A highly reproducible method for the measurement of [6-O-methyl-11 C]diprenorphine and its radio-metabolites based on solid-phase extraction and radio-high-pressure liquid chromatography.

Described here is a method for the measurement of the radio-metabolites of the positron emission tomography radiotracer [6-O-methyl-11 C]diprenorphine ([11 C]diprenorphine) using in-line solid-phase extraction (SPE) combined with radio-high-pressure liquid chromatography analysis. We believe that this method offers a reliable and reproducible approach to [11 C]diprenorphine metabolite analysis. In addition, different SPE stationary phases are assessed for their efficiency for loading, retention and elution of the parent molecule and its metabolites. Having assessed C4, phenyl and C18 stationary phase, we concluded that a C18 SPE was optimal for our method. Finally, in silico predictions of diprenorphine metabolism were compared with in vivo metabolism of [11 C]diprenorphine induced by hepatic microsomal digestion and analysed by matrix-assisted laser desorption/ionisation mass spectrometry. It was found that there was a high degree of agreement between the two methods and in particular the formation of the diprenorphine-3-glucuronide metabolite.

Multiscale 3D Curvature Analysis of Processed Surface Textures of Aluminum Alloy 6061 T6.

The objectives of this paper are to demonstrate the viability, and to validate, in part, a multiscale method for calculating curvature tensors on measured surface topographies with two different methods of specifying the scale. The curvature tensors are calculated as functions of scale, i.e., size, and position from a regular, orthogonal array of measured heights. Multiscale characterization of curvature is important because, like slope and area, it changes with the scale of observation, or calculation, on irregular surfaces. Curvatures can be indicative of the topographically dependent behavior of a surface and, in turn, curvatures are influenced by the processing and use of the surface. Curvatures of surface topographies have not been well- characterized yet. Curvature has been used for calculations in contact mechanics and for the evaluation of cutting edges. Manufactured surfaces are studied for further validation of the calculation method because they provide certain expectations for curvatures, which depend on scale and the degree of curvature. To study a range of curvatures on manufactured surfaces, square edges are machined and honed, then rounded progressively by mass finishing; additionally, a set of surfaces was made by turning with different feeds. Topographic measurements are made with a scanning laser confocal microscope. The calculations use vectors, normal to the measured surface, which are calculated first, then the eigenvalue problem is solved for the curvature tensor. Plots of principal curvatures as a function of position and scale are presented. Statistical analyses show expected interactions between curvature and these manufacturing processes.

'I didn't even know boys could get the vaccine': Parents' reasons for human papillomavirus (HPV) vaccination decision making for their sons.

OBJECTIVE: The study's objective was to examine parents' reasons for their decision to vaccinate their 9-16-year-old sons with the human papillomavirus (HPV) vaccine. METHODS: Using the precaution adoption process model (PAPM), parents were classified according to one of six stages of decision making: unaware, unengaged, undecided, decided not to vaccinate, decided to vaccinate, or vaccinated. Parents responded to an open-ended question: 'What would influence your decision to have your son vaccinated or not against HPV?' RESULTS: Three thousand one hundred and seventeen parents provided 2,874 interpretable narrative responses that were coded using thematic content analyses. The majority of parents were in the earlier precaution adoption process model stages, that is, unaware that the HPV vaccine could be given to boys (57.0%), unengaged (20.9%), or undecided (9.1%). Needing more information, vaccine cost, risks associated with vaccination, and wanting a doctor's recommendation influenced these earlier-staged parents' decisions. Parents who decided not to vaccinate their sons (6.8%) reported their decision was due to the risks, insufficient research, lack of confidence in vaccines, and/or no need for the vaccine (as their sons are not sexually active and/or too young). Parents who had decided to vaccinate their sons (5.0%) or who had vaccinated their sons (1.1%) reported that their decisions were based on protecting their sons' health and preventing disease. CONCLUSION: There are important differences in the factors that influence parents' decision depending on where they are along the decision-making trajectory. Assuring that parents are well informed about the importance of male vaccination, reducing vaccine cost, accurately communicating vaccine safety, and improving patient-provider communication may augment vaccine coverage and prevent HPV-associated cancers in Canada. Copyright © 2015 John Wiley & Sons, Ltd.

Biomechanical comparison of an all-soft suture anchor with a modified Broström-Gould suture repair for lateral ligament reconstruction.

