Extracellular matrix protein composition dynamically changes during murine forelimb development.

The extracellular matrix (ECM) is an integral part of multicellular organisms, connecting different cell layers and tissue types. During morphogenesis and growth, tissues undergo substantial reorganization. While it is intuitive that the ECM remodels in concert, little is known regarding how matrix composition and organization change during development. Here, we quantified ECM protein dynamics in the murine forelimb during appendicular musculoskeletal morphogenesis (embryonic days 11.5-14.5) using tissue fractionation, bioorthogonal non-canonical amino acid tagging, and mass spectrometry. Our analyses indicated that ECM protein (matrisome) composition in the embryonic forelimb changed as a function of development and growth, was distinct from other developing organs (brain), and was altered in a model of disease (osteogenesis imperfecta murine). Additionally, the tissue distribution for select matrisome was assessed via immunohistochemistry in the wild-type embryonic and postnatal musculoskeletal system. This resource will guide future research investigating the role of the matrisome during complex tissue development.

A method for defining tissue injury criteria reveals that ligament deformation thresholds are multimodal.

Soft tissue injuries (such as ligament, tendon, and meniscus tears) are the result of extracellular matrix damage from excessive tissue stretching. Deformation thresholds for soft tissues, however, remain largely unknown due to a lack of methods that can measure and compare the spatially heterogeneous damage and deformation that occurs in these materials. Here, we propose a full-field method for defining tissue injury criteria: multimodal strain limits for biological tissues analogous to yield criteria that exist for crystalline materials. Specifically, we developed a method for defining strain thresholds for mechanically-driven fibrillar collagen denaturation in soft tissues, using regional multimodal deformation and damage data. We established this new method using the murine medial collateral ligament (MCL) as our model tissue. Our findings revealed that multiple modes of deformation contribute to collagen denaturation in the murine MCL, contrary to the common assumption that collagen damage is driven only by strain in the direction of fibers. Remarkably, hydrostatic strain (computed here with an assumption of plane strain) was the best predictor of mechanically-driven collagen denaturation in ligament tissue, suggesting crosslink-mediated stress transfer plays a role in molecular damage accumulation. This work demonstrates that collagen denaturation can be driven by multiple modes of deformation and provides a method for defining deformation thresholds, or injury criteria, from spatially heterogeneous data. STATEMENT OF SIGNIFICANCE: Understanding the mechanics of soft tissue injuries is crucial for the development of new technology for injury detection, prevention, and treatment.  Yet, tissue-level deformation thresholds for injury are unknown, due to a lack of methods that combine full-field measurements of multimodal deformation and damage in mechanically loaded soft tissues. Here, we propose a method for defining tissue injury criteria: multimodal strain thresholds for biological tissues. Our findings reveal that multiple modes of deformation contribute to collagen denaturation, contrary to the common assumption that collagen damage is driven by strain in the fiber direction alone. The method will inform the development of new mechanics-based diagnostic imaging, improve computational modeling of injury, and be employed to study the role of tissue composition in injury susceptibility.

Isolation and Characterization of the Murine Uterosacral Ligaments and Pelvic Floor Organs.

Pelvic organ prolapse (POP) is a condition that affects the integrity, structure, and mechanical support of the pelvic floor. The organs in the pelvic floor are supported by different anatomical structures, including muscles, ligaments, and pelvic fascia. The uterosacral ligament (USL) is a critical load-bearing structure, and injury to the USL results in a higher risk of developing POP. The present protocol describes the dissection of murine USLs and the pelvic floor organs alongside the acquisition of unique data on the USL biochemical composition and function using Raman spectroscopy and the evaluation of mechanical behavior. Mice are an invaluable model for preclinical research, but dissecting the murine USL is a difficult and intricate process. This procedure presents an approach to guide the dissection of murine pelvic floor tissues, including the USL, to enable multiple assessments and characterization. This work aims to aid the dissection of pelvic floor tissues by basic scientists and engineers, thus expanding the accessibility of research on the USL and pelvic floor conditions and the preclinical study of women's health using mouse models.

