Osteoporotic Vertebral Fractures: An Analysis of Readability and Quality of Web-Based information.

Introduction Vertebral compression fractures are among the most common fragility fractures with significant morbidity and mortality. With an aging population, the incidence of these fractures is on the rise. In this age of social and electronic media, there is a plethora of online information available. While access to healthcare information has increased, most of these websites remain beyond the comprehension of their target audience. Objective To assess the readability and quality of online information regarding osteoporotic vertebral fractures. Methods A search for the terms osteoporotic vertebral fractures, osteoporotic spinal fractures, and fragility spinal fractures was performed using the top five search engines. Eighty-three websites were identified and analyzed. Quality assessment was done using the DISCERN and Journal of the American Medical Association (JAMA) tools while readability was analyzed using the Flesch Reading Ease Score (FRES), Flesch Kincaid Grade (FKG), and Gunning Fog Index (GFI). Results The mean DISCERN score was 39.55 while the mean JAMA was 2.2. Readability testing revealed a mean FRES score of 49.26 with 16 websites having a score of > 60, FKG 8.38, and GFI of 9.51. 33 websites had an FKG score of 8 or below 8. Conclusion The above results indicate that web-based information relating to osteoporotic vertebral fractures is of variable quality and readability. Although 40 % of websites are at the eighth grade or below level, only 16 % of websites are above the FRES score of 60, which makes online information difficult to comprehend by an average patient.

The relationship between bronchoalveolar lavage fluid cytology and airway hyper-reactivity in a population of Australian horses presented for poor performance.

OBJECTIVE: To investigate the relationship between bronchoalveolar lavage fluid cytology, particularly mast cells, and airway hyper-reactivity in athletic horses presented for poor performance that included a respiratory tract evaluation in two disparate locations in Australia. DESIGN: Multi-centre, retrospective and prospective cross-sectional study METHODS: Eighty four adult horses underwent both pulmonary function testing and histamine bronchoprovocation with a commercial flowmetric plethysmography system. A bronchoalveolar lavage was performed four to twelve hours later. Bronchoalveolar lavage fluid cytology was categorised using two differing classification systems to define mild equine asthma. Statistical analysis was used to assess associations between bronchoalveolar lavage fluid relative inflammatory cell percentages, and airway hyper-reactivity and their associated categorisations. RESULTS: Sixty four percent (54/84) of horses displayed airway hyper-reactivity, as defined by PC35 < 6 mg/ml of histamine. A relative mastocytosis was the most common bronchoalveolar lavage fluid cytological abnormality. Horses with a sole mast cell response of ≥ 5% within their bronchoalveolar lavage fluid displayed airway hyper-reactivity at a lower dose of nebulized histamine than horses with normal bronchoalveolar lavage fluid cytology. Horses with mixed cell responses (relative mast cell percentage > 2% and/or relative neutrophil percentage > 5% and/or eosinophil relative cell percentage ≥ 1%) displayed airway hyper-reactivity at a lower dose of nebulized histamine than horses with normal bronchoalveolar lavage fluid cytology. CONCLUSION: In the Australian context, recently revised increased bronchoalveolar lavage fluid cytology relative cell percentage cut offs appear appropriate for sole mast cell responses. The historical lower cut offs appear to be appropriate for mixed inflammatory cell responses.

COMPARISON OF PATELLA ALIGNMENT AND CARTILAGE BIOMARKERS IN YOUNG ADULT FEMALES WITH AND WITHOUT PATELLOFEMORAL PAIN: A PILOT STUDY.

