Advances in clinical chemistry patient-based real-time quality control (PBRTQC)

Patient-Based Real-Time Quality Control involves monitoring an assay using patient samples rather than external material. If the patient population does not change, then a shift in the long-term assay population results represents the introduction of a change in the assay. The advantages of this approach are that the sample(s) are commutable, it is inexpensive, the rules are simple to interpret and there is virtually continuous monitoring of the assay. The disadvantages are that the laboratory needs to understand their patient population and how they may change during the day, week or year and the initial change of mindset required to adopt the system. The concept is not new, having been used since the 1960s and widely adopted on hematology analyzers in the mid-1970s. It was not widely used in clinical chemistry as there were other stable quality control materials available. However, the limitations of conventional quality control approaches have become more evident. There is a greater understanding of how to collect and use patient data in real time and a range of powerful algorithms which can identify changes in assays. There are more assays on more samples being run. There is also a greater interest in providing a theoretical basis for the validation and integration of these techniques into routine practice.No Full Tex

Patient-Based Real Time QC

Patient-Based Real-Time Quality Control (PBRTQC) uses the statistical characteristics of a particular patient population(s) served by a laboratory using specific analytical platforms. There is a review of this approach in the August 2019 issue of Clinical Chemistry. The power of these techniques is that they offer exquisite customization to provide very sensitive detection of a change in bias. They are not subject to the impact of noncommutability issues and, once set up, are low cost to maintain. However, they do require knowledge of the characteristics of the laboratory patient population(s) and the analytical methods used. PBRTQC needs to be tailored for each measurand in a population, though this is certainly not a major limitation. Optimization requires access to simulation software and patient data from the Laboratory Information System.No Full Tex

Inhibition of Dopamine Transporter sctivity by G protein beta gamma subunits

Uptake through the Dopamine Transporter (DAT) is the primary mechanism of terminating dopamine signaling within the brain, thus playing an essential role in neuronal homeostasis. Deregulation of DAT function has been linked to several neurological and psychiatric disorders including ADHD, schizophrenia, Parkinson’s disease, and drug addiction. Over the last 15 years, several studies have revealed a plethora of mechanisms influencing the activity and cellular distribution of DAT; suggesting that fine-tuning of dopamine homeostasis occurs via an elaborate interplay of multiple pathways. Here, we show for the first time that the βγ subunits of G proteins regulate DAT activity. In heterologous cells and brain tissue, a physical association between Gβγ subunits and DAT was demonstrated by co-immunoprecipitation. Furthermore, in vitro pull-down assays using purified proteins established that this association occurs via a direct interaction between the intracellular carboxy-terminus of DAT and Gβγ. Functional assays performed in the presence of the non-hydrolyzable GTP analog GTP-γ-S, Gβγ subunit overexpression, or the Gβγ activator mSIRK all resulted in rapid inhibition of DAT activity in heterologous systems. Gβγ activation by mSIRK also inhibited dopamine uptake in brain synaptosomes and dopamine clearance from mouse striatum as measured by high-speed chronoamperometry in vivo. Gβγ subunits are intracellular signaling molecules that regulate a multitude of physiological processes through interactions with enzymes and ion channels. Our findings add neurotransmitter transporters to the growing list of molecules regulated by G-proteins and suggest a novel role for Gβγ signaling in the control of dopamine homeostasis.Jennie Garcia-Olivares, Delany Torres-Salazar, William A. Owens, Tracy Baust, David P. Siderovski, Susan G. Amara, Jun Zhu, Lynette C. Daws, Gonzalo E. Torre