Respiratory-resolved 5D Flow MRI: in-vivo validation and respiratory dependent flow changes in healthy volunteers and patients with congenital heart disease.

BACKGROUND: This study aimed to validate respiratory-resolved 5D flow MRI against real-time 2D phase contrast MRI, assess the impact of number of respiratory states, and measure the impact of respiration on hemodynamics in congenital heart disease (CHD) patients. METHODS: Respiratory-resolved 5D flow MRI derived net and peak flow measurements were compared to real-time 2D phase contrast MRI derived measurements in 10 healthy volunteers. Pulmonary to systemic flow ratios (Qp:Qs) were measured in 19 CHD patients and aortopulmonary collateral burden was measured in 5 Fontan patients. Additionally, the impact of number of respiratory states on measured respiratory-driven net flow changes was investigated in 10 healthy volunteers and 19 CHD patients (shunt physiology, n=11, single ventricle disease (SVD), n=8). RESULTS: There was good agreement between 5D flow MRI and real-time 2D phase contrast derived net and peak flow. Respiratory driven changes had good correlation (rho=0.64, p<0.001). In healthy volunteers, fewer than four respiratory states reduced measured respiratory driven flow changes in veins (5.2mL/cycle, p<0.001) and arteries (1.7mL/cycle, p=0.05). Respiration drove substantial venous net flow changes in SVD (64% change) and shunt patients (57% change). Respiration had significantly greater impact in SVD patients compared to shunt patients in the right and left pulmonary arteries (46% vs 15%, p=0.003 & 59% vs 20%, p=0.002). Qp:Qs varied by 37±24% over respiration in SVD patients and 12±20% in shunt patients. Aortopulmonary collateral burden varied by 118±84% over respiration in Fontan patients. The smallest collateral burden was measured during active inspiration in all patients and the greatest burden was during active expiration in 4 of 5 patients. Reduced respiratory resolution blunted measured flow changes in the caval veins of shunt and SVD patients (p<0.005). CONCLUSIONS: Respiratory-resolved 5D flow MRI measurements agree with real-time 2D phase contrast. Venous measurements are sensitive to number of respiratory states, whereas arterial measurements are more robust. Respiration has substantial impact on caval vein flow, Qp:Qs, and collateral burden in CHD patients.

Interval Changes in 4D Flow-Derived in vivo Hemodynamics Stratify Aortic Growth in Type B Aortic Dissection Patients.

BACKGROUND: Aortic diameter growth in type B aortic dissection (TBAD) is associated with progressive aortic dilation, resulting in increased mortality in patients with both de novo TBAD (dnTBAD) and residual dissection after type A dissection repair (rTAAD). Preemptive thoracic endovascular aortic repair may improve mortality in patients with TBAD, although it is unclear which patients may benefit most from early intervention. In vivo hemodynamic assessment using four-dimensional (4D) flow magnetic resonance imaging (MRI) has been used to characterize TBAD patients with growing aortas. In this longitudinal study, we investigated whether changes over time in 4D flow derived true and false lumen (TL, FL) hemodynamic parameters correlate with aortic growth rate which is a marker of increased risk. METHODS: We retrospectively identified TBAD patients with baseline and follow-up 4D flow MRI at least 120 days apart. Patients with TBAD intervention before baseline or between scans were excluded. 4D flow MRI data analysis included segmentation of the TL and FL, followed by voxel-wise calculation of TL and FL total kinetic energy (KE), maximum velocity (MV), mean forward flow (FF), and mean reverse flow (RF). Changes over time (Δ) were calculated for all hemodynamic parameters. Maximal diameter in the descending aorta was measured from MR angiogram images acquired at the time of 4D flow. Aortic growth rate was defined as the change in diameter divided by baseline diameter and standardized to scan interval. RESULTS: 32 patients met inclusion criteria (age: 56.9±14.1 years, Female: 13, n=19 rTAAD, n=13 dnTBAD). Mean follow up time was 538 days (range: 135-1689). Baseline aortic diameter did not correlate with growth rate. In the entire cohort, Δ FL MV (rho=0.37, p=.04) and Δ FL RF (rho=0.45, p=0.01) correlated with growth rate. In rTAAD only, Δ FL MV (rho=0.48, p=.04) and Δ FL RF (rho=0.51, p=0.03) correlated with growth rate, while in dnTBAD only, Δ TL KE (rho=0.63, p=.02) and Δ TL MV (rho=0.69, p=.01) correlated with growth rate. CONCLUSIONS: 4D flow derived longitudinal hemodynamic changes correlate with aortic growth rate in TBAD and may provide additional prognostic value for risk stratification. 4D flow MRI could be integrated into existing imaging protocols to allow for identification of TBAD patients who would benefit from preemptive surgical or endovascular intervention.

