fear, restlessness, hyperactivity, hiding, staring into space or at the sky, lip licking, and hypersalivation. In two dogs, there were also signs of impaired consciousness. The frequency of the episodes varied, ranging from multiple episodes per day to one episode every 1–2 weeks. No pre-ictal or post-ictal signs were reported. Clinical and neurological examinations were unremarkable in all three cases. The diagnostic workup included EEG using a Natus Xltek system and magnetic resonance imaging (MRI) of the brain with a Siemens 1.5 T scanner. EEG monitoring lasting 1.5–4 h revealed epileptiform discharges predominantly in the parietal and temporal regions. This case series highlights the occurrence of behavioral and autonomic symptoms without typical motor manifestations as a presentation of presumed idiopathic epilepsy in dogs, supported by EEG findings in conjunction with the clinical presentation. All three dogs were treated with antiseizure medication, which resulted in a decrease in both the frequency and intensity of episodes, with complete seizure freedom achieved in two dogs.
Comparison of four yogic breathing techniques regarding their impact on heart-rate variability and breathing frequency
(2026) Epe, Janika; Stark, Rudolf; Walter, Bertram; Ott, Ulrich
Background: The manner of breathing affects the autonomic nervous system (ANS) and corresponding processes in the body. Most studies focus on techniques prolonging breathing. Nevertheless, there are also techniques aimed at accelerating or changing other aspects of the breath. The variety of concepts suggests that different breathing techniques could have different effects on the ANS. This study intended to investigate differential effects of four yogic breathing techniques on heart-rate variability: (1) Ujjayi—relaxation through slowing down the breath; (2) Paced Breathing—enhancement of concentration by following a precise protocol of slowdown breathing; (3) Kapalabhati—raising wakefulness by mild hyperventilation; and (4) Alternate Nostril Breathing—balancing the autonomic nervous system by alternating breath between the nostrils. We hypothesized that there would be (1) differences in the impact on baseline measures after the training weeks, (2) differences between baseline and practice, (3) higher impact with longer performance of the breathing technique, and (4) differences between techniques. Methods: This study was conducted with 36 participants who learned each technique in group sessions and practiced them daily for two weeks of an eight-week program. After practicing each technique, participants performed the breathing technique while HRV, breathing frequency and other psychological variables were recorded; rMSSD was calculated for to interpret the HRV. Results: We found significant differences between calming and accelerating breathing techniques on HRV and breathing frequency, and different effects of the four breathing techniques compared to their baseline and over practice time. Conclusions: Because breathing is such an easily accessible tool for regulation, it seems important to see which breathing technique helps people regulate quickly and effectively in everyday life.
Case report : a multimodal treatment approach combining multi-target rTMS with a modified, largely self-administered, clinician-supervised exposure and response prevention protocol using a verbal termination cue in two patients with severe obsessive-compulsive disorder
(2026) Mari, Mohammad; Stingl, Markus; Hanewald, Bernd; Mulert, Christoph
Background/Objectives: Obsessive–compulsive disorder (OCD) is a chronic and disabling condition. Despite first-line treatment with cognitive behavioral therapy (CBT) including exposure and response prevention (ERP) and selective serotonin reuptake inhibitors (SSRIs), many patients remain severely symptomatic. This highlights the need for further research into innovative multimodal treatment strategies for OCD. We report two inpatients with severe OCD, comorbid depressive symptoms, and insufficient response to previous pharmacological and psychotherapeutic treatments. Methods: Both patients completed 30 multimodal treatment units over six weeks consisting of sequential multi-target repetitive transcranial magnetic stimulation (rTMS) targeting the left dorsolateral prefrontal cortex (DLPFC), right DLPFC, and supplementary motor area (SMA), combined with a modified, largely self-administered, clinician-supervised exposure and response prevention protocol. Results: Both patients demonstrated large observed reductions in OCD symptom severity. Y-BOCS total scores decreased from 29 to 11 in Patient 1 and from 29 to 10 in Patient 2, corresponding to reductions of 62.1% and 65.5%, respectively, with improvements apparent after approximately four weeks and largely maintained at follow-up. Depressive symptoms also improved, and no clinically relevant adverse effects were observed. Conclusions: These cases provide an initial description of the feasibility and tolerability of this specific protocol architecture but do not establish efficacy, long-term durability, or the contribution of individual components. The protocol-level novelty lies in integrating a repeated three-target TBS schedule with a response-limited, cue-terminated exposure procedure, a combination not previously described in the OCD neuromodulation literature.
The patient journey in interstitial lung disease : mobility, independence, and psychological burden
(2025) Krauss, Ekaterina; Tello, Silke; Kuhlewey, Daniel; Mahavadi, Poornima; Scharmer, Claudia; Behr, Juergen; Guenther, Andreas; Huss, Gottfried
Background: Interstitial lung diseases (ILDs) profoundly affect daily life, limiting mobility, independence, and emotional stability. While antifibrotic therapies may slow physiological decline, the living experience—characterized by breathlessness, cough, frailty, and psychological distress—remains insufficiently understood; this study therefore aimed to capture real-world patient perspectives on functional capacity, self-management, and mental health to identify treatable traits beyond conventional physiological measures. Materials and Methods: A cross-sectional quantitative online survey was conducted between September 2024 and January 2025 by Lungenfibrose e.V. in collaboration with the Center for Interstitial and Rare Lung Diseases (ZISL), Universities of Giessen and Marburg Lung Center (Giessen site). Patients with physician-confirmed ILD completed standardized instruments assessing dyspnea (MRC), cough intensity (VAS-Cough), frailty (CFS), and health-related quality of life (EQ-5D-5L). Data were analyzed descriptively across physical, functional, and psychosocial domains. Results: The majority of 69 respondents had idiopathic pulmonary fibrosis (64.7%) with a mean diagnostic delay of 1.4 ± 2.2 years; 69% were diagnosed within two years of symptom onset, and 77% were receiving antifibrotic therapy (nintedanib 57%, pirfenidone 19%). Functional limitations were substantial—55% were mobile for fewer than two hours per day, 73% reported mobility impairment, and oxygen use was common (51% during exertion, 26% at rest). Frailty increased over time (mean CFS 3.2 → 3.8), with 46% classified as fit, 36% vulnerable, and 18% frail. Dyspnea and cough remained burdensome (mean VAS-cough 40 ± 26; 58% moderate–severe), and health-related quality of life was reduced (mean EQ-VAS 56.5 ± 23.7), with high rates of anxiety/depression (78%), limitations in daily activities (76%), and pain/discomfort (74%). Despite overall satisfaction with care (mean 7.1 ± 2.5), respondents frequently reported unmet needs for psychological support and clearer communication about treatment and disease management. Conclusions: Despite antifibrotic therapy and structured specialist care, individuals living with ILD continue to face substantial physical and emotional challenges. Treatable traits—including frailty, dyspnea, inactivity, anxiety, and social isolation—emerge as key determinants of well-being. Multidisciplinary strategies integrating rehabilitation, psychosocial support, and patient education alongside pharmacological therapy are essential to preserve autonomy and improve quality of life in pulmonary fibrosis.