Miniaturized ultra high field asymmetric waveform ion mobility spectrometry combined with mass spectrometry for peptide analysis.

Miniaturized ultra high field asymmetric waveform ion mobility spectrometry (ultra-FAIMS) combined with mass spectrometry (MS) has been applied to the analysis of standard and tryptic peptides, derived from α-1-acid glycoprotein, using electrospray and nanoelectrospray ion sources. Singly and multiply charged peptide ions were separated in the gas phase using ultra-FAIMS and detected by ion trap and time-of-flight MS. The small compensation voltage (CV) window for the transmission of singly charged ions demonstrates the ability of ultra-FAIMS-MS to generate pseudo-peptide mass fingerprints that may be used to simplify spectra and identify proteins by database searching. Multiply charged ions required a higher CV for transmission, and ions with different amino acid sequences may be separated on the basis of their differential ion mobility. A partial separation of conformers was also observed for the doubly charged ion of bradykinin. Selection on the basis of charge state and differential mobility prior to tandem mass spectrometry facilitates peptide and protein identification by allowing precursor ions to be identified with greater selectivity, thus reducing spectral complexity and enhancing MS detection.

Cruciate ligament forces in the human knee during rehabilitation exercises.

OBJECTIVE: To determine the cruciate ligament forces occurring during typical rehabilitation exercises.Design. A combination of non-invasive measurements with mathematical modelling of the lower limb.Background. Direct measurement of ligament forces has not yet been successful in vivo in humans. A promising alternative is to calculate the forces mathematically. METHODS: Sixteen subjects performed isometric and isokinetic or squat exercises while the external forces and limb kinematics were measured. Internal forces were calculated using a geometrical model of the lower limb and the "dynamically determinate one-sided constraint" analysis procedure. RESULTS: During isokinetic/isometric extension, peak anterior cruciate ligament forces, occurring at knee angles of 35-40 degrees, may reach 0.55x body-weight. Peak posterior cruciate ligament forces are lower and occur around 90 degrees. During isokinetic/isometric flexion, peak posterior cruciate forces, which occur around 90 degrees, may exceed 4x body-weight; the anterior cruciate is not loaded. During squats, the anterior cruciate is lightly loaded at knee angles up to 50 degrees, after which the posterior cruciate is loaded. Peak posterior cruciate forces occur near the lowest point of the squat and may reach 3.5x body-weight. CONCLUSIONS: For anterior cruciate injuries, squats should be safer than isokinetic or isometric extension for quadriceps strengthening, though isokinetic or isometric flexion may safely be used for hamstrings strengthening. For posterior cruciate injuries, isokinetic extension at knee angles less than 70 degrees should be safe but isokinetic flexion and deep squats should be avoided until healing is well-advanced. RELEVANCE: Good rehabilitation is vital for a successful outcome to cruciate ligament injuries. Knowledge of ligament forces can aid the physician in the design of improved rehabilitation protocols.

Parameter sensitivity of a mathematical model of the anterior cruciate ligament.

This paper presents the results of an investigation into parameter sensitivity of a mathematical model of the human anterior cruciate ligament (ACL). The model ACL comprised a continuous array of fibres mapped between part-elliptical attachment areas on the femur and tibia. Relative motion of the two bones was controlled by a planar four-bar linkage. Parameter modifications were: (a) an alternative set of values for the coordinates of the four-bar linkage joints; (b) rotation of the attachment areas of the ligament by +/- 30 degrees; and (c) variation of some mechanical properties. The alternative four-bar linkage parameter set produced extremely large changes in ACL force values, up to 130 per cent. Rotating the tibial attachment changed forces by less than 20 per cent, whereas rotating the femoral attachment changed forces by up to 100 per cent. Altering the mechanical parameters produced the smallest differences in force, under 15 per cent. These results highlight the importance, when using a theoretical model, of establishing the values of the parameters defining the model as accurately as possible and of carrying out a parameter sensitivity study. From a clinical viewpoint, they also suggest that, when reconstructing a ruptured ACL, one of the most important considerations must be to position the femoral attachment of the graft as accurately as is feasible.