No Bones About It: Sex Is Binary.

Anthropologists have led the way in formulating techniques that reveal skeletal differences between males and females. Understanding of physical differences in the pelvis related to childbirth, hormonal impacts on bones, and extensive comparative studies have provided anthropologists with an array of traits and measurements that help them estimate sex using just bones. Forensic anthropologists and bioarcheologists are improving their ability to differentiate males and females by increasing research on a variety of postcranial bones and through the use of molecular data, especially new methods called proteomics, to identify sex in prepubescent juveniles. As remains from more cultures and time periods are studied, sex identification will continue to improve, because skeletal sex differences are in large part biologically determined. Yet, anthropologists have also been at the forefront of arguing that sex lies on a spectrum. Anthropologists who view sex as on a spectrum may deter sex identification progress; from their perspective, an individual of an undetermined sex may just be a nonbinary individual. Anthropologists who consider sex is on a spectrum are coming to this conclusion in part because they are looking for anatomical ideals, mistaking pathology for variation, and confusing independent variables with dependent variables. Nonetheless, anthropologists need to continue to improve sex identification techniques to reconstruct the past accurately, which may reveal less strict sex roles than previously presumed and help with the identification of crime victims. Forensic anthropologists should also increase their efforts to identify whether individuals have undergone medical procedures intended to change one's gender due to the current rise in transitioning individuals.

Focused navigation for respiratory-motion-corrected free-running radial 4D flow MRI.

PURPOSE: To validate a respiratory motion correction method called focused navigation (fNAV) for free-running radial whole-heart 4D flow MRI. METHODS: Using fNAV, respiratory signals derived from radial readouts are converted into three orthogonal displacements, which are then used to correct respiratory motion in 4D flow datasets. Hundred 4D flow acquisitions were simulated with non-rigid respiratory motion and used for validation. The difference between generated and fNAV displacement coefficients was calculated. Vessel area and flow measurements from 4D flow reconstructions with (fNAV) and without (uncorrected) motion correction were compared to the motion-free ground-truth. In 25 patients, the same measurements were compared between fNAV 4D flow, 2D flow, navigator-gated Cartesian 4D flow, and uncorrected 4D flow datasets. RESULTS: For simulated data, the average difference between generated and fNAV displacement coefficients was 0.04 ± $$ \pm $$ 0.32 mm and 0.31 ± $$ \pm $$ 0.35 mm in the x and y directions, respectively. In the z direction, this difference was region-dependent (0.02 ± $$ \pm $$ 0.51 mm up to 5.85 ± $$ \pm $$ 3.41 mm). For all measurements (vessel area, net volume, and peak flow), the average difference from ground truth was higher for uncorrected 4D flow datasets (0.32 ± $$ \pm $$ 0.11 cm2 , 11.1 ± $$ \pm $$ 3.5 mL, and 22.3 ± $$ \pm $$ 6.0 mL/s) than for fNAV 4D flow datasets (0.10 ± $$ \pm $$ 0.03 cm2 , 2.6 ± $$ \pm $$ 0.7 mL, and 5.1 ± 0 $$ \pm 0 $$ .9 mL/s, p < 0.05). In vivo, average vessel area measurements were 4.92 ± $$ \pm $$ 2.95 cm2 , 5.06 ± $$ \pm $$ 2.64 cm2 , 4.87 ± $$ \pm $$ 2.57 cm2 , 4.87 ± $$ \pm $$ 2.69 cm2 , for 2D flow and fNAV, navigator-gated and uncorrected 4D flow datasets, respectively. In the ascending aorta, all 4D flow datasets except for the fNAV reconstruction had significantly different vessel area measurements from 2D flow. Overall, 2D flow datasets demonstrated the strongest correlation to fNAV 4D flow for both net volume (r2  = 0.92) and peak flow (r2  = 0.94), followed by navigator-gated 4D flow (r2  = 0.83 and r2  = 0.86, respectively), and uncorrected 4D flow (r2  = 0.69 and r2  = 0.86, respectively). CONCLUSION: fNAV corrected respiratory motion in vitro and in vivo, resulting in fNAV 4D flow measurements that are comparable to those derived from 2D flow and navigator-gated Cartesian 4D flow datasets, with improvements over those from uncorrected 4D flow.