A method for defining tissue injury criteria reveals ligament deformation thresholds are multimodal.

Soft tissue injuries (such as ligament, tendon, and meniscus tears) are the result of extracellular matrix damage from excessive tissue stretching. Deformation thresholds for soft tissues, however, remain largely unknown due to a lack of methods that can measure and compare the spatially heterogeneous damage and deformation that occurs in these materials. Here, we propose a method for defining tissue injury criteria : multimodal strain limits for biological tissues analogous to yield criteria that exist for crystalline materials. Specifically, we developed a method for defining injury criteria for mechanically-driven fibrillar collagen denaturation in soft tissues, using regional multimodal deformation and damage data. We established this new method using the murine medial collateral ligament (MCL) as our model tissue. Our findings revealed that multiple modes of deformation contribute to collagen denaturation in the murine MCL, contrary to the common assumption that collagen damage is driven by strain in the fiber direction alone. Remarkably, our results indicated that hydrostatic strain, or volumetric expansion, may be the best predictor of mechanically-driven collagen denaturation in ligament tissue, suggesting crosslink-mediated stress transfer plays a role in molecular damage accumulation. This work demonstrates that collagen denaturation can be driven by multiple modes of deformation and provides a method for defining deformation thresholds, or injury criteria, from spatially heterogeneous data.

Hyperelastic characterization reveals proteoglycans drive the nanoscale strain-stiffening response in hyaline cartilage.

Degenerative diseases such as osteoarthritis (OA) result in deterioration of cartilage extracellular matrix (ECM) components, significantly compromising tissue function. For measurement of mechanical properties at micron resolution, atomic force microscopy (AFM) is a leading technique in biomaterials research, including in the study of OA. It is common practice to determine material properties by applying classical Hertzian contact theory to AFM data. However, errors are consequential because the application of a linear elastic contact model to tissue ignores the fact that soft materials exhibit nonlinear properties even at small strains, influencing the biological conclusions of clinically-relevant studies. Additionally, nonlinear material properties are not well characterized, limiting physiological relevance of Young's modulus. Here, we probe the ECM of hyaline cartilage with AFM and explore the application of Hertzian theory in comparison to five hyperelastic models: NeoHookean, Mooney-Rivlin, Arruda-Boyce, Fung, and Ogden. The Fung and Ogden models achieved the best fits of the data, but the Fung model demonstrated robust sensitivity during model validation, demonstrating its ideal application to cartilage ECM and potentially other connective tissues. To develop a biological understanding of the Fung nonlinear parameter, we selectively degraded ECM components to target collagens (purified collagenase), hyaluronan (bacterial hyaluronidase), and glycosaminoglycans (chondroitinase ABC). We found significant differences in both Fung parameters in response to enzymatic treatment, indicating that proteoglycans drive the nonlinear response of cartilage ECM, and validating biological relevance of these phenomenological parameters. Our findings add value to the biomechanics community of using two-parameter material models for microindentation of soft biomaterials.

High frame rate deformation analysis of knee cartilage by spiral dualMRI and relaxation mapping.

PURPOSE: Daily activities including walking impose high-frequency cyclic forces on cartilage and repetitive compressive deformation. Analyzing cartilage deformation during walking would provide spatial maps of displacement and strain and enable viscoelastic characterization, which may serve as imaging biomarkers for early cartilage degeneration when the damage is still reversible. However, the time-dependent biomechanics of cartilage is not well described, and how defects in the joint impact the viscoelastic response is unclear. METHODS: We used spiral acquisition with displacement-encoding MRI to quantify displacement and strain maps at a high frame rate (25 frames/s) in tibiofemoral joints. We also employed relaxometry methods (T1 , T1ρ , T2 , T2 *) on the cartilage. RESULTS: Normal and shear strains were concentrated on the bovine tibiofemoral contact area during loading, and the defected joint exhibited larger compressive strains. We also determined a positive correlation between the change of T1ρ in cartilage after cyclic loading and increased compressive strain on the defected joint. Viscoelastic behavior was quantified by the time-dependent displacement, where the damaged joint showed increased creep behavior compared to the intact joint. This technique was also successfully demonstrated on an in vivo human knee showing the gradual change of displacement during varus load. CONCLUSION: Our results indicate that spiral scanning with displacement encoding can quantitatively differentiate the damaged from intact joint using the strain and creep response. The viscoelastic response identified with this methodology could serve as biomarkers to detect defects in joints in vivo and facilitate the early diagnosis of joint diseases such as osteoarthritis.

