Intracellular survival of persistent group A streptococci in cultured epithelial cells

Group A streptococcus (GAS) is the principle etiologic agent of bacterial pharyngotonsillitis and a wide range of other diseases. Failure to eradicate GAS from patients has been documented in 5-30% of patients with pharyngotonsillitis, in spite of the continued sensitivity of GAS to penicillin and other beta-lactams. It was recently proposed that eradication failure might be attributed to the ability of GAS to maintain an intracellular reservoir during antibiotic treatment. We have previously shown that strains derived from patients with bacterial eradication failure, despite antibiotic treatment (persistent strains), adhered to and were internalized by cultured epithelial cells more efficiently than strains that were successfully eradicated. Since, penicillin and other beta-lactams do not penetrate well into mammalian cells, intracellular survival of GAS is crucial in order to persist during prolonged antibiotic treatment. In this study, we compared the survival of GAS strains from cases of eradication failure and eradication success, using an epithelial cell culture model. We found that persistent strains show significantly increased intracellular survival, compared to the 'eradication success' strains. This finding supports the idea that an intracellular reservoir of GAS plays a role in the etiology of antibiotic eradication failure.     

Hybrid positron detection and optical coherence tomography system: design, calibration, and experimental validation with rabbit atherosclerotic models

We evaluate the performance of our novel hybrid optical coherence tomography (OCT) and scintillating probe, demonstrate simultaneous OCT imaging and scintillating detection, and validate the system using an atherosclerotic rabbit model. Preliminary data obtained from the rabbit model suggest that our prototype positron probe detects local uptake of fluorodeoxyglucose (FDG) labeled with 18F positron (beta) radionuclide emitter, and the high-uptake regions correlate with sites of injury and extensive atherosclerosis areas. Preliminary data also suggest that coregistered high-resolution OCT images provide imaging of detailed plaque microstructures, which cannot be resolved by positron detection.     

Predicting Skull Loading: Applying Multibody Dynamics Analysis to a Macaque Skull

Evaluating stress and strain fields in anatomical structures is a way to test hypotheses that relate specific features of facial and skeletal morphology to mechanical loading. Engineering techniques such as finite element analysis are now commonly used to calculate stress and strain fields, but if we are to fully accept these methods we must be confident that the applied loading regimens are reasonable. Multibody dynamics analysis (MDA) is a relatively new three dimensional computer modeling technique that can be used to apply varying muscle forces to predict joint and bite forces during static and dynamic motions. The method ensures that equilibrium of the structure is maintained at all times, even for complex statically indeterminate problems, eliminating nonphysiological constraint conditions often seen with other approaches. This study describes the novel use of MDA to investigate the influence of different muscle representations on a macaque skull model (Macaca fascicularis), where muscle groups were represented by either a single, multiple, or wrapped muscle fibers. The impact of varying muscle representation on stress fields was assessed through additional finite element simulations. The MDA models highlighted that muscle forces varied with gape and that forces within individual muscle groups also varied; for example, the anterior strands of the superficial masseter were loaded to a greater extent than the posterior strands. The direction of the muscle force was altered when temporalis muscle wrapping was modeled, and was coupled with compressive contact forces applied to the frontal, parietal and temporal bones of the cranium during biting. Anat Rec, 291:491–501, 2008. © 2008 Wiley‐Liss, Inc.     

Phenotype-genotype characterization of 10 families with severe a subunit factor XIII deficiency

Factor XIII (FXIII) deficiency is a very rare severe autosomal bleeding disorder with a frequency of 1:2,000,000 in the general population and only a few patients have been genetically characterized so far. We report a phenotype-genotype characterization of 10 unrelated Iranian patients. Two FXIII (transglutaminase) activity assays showed no FXIII activity, except a conserved residual activity in patients receiving prophylactic substitution treatment. FXIII antigen concentrations measured by two immunoassays were comparable. Genotype characterization identified four novel mutations (2 missense and 2 small deletions) and two previously reported missense mutations in the FXIII A subunit gene (F13A). Molecular modeling was carried out to reveal the structural consequences of the missense mutations, that caused the replacement of an arginine residue involved in the formation of structurally important extensive hydrogen-bonded network. The replacements [c.320G>A (p.Arg77His) in the beta-sandwich, c.868C>T (p.Arg260Cys), c.869G>A (p.Arg260His) and c.1236G>T (p.Arg382Ser) in the core domain] resulted in the loss or impairment of such H-bonded network. Energy decomposition analysis demonstrated that this situation leads to the instability and perhaps to the incorrect folding of the A subunit, that would explain the development of severe FXIII deficiency.     

Changes of enzyme activities in ovine fetal fluids and maternal blood serum with gestational age

Our objectives were threefold: (1) to assess the activities of tissue enzymes, aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), and creatine kinase (CK) in the fetal fluids (amniotic and allantoic) collected from the gravid sheep uteri obtained from the abattoir and also in the maternal blood serum at various stages of pregnancy, (2) to compare the enzyme activities of fetal fluids relative to the maternal serum, and (3) to compare the enzyme activities in serum of pregnant ewes to nonpregnant ones. The stages of gestation, viz., stage I (30–60 days), stage II (61–90 days), and stage III (91–120 days) were identified based on the crown anus length of the fetus. As the gestational ages increased, AST significantly (p < 0.01) increased in the amniotic fluid and maternal blood serum but decreased in allantoic fluid; ALT had no changes in fetal fluids and the maternal blood serum; ALP decreased significantly (p < 0.01) in allantoic fluid but had no changes in the amniotic fluid and maternal serum; LDH increased significantly (p < 0.01) in amniotic and allantoic fluids but decreased in maternal serum; CK decreased significantly (p < 0.01) in fetal fluids and maternal serum. The enzyme activities were significantly higher in maternal sera than fetal fluids but were successively less in allantoic and amniotic fluids (p < 0.01). The activity of enzymes in maternal sera of pregnant and nonpregnant ewes were nearly the same. Except for LDH and CK, the greatest activities were found in the maternal serum in stage I and for AST in stage III of pregnancy (p < 0.01). These findings may have appreciable diagnostic significance in prenatal detection of disease status in both the dam and the fetus.