Myocardial deformation imaging by 2D speckle tracking echocardiography for assessment of diastolic dysfunction in murine cardiopathology.

Diastolic dysfunction is increasingly identified as a key, early onset subclinical condition characterizing cardiopathologies of rising prevalence, including diabetic heart disease and heart failure with preserved ejection fraction (HFpEF). Diastolic dysfunction characterization has important prognostic value in management of disease outcomes. Validated tools for in vivo monitoring of diastolic function in rodent models of diabetes are required for progress in pre-clinical cardiology studies. 2D speckle tracking echocardiography has emerged as a powerful tool for evaluating cardiac wall deformation throughout the cardiac cycle. The aim of this study was to examine the applicability of 2D speckle tracking echocardiography for comprehensive global and regional assessment of diastolic function in a pre-clinical murine model of cardio-metabolic disease. Type 2 diabetes (T2D) was induced in C57Bl/6 male mice using a high fat high sugar dietary intervention for 20 weeks. Significant impairment in left ventricle peak diastolic strain rate was evident in longitudinal, radial and circumferential planes in T2D mice. Peak diastolic velocity was similarly impaired in the longitudinal and radial planes. Regional analysis of longitudinal peak diastolic strain rate revealed that the anterior free left ventricular wall is particularly susceptible to T2D-induced diastolic dysfunction. These findings provide a significant advance on characterization of diastolic dysfunction in a pre-clinical mouse model of cardiopathology and offer a comprehensive suite of benchmark values for future pre-clinical cardiology studies.

Treatment of Grade III Acromioclavicular Separations in Professional Baseball Pitchers: A Survey of Major League Baseball Team Physicians.

Despite advancements in surgical technique and understanding of throwing mechanics, controversy persists regarding the treatment of grade III acromioclavicular (AC) joint separations, particularly in throwing athletes. Twenty-eight major league baseball (MLB) orthopedic team physicians were surveyed to determine their definitive management of a grade III AC separation in the dominant arm of a professional baseball pitcher and their experience treating AC joint separations in starting pitchers and position players. Return-to-play outcomes were also evaluated. Twenty (71.4%) team physicians recommended nonoperative intervention compared to 8 (28.6%) who would have operated acutely. Eighteen (64.3%) team physicians had treated at least 1 professional pitcher with a grade III AC separation; 51 (77.3%) pitchers had been treated nonoperatively compared to 15 (22.7%) operatively. No difference was observed in the proportion of pitchers who returned to the same level of play (P = .54), had full, unrestricted range of motion (P = .23), or had full pain relief (P = .19) between the operatively and nonoperatively treated MLB pitchers. The majority (53.6%) of physicians would not include an injection if the injury was treated nonoperatively. Open coracoclavicular reconstruction (65.2%) was preferred for operative cases; 66.7% of surgeons would also include distal clavicle excision as an adjunct procedure. About 90% of physicians would return pitchers to throwing >12 weeks after surgery compared to after 4 to 6 weeks in nonoperatively treated cases. In conclusion, MLB team physicians preferred nonoperative management for an acute grade III AC joint separation in professional pitchers. If operative intervention is required, ligament reconstruction with adjunct distal clavicle excision were the most commonly performed procedures.

Species of Metarhizium anisopliae complex implicated in human infections: retrospective sequencing study.

OBJECTIVES: Fungi belonging to the Metarhizium anisopliae complex comprise ubiquitous arthropod pathogenic moulds used as mycopesticides. Rare cases of human infections due to M. anisopliae have been reported. We hypothesize misidentifications of fungal strains implicated in these cases or used in mycopesticides. METHODS: A review of the literature was conducted to identify previously published cases. We collected some of these previous described strains and reported new cases, and a French mycopesticide containing M. anisopliae. All identifications were performed based on elongation factor-1α gene sequencing. RESULTS: We report eight new cases of Metarhizium infection in humans (three from France and five from Australia). The strains isolated from these cases, and three others from already published cases and reported as M. anisopliae, were molecularly identified based on elongation factor-1α (Ef1-α) gene sequencing as follows: Metarhizium robertsii (six), Metarhizium guizhouense (three), Metarhizium brunneum (one) and Metarhizium pingshaense (one). CONCLUSIONS: In this study, we report new human cases of Metarhizium infections, and, based on Ef-1α gene sequencing, we demonstrate the misidentification of species in case reports. We also correct the species identification of a strain reported as M. anisopliae used in a commercially available mycopesticide. According to our results, none of the strains from the human infection reports reviewed belongs to the species M. anisopliae.