BACKGROUND: Evidence suggests that individuals with patellofemoral pain (PFP) may develop patellofemoral joint osteoarthritis (PFJOA). Limited data exist regarding an absolute association between PFP and PFJOA. Understanding this relationship will support the need for early interventions to manage PFP. HYPOTHESIS/PURPOSE: This study was conducted to determine if females with PFP have a patella position and cartilage biomarkers similar to individuals with PFJOA. It was hypothesized that females with PFP and excessive patella lateralization would have higher cartilage biomarker levels than controls. It also was hypothesized that a significant association would exist between pain and cartilage biomarker levels in subjects with excessive patella lateralization. STUDY DESIGN: Single-occasion, cross-sectional, observational. METHODS: Pain was assessed using a 10-cm visual analog scale (VAS) for activity pain over the previous week. Patella offset position (RAB angle) was measured using diagnostic ultrasound. Urine was collected and cartilage biomarkers quantified by analyzing C-telopeptide fragments of type II collagen (uCTX-II). Independent t-tests were used to determine between-group differences for RAB angle and uCTX-II. Bivariate correlations were used to determine associations between VAS and uCTX-II for females with PFP. RESULTS: Subjects (age range 20 to 30 years) had similar RAB angles (p = 0.21) and uCTX-II (p = 0.91). A significant association only existed between VAS scores and uCTX-II for females with PFP who had a RAB angle > 13 ° (r = 0.86; p = 0.003). Comparison of uCTX-II in the 25-to-30-year-old females with PFP and excessive patella lateralization in the current study to published normative data showed that this cohort had elevated biomarkers. CONCLUSION: These findings support that a certain cohort of individuals with PFP have features similar to individuals with confirmed PFJOA (patella lateralization and elevated biomarkers). Additional studies are needed to determine if interventions can reverse not only pain but biomarker levels. LEVEL OF EVIDENCE: 2b (diagnosis).

Exercise-induced pulmonary haemorrhage in Thoroughbred racehorses: a longitudinal study.

BACKGROUND: Exercise-induced pulmonary haemorrhage (EIPH) is considered a progressive disease based on histopathology, but it is unknown if tracheobronchoscopic EIPH severity worsens over time. OBJECTIVES: The aim of this study was to examine tracheobronchoscopic EIPH changes over time in a population of Thoroughbred racehorses. A secondary aim was to identify factors that affect changes in tracheobronchoscopic EIPH severity between observations. STUDY DESIGN: Prospective, longitudinal, observational cross-sectional study. METHODS: Thoroughbred racehorses were examined with tracheobronchoscopy no earlier than 30 min after racing. Examinations were recorded and graded blindly by experienced veterinarians using a 0-4 scale. Horses with 2 or more observations were included in the analysis. The association between the previous and current EIPH score was investigated using a linear mixed effect model. Factors associated with transitioning from a lower to a high EIPH grade and vice versa were examined using multiple ordinal regression. A semi-parametric regression model was used to examine progression using the number of career starts as a marker for time. Models were adjusted for potential confounding variables. RESULTS: There were 2974 tracheobronchoscopic examinations performed on 747 horses. Blood was detected in over half of all examinations (55.6%). The population prevalence of EIPH increased as the number of examinations for each horse increased. The preceding EIPH score was significantly associated with the current EIPH score. Significant variables associated with moving between EIPH grades were the number of days since last racing, ambient temperature and weight carried. Tracheobronchoscopic EIPH is mildly progressive over the first thirty career starts. MAIN LIMITATIONS: Enrolment was voluntary. Horses were not followed for their entire career. CONCLUSION: Limiting the number of days in the current racing preparation and spacing races for horses with moderate to severe EIPH may be beneficial for reducing tracheobronchoscopic EIPH severity. The association between ambient temperature and EIPH warrants further investigation.

Bronchoalveolar lavage fluid cytology and airway hyper-reactivity in clinically normal horses.

OBJECTIVE: To characterise the relationship between bronchoalveolar lavage fluid (BALF) cytology and pulmonary function testing with histamine bronchoprovocation (HBP) methods in a population of clinically normal horses. DESIGN: Cross-sectional study METHODS: Clinically normal adult horses (n = 33) underwent pulmonary function testing and HBP with a commercial flowmetric plethysmography system. BAL was performed 1-5 days later. Statistical analysis was used to assess associations between BALF cell concentration, relative inflammatory cell percentages and categorisation, and airway hyper-reactivity (AHR). RESULTS: AHR (PC35 ≤ 8 mg/mL) was demonstrated in 17 (52%) of the horses. Using current definitions, BALF cytology was consistent with inflammatory airway disease in 14 (42%) of the horses and 7 of those demonstrated either mastocytic and/or eosinophilic responses. There was no correlation between total inflammatory cell counts or relative percentage and AHR. No statistical association was found between BALF inflammatory cell categories and AHR. CONCLUSION: A direct association between cytological evidence of airway inflammation and AHR was not identified in this population of clinically normal horses. Determining the presence and measuring inflammatory cell mediators in BALF may more accurately reflect AHR. In addition, normal values for cell proportions in BALF may vary between different populations of horses and more appropriate regional reference ranges should be established.