Hemodynamic Evaluation of Type B Aortic Dissection Using Compressed Sensing Accelerated 4D Flow MRI.

BACKGROUND: 4D Flow MRI is a quantitative imaging technique to evaluate blood flow patterns; however, it is unclear how compressed sensing (CS) acceleration would impact aortic hemodynamic quantification in type B aortic dissection (TBAD). PURPOSE: To investigate CS-accelerated 4D Flow MRI performance compared to GRAPP-accelerated 4D Flow MRI (GRAPPA) to evaluate aortic hemodynamics in TBAD. STUDY TYPE: Prospective. POPULATION: Twelve TBAD patients, two volunteers. FIELD STRENGTH/SEQUENCE: 1.5T, 3D time-resolved cine phase-contrast gradient echo sequence. ASSESSMENT: GRAPPA (acceleration factor [R] = 2) and two CS-accelerated (R = 7.7 [CS7.7] and 10.2 [CS10.2]) 4D Flow MRI scans were acquired twice for interscan reproducibility assessment. Voxelwise kinetic energy (KE), peak velocity (PV), forward flow (FF), reverse flow (RF), and stasis were calculated. Plane-based mid-lumen flows were quantified. Imaging times were recorded. TESTS: Repeated measures analysis of variance, Pearson correlation coefficients (r), intraclass correlation coefficients (ICC). P < 0.05 indicated statistical significance. RESULTS: The KE and FF in true lumen (TL) and PV in false lumen (FL) did not show difference among three acquisition types (P = 0.818, 0.065, 0.284 respectively). The PV and stasis in TL were higher, KE, FF, and RF in FL were lower, and stasis was higher in GRAPPA compared to CS7.7 and CS10.2. The RF was lower in GRAPPA compared to CS10.2. The correlation coefficients were strong in TL (r = [0.781-0.986]), and low to strong in FL (r = [0.347-0.948]). The ICC levels demonstrated moderate to excellent interscan reproducibility (0.732-0.989). The FF and net flow in mid-descending aorta TL were significantly different between CS7.7 and CS10.2. CONCLUSION: CS-accelerated 4D Flow MRI has potential for clinical utilization with shorter scan times in TBAD. Our results suggest similar hemodynamic trends between acceleration types, but CS-acceleration impacts KE, FF, RF, and stasis more in FL. EVIDENCE LEVEL: 1 Technical Efficacy: Stage 2.

Systolic reverse flow derived from 4D flow cardiovascular magnetic resonance in bicuspid aortic valve is associated with aortic dilation and aortic valve stenosis: a cross sectional study in 655 subjects.