Targeting a New Safe Zone: A Step in the Development of Patient-Specific Component Positioning for Total Hip Arthroplasty.

Surgeons often target the Lewinnek zone, with its mean (SD) inclination of 40° (10°) and mean (SD) anteversion of 15° (10°), for acetabular orientation during total hip arthroplasty (THA). However, matching native anteversion (20°-25°) may achieve optimal stability. We conducted a study in a large single-surgeon patient cohort to determine the incidence of early postoperative dislocation with increased acetabular anteversion and the accuracy of imageless navigation in achieving target acetabular position. Soft-tissue repair through a posterolateral approach was performed in 553 THAs that met the inclusion criteria. Mean (SD) target acetabular orientation was 40° (10°) of inclination and 25° (10°) of anteversion. Software was used to measure acetabular positioning on postoperative radiographs. Incidence of dislocation within 6 months after surgery was determined. Mean (SD) inclination was 42.2° (4.9°), and mean (SD) anteversion was 23.9° (6.5°). Approximately 82% of cups were placed in the target zone. Variation in anteversion accounted for 67.3% of outliers. Only body mass index was associated with inclination outside the target range (P = .017), and only female sex was associated with anteversion outside the target range (P = .030). Six THAs (1.1%) experienced early dislocation, and 3 (0.54%) of these were revised for multiple dislocations. There was no relationship between dislocation and component placement in either the Lewinnek zone (P = .224) or the target zone (P = .287).

Accelerometer-based, portable navigation vs imageless, large-console computer-assisted navigation in total knee arthroplasty: a comparison of radiographic results.

Computer-assisted surgery (CAS) systems improve alignment accuracy in total knee arthroplasty (TKA) but have not been widely implemented. Eighty knees underwent TKA using an accelerometer-based, portable navigation device (KneeAlign 2; OrthAlign Inc, Aliso Viejo, California), and the radiographic results were compared with 80 knees performed using a large-console, imageless CAS system (AchieveCAS; Smith and Nephew, Memphis, Tennessee). In the KneeAlign 2 cohort, 92.5% of patients had an alignment within 3° of a neutral mechanical axis (vs 86.3% with AchieveCAS, P < .01), 96.2% had a tibial component alignment within 2° of perpendicular to tibial mechanical axis (vs 97.5% with AchieveCAS, P = .8), and 94.9% had a femoral component alignment within 2° of perpendicular to the femoral mechanical axis (vs 92.5% with AchieveCAS, P < .01). The mean tourniquet time in the KneeAlign 2 cohort was 48.1 ± 10.2 minutes vs 54.1 ± 10.5 minutes in the AchieveCAS cohort (P < .01). Accelerometer-based, portable navigation is as accurate as large-console, imageless CAS systems in TKA.

Outcome of a tobacco use cessation randomized trial with high-school students.

This study analyzed quantitative data on tobacco use and dependency for 3,589 high-school students, qualitative data for 448 students, and outcome data for a randomized trial comparing the efficacy of two cessation interventions and a control condition for 337 students. Data were collected from 1988 through 1992 in California and Illinois as part of a larger longitudinal study. Smokeless tobacco users, but not smokers, were more likely than controls to maintain cessation for 4 months: biochemically validated cessation at 4 months was 6.5% versus 3.2% for smokers and 14.3% versus 0.0% for smokeless tobacco users. Implications and limitations are discussed.

Protein-dependent transition states for ribonucleoprotein assembly.

