A maximum auditory steady-state response (ASSR) amplitude is yielded when the ASSR is elicited by an amplitude-modulated tone (f(c)) with a fixed modulation frequency (f(m) = 40 Hz), whereas the maximum distortion product otoacoustic emission (DPOAE) level is yielded when the DPOAE is elicited using a fixed frequency ratio of the primary tones (f2/f1 = 1.2). When eliciting the DPOAE and ASSR by the same tone pair, optimal stimulation is present for either DPOAE or ASSR and thus adequate simultaneous DPOAE/ASSR measurement is not possible across test frequency f2 or f(c), respectively. The purpose of the present study was to determine whether the ASSR and DPOAE can be measured simultaneously without notable restrictions using a DPOAE stimulus setting in which one primary tone is amplitude modulated. A DPOAE of frequency 2f1-f2 and ASSR of modulation frequency 41 Hz were measured in ten normal hearing subjects at a test frequency between 0.5 and 8 kHz (f2 = f(c)). The decrease in the DPOAE level and the loss in ASSR amplitude during hybrid mode stimulation amounted, on average, to only 2.60 dB [standard deviation (SD) = 1.38 dB] and 1.83 dB (SD = 2.38 dB), respectively. These findings suggest simultaneous DPOAE and ASSR measurements to be feasible across all test frequencies when using a DPOAE stimulus setting where the primary tone f2 is amplitude modulated.

Patients with cancer frequently receive immune-checkpoint inhibitors (ICIs), which may modulate immune responses to COVID-19 vaccines. Recently, cytokine release syndrome (CRS) was observed in a patient with cancer who received BTN162b2 vaccination under ICI treatment. Here, we analyzed adverse events and serum cytokines in patients with 23 different tumors undergoing (n = 64) or not undergoing (n = 26) COVID-19 vaccination under ICI therapy in a prospectively planned German single-center cohort study (n = 220). We did not observe clinically relevant CRS (≥grade 2) after vaccination (95% CI 0–5.6%; Common Terminology of Adverse Events v.5.0) in this small cohort. Within 4 weeks after vaccination, serious adverse events occurred in eight patients (12.5% 95% CI 5.6–23%), six patients were hospitalized and two patients died. Despite absence of CRS symptoms, a set of pairwise-correlated CRS-associated cytokines, including CXCL8 and interleukin-6 was >1.5-fold upregulated in 40% (95% CI 23.9–57.9%) of patients after vaccination. Hence, elevated cytokine levels are common and not sufficient to establish CRS diagnosis.

In research models of liver cancer, regeneration, inflammation, and fibrosis, flexible systems for in vivo gene expression and silencing are highly useful. Hydrodynamic tail vein injection of transposon-based constructs is an efficient method for genetic manipulation of hepatocytes in adult mice. In addition to constitutive transgene expression, this system can be used for more advanced applications, such as shRNA-mediated gene knock-down, implication of the CRISPR/Cas9 system to induce gene mutations, or inducible systems. Here, the combination of constitutive CreER expression together with inducible expression of a transgene or miR-shRNA of choice is presented as an example of this technique. We cover the multi-step procedure starting from the preparation of sleeping beauty-transposon constructs, to the injection and treatment of mice, and the preparation of liver tissue for analysis by immunostaining. The system presented is a reliable and efficient approach to achieve complex genetic manipulations in hepatocytes. It is specifically useful in combination with Cre/loxP-based mouse strains and can be applied to a variety of models in the research of liver disease.