ABSTRACT
This review is focused on methods for detecting small molecules and, in particular, the characterisation of their interaction with natural proteins (e.g. receptors, ion channels). Because there are intrinsic advantages to using label-free methods over labelled methods (e.g. fluorescence, radioactivity), this review only covers label-free techniques. We briefly discuss available techniques and their advantages and disadvantages, especially as related to investigating the interaction between small molecules and proteins. The reviewed techniques include well-known and widely used standard analytical methods (e.g. HPLC-MS, NMR, calorimetry, and X-ray diffraction), newer and more specialised analytical methods (e.g. biosensors), biological systems (e.g. cell lines and animal models), and in-silico approaches.
Subject(s)
Proteins/chemistry , Animals , Biosensing Techniques , Chromatography, Liquid , Humans , Ligands , Mass Spectrometry , Protein Binding , X-Ray DiffractionABSTRACT
Although surgical division of the transverse carpal ligament is the operative treatment of choice for carpal tunnel syndrome (CTS), controversy exists about the immediate postoperative treatment regimen. Splinting for up to 6 weeks after surgery is recommended by some investigators. We therefore evaluated effectiveness of splinting after open carpal tunnel surgery by a randomized, controlled trial. Fifty consecutive patients with clinically and electrophysiologically confirmed idiopathic CTS were assigned to open carpal tunnel release and randomized to receiving a light bandage (25 patients) or a bulky dressing with a volar splint (25 patients) for 2 days each. All patients were followed up at 3 months. Parameters retrieved were pain as measured using a visual analog scale, two-point discrimination, and grip strength, and nerve conduction studies. At follow-up, all patients reported definite improvement of symptoms, but there was no statistically significant difference between the two groups for any of our outcome measures. Thus, postoperative splinting after open carpal tunnel release does not yield any benefit to eventual outcome. In fact, it adds to the overall operating time and can safely be abandoned.
Subject(s)
Carpal Tunnel Syndrome/surgery , Decompression, Surgical/methods , Postoperative Period , Splints , Carpal Tunnel Syndrome/physiopathology , Follow-Up Studies , Humans , Neurologic Examination/methods , Pain Measurement/methods , Retrospective Studies , Treatment OutcomeABSTRACT
T-cell-mediated loss of pancreatic beta-cells is the crucial event in the development of type 1 diabetes. The phenotypic characteristics of disease-associated T-cells in type 1 diabetes have not yet been defined. The negative results from two intervention trials (the Diabetes Prevention Trial-Type 1 Diabetes and the European Nicotinamide Diabetes Intervention Trial) illustrate the need for technologies to specifically monitor ongoing autoimmune reactions. We used fluorescence-activated cell sorter analysis to study surface marker expression on T-cell lines specific for two major type 1 diabetes autoantigens, GAD65 and proinsulin. We then applied this knowledge in a cross-sectional approach to delineate the phenotype of circulating memory T-cells. The autoreactive T-cells of patients could be distinguished from those of control subjects by their coexpression of CD25 and CD134. Autoantigen-specific T-cells that recognized multiple GAD65- and preproinsulin-derived peptides and coexpressed CD25(+)CD134(+) were confined to patients (n = 32) and pre-diabetic probands (n = 5). Autoantigen-reactive T-cells in control subjects (n = 21) were CD25(+)CD134(-) and recognized fewer autoantigen-derived peptides. Insulin therapy did not induce CD25(+)CD134(+) T-cells in type 2 diabetic patients. The coexpression of CD25 and the costimulatory molecule CD134 on memory T-cells provides a novel marker for type 1 diabetes-associated T-cell immunity. The CD134 costimulatory molecule may also provide a novel therapeutic target in type 1 diabetes.