BACKGROUND: Bicuspid aortic valve (BAV) disease is associated with increased risk of aortopathy. In addition to current intervention guidelines, BAV mediated changes in aortic 3D hemodynamics have been considered as risk stratification measures. We aimed to evaluate the association of 4D flow cardiovascular magnetic resonance (CMR) derived voxel-wise aortic reverse flow with aortic dilation and to investigate the role of aortic valve regurgitation (AR) and stenosis (AS) on reverse flow in systole and diastole. METHODS: 510 patients with BAV (52 ± 14 years) and 120 patients with trileaflet aortic valve (TAV) (61 ± 11 years) and mid-ascending aorta diameter (MAAD) > 35 mm who underwent CMR including 4D flow CMR were retrospectively included. An age and sex-matched healthy control cohort (n = 25, 49 ± 12 years) was selected. Voxel-wise reverse flow was calculated in the aorta and quantified by the mean reverse flow in the ascending aorta (AAo) during systole and diastole. RESULTS: BAV patients without AS and AR demonstrated significantly increased systolic and diastolic reverse flow (222% and 13% increases respectively, p < 0.01) compared to healthy controls and also had significantly increased systolic reverse flow compared to TAV patients with aortic dilation (79% increase, p < 0.01). In patients with isolated AR, systolic and diastolic AAo reverse flow increased significantly with AR severity (c = - 83.2 and c = - 205.6, p < 0.001). In patients with isolated AS, AS severity was associated with an increase in both systolic (c = - 253.1, p < 0.001) and diastolic (c = - 87.0, p = 0.02) AAo reverse flow. Right and left/right and non-coronary fusion phenotype showed elevated systolic reverse flow (> 17% increase, p < 0.01). Right and non-coronary fusion phenotype showed decreased diastolic reverse flow (> 27% decrease, p < 0.01). MAAD was an independent predictor of systolic (p < 0.001), but not diastolic, reverse flow (p > 0.1). CONCLUSION: 4D flow CMR derived reverse flow associated with BAV was successfully captured even in the absence of AR or AS and in comparison to TAV patients with aortic dilation. Diastolic AAo reverse flow increased with AR severity while AS severity strongly correlated with increased systolic reverse flow in the AAo. Additionally, increasing MAAD was independently associated with increasing systolic AAo reverse flow. Thus, systolic AAo reverse flow may be a valuable metric for evaluating disease severity in future longitudinal outcome studies.

Impact of pulmonary artery flow distribution on Fontan hemodynamics and flow energetics.

BACKGROUND: With improved life expectancy following Fontan palliation, there is an increasing population of patients with a total cavopulmonary connection. However, there is a poor understanding of which patients will experience Fontan failure and when. 4D flow MRI has identified several metrics of clinical interest, but longitudinal studies investigating hemodynamics in Fontan patients are lacking. OBJECTIVE: We aimed to investigate the relationship between flow distribution to the pulmonary arteries and regional hemodynamic metrics in a unique cohort with follow-up 4D flow MRI. MATERIALS AND METHODS: Patients with > 6 months of 4D flow MRI follow-up were included. Flow distribution from the caval veins to pulmonary arteries was measured in addition to regional measures of peak velocity, viscous energy loss (ELmean and ELtot), and kinetic energy. RESULTS: Ten patients with total cavopulmonary connection (17.7 ± 8.8 years at baseline, follow-up: 4.4 ± 2.6 years) were included. Five subjects had unequal flow distribution from the IVC to the pulmonary arteries at baseline. Over time, these subjects tended to have larger increases in peak velocity (39.2% vs 6.6%), ELmean (11.6% vs -38.3%), ELtot (9.5% vs -36.2%), and kinetic energy (96.1% vs 36.3%) in the IVC. However, these differences were statistically insignificant. We found that changes in ELmean and ELtot were significantly associated with changes in peak velocity in the caval veins (R2 > 0.5, P < 0.001). CONCLUSION: Unequal flow distribution from the IVC may drive increasing peak velocities and viscous energy losses, which have been associated with worse clinical outcomes. Changes in peak velocity may serve as a surrogate measure for changes in viscous energy loss.

