Liquid profiling of circulating tumor DNA in colorectal cancer: steps needed to achieve its full clinical value as standard care

The analysis of circulating tumor DNA (ctDNA) is at the threshold of implementation into standard care for colorectal cancer (CRC) patients. However, data about the clinical utility of liquid profiling (LP), its acceptance by clinicians, and its integration into clinical workflows in real‐world settings remain limited. Here, LP tests requested as part of routine care since 2016 were retrospectively evaluated. Results show restrained request behavior that improved moderately over time, as well as reliable diagnostic performance comparable to translational studies, with an overall agreement of 91.7%. Extremely low ctDNA levels at < 0.1% in over 20% of cases, a high frequency of concomitant driver mutations (in up to 14% of cases), and ctDNA levels reflecting the clinical course of disease were revealed. However, certain limitations hampering successful translation of ctDNA into clinical practice were uncovered, including the lack of clinically relevant ctDNA thresholds, appropriate time points of LP requests, and integrative evaluation of ctDNA, imaging, and clinical findings. In conclusion, these results highlight the potential clinical value of LP for CRC patient management and demonstrate issues that need to be addressed for successful long‐term implementation in clinical workflows.     

Riluzole therapy in cervical dystonia.

We conducted a 6-week open-label pilot study with blinded video rating of riluzole (50 mg twice a day) in six patients with cervical dystonia (CD) refractory to botulinum toxin A and oral pharmacological treatment. The Tsui rating scale served as primary efficacy measure and improved significantly under riluzole (P = 0.002). In three of six patients, the Tsui score improved by more than 30% with a greater 50% reduction in the head tremor/jerk subscore of the Tsui scale. These data suggest that riluzole may be helpful in a subgroup of patients with disabling CD refractory to other therapies.     

VerstÄrkung der radiotherapeutischen wirkung auf HeLa-Zellen durch gemcitabine

BACKGROUND: Gemcitabine (2'.2'-difluorodeoxycytidine; dFdC) is a new nucleoside analog with promising activity in different solid tumors in vivo and in vitro. As published up to now, combined with irradiation dFdC demonstrates a radiosensitizing effect on pancreas and colon carcinoma cell lines. We investigated the influence of dFdC on the radiosensitization of human squamous carcinoma cells of the cervix (HeLa-cells, ATCC CCL-2). MATERIAL AND METHODS: Under standardized conditions monolayer cultures of HeLa-cells were incubated in medium with dFdC for different times (4 to 24 hours) and exposed to different concentrations (0.003, 0.01 and 0.03 mumol/l). Irradiation (2 to 6 Gy, electron beam) followed immediately or 12 hours after dFdC-exposure. Cell survival was determined by colony forming assay. Using the linear-quadratic model cell survival curves were fit after correction for drug-induced cytotoxicity and the mean inactivation dose (MID) was calculated. Radiation enhancement was defined as the ratio MIDRT(= Control)/MIDRT + dFdC > 1. RESULTS: Exposed to gemcitabine for 4 and 8 hours and followed by immediate irradiation the radiation enhancement ratio (Table 1) is 1.07 to 1.14 and 1.04 to 1.22, respectively, if dFdC concentration is > or = 0.01 to 0.03 mumol/l. Further increase of the irradiation effect is demonstrated in cells exposed to > or = 0.003 to 0.03 mumol/l dFdC for 16 and 24 hours (radiation enhancement ratio 1.08 to 2.0 and 1.08 to 2.48, respectively) (Figure 3). If irradiation is applied 12 hours after 24-hour-exposure (0.01 and 0.03 mumol/l) the enhancement ratio was 1.18 and 1.7, respectively (Figure 4). CONCLUSIONS: In cell cultures the assays combining irradiation with dFdC demonstrate that dFdC is a potent radiation sensitizer of HeLa-cells. The effect of irradiation on cells pre-treated with non- and hardly cytotoxic concentrations of dFdC is increased in dependence of dose and time of exposure.     

The preparation of clinical grade 5-[123I]iodo-2'-deoxyuridine and 5-[125I]iodo-2'-deoxyuridine with high in vitro stability and the potential for early proliferation scintigraphy

BACKGROUND AND METHODS: 5-Iodo-2'-deoxyuridine (IdUrd) radiolabelled with the positron emitter I or with the gamma and Auger electron emitters I or I has been proposed for cancer diagnosis and therapy. We modified the synthesis to reliably obtain [I]IdUrd and [I]IdUrd by using an Iodogen supported destannylation reaction of 5-(tri-n-butylstannyl)-2'-deoxyuridine (Bu3SndUrd) which meets the requirements for good laboratory practice (GLP) and good clinical practice (GCP). A method of purification was developed to eliminate by-products as well as any unreacted starting material. RESULTS: [I]IdUrd, which originated from a trace of iodide in the Bu3SndUrd precursor, was identified as the unknown by-product reported for this method. This trace could be eliminated by modified purification of Bu3SndUrd. Stabilization of pH was essential for unequivocal identification of radiolabelled IdUrd and possible degradation products in the different systems tested for quality control. Biodistribution in tumour bearing nude mice was measured as early as 3 and 6 h after i.v. injection of [I]IdUrd. This compound showed high and specific activity uptake in tumour and dividing tissues when combined with 5-fluoro-2'-deoxyuridine pre-treatment. Uptake was specifically inhibited by injection of excess thymidine.     

Emergence of Usutu virus,an African mosquito-borne flavivirus of the Japanese encephalitis virus group,central Europe

During late summer 2001 in Austria, a series of deaths in several species of birds occurred, similar to the beginning of the West Nile virus (WNV) epidemic in the United States. We necropsied the dead birds and examined them by various methods; pathologic and immunohistologic investigations suggested a WNV infection. Subsequently, the virus was isolated, identified, partially sequenced, and subjected to phylogenetic analysis. The isolates exhibited 97% identity to Usutu virus (USUV), a mosquito-borne Flavivirus of the Japanese encephalitis virus group; USUV has never previously been observed outside Africa nor associated with fatal disease in animals or humans. If established in central Europe, this virus may have considerable effects on avian populations; whether USUV has the potential to cause severe human disease is unknown.     

Toilet training in individuals with Angelman syndrome: A case series

Objective: To assess if adapted versions of the response restriction toilet training protocol, based on the behavioral phenotype of Angelman syndrome (AS), were successful in fostering urinary continence in seven individuals with AS.

Method: Data were collected in AB-designs during baseline, training, generalization and follow-up. The response restriction protocol was adapted: individuals were trained in their natural environment, were prompted to void and along with improving continence, the interval between voids was prolonged and time-on-toilet decreased.

Results: During generalization five individuals had less than two accidents and one to six correct voids per day; during baseline more accidents and/or less correct voids occurred. In two participants correct voids increased, but several accidents still occurred. Three participants maintained positive results after 3–18 months.

Conclusion: Despite their intellectual and behavioral challenges, urinary continence can be acquired in AS. Several indications of voiding dysfunctions were found; further research is indicated.