Using Relational Training to Improve Performance During Acceptance and Commitment Training Sessions.

The current study investigated the effect of relational training on improving engagement during acceptance and commitment training (ACT) sessions that aimed to reduce maladaptive behavior. Three female children diagnosed with autism participated in the current research. The relational training protocol involved eight programs designed to improve participants' ability to engage in relational responding. A concurrent multiple-baseline across-participants design was used, and participants' maladaptive behavior during ACT sessions was recorded. Results indicated a decrease in maladaptive behavior for one of the three participants, with a medium to large treatment effect. A secondary measure also showed improvement in all three participants' performance during ACT sessions. The percentage of nonoverlapping data and Cohen's d suggest a small to moderate treatment effect. Implications for implementing ACT with individuals diagnosed with autism are discussed.

A Novel and Inexpensive Technique for High-Tension Tendon Clamping With Expandable Mesh Sleeving for In Vitro Foot Biomechanics Testing.

In vitro biomechanical testing is common in the field of orthopedics when novel devices are investigated prior to human trials. It is typically necessary to apply loads through tendons to simulate normal activities, such as walking during a foot and ankle study. However, attachment of tendons to linear actuators has proven challenging because of the tendency of clamps to either slip off or rupture the tendon. Various techniques have been utilized. Freeze clamping is generally accepted as the gold standard for very high load testing in excess of 3000 N, but is expensive, time-consuming, and requires significant ancillary equipment. Purely mechanical solutions such as metal jaw clamps, wire meshes, and others have been explored, but these techniques are either costly, have low load capacities, or have not proven to be reproducible. We have developed a novel tendon clamping technique that utilizes a slip-resistant polyester mesh sleeving that encases the tendon and is fixated at the bottom of the tendon/sleeve interaction with a giftbox suture. The loose end of the sleeving can then be tied in to the linear actuator or load cell apparatus using a timber hitch knot. The sleeving technique allows for loads of 2000-2500 N on the Achilles tendon, and is inexpensive, reproducible, and can be modified to apply loads to smaller tendons as well, though a length of tendon/sleeve overlap of at least 16 cm is required to reach maximum loads. This technique should assist researchers in integrating muscle forces into future biomechanical study designs.

Assessment of L5-S1 anterior lumbar interbody fusion stability in the setting of lengthening posterior instrumentation constructs: a cadaveric biomechanical study.

OBJECTIVE: Excessive stress and motion at the L5-S1 level can lead to degenerative changes, especially in patients with posterior instrumentation suprajacent to L5. Attention has turned to utilization of L5-S1 anterior lumbar interbody fusion (ALIF) to stabilize the lumbosacral junction. However, questions remain regarding the effectiveness of stand-alone ALIF in the setting of prior posterior instrumented fusions terminating at L5. The purpose of this study was to assess the biomechanical stability of an L5-S1 ALIF with increasing lengths of posterior thoracolumbar constructs. METHODS: Seven human cadaveric spines (T9-sacrum) were instrumented with pedicle screws from T10 to L5 and mounted to a 6 degrees-of-freedom robot. Posterior fusion construct lengths (T10-L5, T12-L5, L2-5, and L4-5) were instrumented to each specimen, and torque-fusion level relationships were determined for each construct in flexion-extension, axial rotation, and lateral bending. A stand-alone L5-S1 ALIF was then instrumented, and L5-S1 motion was measured as increasing pure moments (2 to 12 Nm) were applied. Motion reduction was calculated by comparing L5-S1 motion across the ALIF and non-ALIF states. RESULTS: The average motion at L5-S1 in axial rotation, flexion-extension, and lateral bending was assessed for each fusion construct with and without ALIF. After adding ALIF to a posterior fusion, L5-S1 motion was significantly reduced relative to the non-ALIF state in all but one fused surgical condition (p < 0.05). Longer fusions with ALIF produced larger L5-S1 motions, and in some cases resulted in motions higher than native state motion. CONCLUSIONS: Posterior fusion constructs up to L4-5 could be appropriately stabilized by a stand-alone L5-S1 ALIF when using a nominal threshold of 80% reduction in native motion as a potential positive indicator of fusion. The results of this study allow conclusions to be drawn from a biomechanical standpoint; however, the clinical implications of these data are not well defined. These findings, when taken in appropriate clinical context, can be used to better guide clinicians seeking to treat L5-S1 pathology in patients with prior posterior thoracolumbar constructs.