Biological distance at the Ryan Mound site.

OBJECTIVES: The Ryan Mound site in California spans 2000 years and has been utilized in over 200 studies. The Ryan Mound has been assumed to be a culturally and, therefore, a biologically continuous population over time. This study attempts to determine whether adults at the Ryan Mound consisted of a continuous population over the span of three temporal periods by using nonmetric skeletal traits. MATERIALS AND METHODS: Thirty-eight nonmetric cranial traits and four nonmetric post-cranial traits were scored on adults. Trait correlations were assessed for sex and age using chi-square and Fisher's exact tests. For bilateral traits, data were recorded for both sides, but only results from the left side are reported. Most data were recorded as present or absent. Twelve traits had scores that had more than two nominal categories, which were converted to binary values to enable mean measure of divergence (MMD) statistical analyses. RESULTS: After data reduction, 36 traits remained. Using these traits, standardized MMD analyses revealed that the oldest temporal period and the most recent temporal period individuals were significantly different. CONCLUSION: This study illustrates the importance of testing for biological continuity. Nonmetric studies provide a way to examine relationships within a sample to determine biological continuity. For the Ryan Mound, new populations may have moved into the region early on as part of the Meganos intrusion, or new populations may have moved into the region later, between AD 1500 and AD 1800. Comparative data from nearby sites further support the Meganos intrusion theory.

Knee osteoarthritis, body mass index and pain: data from the Osteoarthritis Initiative.

OBJECTIVE: This study aimed to determine whether BMI increases knee pain as measured from self-reported surveys even when controlling for OA severity as measured by osteophytes and joint space narrowing visible on X-rays. METHODS: Data available through the Osteoarthritis Initiative (OAI) were analysed, which included a sample of 4769 individuals, to answer the above question regarding OA, excess weight and pain. OA severity was assessed through baseline X-rays on right knees that were scored on a composite quasi-Kellgren and Lawrence grade. Weight was assessed through BMI. Pain was assessed through self-reports of the WOMAC pain subset as well as a 30-day pain severity question based on a 0-10 scale. Data were analysed using SPSS and analyses of covariance (ANCOVAs) were run to examine models adjusted for age, smoking, prior injury, pain medication and Heberden's nodes. Critical alpha levels were set at 0.05. RESULTS: The results reported here confirm that knee pain does increase with OA severity. However, ANCOVA multiple regressions with controls reveal that even when taking into account OA severity, individuals with higher BMIs experience greater pain than individuals with lower BMIs. CONCLUSION: Weight loss may reduce knee OA pain even if the osteological symptoms are not treated.

Attachment style and coping in relation to posttraumatic stress disorder symptoms among adults living with HIV/AIDS.

Research indicates that a significant proportion of people living with HIV/AIDS report symptoms of posttraumatic stress disorder (PTSD). Moreover, attachment style has been associated with psychological and behavioral outcomes among persons living with HIV/AIDS. Attachment style may influence the ability to cope with traumatic stress and affect PTSD symptoms. To examine the association between attachment style and coping with PTSD symptoms, we assessed 94 HIV-positive adults on self-report measures of posttraumatic stress, coping, and attachment style. In multiple regression analysis, avoidant attachment and emotion-focused coping were positively and significantly associated with greater PTSD symptomatology. Support was also found for the moderating effects of avoidant and insecure attachment styles on emotion-focused coping in relation to greater PTSD symptoms. Taken altogether, these results suggest that interventions that develop adaptive coping skills and focus on the underlying construct of attachment may be particularly effective in reducing trauma-related symptoms in adults living with HIV/AIDS.

Clinical Applications of Four-Dimensional Flow MRI.

