Accuracy and reproducibility of semi-automated late gadolinium enhancement quantification techniques in patients with hypertrophic cardiomyopathy.

BACKGROUND: The presence and extent of late gadolinium enhancement (LGE) has been associated with adverse events in patients with hypertrophic cardiomyopathy (HCM). Signal intensity (SI) threshold techniques are routinely employed for quantification; Full-Width at Half-Maximum (FWHM) techniques are suggested to provide greater reproducibility than Signal Threshold versus Reference Mean (STRM) techniques, however the accuracy of these approaches versus the manual assignment of optimal SI thresholds has not been studied. In this study, we compared all known semi-automated LGE quantification techniques for accuracy and reproducibility among patients with HCM. METHODS: Seventy-six HCM patients (51 male, age 54 ± 13 years) were studied. Total LGE volume was quantified using 7 semi-automated techniques and compared to expert manual adjustment of the SI threshold to achieve optimal segmentation. Techniques tested included STRM based thresholds of >2, 3, 4, 5 and 6 SD above mean SI of reference myocardium, the FWHM technique, and the Otsu-auto-threshold (OAT) technique. The SI threshold chosen by each technique was recorded for all slices. Bland-Altman analysis and intra-class correlation coefficients (ICC) were reported for each semi-automated technique versus expert, manually adjusted LGE segmentation. Intra- and inter-observer reproducibility assessments were also performed. RESULTS: Fifty-two of 76 (68%) patients showed LGE on a total of 202 slices. For accuracy, the STRM >3SD technique showed the greatest agreement with manual segmentation (ICC = 0.97, mean difference and 95% limits of agreement = 1.6 ± 10.7 g) while STRM >6SD, >5SD, 4SD and FWHM techniques systematically underestimated total LGE volume. Slice based analysis of selected SI thresholds similarly showed the STRM >3SD threshold to most closely approximate manually adjusted SI thresholds (ICC = 0.88). For reproducibility, the intra- and inter-observer reproducibility of the >3SD threshold demonstrated an acceptable mean difference and 95% limits of agreement of -0.5 ± 6.8 g and -0.9 ± 5.6 g, respectively. CONCLUSIONS: FWHM segmentation provides superior reproducibility, however systematically underestimates total LGE volume compared to manual segmentation in patients with HCM. The STRM >3SD technique provides the greatest accuracy while retaining acceptable reproducibility and may therefore be a preferred approach for LGE quantification in this population.

N-Acetylcysteine Prevents Retrograde Motor Neuron Death after Neonatal Peripheral Nerve Injury.

BACKGROUND: Neuronal death may be an overlooked and unaddressed component of disability following neonatal nerve injuries, such as obstetric brachial plexus injury. N-acetylcysteine and acetyl-L-carnitine improve survival of neurons after adult nerve injury, but it is unknown whether they improve survival after neonatal injury, when neurons are most susceptible to retrograde neuronal death. The authors' objective was to examine whether N-acetylcysteine or acetyl-L-carnitine treatment improves survival of neonatal motor or sensory neurons in a rat model of neonatal nerve injury. METHODS: Rat pups received either a sciatic nerve crush or transection injury at postnatal day 3 and were then randomized to receive either intraperitoneal vehicle (5% dextrose), N-acetylcysteine (750 mg/kg), or acetyl-L-carnitine (300 mg/kg) once or twice daily. Four weeks after injury, surviving neurons were retrograde-labeled with 4% Fluoro-Gold. The lumbar spinal cord and L4/L5 dorsal root ganglia were then harvested and sectioned to count surviving motor and sensory neurons. RESULTS: Transection and crush injuries resulted in significant motor and sensory neuron loss, with transection injury resulting in significantly less neuron survival. High-dose N-acetylcysteine (750 mg/kg twice daily) significantly increased motor neuron survival after neonatal sciatic nerve crush and transection injury. Neither N-acetylcysteine nor acetyl-L-carnitine treatment improved sensory neuron survival. CONCLUSIONS: Proximal neonatal nerve injuries, such as obstetric brachial plexus injury, produce significant retrograde neuronal death after injury. High-dose N-acetylcysteine significantly increases motor neuron survival, which may improve functional outcomes after obstetrical brachial plexus injury.

Contribution of mitral valve leaflet length and septal wall thickness to outflow tract obstruction in patients with hypertrophic cardiomyopathy.