BACKGROUND: Anatomic repair is indicated for patients who have recurrent lateral ankle instability despite nonoperative measures. HYPOTHESIS: There is no difference in repair stiffness, failure torque, or failure angle between specimens repaired with all-soft suture anchors versus the modified Broström-Gould technique with sutures only. STUDY DESIGN: Controlled laboratory study. METHODS: In 10 matched pairs of human cadaveric ankles, the anterior talofibular ligament (ATFL) was incised from its origin on the fibula. After randomization, 1 ankle was repaired to its anatomic insertion using two 1.4-mm JuggerKnot all-soft suture anchors; the other ankle was repaired with a modified Broström-Gould technique using 2-0 FiberWire. All were augmented using the inferior extensor retinaculum. All ankles were mounted to the testing machine in 20° of plantar flexion and 15° of internal rotation and loaded to failure after the repair. Stiffness, failure torque, and failure angle were recorded and compared using a paired Student t test with a significance level set at P < .05. RESULTS: There was no significant difference in failure torque, failure angle, or stiffness. No anchors pulled out of bone. The primary mode of failure was pulling through the ATFL tissue. CONCLUSION: There was no statistical difference in strength or stiffness between a 1.4-mm all-soft suture anchor and a modified Broström-Gould repair with 2-0 FiberWire. The primary mode of failure was at the tissue level rather than knot failure or anchor pullout. CLINICAL RELEVANCE: The particular implant choice (suture only, tunnel, anchor) in repairing the lateral ligament complex may not be as important as the time to biological healing. The suture-only construct as described in the Broström-Gould repair was as strong as all-soft suture anchors, and the majority of the ankles failed at the tissue level. For those surgeons whose preference is to use anchor repair, this novel all-soft suture anchor may be an alternative to other larger anchors, as none failed by pullout.

When the brain expects pain: common neural responses to pain anticipation are related to clinical pain and distress in fibromyalgia and osteoarthritis.

Supraspinal processes in humans can have a top-down enhancing effect on nociceptive processing in the brain and spinal cord. Studies have begun to suggest that such influences occur in conditions such as fibromyalgia (FM), but it is not clear whether this is unique to FM pain or common to other forms of chronic pain, such as that associated with osteoarthritis (OA). We assessed top-down processes by measuring anticipation-evoked potentials and their estimated sources, just prior (< 500 ms) to laser heat pain stimulation, in 16 patients with FM, 16 patients with OA and 15 healthy participants, by using whole-brain statistical parametric mapping. Clinical pain and psychological coping factors (pain catastrophizing, anxiety, and depression) were well matched between the patient groups, such that these did not confound our comparisons between FM and OA patients. For the same level of heat pain, insula activity was significantly higher in FM patients than in the other two groups during anticipation, and correlated with the intensity and extent of reported clinical pain. However, the same anticipatory insula activity also correlated with OA pain, and with the number of tender points across the two patient groups, suggesting common central mechanisms of tenderness. Activation in the dorsolateral prefrontal cortex was reduced during anticipation in both patient groups, and was related to less effective psychological coping. Our findings suggest common neural correlates of pain and tenderness in FM and OA that are enhanced in FM but not unique to this condition.

Prevalence of abnormal patellofemoral congruence in elite American football players and association with quadriceps isokinetic testing.

Abnormal patellofemoral joint alignment has been discussed as a potential risk factor for patellofemoral disorders and can impact the longevity of any elite athlete's career. The prevalence of abnormal patellofemoral congruence in elite American football athletes is similar to the general population and does not have a relationship with quadriceps isokinetic testing. A total of 125 athletes (220 knees) from the 2011 National Football League (NFL) Combine database who had radiographic and isokinetic studies were reviewed. Congruence angles (CA) and lateral patellofemoral angles (LPA) were calculated on a Merchant radiographic view. Isokinetic testing was used to determine quadriceps-to-hamstring strength (Q/H) ratio and side-to-side deficits. The relationships between abnormal CA and LPA with Q/H ratios, side-to-side deficits, and body mass index (BMI) were examined in separate logistic regression models. A Chi-square test was used to examine the association between CA and player position. Of all, 26.8% of the knees (95% CI: 21.1-33.2%) had an abnormal CA. Knees with normal CA (n = 161) did not significantly differ from those with an abnormal CA (n = 59) in Q/H ratios (mean: 0.699 vs. 0.728, p = 0.19) or side-to-side quadriceps deficits (mean: 4.0 vs. 1.24, p = 0.45). For each point increase in BMI, the odds ratio (OR) of abnormal congruence increased by 11.4% (p = 0.002). Of all the knees, 4.1% (95% CI: 1.9-7.6%) had an abnormal LPA, and this was not associated with Q/H ratios (p = 0.13). For each point increase in BMI, the odds of abnormal LPA increased by 16% (p = 0.036). CA abnormality had much higher odds of having an abnormal LPA (OR: 5.96, p = 0.014). We found that abnormal patellofemoral radiographic alignment in elite American football players is relatively common and there was no association with isokinetic testing.

