Investigations have been carried out on rats to obtain information about the selenium-containing proteins present in the microsomal
fraction and especially in the endoplasmatic reticulum (ER) of the kidney. For the determination of the selenium levels instrumental
neutron activation analysis via 75Se was used. After labeling of rats in vivo with 75Se-selenite and separation of the proteins in the renal homogenate and cell compartments by electrophoretic methods, the 75Se-containing proteins were detected by autoradiography. In this way, six selenium-containing proteins with molecular masses
of 15, 16, 20, 23-25, 40-42 and 58-60 were found in the endoplasmatic reticulum. All of those were characterized as selenocysteine-containing
Caveolin-3 (cav-3) mutations are linked to the long-QT syndrome (LQTS) causing distinct clinical symptoms. Hyperpolarization-activated cyclic nucleotide channel 4 (HCN4) underlies the pacemaker current If. It associates with cav-3 and both form a macromolecular complex.
To examine the effects of human LQTS-associated cav-3 mutations on HCN4-channel function, HEK293-cells were cotransfected with HCN4 and wild-type (WT) cav-3 or a LQTS-associated cav-3 mutant (T78M, A85T, S141R, or F97C). HCN4 currents were recorded using the whole-cell patch-clamp technique.
WT cav-3 significantly decreased HCN4 current density and shifted midpoint of activation into negative direction. HCN4 current properties were differentially modulated by LQTS-associated cav-3 mutations. When compared with WT cav-3, A85T, F97C, and T78M did not alter the specific effect of cav-3, but S141R significantly increased HCN4 current density. Compared with WT cav-3, no significant modifications of voltage dependence of steady-state activation curves were observed. However, while WT cav-3 alone had no significant effect on HCN4 current activation, all LQTS-associated cav-3 mutations significantly accelerated HCN4 activation kinetics.
Our results indicate that HCN4 channel function is modulated by cav-3. LQTS-associated mutations of cav-3 differentially influence pacemaker current properties indicating a pathophysiological role in clinical manifestations.
Rapid detection of methicillin-resistant Staphylococcus aureus (MRSA) colonization status facilitates isolation and decolonization and reduces MRSA infections. Liquid but not dry swabs allow fully automated detection methods. However, the accuracy of culture and polymerase chain reaction (PCR) using liquid and dry swabs has not been analyzed. We compared different swab collection systems for routine nasal–throat MRSA screening in patients admitted to a tertiary care trauma center in Germany. Over 3 consecutive months, dry swabs (month 1), ESwabs (month 2), or MSwabs (month 3) were processed using Cepheid GeneXpert, Roche cobas and BD-MAX™ MRSA tests compared to chromogenic culture. Among 1680 subjects, the MRSA detection rate using PCR methods did not differ significantly between dry swabs, ESwab, and MSwab (6.0%, 6.2%, and 5.3%, respectively). Detection rates using chromogenic culture were 2.9%, 3.9%, and 1.9%, using dry, ESwab, and MSwab, respectively. Using chromogenic culture as the “gold standard”, negative predictive values for the PCR tests ranged from 99.2–100%, and positive predictive values from 33.3–54.8%. Thus, efficient and accurate MRSA screening can be achieved using dry, as well as liquid E- or MSwab, collection systems. Specimen collection using ESwab or MSwab facilitates efficient processing for chromogenic culture in full laboratory automation while also allowing molecular testing in automated PCR systems.