We sought to examine whether elongation of the mitral valve leaflets in patients with hypertrophic cardiomyopathy (HCM) is synergistic to septal wall thickness (SWT) in the development of left ventricular outflow tract obstruction (LVOTO). HCM is a common genetic cardiac disease characterized by asymmetric septal hypertrophy and predisposition towards LVOTO. It has been reported that elongation of the mitral valve leaflets may be a primary phenotypic feature and contribute to LVOTO. However, the relative contribution of this finding versus SWT has not been studied. 152 patients (76 with HCM and 76 non-diseased age, race and BSA-matched controls) and 18 young, healthy volunteers were studied. SWT and the anterior mitral valve leaflet length (AMVLL) were measured using cine MRI. The combined contribution of these variables (SWT × AMVLL) was described as the Septal Anterior Leaflet Product (SALP). Peak LVOT pressure gradient was determined by Doppler interrogation and defined as "obstructive" if ≥ 30 mmHg. Patients with HCM were confirmed to have increased AMVLL compared with controls and volunteers (p < 0.01). Among HCM patients, both SWT and SALP were significantly higher in patients with LVOTO (N = 17) versus without. SALP showed modest improvement in predictive accuracy for LVOTO (AUC = 0.81) among the HCM population versus SWT alone (AUC = 0.77). However, in isolated patients this variable identified patients with LVOTO despite modest SWT. Elongation of the AMVLL is a primary phenotypic feature of HCM. While incremental contributions to LVOTO appear modest at a population level, specific patients may have dominant contribution to LVOTO. The combined marker of SALP allows for maintained identification of such patients despite modest increases in SWT.

Ventricular pacing site separation by cardiac computed tomography: validation for the prediction of clinical response to cardiac resynchronization therapy.

Cardiac Resynchronization Therapy (CRT) fails to provide benefit in up to one-third of patients. Maximizing the geographic separation of right and left ventricular pacing lead sites has been suggested as one way to improve response. Cardiac CT provides an opportunity to explore 3-dimensional inter-lead distance (ILD) measures for the prediction of CRT response. The objective of this study was to investigate associations between standardized measures of ILD by cardiac CT and echocardiographic response to CRT. Forty-two consecutive patients undergoing CRT had serial clinical and echocardiographic evaluations performed in addition to a post-procedural cardiac-gated CT with blinded measurement of direct and circumferential (via the myocardium) ILD measures. Clinical response to CRT, the primary clinical outcome, was defined as a ≥15% reduction in LVESV using echocardiography at 6-months. The mean age and ejection fraction was 63.6 ± 8.9 years and 25.2 ± 7.8%, respectively. The primary outcome occurred in 35 of 42 patients (83%). Both direct and circumferential CT-based ILD measures were associated with the primary outcome by univariate analysis. Receiver Operator Characteristic analysis identified Circumferential ILD to have the strongest predictive accuracy (AUC 0.78). Inter- and intra-observer reproducibility of CT-derived ILD measures was excellent. Circumferential ILD measures on cardiac CT are predictive of clinical response to CRT. Incorporation of these measures into the selection of optimal pacing targets, particularly from pre-procedural CT coronary vein imaging may be of therapeutic benefit and warrants further investigation.

Retrograde labeling of regenerating motor and sensory neurons using silicone caps.

BACKGROUND: Retrograde labeling permits the investigation of the number, distribution and axonal projections of neurons in the peripheral nervous system. The well technique for labeling peripheral nerves consists of incubating the exposed peripheral nerve in a well for one hour, a time intensive technique. However, other techniques that inject tracers directly into the nerve or muscle may result in variable labeling depending on nerve preparation and location of injection. NEW METHOD: We describe a method of retrograde labeling peripheral nerves that increases tracer uptake and improves labeling efficiency. This technique utilizes a silicone cap over the nerve that is kept in place with fibrin glue, permitting closure of the incision with the cap in place, mitigating the need to wait one hour for back-labeling as with the standard well technique. RESULTS: In the rat common peroneal nerve, the new silicone cap technique, compared to the standard well technique, labeled 405±11 (SEM) vs. 378±21 motoneurons and 953±40 vs. 948±57 sensory neurons. These counts were not statistically different. Labeling intensity was greater in DRG neurons with the silicone cap technique, but this difference was not evident in motoneurons. COMPARISON WITH EXISTING METHOD: Retrograde-labeling with silicone caps labels an equal number of motor and sensory neurons in comparison with the standard well technique and labels sensory neurons with greater intensity. CONCLUSIONS: Retrograde-labeling with silicone caps reliably labels neurons and significantly decreases the time required for labeling, reducing anesthetic exposure and improving the efficiency of the technique.