Effects of Barometric Pressure and Temperature on Acute Ischemic Stroke Hospitalization in Augusta, GA.

Several studies worldwide have demonstrated significant relationships between meteorological parameters and stroke events. However, authors often reported discordant effects of both barometric pressure and air temperature on stroke occurrence. The present study investigated whether there was an association between weather parameters (barometric pressure and temperature) and ischemic stroke hospitalization. The aim of the study was to find out whether daily barometric pressure may be used as a prognostic variable to evaluate the workload change of a neurological intensive care unit. We conducted a retrospective review study in which we collected the independent (barometric pressure and temperature) and dependent variables (stroke hospitalization) every 24 h for the periods 10/1/2016-4/30/2017 at Augusta University Medical Center of Augusta, GA. We analyzed the data with zero-inflated Poisson model to assess the relationship between the barometric pressure, temperature, and daily stroke hospitalization. The results showed that there was a significantly correlation between daily barometric pressure variation and daily stroke hospitalization, especially on elder male patients (≥ 65). Stroke events were more likely to occur in the patients with risk factors than in those without risk factors when exposed to barometric pressure and temperature changes. Decreased barometric pressure and increased temperature were associated with increased daily stroke hospitalization. Furthermore, there was a potential delayed effect of increased stroke events after cold temperature exposure. Barometric pressure and temperature changes over the preceding 24 h are associated with daily stroke hospitalization. These findings may enhance our understanding of relationship between stroke and weather and maybe used in the development of public health strategies to minimize the weather-related stroke risk.

Hypertrophic cardiomyopathy in a Clydesdale gelding.

CASE REPORT: A 7-year-old Clydesdale gelding was referred with a history of progressive lethargy and ventral oedema. The horse developed diarrhoea after treatment with antibiotics by the referring veterinarian. History, clinical signs, imaging and laboratory findings were consistent with hypertrophic cardiomyopathy complicated by antibiotic-associated colitis. Progression of clinical signs prompted euthanasia and the antemortem diagnosis of hypertrophic cardiomyopathy was confirmed at postmortem examination. CONCLUSION: Primary HCM is reported as rare in horses, but specific descriptions are lacking. The history, clinical, laboratory and postmortem findings in this case supported a diagnosis of HCM, complicated by antibiotic-associated colitis.

Complete tendon Achilles rupture following injection of Aethoxysklerol (polidocanol) for the treatment of chronic Achilles tendinopathy.

Achilles tendinopathy can be a chronic disabling condition affecting both athletic and sedentary patients. Multiple new treatment approaches have developed, including shock wave therapy and various types of injection. One of the novel treatment methods used is the injection of Aethoxysklerol or polidocanol, a sclerosing substance injected under ultrasound guidance targeting areas of neovascularisation. We report the case of a 78-year-old lady who suffered a complete Achilles tendon rupture following injection of Aethoxysklerol. This is the first case of Achilles tendon rupture following Aethoxysklerol injection in isolation to our knowledge in the literature and the first published complication of this treatment.

Obtaining local bone graft for Evans calcaneal osteotomy in pes planovalgus deformity correction.

Lateral column lengthening osteotomy is very useful surgical technique in treating stage II of pes planovalgus deformity of the foot. Either autograft from the iliac crest or allograft can be used for this purpose. In our technique we describe a novel method of obtaining bone graft locally from the calcaneus and therefore avoiding complications and morbidity associated with iliac crest graft.

Polyionic complexes of butyrylcholinesterase and poly-l-lysine-g-poly(ethylene glycol): Comparative kinetics of catalysis and inhibition and in vitro inactivation by proteases and heat.

