Reliability of the Clinical Application of a Mechanical Inclinometer in Measuring Glenohumeral Motion

Objective: Establish intra- and inter-examiner reliability of glenohumeral range of motion (ROM) measures taken by a single-clinician using a mechanical inclinometer. Design: A single-session, repeated-measure, randomized, counterbalanced design. Setting: Athletic Training laboratory. Participants: Ten college-aged volunteers (9 right-hand dominant; 4 males, 6 females; age=23.2±2.4y, mass=73±16kg, height=170±8cm) without shoulder or neck injuries within one year. Interventions: Two Certified Athletic Trainers separately assessed passive glenohumeral (GH) internal (IR) and external (ER) rotation bilaterally. Each clinician secured the inclinometer to each subject’s distal forearm using elastic straps. Clinicians followed standard procedures for assessing ROM, with the participants supine on a standard treatment table with 90° of elbow flexion. A second investigator recorded the angle. Clinicians measured all shoulders once to assess inter-clinician reliability and eight shoulders twice to assess intra-clinician reliability. We used SPSS 14.0 (SPSS Inc., Chicago, IL) to calculate standard error of measure (SEM) and Intraclass Correlation Coefficients (ICC) to evaluate intra- and inter-clinician reliability. Main Outcome Measures: Dependent variables were degrees of IR, ER, glenohumeral internal rotation deficit (GIRD) and total arc of rotation. We calculated GIRD as the bilateral difference in IR (nondominant–dominant) and total arc for each shoulder (IR+ER). Results: Intra-clinician reliability for each examiner was excellent (ICC[1,1] range=0.90-0.96; SEM=2.2°-2.5°) for all measures. Examiners displayed excellent inter-clinician reliability (ICC[2,1] range=0.79-0.97; SEM=1.7°-3.0°) for all measures except nondominant IR which had good reliability(0.72). Conclusions: Results suggest that clinicians can achieve reliable measures of GH rotation and GIRD using a single-clinician technique and an inexpensive, readily available mechanical inclinometer.

Specific binding of the mammalian high mobility group protein AT-hook 2 to the minor groove of AT -rich DNAs: Thermodynamic and specificity studies

The mammalian high mobility group protein AT-hook 2 (HMGA2) is a small transcriptional factor involved in cell development and oncogenesis. It contains three "AT-hook" DNA binding domains, which specifically recognize the minor groove of AT-rich DNA sequences. It also has an acidic C-terminal motif. Previous studies showed that HMGA2 mediates all its biological effects through interactions with AT-rich DNA sequences in the promoter regions. In this dissertation, I used a variety of biochemical and biophysical methods to examine the physical properties of HMGA2 and to further investigate HMGA2's interactions with AT-rich DNA sequences. The following are three avenues perused in this study: (1) due to the asymmetrical charge distribution of HMGA2, I have developed a rapid procedure to purify HMGA2 in the milligram range. Preparation of large amounts of HMGA2 makes biophysical studies possible; (2) Since HMGA2 binds to different AT-rich sequences in the promoter regions, I used a combination of isothermal titration calorimetry (ITC) and DNA UV melting experiment to characterize interactions of HMGA2 with poly(dA-dT) 2 and poly(dA)poly(dT). My results demonstrated that (i) each HMGA2 molecule binds to 15 AT bp; (ii) HMGA2 binds to both AT DNAs with very high affinity. However, the binding reaction of HMGA2 to poly(dA-dT) 2 is enthalpy-driven and the binding reaction of HMGA2 with poly(dA)poly(dT) is entropy-driven; (iii) the binding reactions are strongly depended on salt concentrations; (3) Previous studies showed that HMGA2 may have sequence specificity. In this study, I used a PCR-based SELEX procedure to examine the DNA binding specificity of HMGA2. Two consensus sequences for HMGA2 have been identified: 5'-ATATTCGCGAWWATT-3' and 5'-ATATTGCGCAWWATT-3', where W represents A or T. These consensus sequences have a unique feature: the first five base pairs are AT-rich, the middle four to five base pairs are GC-rich, and the last five to six base pairs are AT-rich. All three segments are critical for high affinity binding. Replacing either one of the AT-rich sequences to a non-AT-rich sequence causes at least 100-fold decrease in the binding affinity. Intriguingly, if the GC-segment is substituted by an AT-rich segment, the binding affinity of HMGA2 is reduced approximately 5-fold. Identification of the consensus sequences for HMGA2 represents an important step towards finding its binding sites within the genome.