Comparison of semi-automated scar quantification techniques using high-resolution, 3-dimensional late-gadolinium-enhancement magnetic resonance imaging.

The quantification and modeling of myocardial scar is of expanding interest for image-guided therapy, particularly in the field of arrhythmia management. Migration towards high-resolution, three-dimensional (3D) MRI techniques for spatial mapping of myocardial scar provides superior spatial registration. However, to date no systematic comparison of available approaches to 3D scar quantification have been performed. In this study we compare the reproducibility of six 3D scar segmentation algorithms for determination of left ventricular scar volume. Additionally, comparison to two-dimensional (2D) scar quantification and 3D manual segmentation is performed. Thirty-five consecutive patients with ischemic cardiomyopathy were recruited and underwent conventional 2D late gadolinium enhancement (LGE) and 3D isotropic LGE imaging (voxel size 1.3 mm(3)) using a 3 T scanner. 3D LGE datasets were analyzed using six semi-automated segmentation techniques, including the signal threshold versus reference mean (STRM) technique at >2, >3, >5 and >6 standard deviations (SD) above reference myocardium, the full width at half maximum (FWHM) technique, and an optimization-based technique called hierarchical max flow (HMF). The mean ejection fraction was 32.1 ± 12.7 %. Reproducibility was greatest for HMF and FWHM techniques with intra-class correlation coefficient values ≥0.95. 3D scar quantification and modeling is clinically feasible in patients with ischemic cardiomyopathy. While several approaches show acceptable reproducibility, HMF appears superior due to maintenance of accuracy towards manual segmentations.

Restoration of Trigeminal Cutaneous Sensation with Cross-Face Sural Nerve Grafts: A Novel Approach to Facial Sensory Rehabilitation.

UNLABELLED: Although treating facial palsy is considered debilitating for patients, trigeminal nerve palsy and sensory deficits of the face are overlooked components of disability. Complete anesthesia leaves patients susceptible to occult injury, and facial sensation is an important component of interaction and activities of daily living. Sensory reconstruction is well established in the restoration of hand sensation; however, only one previous report proposed a surgical strategy for sensory nerve reconstruction of the face with use of nerve transfers. Nerve transfers, when used alone, have limited application because of their restricted arc of rotation in the face; extending their arc by adding nerve grafts greatly expands their utility. The following cases demonstrate the early results after V2 and V3 reconstruction with cross-face nerve grafts in three patients with acquired trigeminal nerve palsy. Cross-face nerve grafts using the sural nerve permit more proximal reconstruction of the infraorbital and mental nerves, which allows reinnervation of their entire cutaneous distribution. All patients demonstrated improved sensation in the reconstructed dermatomes, and no patients reported donor-site abnormalities. Cross-face nerve grafts result in minimal donor-site morbidity and are promising as a surgical strategy to address sensory deficits of the face. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, V.

Rapid and sensitive detection of respiratory virus infections for directed antiviral treatment using R-Mix cultures.

