3,3',5-Triiodothyronine (T3) is an important diagnostic marker for thyroid function. A reference measurement procedure (RMP) for total T3 in serum involving isotope dilution coupled with liquid chromatography-tandem mass spectrometry (LC/MS/MS) has been developed and critically evaluated. The method uses solid-phase extraction with mixed-mode retention mechanisms of reversed phase and ion exchange prior to reversed-phase LC/MS/MS. In addition to a labeled T3 internal standard (T3-13C9), labeled thyroxine (T4-d5) is also added to serum samples in order to monitor the degradation of T4 to T3. The accuracy of the measurement was evaluated by a recovery study for added T3 and was supported by a comparison study with the other RMP. The recovery of the added T3 ranged from 98.9% to 99.4%. The results of this method and the other RMP agreed to within 1%. Samples of frozen serum pools were prepared and measured in three separate sets. Excellent reproducibility was obtained with within-set coefficients of variation (CVs) ranging from 0.8% to 1.6% and between-set CVs ranging from 1.9% to 2.6%. Excellent linearity was also obtained with correlation coefficients of all linear regression lines (measured intensity ratios vs mass ratios) ranging from 0.9995 to 0.9996. The detection limit at a signal-to-noise ratio of approximately 3 was 1 pg of T3. The T4 degradation during sample preparation was minimized to a small percentage (no more than 3% of the T3 values) by use of antioxidants (ascorbic acid, dithiothreitol, citric acid) and can be accounted for in the T3 measurement process. This well-characterized LC/MS/MS method for total serum T3, which demonstrates good accuracy and precision, low susceptibility to interferences, accountability of the conversion of T4 to T3, and comparability with the other RMP, qualifies as a reference measurement procedure and can be used to provide an accuracy base to which routine methods for T3 can be compared.
Bacterial analysis by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry has been demonstrated in numerous laboratories, and a few attempts have been made to compare results from different laboratories on the same organism. It has been difficult to understand the causes behind the observed differences between laboratories when different instruments, matrices, solvents, etc. are used. In order to establish this technique as a useful tool for bacterial identification, additional efforts in standardizing the methods by which MALDI mass spectra are obtained and comparisons of spectra from different instruments with different operators are needed. Presented here is an extension of our previous single-laboratory reproducibility study with three different laboratories in a controlled experiment with aliquots of the same bacterial culture, matrix stock solution, and calibrant standards. Using automated spectral collection of whole-cell bacteria and automated data processing and analysis algorithms, fingerprints from three different laboratories were constructed and compared. Nine of the ions appeared reproducibly within all three laboratories, with additional unique ions observed within each of the laboratories. An initial evaluation of the ability to use a fingerprint generated within one laboratory for bacterial identification of a sample from another laboratory is presented, and strategies for improving identification rates between laboratories is discussed. has been used to analyze intact, cultured microorganisms with minimal sample handling. Two recent review articles, which include the capabilities and current limitations that need to be addressed, provide an excellent overview of this emerging research field [1,2]. The MALDI-TOF MS technique for identifying biomolecules provides rapid analysis time (Ͻ1 min per sample analysis), low sample-volume requirements (Ͻ1 L fluid), and the highly selective nature of mass-spectrometric analysis based on relative molecular masses. The m/z values for mass spectral peaks and the patterns with which they are observed provide very specific and unbiased analysis, as they indicate molecular weights of true components of the sample. Bacterial cells have been identified by comparing MALDI-TOF spectra obtained from cultured bacterial cells and simple microbial mixtures against a library of known MALDI-TOF spectral fingerprints obtained from intact bacterial cells [3,4] or from comparison with masses predicted from a proteomic database [5,6]. The proteomic approach has been demonstrated to correctly identify bacteria from spectra originating at different laboratories [5], however, this approach is currently suffering from an incomplete protein database for many of the organisms that are of interest. As the proteomic database becomes more populated with organisms of concern, this approach will become more feasible for bacterial identification, at
A computed tomographic (CT) guided transgluteal approach through the greater sciatic foramen was used to drain pelvic abscesses and fluid collections in 21 patients. Ideal catheter placement should traverse the lower portion of the greater sciatic foramen at the level of the sacrospinous ligament. This avoids the vascular and neural elements that are located slightly cephalad at the level of the piriformis muscle. Percutaneous drainage through this approach was successful in avoiding surgery in 17 patients (81%). Pain was the most common complication and was generally associated with a more cephalad approach, transgressing the piriformis and the sacral plexus. CT-guided percutaneous drainage of pelvic abscesses through the greater sciatic foramen should be used when the more standard transperitoneal approach is not possible.
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