Evaluating stability of the craniovertebral junction after unilateral C1 lateral mass resection: implications for the direct lateral approach.

OBJECTIVE: The direct lateral approach is an alternative to the transoral or endonasal approaches to ventral epidural lesions at the lower craniocervical junction. In this study, the authors performed, to their knowledge, the first in vitro biomechanical evaluation of the craniovertebral junction after sequential unilateral C1 lateral mass resection. The authors hypothesized that partial resection of the lateral mass would not result in a significant increase in range of motion (ROM) and may not require internal stabilization. METHODS: The authors performed multidirectional in vitro ROM testing using a robotic spine testing system on 8 fresh cadaveric specimens. We evaluated ROM in 3 primary movements (axial rotation [AR], flexion/extension [FE], and lateral bending [LB]) and 4 coupled movements (AR+E, AR+F, LB + left AR, and LB + right AR). Testing was performed in the intact state, after C1 hemilaminectomy, and after sequential 25%, 50%, 75%, and 100% C1 lateral mass resection. RESULTS: There were no significant increases in occipital bone (Oc)-C1, C1-2, or Oc-C2 ROM after C1 hemilaminectomy and 25% lateral mass resection. After 50% resection, Oc-C1 AR ROM increased by 54.4% (p = 0.002), Oc LB ROM increased by 47.8% (p = 0.010), and Oc-C1 AR+E ROM increased by 65.8% (p < 0.001). Oc-C2 FE ROM increased by 7.2% (p = 0.016) after 50% resection; 75% and 100% lateral mass resection resulted in further increases in ROM. CONCLUSIONS: In this cadaveric biomechanical study, the authors found that unilateral C1 hemilaminectomy and 25% resection of the C1 lateral mass did not result in significant biomechanical instability at the occipitocervical junction, and 50% resection led to significant increases in Oc-C2 ROM. This is the first biomechanical study of lateral mass resection, and future studies can serve to validate these findings.

Tissue-specific parameters for the design of ECM-mimetic biomaterials.

The extracellular matrix (ECM) is a complex network of biomolecules that mechanically and biochemically directs cell behavior and is crucial for maintaining tissue function and health. The heterogeneous organization and composition of the ECM varies within and between tissue types, directing mechanics, aiding in cell-cell communication, and facilitating tissue assembly and reassembly during development, injury and disease. As technologies like 3D printing rapidly advance, researchers are better able to recapitulate in vivo tissue properties in vitro; however, tissue-specific variations in ECM composition and organization are not given enough consideration. This is in part due to a lack of information regarding how the ECM of many tissues varies in both homeostatic and diseased states. To address this gap, we describe the components and organization of the ECM, and provide examples for different tissues at various states of disease. While many aspects of ECM biology remain unknown, our goal is to highlight the complexity of various tissues and inspire engineers to incorporate unique components of the native ECM into in vitro platform design and fabrication. Ultimately, we anticipate that the use of biomaterials that incorporate key tissue-specific ECM will lead to in vitro models that better emulate human pathologies. STATEMENT OF SIGNIFICANCE: Biomaterial development primarily emphasizes the engineering of new materials and therapies at the expense of identifying key parameters of the tissue that is being emulated. This can be partially attributed to the difficulty in defining the 3D composition, organization, and mechanics of the ECM within different tissues and how these material properties vary as a function of homeostasis and disease. In this review, we highlight a range of tissues throughout the body and describe how ECM content, cell diversity, and mechanical properties change in diseased tissues and influence cellular behavior. Accurately mimicking the tissue of interest in vitro by using ECM specific to the appropriate state of homeostasis or pathology in vivo will yield results more translatable to humans.