Four-dimensional flow MRI is a powerful phase contrast technique used for assessing three-dimensional (3D) blood flow dynamics. By acquiring a time-resolved velocity field, it enables flexible retrospective analysis of blood flow that can include qualitative 3D visualization of complex flow patterns, comprehensive assessment of multiple vessels, reliable placement of analysis planes, and calculation of advanced hemodynamic parameters. This technique provides several advantages over routine two-dimensional flow imaging techniques, allowing it to become part of clinical practice at major academic medical centers. In this review, we present the current state-of-the-art cardiovascular, neurovascular, and abdominal applications.

Brief communication: radiographic study of metatarsal one basal epiphyseal fusion: a note of caution on age determination.

This study examines radiographs of first metatarsals of 131 individuals from age 17-88 years to determine whether internal basal epiphyseal lines may be visible past the age of metatarsal fusion, which usually occurs between 14 and 16 years of age (Scheuer and Black: The juvenile skeleton. San Diego: Elsevier Academic Press,2004). In 29% (38 out of 131) of the radiographed first metatarsals (MT1s) the basal epiphyseal scar is visible, including in one individual who was 80 years old. Statistically, there was no relationship between the loss of the epiphyseal scar and age. Thus, the presence of the epiphyseal scar does not necessarily indicate subadult age. These data suggest that OH 8's radiographically visible basal epiphyseal line has no bearing on whether it is a subadult or not.

Case study of a rare occipital osteoma related to cranial trauma.

Burial 280 is a 31- to 40-year-old male Native American individual who lived in a 10th to 16th century coastal foraging society in California. His remains display a large ectocranial elevation on the occipital, measuring 23.3 mm anteroposteriorly, 25.3 mm mediolaterally, and is elevated to 20.1 mm. It is located along the superior nuchal line on the left side of the occipital and represents an extremely rare occipital localization of a particular type of trauma-related osteoma. In our prior study of Burial 280, trauma etiology was not considered; herein, we present evidence for such a cause. Such trauma-related neoplasms are sometimes called ballooned (or giant, depending on size) osteomas. The presence of this tumor appears to have resulted in asymmetrical cranial muscle use and consequent skeletal changes. This case study illustrates that cranial trauma may sometimes induce benign tumor elevations, rather than a depression or pronounced fracture.

Baseline 4D Flow-Derived in vivo Hemodynamic Parameters Stratify Descending Aortic Dissection Patients With Enlarging Aortas.

Purpose: The purpose of our study was to assess the value of true lumen and false lumen hemodynamics compared to aortic morphological measurements for predicting adverse-aorta related outcomes (AARO) and aortic growth in patients with type B aortic dissection (TBAD). Materials and Methods: Using an IRB approved protocol, we retrospectively identified patients with descending aorta (DAo) dissection at a large tertiary center. Inclusion criteria includes known TBAD with ≥ 6 months of clinical follow-up after initial presentation for TBAD or after ascending aorta intervention for patients with repaired type A dissection with residual type B aortic dissection (rTAAD). Patients with prior descending aorta intervention were excluded. The FL and TL of each patient were manually segmented from 4D flow MRI data, and 3D parametric maps of aortic hemodynamics were generated. Groups were divided based on (1) presence vs. absence of AARO and (2) growth rate ≥ vs. < 3 mm/year. True and false lumen kinetic energy (KE), stasis, peak velocity (PV), reverse/forward flow (RF/FF), FL to TL KE ratio, as well as index aortic diameter were compared between groups using the Mann-Whitney U or independent t-test. Results: A total of n = 51 patients (age: 58.4 ± 15.0 years, M/F: 31/20) were included for analysis of AARO. This group contained n = 26 patients with TBAD and n = 25 patients with rTAAD. In the overall cohort, AARO patients had larger baseline diameters, lower FL-RF, FL stasis, TL-KE, TL-FF and TL-PV. Among patients with de novo TBAD, those with AAROs had larger baseline diameter, lower FL stasis and TL-PV. In both the overall cohort and in the subgroup of de novo TBAD, subjects with aortic growth ≥ 3mm/year, patients had a higher KE ratio. Conclusion: Our study suggests that 4D flow MRI is a promising tool for TBAD evaluation that can provide information beyond traditional MRA or CTA. 4D flow has the potential to become an integral aspect of TBAD work-up, as hemodynamic assessment may allow earlier identification of at-risk patients who could benefit from earlier intervention.