We previously reported that recombinant human butyrylcholinesterase (rhBChE) complexed with a series of copolymers of poly-l-lysine (PLL) with grafted (polyethylene) glycol (PEG) (i.e., PLL-g-PEG) showed reduced catalytic activity but relatively similar concentration-dependent inactivation of the organophosphorus inhibitor paraoxon. Herein, we compared the kinetics of catalysis (using butyrylthiocholine as the substrate) and inhibition (using four different inhibitors) of free and copolymer-complexed rhBChE. Using scanning electron microscopy, polyionic complexes of rhBChE with three different PLL-g-PEG copolymers (based on PLL size) appeared as spheroid-shaped particles with relatively similar particle sizes (median diameter = 35 nm). Relatively similar particle sizes were also noted using dynamic light scattering (mean = 26-35 nm). The three copolymer-complexed enzymes exhibited reduced kcat (30-33% reduction), but no significant changes in Km. Inhibitory potency (as reflected by the bimolecular rate constant, ki) was similar among the free and copolymer-complexed enzymes when paraoxon was the inhibitor, whereas statistically significant reductions in ki (16-60%) were noted with the other inhibitors. Sensitivity to inactivation by proteases and heat was also compared. Copolymer-complexed enzymes showed lesser time-dependent inactivation by the proteases trypsin and pronase and by heat compared to the free enzyme. Understanding the unique properties of PLL-g-PEG-BChE complexes may lead to enhanced approaches for use of BChE and other protein bioscavengers.

The association between exercise-induced pulmonary haemorrhage and race-day performance in Thoroughbred racehorses.

BACKGROUND: Exercise-induced pulmonary haemorrhage (EIPH) is commonly implicated as a cause of poor athletic performance but there is limited and conflicting evidence for this association. OBJECTIVES: The aim of this study was to determine if EIPH, based on endoscopic examination after racing, is associated with a variety of novel and established performance parameters. STUDY DESIGN: Prospective, observational cross-sectional study. METHODS: Thoroughbred racehorses competing between 2012 and 2015 were examined on-course no earlier than 30 min after racing. Examinations were recorded and graded blindly by experienced veterinarians using a 0-4 scale. Linear mixed effect models were used for analysis of continuous response variables with horse name incorporated as a random effect to account for repeated sampling and horse variability. Generalised estimating equations were used for analysis of binary responses. Performance variables were examined in 2 models, comparing EIPH grade 0 to grades 1-4, and EIPH grade ≤2 compared with EIPH grades ≥3, controlling race factors that could influence performance. RESULTS: There were 3794 observations collected from 1567 horses. EIPH was detected in 55.1% of observations. Horses with grade 4 EIPH were significantly more likely to have a lower finishing position and finish further behind the winner, less likely to place in the first 3 positions and collect race earnings, collected less earnings per race start and were slower over the last 600 m of the race than horses without EIPH (grade 0). Similar associations were seen in Model 2, with horses with EIPH grade ≥3 having inferior performance when compared to horses with EIPH ≤2. MAIN LIMITATIONS: Enrolment was voluntary. CONCLUSION: Mild to moderate haemorrhage was not associated with inferior race day performance in this population of Thoroughbred racehorses.

The effect of dobutamine and bolus crystalloid fluids on the cardiovascular function of isoflurane-anaesthetised horses.