Fiber splay precludes the direct identification of ligament material properties: Implications for ACL graft selection.

Anterior cruciate ligament (ACL) injuries typically require surgical reconstruction to restore adequate knee stability. The middle third of an injured patient's patellar tendon (PT) is a commonly used graft for ACL reconstruction. However, many clinicians and researchers question whether it is the best option, as several studies have suggested that it is a stiffer material than the ACL. Still, there is little to no consensus on even the most basic material property of ligaments/tendons: the tangent modulus in the fiber direction, or slope of the linear portion of the uniaxial stress-strain curve. In this study, we investigate the effect of fiber splay (the tendency of collagen fibers to spread out near the enthesis) on the apparent tangent modulus. Using a simplified theoretical model, we establish a quantity we call the splay ratio, which describes the relationship between splay geometry and the apparent tangent modulus. We then more rigorously investigate the effect of the splay ratio on the apparent tangent modulus of the ovine PT and anteromedial and posterolateral regions of the ACL using experimental and computational methods. Both approaches confirmed that splay geometry significantly affects the apparent material behavior. Because true material properties are independent of geometry, we conclude that the macroscopic response of ligaments and tendons is not sufficient for the characterization of their material properties, but rather is reflective of both material and structural properties. We further conclude that the PT is probably not a stiffer material than ACL, but that the PT graft is likely a stiffer structure than either ACL region.

Development of a novel in vitro cadaveric model for analysis of biomechanics and surgical treatment of Bertolotti syndrome.