Effects of N-acetylcysteine on renal hemodynamics in contrast media-induced nephropathy.

BACKGROUND: N-acetylcysteine (NAC) has been proposed to prevent radiocontrast nephropathy in high-risk patients. METHODS: The effect of single-dose and prolonged administration of NAC before application of either the ionic, high-osmolar radiocontrast agent diatrizoate sodium (DTZ) or the nonionic, low-osmolar radiocontrast agent iohexol (IOH) in a rat model combining uninephrectomy, salt depletion, and administration of indomethacin was explored. Arterial blood pressure and total, cortical, and medullary blood flow were continuously recorded in anesthetized Sprague-Dawley rats. RESULTS: NAC had no effect on renal hemodynamics in control rats. Both DTZ and IOH induced biphasic changes in renal blood flow and cortical renal blood flux and persistently reduced medullary blood flux. Neither single-dose nor prolonged administration of NAC prevented the hemodynamic changes following administration of DTZ or IOH, respectively. Acute prophylactic administration of NAC prevented increased urinary ET excretion after injection of IOH and, to a smaller degree, of DTZ. Both an ionic, high-osmolar (DTZ) and a nonionic, low-osmolar (IOH) radiocontrast agent induce marked changes in renal hemodynamics in salt-depleted rats treated with indomethacin. CONCLUSIONS: Renal perfusion is not affected by NAC application in a model of experimental contrast nephropathy in rats. Other effects of NAC might thus account for the presumed renoprotective properties.

Peptides with dual binding specificity for HLA-A2 and HLA-E are encoded by alternatively spliced isoforms of the antioxidant enzyme peroxiredoxin 5.

Peptides with dual binding specificity for classical HLA class I and non-classical HLA-E molecules have been identified in virus-encoded proteins, but not in cellular proteins from normal or neoplastic cells. Expression screening of a melanoma cDNA library with a CTL clone recognizing an HLA-A2-restricted tumor-specific epitope encoded by mutant peroxiredoxin 5 (Prdx5), a stress-inducible peroxidase, led to the identification of two alternatively spliced isoforms of the same gene. These isoforms, which lack the catalytic cysteine fundamental for enzymatic activity, showed widespread expression in neoplastic and normal tissues but were unstable at the protein level, being detectable, following transient transfection, only after lactacystin treatment to inhibit proteasomal degradation. Isoform-specific sequences which formed, respectively, as result of exon 1 splicing to either exon 3 or 4, encoded two distinct nonapeptides (AMAPIKTHL and AMAPIKVRL, not present in the full-length protein) with anchor residues for HLA-A2 and HLA-E molecules and able to stabilize HLA-A2 and HLA-E cell surface expression. HLA-E+ targets, loaded with these peptides, were not recognized by NK cells expressing CD94/NKG2A inhibitory or CD94/NKG2C activatory receptors. However, both peptides were recognized, although with low avidity, by HLA-E-restricted CD8+ CTL. The nonapeptide AMAPIKVRL was used to elicit HLA-A2-restricted CTL clones that killed peptide-pulsed lymphoblastoid cell lines and melanoma cells expressing the corresponding Prdx5 isoform. Our results suggest that alternatively spliced isoforms of Prdx5, through the generation of HLA-E- and HLA-A2-restricted peptides may be part of immune-mediated stress response contributing to the detection and elimination of damaged normal or neoplastic cells.

Optimized elution of daptomycin from polymethylmethacrylate beads.