BACKGROUND: The development of new anti-influenza drugs has led to concerns regarding the impact on healthcare costs if they are used indiscriminately. Restricting their use to proven influenza virus infections has the potential to overcome costly inappropriate therapy. However, conventional culture (CC) does not generate results quickly enough to facilitate the timely initiation of treatment, and rapid detection tests have suboptimal sensitivity. We therefore investigated a new rapid culture system (R-Mix) that contains a mixture of two cell lines and detects respiratory viruses within 24 h. OBJECTIVES: To compare the analytical sensitivity of R-Mix with CC and rapid detection methods, for the detection of influenza and other respiratory viruses. To compare the clinical sensitivity of R-Mix with CC and direct antigen detection for the detection of respiratory viruses in primary and acute care settings. STUDY DESIGN: Stock cultures of influenza virus were titrated and tested by R-Mix, ZstatFlu and FLU OIA. Stock cultures of adenovirus and parainfluenza virus type 3 were titrated and tested by R-Mix and CC. Specimens, which had previously tested positive for influenza viruses, were titrated and tested by R-Mix and CC. In symptomatic patients, the majority of whom were from primary care settings, 124 sequential specimens were tested for influenza viruses by immunofluorescent direct antigen detection and R-Mix. A separate set of 111 sequential specimens, from various symptomatic patient groups, were tested for influenza viruses by CC and R-Mix. Additionally, in acute care patients being surveillance tested during periods of immunosuppression, 155 specimens were tested for respiratory viruses (influenza A and B, parainfluenza 1-3, adenovirus and respiratory syncytial virus (RSV)) by CC and R-Mix. RESULTS: With titrated stock cultures, R-Mix showed an analytical limit of detection of ten infectious virus particles per vial for influenza A, compared with 100,000 particles per test for FLU OIA and 1,000,000 for ZstatFlu. R-Mix also showed a 100-fold greater sensitivity for the detection of influenza A and equivalent sensitivity for the detection of influenza B when compared with CC in titrated known positive specimens. Further, it showed equivalent sensitivity to CC for the detection of adenovirus and parainfluenza virus type 3 in titrated stock cultures. Among prospective specimens from symptomatic patients, the sensitivity of R-Mix, CC and direct antigen detection tests (DAT) for influenza virus detection, was 100, 67 and 66%, respectively, and the specificity was 100, 100 and 98%, respectively. In surveillance specimens from immunosuppressed patients, the sensitivities of R-Mix and CC for respiratory virus detection were equivalent. Moreover, R-Mix results were available within 24 h, and by altering the antibody staining reagents either influenza viruses, or all seven major respiratory viruses, could be detected and distinguished in a single test. CONCLUSIONS: R-Mix is a simple, rapid and sensitive system for the detection of influenza viruses that facilitates the restriction of antiviral drugs to patients with culture-confirmed infections.

Degradation of Quillaja saponaria Molina saponins: loss of the protective effects of a herpes simplex virus 1 subunit vaccine.

Quillaja saponins (Q. saponins) are readily hydrolyzed at neutral pH to yield degraded deacylated saponins (DS-saponins). Degradation of Q. saponins resulted in some reduction of their capacity to elicit IgG1, IgG2a and IgG2b isotypes against the highly immunogenic envelope glycoprotein D (gD) from herpes simplex virus, type 1 (HSV-1). Addition to gD of a dose of DS-saponins tenfold higher than the original Q. saponins dose stimulated lower IgG2a and IgG2b titers than those obtained with gD alone or combined with native saponins. However, the IgG1 response was somewhat similar in all the groups. In contrast, Q. saponins' deacylation resulted in a significant reduction in both the production of HSV-1 neutralizing antibodies and survival rates after viral challenge. Vaccination with gD alone did not protect mice against a lethal challenge with HSV-1, while the addition of Q. saponins to gD resulted in protection against HSV-1. Vaccines containing partially deacylated saponins yielded lower survival rates, while vaccines containing DS-saponins did not protect mice against HSV-1. Increasing the dose of DS-saponins tenfold resulted in a marginal increase in protection. These results show that degradation of Q. saponins during storage can have a deleterious effect on vaccines' efficacies.

Model-based navigation of left and right ventricular leads to optimal targets for cardiac resynchronization therapy: a single-center feasibility study.

BACKGROUND: Left ventricular (LV) and right ventricular pacing site characteristics have been shown to influence response to cardiac resynchronization therapy (CRT). This study aimed to determine the clinical feasibility of image-guided lead delivery using a 3-dimensional navigational model displaying both LV and right ventricular (RV) pacing targets. Serial echocardiographic measures of clinical response and procedural metrics were evaluated. METHODS AND RESULTS: Thirty-one consecutive patients underwent preimplant cardiac MRI with the generation of a 3-dimensional navigational model depicting optimal segmental targets for LV and RV leads. Lead delivery was guided by the model in matched views to intraprocedural fluoroscopy. Blinded assessment of final lead tip location was performed from postprocedural cardiac computed tomography. Clinical and LV remodeling response criteria were assessed at baseline, 3 months, and 6 months using a 6-minute hall walk, quality of life questionnaire, and echocardiography. Mean age and LV ejection fraction was 66 ± 8 years and 26 ± 8%, respectively. LV leads were successfully delivered to a target or adjacent segment in 30 of 31 patients (97%), 68% being nonposterolateral. RV leads were delivered to a target or adjacent segment in 30 of 31 patients (97%), 26% being nonapical. Twenty-three patients (74%) met standard criteria for response (LV end-systolic volume reduction ≥ 15%), 18 patients (58%) for super-response (LV end-systolic volume reduction ≥ 30%). LV ejection fraction improved at 6 months (31 ± 8 versus 26 ± 8%, P=0.04). CONCLUSIONS: This study demonstrates clinical feasibility of dual cardiac resynchronization therapy lead delivery to optimal targets using a 3-dimensional navigational model. High procedural success, acceptable procedural times, and a low rate of early procedural complications were observed. CLINICAL TRIAL REGISTRATION URL: http://www.clinicaltrials.gov. Unique identifier: NCT01640769.