REASONS FOR PERFORMING STUDY: Cardiac output does not always increase with dobutamine administration in anaesthetised horses and information on peripheral perfusion is lacking. OBJECTIVES: To determine the effect of intravenous (i.v.) dobutamine infusion with and without a concurrent 20 mL/kg bodyweight (bwt) bolus of crystalloid fluids on the cardiovascular function of acepromazine premedicated, hypotensive, isoflurane-anaesthetised horses. STUDY DESIGN: Randomised, cross-over experiment. METHODS: A total of 6 horses aged 5-13 years, weighing 464-578 kg were premedicated with acepromazine 0.02 mg/kg bwt and then sedated with xylazine 0.8 mg/kg bwt i.v. Anaesthesia was induced with ketamine 2.2 mg/kg bwt and diazepam 0.08 mg/kg bwt i.v. and maintained with isoflurane, adjusted to achieve a target mean arterial pressure (MAP) (60 mmHg ± 5%) 60 min post-induction of anaesthesia (T0). One of 2 treatments was then given. In treatment D, dobutamine was initially infused at 0.5 µg/kg bwt/min and adjusted to achieve a target MAP (80 mmHg ± 5%) within 30 min of infusion initiation. In treatment D+F dobutamine was administered as described for treatment D, with 20 mL/kg bwt Hartmann's solution infused i.v. over 20 min. Cardiac index (CI), haemoglobin concentration ([Hb]), arterial oxygen content (CaO2 ), oxygen delivery index (DO2 I) and bilateral femoral arterial blood flow (FBF) were recorded at T0, 30 min following dobutamine initiation (T1) and 15 min following dobutamine cessation (T2). Data were analysed using a mixed-effect linear model (P<0.05 considered significant). RESULTS: A significant increase in DO2 I (P = 0.008, T0/T1), CaO2 (P = 0.0002, T0/T1) and [Hb] (P<0.0001, T0/T1) and in CaO2 (P = 0.0005, T1/T2) and [Hb] (P = 0.002,T1/T2) occurred during treatment D. A significant increase in FBF (P = 0.005, upper limb; P = 0.042 lower limb, T0/T1) occurred during treatment D+F. Significant differences between treatments were recorded at T1 ([Hb] P = 0.0001, CaO2 P = 0.0003) and T2 ([Hb] P = 0.013). There was no change in CI during either treatment. CONCLUSIONS: The increase in FBF seen with co-administration of fluids and dobutamine may provide a beneficial effect on muscle compared with the use of dobutamine alone.

Equine Pulmonary Cryptococcosis: A Comparative Literature Review and Evaluation of Fluconazole Monotherapy.

Cryptococcus is the most common fungal respiratory pathogen in Australian horses, manifesting primarily as pulmonary granulomas. Disease severity at presentation is dependent on the athletic use of the horse. The diagnosis and estimation of disease severity are centred around clinical findings, cytological evaluation of respiratory tract secretions, diagnostic imaging, and antigen titre testing. Both the lateral flow assay and the latex cryptococcal antigen titre are used, and important similarities and differences between species are discussed. Cryptococcus gattii occurs with greater frequency than Cryptococcus neoformans in equine pulmonic cryptococcosis and can be successfully treated with enteral fluconazole monotherapy, with disease severity determining treatment length.

Bar shoes and ambient temperature are risk factors for exercise-induced pulmonary haemorrhage in Thoroughbred racehorses.

REASONS FOR PERFORMING STUDY: Ambient temperature has been identified as a risk factor for exercise-induced pulmonary haemorrhage (EIPH) in racing Thoroughbreds. This warranted a more expansive investigation of climatic conditions on the incidence and severity of EIPH. The impact of other variables such as the type of bit used, tongue ties and nonstandard shoes has not been reported and also warrant investigation. OBJECTIVES: To examine the effect of various climatic variables as contributing risk factors for EIPH. Other previously uninvestigated variables as well as standard track and population factors will also be examined. STUDY DESIGN: Cross-sectional study. METHODS: Thoroughbred racehorses competing at metropolitan racetracks in Perth, Western Australia were examined 30-200 min post race with tracheobronchoscopy. Examination took place at 48 race meetings over a 12 month period. Examinations were graded (0-4), independently by two experienced veterinarians. Univariable analyses were performed and variables with a P<0.25 were entered into a multivariable logistic regression analysis. The analysis was performed twice using the presence of blood (EIPH grade 0 vs. grades ≥1) and EIPH grades ≤1 vs. EIPH grades ≥2 as dependent variables. RESULTS: Exercise-induced pulmonary haemorrhage was diagnosed in 56.6% of observations. Lower ambient temperature was significantly associated with EIPH grades ≥1 (OR 0.95; 95% CI 0.93-0.98) and EIPH grades ≥2 (OR 0.97; 95% CI 0.94-1.0). Bar shoes were significantly associated with EIPH grades ≥1 (OR 6.35; 95% CI 2.17-18.54) and EIPH grades ≥2 (OR 2.72; 95% CI 1.3-5.68). Increasing race distance was significantly associated with EIPH grade ≥1 and increasing lifetime starts was significantly associated with EIPH grade ≥2. CONCLUSIONS: Ambient temperature is a risk factor for EIPH in Thoroughbred racehorses, with lower temperatures associated with increased risk. Bar shoes are a novel risk factor for EIPH in this population.