We demonstrate that xylitol can be added to polymethylmethacrylate (PMMA) bone cement to enhance the elution of daptomycin in terms of both the peak and sustained release of antibiotic. We also demonstrate that a PMMA-xylitol formulation optimized for daptomycin can be used to enhance the elution of both vancomycin and gentamicin.

Impact of sarA on antibiotic susceptibility of Staphylococcus aureus in a catheter-associated in vitro model of biofilm formation.

Mutation of the staphylococcal accessory regulator (sarA) in Staphylococcus aureus limits but does not abolish the capacity of the organism to form a biofilm. As a first step toward determining whether this limitation is therapeutically relevant, we carried out in vitro studies comparing the relative susceptibility of an S. aureus clinical isolate (UAMS-1) and its isogenic sarA mutant (UAMS-929) in the specific context of a catheter-associated biofilm. The antibiotics tested were daptomycin, linezolid, and vancomycin, all of which were evaluated by using concentrations based on the MIC defined as the breakpoint for a susceptible strain of S. aureus (< or = 1.0, < or = 2.0, and < or = 4.0 microg/ml for daptomycin, vancomycin, and linezolid, respectively). Mutation of sarA had no significant impact on the MIC of UAMS-1 for any of the targeted antibiotics, as defined by Etest antimicrobial susceptibility testing. However, mutation of sarA did result in a significant increase in antimicrobial susceptibility to all targeted antibiotics when they were tested in the specific context of a biofilm. Additionally, whether susceptibility was assessed by using UAMS-1 or its sarA mutant, daptomycin was found to be more effective against established S. aureus biofilms than either linezolid or vancomycin.

Impact of sarA on daptomycin susceptibility of Staphylococcus aureus biofilms in vivo.

We used a murine model of catheter-associated biofilm formation to determine whether the mutation of the staphylococcal accessory regulator (sarA) has an impact on the susceptibility of established Staphylococcus aureus biofilms to treatment with daptomycin in vivo. The experiments were done with two clinical isolates, one of which (UAMS-1) was obtained from the bone of a patient suffering from osteomyelitis, while the other (UAMS-1625) is an isolate of the USA300 clonal lineage of community-acquired methicillin (meticillin)-resistant S. aureus. UAMS-1625 had a reduced capacity to form a biofilm in vivo compared to that of UAMS-1 (P = 0.0015), but in both cases the mutation of sarA limited biofilm formation compared to that of the corresponding parent strain (P < or = 0.001). The mutation of sarA did not affect the daptomycin MIC for either strain, but it did result in increased susceptibility in vivo in the context of an established biofilm. Specifically, daptomycin treatment resulted in the clearance of detectable bacteria from <10% of the catheters colonized with the parent strains, while treatment with an equivalent daptomycin concentration resulted in the clearance of 46.4% of the catheters colonized with the UAMS-1 sarA mutant and 69.1% of the catheters colonized with the UAMS-1625 sarA mutant. In the absence of daptomycin treatment, mice with catheters colonized with the UAMS-1625 parent strain also developed skin lesions in the region adjacent to the implanted catheter. No such lesions were observed in any other experimental group, including untreated mice containing catheters colonized with the UAMS-1625 sarA mutant.

Sex differences in humeral bilateral asymmetry in two hunter-gatherer populations: California Amerinds and British Columbian Amerinds.

This study uses two prehistoric Amerindian populations of hunter-gatherer subsistence patterns to determine whether levels of sexual dimorphism in humeral bilateral cross-sectional asymmetry are related to sex-specific differences in activities among these populations. Results confirmed that males of the California Amerind population who engaged in the more unimanual activities of spear hunting and warfare were more asymmetrical than were their female counterparts who engaged in the more bimanual activities of grinding acorns. California Amerind males were also more asymmetrical than British Columbian Amerind males who rowed (using both arms) extensively. Sex differences within British Columbian Amerinds were not statistically significant, nor were female differences between populations. In general, levels of humeral asymmetry appear to be more dependent on sex and population-specific behaviors rather than broad subsistence patterns.