Myocardial fibrosis and quality of life in patients with non-ischemic cardiomyopathy: a cardiovascular magnetic resonance imaging study.

Myocardial fibrosis (MF) is a common pathophysiologic endpoint in non-ischemic cardiomyopathy and may be identified by late gadolinium enhancement (LGE) MRI. While associated with future cardiovascular events in hypertrophic cardiomyopathy (HCM) and dilated cardiomyopathy (DCM) the influence of MF on interim quality of life (QOL) has not been explored. In this study we investigate for associations between MF and validated indices of QOL in patients with HCM and DCM. Ninety-eight patients with known cardiomyopathy (n = 56-HCM/n = 42-DCM) underwent LGE-MRI in addition to standardized testing for QOL using the disease-specific Minnesota Living With Heart Failure (MLWHF) and the generic SF-12 questionnaires. LGE-MRI images were blindly analyzed for the presence and volume of MF using validated techniques. All analyses were stratified according to cardiomyopathy sub-type. The mean age of the population was 56.8 ± 12.9 years. MF was identified in 82 % of patients with HCM and 74 % of patients with DCM with respective mean MF burdens of 20.0 and 13.7 % of the left ventricular mass (p = 0.008). QOL scores for those with HCM or DCM, as assessed by both MLWHF and SF-12, were not significantly different between those with versus those without MF, and showed no association with MF burden by quantitative signal analysis. In this study we identified no association between QOL and MF burden by LGE-MRI in patients with HCM or DCM. Therefore, the severity of underlying myocardial tissue disease, a recognized substrate for ventricular arrhythmia, cannot and should not be inferred from the patient's symptom status or QOL.

Stress hypoperfusion and tissue injury in hypertrophic cardiomyopathy: spatial characterization using high-resolution 3-tesla magnetic resonance imaging.

BACKGROUND: Ischemia and tissue injury are common in patients with hypertrophic cardiomyopathy. Cardiovascular magnetic resonance imaging offers combined evaluations of each phenomenon at sufficiently high resolution to examine transmural spatial distribution. In this prospective cohort study, we examine the spatial distribution of stress perfusion abnormalities and tissue injury in patients with hypertrophic cardiomyopathy. METHODS AND RESULTS: One hundred consecutive patients with hypertrophic cardiomyopathy underwent cardiovascular magnetic resonance imaging. Cine, stress perfusion, late gadolinium enhancement, and T2-weighted imaging techniques were used. Each was spatially coregistered according to predefined segmental and subsegmental models and was blindly analyzed for abnormalities using validated techniques. Spatial associations among stress perfusion, late gadolinium enhancement, and T2 imaging were made at segmental and subsegmental levels. Of the 100 patients studied, the phenotype was septal in 86 and apical in 14. Late gadolinium enhancement imaging was abnormal in 79 patients (79%). Eighty-six patients met prespecified safety criteria to undergo stress perfusion, and ischemia was identified in 46 patients (57%). T2 imaging was available in 81 patients and was abnormal in 19 (29%). The dominant distribution of all 3 findings was to segment with hypertrophy. Subsegmental analysis revealed geographic dominance of ischemia within the subendocardial zones. However, this zone was most commonly spared from late gadolinium enhancement and T2 abnormalities, typically seen in midwall and subepicardial zones. CONCLUSIONS: Inducible hypoperfusion is a common finding in hypertrophic cardiomyopathy and is typically identified within segments exhibiting imaging markers of tissue injury. However, the respective transmural dominance of these phenomena seems distinct. Alternate factors contributing to a regional susceptibility to tissue injury are deserving of further study.