Native folding and splicing by the Saccharomyces cerevisiae mitochondrial bI5 group I intron RNA is facilitated by both the S. cerevisiae CBP2 and Neurospora crassa CYT-18 protein cofactors. Both protein-bI5 RNA complexes splice at similar rates, suggesting that the RNA active site structure is similar in both ribonucleoproteins. In contrast, the two proteins assemble with the bI5 RNA by distinct mechanisms and bind opposing, but partially overlapping, sides of the group I intron catalytic core. Assembly with CBP2 is limited by a slow, unimolecular RNA folding step characterized by a negligible activation enthalpy. We show that assembly with CYT-18 shows four distinctive features. (1) CYT-18 binds stably to the bI5 RNA at the diffusion controlled limit, but assembly to a catalytically active RNA structure is still limited by RNA folding, as visualized directly using time-resolved footprinting. (2) This mechanism of rapid stable protein binding followed by subsequent assembly steps has a distinctive kinetic signature: the apparent ratio of k(off) to k(on), determined in a partitioning experiment, differs from the equilibrium K(d) by a large factor. (3) Assembly with CYT-18 is characterized by a large activation enthalpy, consistent with a rate limiting conformational rearrangement. (4) Because assembly from the kinetically trapped state is faster at elevated temperature, we can identify conditions where CYT-18 accelerates (catalyzes) bI5 RNA folding relative to assembly with CBP2.

Visualizing induced fit in early assembly of the human signal recognition particle.

Assembly of almost all ribonucleoprotein complexes involves induced fit in the RNA and, thus, formation of one or more intermediate states. In assembly of the human signal recognition particle (SRP), we show that SRP19 binding to SRP RNA involves obligatory intermediates. An apparent discrepancy exists between the ratio of dissociation and association rate constants, determined in a partitioning experiment, and the equilibrium binding constant; this kinetic signature reflects formation of a stable intermediate in assembly of the ribonucleoprotein complex. Assembly intermediates were observed directly by time-resolved footprinting. SRP19 binds rapidly to SRP RNA to form an initial labile, but structurally specific, encounter complex involving both helices III and IV. Two subsequent steps of structural consolidation yield the native RNA-protein interface. SRP19 binding stabilizes helix IV in the region recognized by SRP54, consistent with protein-protein cooperativity mediated in part by mutual recognition of similar RNA structures. This mechanism illustrates principles general to ribonucleoprotein assembly reactions that rely on recruitment of architectural RNA binding proteins.

A collapsed state functions to self-chaperone RNA folding into a native ribonucleoprotein complex.

Most large RNAs achieve their active, native structures only as complexes with one or more cofactor proteins. By varying the Mg(2+) concentration, the catalytic core of the bI5 group I intron RNA can be manipulated into one of three states, expanded, collapsed or native, or into balanced equilibria between these states. Under near-physiological conditions, the bI5 RNA folds rapidly to a collapsed but non-native state. Hydroxyl radical footprinting demonstrates that assembly with the CBP2 protein cofactor chases the RNA from the collapsed state to the native state. In contrast, CBP2 also binds to the RNA in the expanded state to form many non-native interactions. This structural picture is reinforced by functional splicing experiments showing that RNA in an expanded state forms a non-productive, kinetically trapped complex with CBP2. Thus, rapid folding to the collapsed state functions to self-chaperone bI5 RNA folding by preventing premature interaction with its protein cofactor. This productive, self-chaperoning role for RNA collapsed states may be especially important to avert misassembly of large multi-component RNA-protein machines in the cell.

van't Hoff enthalpies without baselines.

Analysis of thermal melting curves represents one important approach for evaluating protein stability and the consequences of amino acid substitution on protein structure. By use of the van't Hoff relationship, the differential melting curve can be robustly fit to only three parameters, two of which are the underlying physical constants of melting temperature (Tm) and van't Hoff enthalpy (deltaHvH). Calculated Tm and deltaHvH values are insensitive to the choice of pre- and post-transition baselines. Consequently, the method accurately computes Tm and deltaHvH for extremely truncated data sets, in the complete absence of baseline information, and for proteins with low melting temperatures, where the traditional direct approach routinely fails. Moreover, agreement between deltaHvH values obtained using points derived from pre- vs. post-transition data provide an independent method for detecting some classes of non-two-state transitions. Finally, fitting of the differential denaturation curve should prove useful for analysis of abbreviated data sets obtained from high throughput array analysis of protein stability.

Carcinoma of the uterine cervix: a 3D - CT analysis of dose to the internal, external and common iliac nodes in tandem and ovoid applications.