BACKGROUND CONTEXT: Bertolotti syndrome (BS) is caused by pseudoarticulation between an aberrant L5 transverse process and the sacral ala, termed a lumbosacral transitional vertebra (LSTV). BS is thought to cause low back pain and is treated with resection or fusion, both of which have shown success. Acquiring cadavers with BS is challenging. Thus, we combined 3D printing, based on BS patient CT scans, with normal cadaveric spines to create a BS model. We then performed biomechanical testing to determine altered kinematics from LSTV with surgical interventions. Force sensing within the pseudojoint modeled nociception for different trajectories of motion and surgical conditions. PURPOSE: This study examines alterations in spinal biomechanics with LSTVs and with various surgical treatments for BS in order to learn more about pain and degeneration in this condition, in order to help optimize surgical decision-making. In addition, this study evaluates BS histology in order to better understand the pathology and to help define pain generators-if, indeed, they actually exist. STUDY DESIGN/SETTING: Model Development: A retrospective patient review of 25 patients was performed to determine the imaging criteria that defines the classical BS patient. Surgical tissue was extracted from four BS patients for 3D-printing material selection. Biomechanical Analysis. This was a prospective cadaveric biomechanical study of seven spines evaluating spinal motions, and loads, over various surgical conditions (intact, LSTV, and LSTV with various fusions). Additionally, forces at the LSTV joint were measured for the LSTV and LSTV with fusion condition. Histological Analysis: Histologic analysis was performed prospectively on the four surgical specimens from patients undergoing pseudoarthrectomy for BS at our institution to learn more about potential pain generators. PATIENT SAMPLE: The cadaveric portion of the study involved seven cadaveric spines. Four patients were prospectively recruited to have their surgical specimens assessed histologically and biomechanically for this study. Patients under the age of 18 were excluded. OUTCOME MEASURES: Physiological measures recorded in this study were broken down into histologic analysis, tissue biomechanical analysis, and joint biomechanical analysis. Histologic analysis included pathologist interpretation of Hematoxylin and Eosin staining, as well as S-100 staining. Tissue biomechanical analysis included stiffness measurements. Joint biomechanical analysis included range of motion, resultant torques, relative axis angles, and LSTV joint forces. METHODS: This study received funding from the American Academy of Neurology Medical Student Research Scholarship. Three authors hold intellectual property rights in the simVITRO robotic testing system. No other authors had relevant conflicts of interest for this study. CT images were segmented for a representative BS patient and cadaver spines. Customized cutting and drilling guides for LSTV attachment were created for individual cadavers. 3D-printed bone and cartilage structural properties were based on surgical specimen stiffness, and specimens underwent histologic analysis via Hematoxylin and Eosin, as well as S-100 staining. Joint biomechanical testing was performed on the robotic testing system for seven specimens. Force sensors detected forces in the LSTV joint. Kruskal-Wallis tests and Dunnett's tests were used for statistical analysis with significance bounded to p<.05. RESULTS: LSTV significantly reduces motion at the L5-S1 level, particularly in lateral bending and axial rotation. Meanwhile, the LSTV increases adjacent segment motion significantly at the L2-L3 level, whereas other levels have nonsignificant trends toward increased motion with LSTV alone. Fusion involving L4-S1 (L4-L5 and L5-S1) to treat adjacent level degeneration associated with an LSTV is associated with a significant increase in adjacent segment motion at all levels other than L5-S1 compared to LSTV alone. Fusion of L5-S1 alone with LSTV significantly increases L3-L4 adjacent segment motion compared to LSTV alone. Last, ipsilateral lateral bending with or without ipsilateral axial rotation produces the greatest force on the LSTV, and these forces are significantly reduced with L5-S1 fusion. CONCLUSIONS: BS significantly decreases L5-S1 mobility, and increases some adjacent segment motion, potentially causing patient activity restriction and discomfort. Ipsilateral lateral bending with or without ipsilateral axial rotation may cause the greatest discomfort overall in these patients, and fusion of the L5-S1 or L4-S1 levels may reduce pain associated with these motions. However, due to increased adjacent segment motion with fusions compared to LSTV alone, resection of the joint may be the better treatment option if the superior levels are not unstable preoperatively. CLINICAL SIGNIFICANCE: This study's results indicate that patients with BS have significantly altered spinal biomechanics and may develop pain due to increased loading forces at the LSTV joint with ipsilateral lateral bending and axial rotation. In addition, increased motion at superior levels when an LSTV is present may lead to degeneration over time. Based upon results of LSTV joint force testing, these patients' pain may be effectively treated surgically with LSTV resection or fusion involving the LSTV level if conservative management fails. Further studies are being pursued to evaluate the relationship between in vivo motion of BS patients, spinal and LSTV positioning, and pain generation to gain a better understanding of the exact source of pain in these patients. The methodologies utilized in this study can be extrapolated to recreate other spinal conditions that are poorly understood, and for which few native cadaveric specimens exist.

Robust high resolution strain imaging by alternating pulsed field gradient stimulated echo imaging (APGSTEi) at 7 Tesla.

A novel displacement-encoding spin-echo-stimulated-echo MRI sequence (APGSTEi) was used to obtain full-volume 3D strain fields in samples of two soft materials, a silicone elastomer and an ovine ligament. The samples were stretched cyclically and imaged synchronously. The multi-slice imaging sequence employed a combination of hard and soft spin-echos with bipolar gradient pulses for spatial encoding and decoding, combined with rapid multi-slice spin echo readouts. The sequence minimized undesirable signal loss due to T2∗ and T2 decays, which occur in polymeric materials or in the presence of appreciable air-solid susceptibility contrast, a particular concern for irregularly shaped samples in high magnetic fields. The images' magnitudes were T1-weighted; their phase encoded displacements which occurred during a Δ = 400 ms storage interval separating encoding and decoding pulses. Unwanted residual signals were filtered using a Gaussian filter tailored to attain the desired noise floor. The experiments measured 3D deformation with a nominal resolution of 290 µm × 250 µm × 250 µm in a sample volume of 5.6 cm × 1.6 cm × 1.6 cm, in less than an hour.