Influence of pacing site characteristics on response to cardiac resynchronization therapy.

BACKGROUND: Transmural scar occupying left ventricular (LV) pacing regions has been associated with reduced response to cardiac resynchronization therapy (CRT). However, spatial influences of lead tip delivery relative to scar at both pacing sites remain poorly explored. This study evaluated scar distribution relative to LV and right ventricular (RV) lead tip placement through coregistration of late gadolinium enhancement MRI and cardiac computed tomographic (CT) findings. Influences on CRT response were assessed by serial echocardiography. METHODS AND RESULTS: Sixty patients receiving CRT underwent preimplant late gadolinium enhancement MRI, postimplant cardiac CT, and serial echocardiography. Blinded segmental evaluations of mechanical delay, percentage scar burden, and lead tip location were performed. Response to CRT was defined as a reduction in LV end-systolic volume ≥15% at 6 months. The mean age and LV ejection fraction were 64±9 years and 25±7%, respectively. Mean scar volume was higher among CRT nonresponders for both the LV (23±23% versus 8±14% [P=0.01]) and RV pacing regions (40±32% versus 24±30% [P=0.04]). Significant pacing region scar was identified in 13% of LV pacing regions and 37% of RV pacing regions. Absence of scar in both regions was associated with an 81% response rate compared with 55%, 25%, and 0%, respectively, when the RV, LV, or both pacing regions contained scar. LV pacing region dyssynchrony was not predictive of response. CONCLUSIONS: Myocardial scar occupying the LV pacing region is associated with nonresponse to CRT. Scar occupying the RV pacing region is encountered at higher frequency and seems to provide a more intermediate influence on CRT response.

HSV IgG antibody inhibits virus detection in CSF.

BACKGROUND: Before PCR testing of cerebrospinal fluid (CSF), laboratory diagnosis of herpes encephalitis (HSE) was based on virus isolation from brain biopsy. Viral isolation from CSF has limited clinical value due to low virus recovery; the cause for which has not been demonstrated. OBJECTIVE: To investigate the role of anti-HSV antibodies on recovery of HSV from CSF via cell culture. STUDY DESIGN: HSV-positive CSF samples were evaluated for their ability to neutralize HSV in cell culture. The presence of HSV-specific IgG and IgM antibodies were analyzed using HSV-infected cells. To identify whether HSV-specific IgG is the cause of viral inhibition, IgG was removed using anti-human IgG magnetic beads. Viral inhibition from CSF originating from asymptomatic patients was examined as a comparison. RESULTS: CSF from 13 patients with acute HSV CNS disease was analyzed. All displayed high levels of viral neutralization to both HSV-1 and HSV-2 regardless of the infecting subtype. Interestingly, all the CSF samples stained strongly for anti-IgG antibody but none for anti-IgM antibody. Removal of IgG from CSF eliminated the viral inhibitory activity. Neutralizing IgG antibody was also found to be common in CSF of most patients, even in the absence of HSV disease. CONCLUSIONS: Viral specific IgG is the major determinant of viral inhibition in CSF and prevents virus recovery in cell culture. In CSF from HSE un-infected patients, viral inhibitory IgG originates from circulating serum antibody and is commonly present in CSF. However, this inhibitory IgG is not protective for the development of HSV disease.

Validation of a novel modified wall motion score for estimation of left ventricular ejection fraction in ischemic and non-ischemic cardiomyopathy.

BACKGROUND: Visual determination of left ventricular ejection fraction (LVEF) by segmental scoring may be a practical alternative to volumetric analysis of cine magnetic resonance imaging (MRI). The accuracy and reproducibility of this approach for has not been described. The purpose of this study was to validate a novel segmental visual scoring method for LVEF estimation using cine MRI. METHODS: 362 patients with known or suspected cardiomyopathy were studied. A modified wall motion score (mWMS) was used to blindly score the wall motion of all cardiac segments from cine MRI imaging. The same datasets were subjected to blinded volumetric analysis using endocardial contour tracing. The population was then separated into a model cohort (N=181) and validation cohort (N=181), with the former used to derive a regression equation of mWMS versus true volumetric LVEF. The validation cohort was then used to test the accuracy of this regression model to estimate the true LVEF from a visually determined mWMS. Reproducibility testing of mWMS scoring was performed upon a randomly selected sample of 20 cases. RESULTS: The regression equation relating mWMS to true LVEF in the model cohort was: LVEF=54.23-0.5761×mWMS. In the validation cohort this equation produced a strong correlation between mWMS-derived LVEF and true volumetric LVEF (r=0.89). Bland and Altman analysis showed no systematic bias in the LVEF estimated using the mWMS (-0.3231%, 95% limits of agreement -12.22% to 11.58%). Inter-observer and intra-observer reproducibility was excellent (r=0.93 and 0.97, respectively). CONCLUSION: The mWMS is a practical tool for reporting regional wall motion and provides reproducible estimates of LVEF from cine MRI.

