Mitochondrial Mutations in Adenoid Cystic Carcinoma of the Salivary Glands

Background: The MitoChip v2.0 resequencing array is an array-based technique allowing for accurate and complete sequencing of the mitochondrial genome. No studies have investigated mitochondrial mutation in salivary gland adenoid cystic carcinomas. Methodology: The entire mitochondrial genome of 22 salivary gland adenoid cystic carcinomas (ACC) of salivary glands and matched leukocyte DNA was sequenced to determine the frequency and distribution of mitochondrial mutations in ACC tumors. Principal Findings: Seventeen of 22 ACCs (77%) carried mitochondrial mutations, ranging in number from 1 to 37 mutations. A disproportionate number of mutations occurred in the D-loop. Twelve of 17 tumors (70.6%) carried mutations resulting in amino acid changes of translated proteins. Nine of 17 tumors (52.9%) with a mutation carried an amino acid changing mutation in the nicotinamide adenine dinucleotide dehydrogenase (NADH) complex. Conclusions/Significance: Mitochondrial mutation is frequent in salivary ACCs. The high incidence of amino acid changing mutations implicates alterations in aerobic respiration in ACC carcinogenesis. D-loop mutations are of unclear significance

SYSTEMIC BLOOD ACTIVATION DURING AND AFTER AUTOTRANSFUSION

To evaluate the extent of shed blood activation in two autotransfusion systems and the effect of circulating blood activation upon autotransfusion, we performed a prospective study in 18 patients undergoing internal mammary artery bypass operation and a control group of 10 patients. The autotransfusion systems were from Sorin (n = 9) consisting of a hard shell reservoir with a filter having a small contact area (0.32 m(2)), and from Dideco (n = 9) consisting of a hard shell reservoir with a filter having a larger contact area (4.64 m(2)). We found high concentrations of thromboxane, fibrinogen degradation products, complement split product C3a, and elastase in the shed blood and, with the exception of C3a, in the circulating blood of autotransfused patients. There was no such activation in control patients. The degree of the systemic inflammatory reaction was determined by the type of autotransfusion system and by the amount of infused shed blood. The Dideco system provoked more inflammatory response than did the Sorin. This was reflected by the larger shed blood loss during autotransfusion in the Dideco patients than in Sorin patients, resulting in infusion of more shed blood (means, 737 mL versus 566 mL; not significant). After autotransfusion, Dideco patients shed significantly more blood than did Sorin or control patients (p <0.05). Dideco patients also needed more colloid/crystalloid solution per 24 hours than Sorin patients (p <0.05). This became clinically relevant only after infusion of more than 800 mL of shed blood (p <0.001): hemodilution indicated the need for packed cells in 4 Dideco patients and in 1 Sorin patient. Dideco patients required a similar amount of blood products (0.8 +/- 0.4 unit) to the control patients. In contrast, Sorin patients required a mean of 0.2 +/- 0.2 unit, whereas blood products were avoided in 89% of them, versus 42% of the Dideco and control patients (not significant). In summary, we recommend autotransfusion of a limited amount (less than 800 mL) of shed blood with a reservoir that has the smallest possible contact area. Infusion of more than 800 mL of shed blood provokes derangement of hemostasis and hemodynamics by deleterious systemic blood activation, nullifying blood saving by autotransfusion

Influenza virus in human exhaled breath

Background: Recent studies suggest that humans exhale fine particles during tidal breathing but little is known about where the particles are generated or their role in infection transmission. We conducted a study of influenza infected patients to characterize particle and influenza virus concentrations in their exhaled breath. Methods: We recruited patients with influenza-like illness presenting for medical care at three clinics in Hong Kong, China. We collected two nasal swabs per subject, one for rapid testing and a second one for analysis via quantitative PCR (qPCR). Patients breathed with a steady regular pattern through a mouthpiece supplied with HEPA filtered air. Exhaled breath flowed through a 22 mm diameter and 40 cm long tube to an Exhalair (Pulmatrix, Inc, Lexington, MA), which monitored flow rate, and counted particles between 0.3 and 5 um in diameter using an optical particle counter. After three minutes of particle counting, we collected exhaled breath particles by sampling for 20 minutes on Teflon filters. We assayed each filter for influenza A and B using qPCR. Results: Thirteen of the 51 screened patients tested positive for influenza using the QuickVue rapid test (7 for influenza B, 6 for influenza A). Twelve rapid test positive patients completed the exhaled breath test (7 influenza B subjects and 5 influenza A subjects) and we recovered influenza virus in the exhaled breath of 4 (25%) subjects. Three (60%) of the five patients with influenza A infection and one (14%) of the seven with influenza B infection had detectable influenza virus in their exhaled breath. Exhaled breath virus concentrations ranged between 21 and 312 virus copies per sample, corresponding to a generation rate between 1 and 16 virus particles per minute. Preliminary particle data analysis indicated that 50% of subjects exhaled more than 500 particles per liter of air, a suggested threshold for identification of high particle producers. Conclusions: We recovered influenza virus from the exhaled breath of 4 out of 12 influenza patients, three of whom tested positive for influenza A. The results provide evidence that influenza virus is contained in fine particles generated during tidal breathing

GC-MS DETERMINATION OF RATIOS OF STABLE-ISOTOPE LABELED TO NATURAL UREA USING [(CN2)-C-13-N-15]UREA FOR STUDYING UREA KINETICS IN SERUM AND AS A MEANS TO VALIDATE ROUTINE METHODS FOR THE QUANTITATIVE ASSAY OF UREA IN DIALYSATE

A GC-MS determination of urea in serum or spent dialysate is described, using (CN2)-C-13-N-15-labelled urea and assaying the area ratio of labelled to natural urea by mass fragmentographic monitoring of fragments m/e 153 and 156, after its eventual conversion into the trimethylsilylether-derivative of 2-hydroxypyrimidine. The procedure can be successfully applied in the follow-up of the disappearance of labelled urea in serum after intravenous injection in man, enabling kinetic parameters of urea to be established, e.g. for purposes of studying the effectiveness of dialysis procedures. Furthermore the method can be used for validation of routine methods for measuring urea in other fluids, in particular dialysate. Examples are given of both applications of the GC-MS method described

GC-MS DETERMINATION OF RATIOS OF STABLE-ISOTOPE LABELED TO NATURAL UREA USING [(CN2)-C-13-N-15]UREA FOR STUDYING UREA KINETICS IN SERUM AND AS A MEANS TO VALIDATE ROUTINE METHODS FOR THE QUANTITATIVE ASSAY OF UREA IN DIALYSATE

A GC-MS determination of urea in serum or spent dialysate is described, using (CN2)-C-13-N-15-labelled urea and assaying the area ratio of labelled to natural urea by mass fragmentographic monitoring of fragments m/e 153 and 156, after its eventual conversion into the trimethylsilylether-derivative of 2-hydroxypyrimidine. The procedure can be successfully applied in the follow-up of the disappearance of labelled urea in serum after intravenous injection in man, enabling kinetic parameters of urea to be established, e.g. for purposes of studying the effectiveness of dialysis procedures. Furthermore the method can be used for validation of routine methods for measuring urea in other fluids, in particular dialysate. Examples are given of both applications of the GC-MS method described