Quantum-Induced Symmetry Breaking in the Deuterated Dihydroanthracenyl Radical.

The hydrogen-atom adduct with anthracene, 9-dihydroanthracenyl radical (C14H11), and its deuterated analogue have been identified by laser spectroscopy coupled to time-of-flight mass spectrometry, supported by time-dependent density functional theory calculations. The electronic spectrum of 9-dihydroanthracenyl radical exhibits an origin band at 19115 cm-1 and its ionization energy was determined to be 6.346(1) eV. The spectra reveal a low-frequency vibrational progression corresponding to a mode described by a butterfly inversion. In the deuterated analogue, a zero-point-energy imbalance along this coordinate is found to lead to a doubling of the observed spectral lines in the progression. This is attributed to quantum-induced symmetry breaking as previously observed in isotopologues of CH5+.

Jet-Cooled Spectroscopy of ortho-Hydroxycyclohexadienyl Radicals.

The electronic spectra of the ortho-hydroxycyclohexadienyl radical have been observed following the supersonic expansion of the electric discharge products of phenol and water. Hydrogen atoms, split from water, add to the phenol ring at the ortho position, generating syn and anti rotamers with respect to the hydroxyl group. The D1 ← D0 transitions were recorded by resonance-enhanced multiphoton ionization spectroscopy. The spectrum of each isomer was isolated through hole-burning spectroscopy. The assignment and symmetry of the excited state are evaluated through ab initio calculations and are employed to assign each spectrum. Both rotamers are calculated to have a puckered ring in the excited state, leading to C1 symmetry. The spectrum of the anti isomer is assigned well using this symmetry; however, the syn isomer is assigned better in the C s symmetry of the ground state. We use Duschinsky matrices as a tool for the spectroscopist to determine which point group to use when ab initio calculations are ambiguous.

Femoral entheseal shape and attachment angle as potential risk factors for anterior cruciate ligament injury.

Although non-contact human ACL tears are a common knee injury, little is known about why they usually fail near the femoral enthesis. Recent histological studies have identified a range of characteristic femoral enthesis tidemark profiles and ligament attachment angles. We tested the effect of the tidemark profile and attachment angle on the distribution of strain across the enthesis, under a ligament stretch of 1.1. We employed a 2D analytical model followed by 3D finite element models using three constitutive forms and solved with ABAQUS/Standard. The results show that the maximum equivalent strain was located in the most distal region of the ACL femoral enthesis. It is noteworthy that this strain was markedly increased by a concave (with respect to bone) entheseal profile in that region as well as by a smaller attachment angle, both of which are features more commonly found in females. Although the magnitude of the maximum equivalent strain predicted was not consistent among the constitutive models used, it did not affect the relationship observed between entheseal shape and maximum equivalent strain. We conclude that a concave tidemark profile and acute attachment angle at the femoral ACL enthesis increase the risk for ACL failure, and that failure is most likely to begin in the most distal region of that enthesis.

Interconversion of Methyltropyl and Xylyl Radicals: A Pathway Unavailable to the Benzyl-Tropyl Rearrangement.

The products of an electrical discharge containing toluene are interrogated using resonance-enhanced multiphoton ionization and laser-induced fluorescence spectroscopies. A previously unreported electronic spectrum recorded at m/z = 105, with a putative origin band at 26053 cm-1, is assigned to methyltropyl radical, which appears to be a major product of the toluene discharge, plausibly arising from CH insertion. All three o-, m-, and p-xylyl isomers are also identified. These isomers are detected in electrical discharges containing various xylenes, where it is also found that interconversion occurs: A discharge of o-xylene produces some m-xylyl; a discharge of m-xylene produces some o-xylyl; and a discharge of p-xylene produces all three isomers. No α-methylbenzyl was detected, but styrene was. These observations are supported by state-of-the-art quantum chemical calculations, which reveal an isomerization pathway between methyltropyl and xylyl radicals for which there is no analogue in the canonical tropyl-benzyl isomerization.