ACL reconstruction in patients aged 40 years and older: a systematic review and introduction of a new methodology score for ACL studies.

BACKGROUND: Treatment of the anterior cruciate ligament (ACL)-deficient knee in older patients remains a core debate. PURPOSE: To perform a systematic review of studies that assessed outcomes in patients aged 40 years and older treated with ACL reconstruction and to provide a new methodological scoring system that is directed at critical assessment of studies evaluating ACL surgical outcomes: the ACL Methodology Score (AMS). STUDY DESIGN: Systematic review. METHODS: A comprehensive literature search was performed from 1995 to 2012 using MEDLINE, EMBASE, and Scopus. Inclusion criteria for studies were primary ACL injury, patient age of 40 years and older, and mean follow-up of at least 21 months after reconstruction. Nineteen studies met the inclusion criteria from the 371 abstracts from MEDLINE and 880 abstracts from Scopus. Clinical outcomes (International Knee Documentation Committee [IKDC], Lysholm, and Tegner activity scores), joint stability measures (Lachman test, pivot-shift test, and instrumented knee arthrometer assessment), graft type, complications, and reported chondral or meniscal injury were evaluated in this review. A new methodology scoring system was developed to be specific at critically analyzing ACL outcome studies and used to examine each study design. RESULTS: Nineteen studies describing 627 patients (632 knees; mean age, 49.0 years; range, 42.6-60.0 years) were included in the review. The mean time to surgery was 32.0 months (range, 2.9-88.0 months), with a mean follow-up of 40.2 months (range, 21.0-114.0 months). The IKDC, Lysholm, and Tegner scores and knee laxity assessment indicated favorable results in the studies that reported these outcomes. Patients did not demonstrate a significant difference between graft types and functional outcome scores or stability assessment. The mean AMS was 43.9 ± 7.2 (range, 33.5-57.5). The level of evidence rating did not positively correlate with the AMS, which suggests that the new AMS system may be able to detect errors in methodology or reporting that may not be taken into account by the classic level of evidence rating. CONCLUSION: Patients aged 40 years and older with an ACL injury can have satisfactory outcomes after reconstruction. However, the quality of currently available data is still limited, such that further well-designed studies are needed to determine long-term efficacy and to better inform our patients with regard to expected outcomes.

Clinical benefits of intra-articular anakinra for arthrofibrosis.

Postoperative inflammation and stiffness, as well as the struggle to achieve full range of motion (ROM), following knee surgery is a significant clinical challenge. Interleukin-1 is a crucial mediator of the inflammatory response and development of pathological conditions leading to chronic inflammation. We hypothesized that intra-articular injection of intra-articular anakinra, an IL-1 antagonist, would result in sustained improvements of chronic refractory arthrofibrosis and limited arthrofibrosis of the knee joint. We retrospectively reviewed 8 patients who underwent injection of intra-articular anakinra, 200 mg. Four patients (3 women, 1 man) had intra-articular anakinra for treatment of chronic refractory arthrofibrosis, and 4 patients (4 women) had intra-articular anakinra for limited arthrofibrosis. All 4 of the refractory arthrofibrosis patients had failed conservative treatment with intensive physical therapy, corticosteroid injections, and anti-inflammatory medication. Three of the 4 patients had failed a prior manipulation under anesthesia with lysis of adhesions. All 4 reported improvement in ROM (10°-45°) and swelling, with 75% reporting improvement in pain. Seventy-five percent of these patients returned to prior activity level. All 4 of the limited arthrofibrosis also failed similar attempts at conservative treatment, and 2 of the 4 had failed a prior manipulation under anesthesia with lysis of adhesions. After intra-articular anakinra, all 4 reported improvement in ROM (20°-45°) and swelling, with 80% reporting improvement in pain. Seventy-five percent of these patients were able to return to prior activity level. We found intra-articular anakinra to be effective in this small cohort of patients with refractory arthrofibrosis and limited arthrofibrosis.

Psychophysical properties of low-frequency hearing: implications for perceiving speech and music via electric and acoustic stimulation.