The shift-invariant discrete wavelet transform and application to speech waveform analysis.

The discrete wavelet transform may be used as a signal-processing tool for visualization and analysis of nonstationary, time-sampled waveforms. The highly desirable property of shift invariance can be obtained at the cost of a moderate increase in computational complexity, and accepting a least-squares inverse (pseudoinverse) in place of a true inverse. A new algorithm for the pseudoinverse of the shift-invariant transform that is easier to implement in array-oriented scripting languages than existing algorithms is presented together with self-contained proofs. Representing only one of the many and varied potential applications, a recorded speech waveform illustrates the benefits of shift invariance with pseudoinvertibility. Visualization shows the glottal modulation of vowel formants and frication noise, revealing secondary glottal pulses and other waveform irregularities. Additionally, performing sound waveform editing operations (i.e., cutting and pasting sections) on the shift-invariant wavelet representation automatically produces quiet, click-free section boundaries in the resulting sound. The capabilities of this wavelet-domain editing technique are demonstrated by changing the rate of a recorded spoken word. Individual pitch periods are repeated to obtain a half-speed result, and alternate individual pitch periods are removed to obtain a double-speed result. The original pitch and formant frequencies are preserved. In informal listening tests, the results are clear and understandable.

Discovery of novel human and animal cells infected by the severe acute respiratory syndrome coronavirus by replication-specific multiplex reverse transcription-PCR.

The severe acute respiratory syndrome coronavirus (SARS-CoV) is the causative agent of the recent outbreak of severe acute respiratory syndrome. VeroE6 cells, fetal rhesus monkey kidney cells, and human peripheral blood mononuclear cells were the only cells known to be susceptible to SARS-CoV. We developed a multiplex reverse transcription-PCR assay to analyze the susceptibility of cells derived from a variety of tissues and species to SARS-CoV. Additionally, productive infection was determined by titration of cellular supernatants. Cells derived from three species of monkey were susceptible to SARS-CoV. However, the levels of SARS-CoV produced differed by 4 log(10). Mink lung epithelial cells (Mv1Lu) and R-Mix, a mixed monolayer of human lung-derived cells (A549) and mink lung-derived cells (Mv1Lu), are used by diagnostic laboratories to detect respiratory viruses (e.g., influenza virus); they were also infected with SARS-CoV, indicating that the practices of diagnostic laboratories should be examined to ensure appropriate biosafety precautions. Mv1Lu cells produce little SARS-CoV compared to that produced by VeroE6 cells, which indicates that they are a safer alternative for SARS-CoV diagnostics. Evaluation of cells permissive to other coronaviruses indicated that these cell types are not infected by SARS-CoV, providing additional evidence that SARS-CoV binds an alternative receptor. Analysis of human cells derived from lung, kidney, liver, and intestine led to the discovery that human cell lines were productively infected by SARS-CoV. This study identifies new cell lines that may be used for SARS-CoV diagnostics and/or basic research. Our data and other in vivo studies indicate that SARS-CoV has a wide host range, suggesting that the cellular receptor(s) utilized by SARS-CoV is highly conserved and is expressed by a variety of tissues.

ELVIRA HSV, a yield reduction assay for rapid herpes simplex virus susceptibility testing.

A colorimetric yield reduction assay, ELVIRA (enzyme-linked virus inhibitor reporter assay) HSV, was developed to determine the antiviral drug susceptibilities of herpes simplex virus (HSV). It uses an HSV-inducible reporter cell line. This simple and rapid assay has an objective readout, low inoculum size, and good reproducibility. The results correlate well with those of the plaque reduction assay.