PURPOSE: To describe external, internal and common iliac dose rates estimated with 3D-computed tomography (CT) based dose calculations in tandem and ovoid brachytherapy. MATERIALS AND METHODS: Thirty patients with carcinoma of the uterine cervix received low dose rate brachytherapy with a CT-compatible Fletcher-Suit-Deldos device. A total of 36 implants were performed with axial CT images used to identify internal iliac, external iliac, and common iliac vessels. Dose rates on the surfaces of these vessels were calculated for the purpose of estimating the dose to their associated lymph nodes. RESULTS: In 22 out of 72 comparisons, point B overestimated the maximum dose with the external iliac nodes. In 21 out of 72 comparisons, point B overestimated the maximum dose with the internal iliac nodes. In all cases, Point B overestimated the minimum dose to the internal and external iliac nodal chains. CONCLUSION: It was found that Point B dose is similar to the maximum common iliac nodal dose. Patient to patient variability, of Point B dose, warrants further study of dose distributions to the nodal chains. The minimum dose to the external iliac nodal chain at the bifurcation of the nodal chains may provide a useful measure of 'pelvic side wall dose' and deserves further study to see if it can be correlated with pelvic side wall control and complications.

Tagging DNA mismatches by selective 2'-amine acylation.

BACKGROUND: Widespread characterization of genetic variation and disease at the gene-sequence level has inaugurated a new era in human biology. Techniques for the molecular analysis of these variations and their linkage with measurable phenotypes will profoundly affect diverse fields of biological chemistry and biology. RESULTS: A chemical tagging method has been developed to detect point mutations and other defects in nucleic acid sequences. The method employs oligodeoxynucleotide probes in which one 2'-ribose position (-H) is substituted with an amine (-NH(2)) group. 2'-Amine-substituted nucleotides are specifically acylated by succinimidyl esters to form a 2'-amide product. The mutation detection method exploits our observation that 2'-amine groups at the site of a mismatch are acylated more rapidly than amine substitutions at base-paired nucleotides. 2'-Amine acylation is governed primarily by local, rather than global, differences in nucleotide dynamics, such that site-specific tagging of DNA mismatches does not require discriminatory hybridization conditions to be determined. CONCLUSIONS: 2'-Amine mismatch tagging offers an approach for chemically interrogating the base-paired state of individual nucleotides in a hybridized duplex and for quantifying nucleicacid hybridization with single-base specificity.

A collapsed non-native RNA folding state.

At physiological Mg2+ concentrations, the catalytic core of the bI5 group I intron does not fold into its native structure. In contrast, as judged by the global size, this RNA undergoes structural collapse at Mg 2+ concentrations much lower than required to drive folding of the RNA completely to the native state. The bI5 RNA therefore exists in equilibrium between expanded and collapsed non-native states. The activation energy of RNA folding from the collapsed state to the native state is negligible and the reaction is not accelerated by the addition of urea. This collapsed state is thus distinct from the kinetic traps observed during folding of other large RNAs. The collapsed non-native state forms readily in the case of bI5 RNA and may exist generically prior to assembly of other ribonucleoprotein holoenzymes, such as the ribosome.

Method for determining photonuclear production of radioisotopes using high-energy electron beams.

Linear accelerators can produce electrons at high power and energy. These electrons can be targeted at materials to produce radionuclides. Monte Carlo simulation is used to follow the path of the linac electrons (15-35 MeV) through materials. The production of photons and their passage through the material is modeled. The method of using this Monte Carlo information to calculate activation in the materials is presented. It is found that kilowatt power levels can produce mCi amounts of radioactivity in minutes. This work permits systematic evaluation of the potential for designing linear accelerators for in-house production of radionuclides.

Frequency of the codon 807 mutation in the cGMP phosphodiesterase beta-subunit gene in Irish setters and other dog breeds with hereditary retinal degeneration.

Rod-cone dysplasia 1 (rcd1) in Irish setters is caused by a nonsense mutation in the cGMP phosphodiesterase beta-subunit gene (PDE6B). We examined the frequency of the mutant allele in the Irish setter population and determined if the defect is present in dogs of other breeds which are affected with other inherited photoreceptor diseases. Between 1994 and 1997, samples were obtained from 436 clinically normal Irish setters, a red wolf, and dogs from 23 different breeds. The mutation in codon 807 of PDE6B was detected in genomic DNA by heteroduplex analysis, allele-specific PCR, or restriction enzyme digestion. Of the 436 samples from clinically normal setters, 34 contained the mutation in one of the two PDE6B alleles (carrier rate = 7.8%). In contrast, the same mutation was not found in the red wolf or dogs of other breeds affected with PRA or inherited photoreceptor diseases. The high percentage of tested carriers, however, is not representative of the number of carriers in the population since some dogs tested were closely related and did not represent a random sample of the Irish setter breed.