We have investigated the psychophysical properties of low-frequency hearing, both before and after implantation, to see if we can account for the benefit to speech understanding and melody recognition of adding acoustic stimulation to electric stimulation. In this paper, we review our work and the work of others and describe preliminary results not previously published. We show (a) that it is possible to preserve normal or near-normal nonlinear cochlear processing in the implanted ear following electric and acoustic stimulation surgery - though this is not the typical outcome; (b) that although low-frequency frequency selectivity is generally disrupted following implantation, some degree of frequency selectivity can be preserved, and (c) that neither nonlinear cochlear processing nor frequency selectivity in the acoustic hearing ear is correlated with the gain in speech understanding afforded by combined electric and acoustic stimulation. In another set of experiments, we show that the value of preserving hearing in the implanted ear is best seen in complex listening environments in which binaural cues can play a role in perception.

Interrelationship of growth differentiation factor 9 and inhibin in early folliculogenesis and ovarian tumorigenesis in mice.

To investigate the interrelationship of inhibin alpha and growth differentiation factor 9 (GDF9) during early folliculogenesis, we generated mice lacking both inhibin alpha and GDF9. Our findings on these Inha Gdf9 double-mutant mice are as follows: 1). females develop ovarian tumors and a cachexia-like wasting syndrome, resembling mice lacking inhibin alpha alone. This indicates that the granulosa cells are competent to proliferate despite the lack of GDF9; 2). follicular development progresses to multiple-layer follicle stages before tumorigenesis. This demonstrates that the up-regulation of inhibin alpha in the Gdf9 knockout ovary directly prevents the proliferation of the granulosa cells at the primary follicle stage, an effect that is released in the absence of inhibin alpha; 3). a morphological theca forms around the preantral follicles with no detectable selective theca markers [i.e. 17alpha-hydroxylase (Cyp17), LH receptor (Lhr), and Kit]. These results indicate that the theca recruitment can occur independently of GDF9, but the differentiation of thecal cells is blocked; and 4). inhibin/activin subunits betaA, betaB, and Kit ligand (Kitl) mRNA are highly up-regulated, suggesting that the increased activins and KITL play functional roles in early folliculogenesis. Thus, GDF9 appears to function indirectly to regulate early granulosa cell proliferation and theca recruitment in vivo.

Zygote arrest 1 (Zar1) is an evolutionarily conserved gene expressed in vertebrate ovaries.

Zygote arrest 1 (ZAR1) is an ovary-specific maternal factor that plays essential roles during the oocyte-to-embryo transition. In mice, the Zar1 mRNA is detected as a 1.4-kilobase (kb) transcript that is synthesized exclusively in growing oocytes. To further understand the functions of ZAR1, we have cloned the orthologous Zar1 cDNA and/or genes for mouse, rat, human, frog, zebrafish, and pufferfish. The entire mouse Zar1 gene and a related pseudogene span approximately 4.0 kb, contain four exons, and map to adjacent loci on mouse chromosome 5. The human ZAR1 orthologous gene similarly consists of four exons and resides on human chromosome 4p12, which is syntenic with the mouse Zar1 chromosomal locus. Rat (Rattus norvegicus) and pufferfish (Fugu rubripes) Zar1 genes were recognized by database mining and deduced protein alignment analysis. The rat Zar1 gene also maps to a region that is syntenic with the mouse Zar1 gene locus on rat chromosome 14. Frog (Xenopus laevis) and zebrafish (Danio rerio) Zar1 orthologs were cloned by reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends analysis of ovarian mRNA. Unlike mouse and human, the frog Zar1 is detected in multiple tissues, including lung, muscle, and ovary. The Zar1 mRNA appears in the cytoplasm of oocytes and persists until the tailbud stage during frog embryogenesis. Mouse, rat, human, frog, zebrafish, and pufferfish Zar1 genes encode proteins of 361, 361, 424, 295, 329, and 320 amino acids, respectively, and share 50.8%-88.1% amino acid identity. Regions of the N-termini of these ZAR1 orthologs show high sequence identity among these various proteins. However, the C-terminal 103 amino acids of these proteins, encoded by exons 2-4, contain an atypical eight-cysteine Plant Homeo Domain motif and are highly conserved, sharing 80.6%-98.1% identity among these species. These findings suggest that the carboxyl-termini of these ZAR1 proteins contain an important functional domain that is conserved through vertebrate evolution and that may be necessary for normal female reproduction in the transition from oocyte to embryonic life.