Real-Time Cellular Thermal Shift Assay to Monitor Target Engagement.

Determining a molecule's mechanism of action is paramount during chemical probe development and drug discovery. The cellular thermal shift assay (CETSA) is a valuable tool to confirm target engagement in cells for a small molecule that demonstrates a pharmacological effect. CETSA directly detects biophysical interactions between ligands and protein targets, which can alter a protein's unfolding and aggregation properties in response to thermal challenge.

High-Throughput Detection of Ligand-Protein Binding Using a SplitLuc Cellular Thermal Shift Assay.

The confirmation of a small molecule binding to a protein target can be challenging when switching from biochemical assays to physiologically relevant cellular models. The cellular thermal shift assay (CETSA) is an approach to validate ligand-protein binding in a cellular environment by examining a protein's melting profile which can shift to a higher or lower temperature when bound by a small molecule. Traditional CETSA uses SDS-PAGE and Western blotting to quantify protein levels, a process that is both time consuming and low-throughput when screening multiple compounds and concentrations.

A widely-applicable high-throughput cellular thermal shift assay (CETSA) using split Nano Luciferase.

Assessment of the interactions between a drug and its protein target in a physiologically relevant cellular environment constitutes a major challenge in the pre-clinical drug discovery space. The Cellular Thermal Shift Assay (CETSA) enables such an assessment by quantifying the changes in the thermal stability of proteins upon ligand binding in intact cells. Here, we present the development and validation of a homogeneous, standardized, target-independent, and high-throughput (384- and 1536-well formats) CETSA platform that uses a split Nano Luciferase approach (SplitLuc CETSA).

Acquired platelet dysfunction may be an aetiologic factor in Heyde's syndrome--normalization of bleeding time after aortic valve replacement.

Background: There is a well-documented relationship, with unknown aetiology, between aortic valve stenosis and occult gastrointestinal bleeding in elderly patients. Despite several studies attempting to determine the prevalence and to discuss the aetiology, there are still many unanswered questions.

Methods: A total of 288 consecutive patients with valvular aortic stenosis--mean age 73 +/- 9 years aortic stenosis group (ASG)--were compared with 129 pacemaker-treated patients, mean age 73 +/- 9 years control group (CG).

Later potentials on signal-averaged electrocardiograms in children after right ventriculotomy.

After corrective surgery for congenital heart defects, scars may create fractionation and delay of the electrical signals in the heart muscle, providing a substrate for arrhythmias. Signal-averaged electrocardiograms (SAECGs) were obtained from 33 children after right ventriculotomy, on average 6 years after surgery, and from 38 healthy controls of the same age. The duration of the filtered QRS complex (fQRS), the duration of the low amplitude signal (< 40 microV) in the terminal QRS complex (LAS40), and the root mean square amplitude of the terminal 40 ms of the QRS complex (RMS40) were determined.

Isolated congenital heart block. Long-term outcome of mothers and characterization of the immune response to SS-A/Ro and to SS-B/La.

Objective: To study the long-term outcome of mothers of children with isolated congenital heart block (CHB) and to characterize the maternal autoantibody response to SS-A/Ro and SS-B/La.

Methods: A retrospective clinical study of 33 mothers a mean of 11.2 years (SD 9.

Value of absent or retrograde end-diastolic flow in fetal aorta and umbilical artery as a predictor of perinatal outcome in pregnancy-induced hypertension.

There is insufficient data on the value of absent or retrograde end-diastolic flow (AREDF) in the fetal umbilical artery, descending aorta and aortic arch to predict perinatal outcome. In this prospective investigation, 65 pregnant women between 24 and 34 weeks' gestation with pregnancy-induced hypertension were studied by color Doppler echocardiography. Pregnancies leading to birth at or before 34.

Isolated congenital heart block: fetal and infant outcome and familial incidence of heart block.

Objective: To determine the 1-year outcome of infants with isolated congenital heart block, the risk of fetal loss in mothers of affected infants, and the risk of recurrence of congenital heart block.

Methods: The outcomes of 34 infants with isolated congenital heart block and of the 109 pregnancies in the 32 mothers of these infants were analyzed retrospectively. A control group consisted of 170 pregnancies in 64 women individually matched for age, parity, and socioeconomic status.

Atrial natriuretic peptide and atrial pressures in newborns with transposition of the great arteries.

The relation of atrial pressure to atrial plasma concentration of atrial natriuretic peptide (ANP) was studied in four newborns undergoing catheterization because of transposition of the great arteries. In three patients, mean left atrial pressure clearly exceeded right atrial pressure (12 +/- 7 vs. 2 +/- 3 mmHg; mean +/- SD).

Fibrosarcoma in a girl with celiac disease and IgA deficiency.

A case of abdominal wall fibrosarcoma in a 17-year-old girl with celiac disease and IgA deficiency is described. Celiac disease was putatively diagnosed with intestinal biopsy at the age of 15 years when she came for hospital investigations because of IgA deficiency and recurrent respiratory infections. The tumor occurred at the age of 17 years during the gluten challenge performed for final diagnosis of celiac disease.

Extracardiac malformations associated with congenital heart disease.

The incidence of significant extracardiac malformations was determined in a combined clinical and autopsy study comprising 1000 infants and children with congenital heart disease treated and lost at the Children's Hospital in Helsinki. There were 567 boys and 433 girls. 1/4 of the children had a birthweight of 2500 g or less.

Effect of seasonal variation in daylight on bilirubin level in premature infants.

The effect of daylight on the serum bilirubin level of preterm infants (birthweight less than 2500 g) during the first 10 days of life was studied in a district of Finland (Oulu, latitude 65 degrees ĺ) with large seasonal variations in the length of daylight (range 3 to 22 h). 86 preterm infants born consecutively during one calendar year were studied in incubators where they were completely exposed except for nappies. A significantly lower bilirubin value from the fifth day of life onwards was recorded ih the group of infants born during the light half of the year, compared with the infants born during the dark half of the year.