The thresholds of change model: an approach to analyzing stages of change data.

Stage models are prominent in research describing health behavior change. Since stage models often propose that different factors have varying influences on membership in the different stage, statistical methods that can estimate the thresholds that separate the stages and the relative value of variables in influencing these thresholds are useful. This article describes use of a "thresholds of change" model for analyzing the thresholds separating stages and specifically for examining the effects of explanatory variables on these thresholds using a generalization of an ordinal logistic (or probit) regression model. Data from a skin cancer prevention study (N=3,185) in which participants were grouped into three stages for sunscreen use (precontemplation, contemplation, and action) are used to illustrate the Thresholds of Change Model. For this example, two thresholds exist: a contemplation (between precontemplation and contemplation) and an action threshold (between contemplation and action). Variables examined include gender, skin type, perceived susceptibility to sunburn, worry about skin cancer, and sun protection self-efficacy. We examine models that assume that the effects of these variables are the same across thresholds, and then allow the effects of these variables to vary across thresholds. Results indicate that perceived susceptibility has an equal effect on both thresholds, but that worry and self-efficacy have differential effects: worry exerts a greater influence on the contemplation threshold, whereas self-efficacy has a significantly stronger effect on the action threshold. Gender also has a stronger effect on the action threshold; males were less likely to be classified in the action stage than females. This analytic approach has broad applications to many types of stage data.

Production of radioisotopes by direct electron activation.

High-energy electrons bombarded on materials can induce radioactivity by either directly knocking out neutrons or by first converting a fraction of the electron kinetic energy into electromagnetic energy, with subsequent neutron emission induced by the photons produced. The purpose of this paper was to develop a calculation method for estimating neutron emission and radionuclide production by high-energy (15-25 MeV) electrons directly interacting with a nucleus. The reaction (e,n) is considered using the method of virtual photons. The cross section for electron bombardment of lead, tantalum, rhenium, and tungsten targets is calculated. The electron cross sections are roughly 100 times less than the corresponding photon cross sections. The cross section increases monotonically with incident energy. A traveling wave linear accelerator was used for a qualitative test of the magnitude and energy dependence of the calculated cross sections. Tantalum was bombarded with electrons and the resultant emission of neutrons was inferred from the induced activation of 180Ta. The energy dependence and magnitude of the calculated electron cross sections agree with experiment within experimental uncertainties. It is concluded that accurate estimates of electron activation via the direct process is possible.

Computed axial tomography tandem and ovoids (CATTO) dosimetry: three-dimensional assessment of bladder and rectal doses.

The purpose of this work is to compare bladder and rectal dose rates in brachytherapy for carcinoma of the cervix using two different dosimetry systems: traditional orthogonal radiograph-based dosimetry vs. computed axial tomography tandem and ovoids (CATTO) dosimetry. Twenty-two patients with carcinoma of the uterine cervix received the brachytherapy component of their radiotherapy with a computed-tomography compatible Fletcher-Suit-Delclos device. A total of 27 implants were performed. The average maximum bladder dose (Bmax) for the implants was 85.8 cGy/hr using the CATTO system as compared to 42.6 cGy/hr using traditional dosimetry, (P < 0.005). The average maximum rectal dose (R.) using the CATTO system was 59.2 cGy/hr as compared with 46.3 cGy/hr using the traditional system (P < 0.05). The traditional methods for choosing points to determine bladder and rectal dose rates underestimated the true Bmax in all cases and the R. in most. Based on the complication rates published in the literature, it is likely that the maximum tolerance dose of both the rectum and bladder, but especially the bladder, is higher than previously thought.

Monte Carlo dose calculations for a new ovoid shield system for carcinoma of the uterine cervix.

The dose distribution for an ovoid with a new tungsten shielding design was determined using Monte Carlo simulation. Standard Cesium-137 tube sources, tungsten shielding, and aluminum ovoid applicator were each modeled as a collection of solid objects. Dose was calculated in planes above, below, in front of, and on the sides of the colpostat. The Monte Carlo results were compared with the results from a parametrized calculation algorithm and good agreement was obtained. The dose distribution matrix derived from the parametrized algorithm can be used for clinical treatment planning.