The Use of Darbepoetin to Stimulate Erythropoiesis in the Treatment of Anemia of Chronic Kidney Disease in Dogs.

BACKGROUND: Darbepoetin alfa (darbepoetin) is an erythropoiesis-stimulating agent used for the treatment of anemia secondary to chronic kidney disease (CKD) in dogs, but reports describing response are lacking. HYPOTHESIS/OBJECTIVES: To evaluate the effectiveness of darbepoetin in dogs with anemia secondary to CKD, dosing protocols, and adverse events. ANIMALS: Thirty-three client-owned dogs with naturally occurring CKD, including 26 with comorbidities. METHODS: Multi-institutional retrospective study. RESULTS: The median starting dosage and highest dosage of darbepoetin administered were 0.5 and 0.8 µg/kg SC once weekly, respectively. Response to treatment was defined as achieving a packed cell volume (PCV) ≥30% or an increase in PCV ≥10%. Twenty-eight of 33 dogs (85%) achieved a PCV ≥30% and 22 of 33 (67%) dogs achieved an increase in PCV ≥10%. Median time to achieve a PCV ≥30% was 29 days. A higher starting dosage was associated with achieving an increase in PCV ≥10% (P = .01). No dog sustained a response at a dosing interval >q21d. Potential adverse events included increased blood pressure requiring treatment (n = 12), seizures (n = 5), vomiting (n = 3), diarrhea (n = 3), and possible pure red cell aplasia (PRCA) (n = 2). CONCLUSIONS AND CLINICAL IMPORTANCE: Darbepoetin, when combined with treatment of comorbidities, is an effective treatment for anemia secondary to CKD in dogs. A dosing interval >q21d was ineffective at maintaining a response to treatment. PRCA was a possible adverse event in 2 of 33 dogs (6%).

Reliability of the English version of the painDETECT questionnaire.

BACKGROUND: The painDETECT questionnaire (PD-Q) has been used widely for the identification of neuropathic pain (NeP); however, the reliability of the English version of the PD-Q has never been investigated. OBJECTIVE: This study aimed to determine the reliability of the PD-Q pre- (T0) and immediately post- (T1) clinical consultation and at one-week follow-up (T2). METHODS: We recruited 157 patients attending a Neurosurgery Spinal Clinic and Pain Management Department. Minor changes to PD-Q instructions were made to facilitate patient understanding; however, no changes to individual items or scoring were made. Intraclass correlation coefficients (ICCs) were used to assess the reliability of PD-Q total scores between T0-T1 and T0-T2; weighted kappa (κ) was used to assess the agreement of PD-Q classifications (unlikely NeP, ambiguous, likely NeP) between all time-points. To ensure stability of clinical pain, patients scoring ≤2 or ≥6 on the Patient Global Impression Scale (PGIC) at T2 were excluded from the T0-T2 analysis. RESULTS: Accounting for missing data and exclusions (change in PGIC score), data for 136 individuals (mean [SD] age: 56.8 [15.2]; 54% male) was available, of whom n = 129 were included in the T0-T1 and n = 69 in the T0-T2 comparisons. There was almost perfect agreement between the PD-Q total scores at T0-T1 time-points (ICC 0.911; 95% CI: 0.882-0.941) and substantial agreement at T0-T2 (ICC 0.792; 95% CI: 0.703-0.880). PD-Q classifications demonstrated substantial agreement for T0-T1 (weighted κ: 0.771; 95% CI: 0.683-0.858) and for T0-T2 (weighted κ: 0.691; 95% CI: 0.553-0.830). Missing data was accounted in 13% of our cohort and over 42% of our patients drew multiple pain areas on the PD-Q body chart. CONCLUSION: The English version of the PD-Q is reliable as a screening tool for NeP. The validity of the questionnaire is still in question and has to be investigated in future studies.