Retrospective survey of bronchoalveolar lavage fluid cytology in Western Australian horses presented for evaluation of the respiratory tract: effect of season on relative cell percentages.

OBJECTIVE: To characterise the cytological changes in equine bronchoalveolar lavage (BAL) samples over multiple years to determine if the prevalence of a relative mast cell response was influenced by season. METHODS: Medical records of 228 horses with clinical signs consistent with non-infectious respiratory disease or poor performance where a BAL was performed were reviewed retrospectively. BAL fluid cytology and categorised clinical variables were analysed using a Chi-square test to determine associations. RESULTS: The predominant signalment was a racing horse between 2 and 6 years of age, and poor athletic performance was the most common presenting complaint. Based on published criteria, 69.7% of horses had abnormal BAL cytology. The presence of nasal discharge was the only clinical finding that differed significantly between horses with abnormal and normal cytology. The most common cytological derangement was a mixed cell response (26.7%), the majority of which comprised elevated percentages of neutrophils and mast cells. A solely neutrophilic response or mast cell response occurred with equal frequency (18% and 18.95%, respectively), and an isolated eosinophilic response was noted in 6.1% of cases. Of the horses with cytology consistent with non-infectious lower airway inflammation, 58.4% had increases in the relative percentage of mast cells. Cell profiles were significantly associated with season, with mastocytosis occurring more commonly in spring, whereas eosinophilic and neutrophilic responses were more common in summer. CONCLUSION: In this study, a relative mast cell increase was the most common cytological profile in horses with lower airway inflammation and was associated with season.

The accurate assessment of small-angle X-ray scattering data.

Small-angle X-ray scattering (SAXS) has grown in popularity in recent times with the advent of bright synchrotron X-ray sources, powerful computational resources and algorithms enabling the calculation of increasingly complex models. However, the lack of standardized data-quality metrics presents difficulties for the growing user community in accurately assessing the quality of experimental SAXS data. Here, a series of metrics to quantitatively describe SAXS data in an objective manner using statistical evaluations are defined. These metrics are applied to identify the effects of radiation damage, concentration dependence and interparticle interactions on SAXS data from a set of 27 previously described targets for which high-resolution structures have been determined via X-ray crystallography or nuclear magnetic resonance (NMR) spectroscopy. The studies show that these metrics are sufficient to characterize SAXS data quality on a small sample set with statistical rigor and sensitivity similar to or better than manual analysis. The development of data-quality analysis strategies such as these initial efforts is needed to enable the accurate and unbiased assessment of SAXS data quality.

Nematic director reorientation at solid and liquid interfaces under flow: SAXS studies in a microfluidic device.

In this work we investigate the interplay between flow and boundary condition effects on the orientation field of a thermotropic nematic liquid crystal under flow and confinement in a microfluidic device. Two types of experiments were performed using synchrotron small-angle X-ray-scattering (SAXS). In the first, a nematic liquid crystal flows through a square-channel cross section at varying flow rates, while the nematic director orientation projected onto the velocity/velocity gradient plane is measured using a 2D detector. At moderate-to-high flow rates, the nematic director is predominantly aligned in the flow direction, but with a small tilt angle of ∼±11° in the velocity gradient direction. The director tilt angle is constant throughout most of the channel width but switches sign when crossing the center of the channel, in agreement with the Ericksen-Leslie-Parodi (ELP) theory. At low flow rates, boundary conditions begin to dominate, and a flow profile resembling the escaped radial director configuration is observed, where the director is seen to vary more smoothly from the edges (with homeotropic alignment) to the center of the channel. In the second experiment, hydrodynamic focusing is employed to confine the nematic phase into a sheet of liquid sandwiched between two layers of Triton X-100 aqueous solutions. The average nematic director orientation shifts to some extent from the flow direction toward the liquid boundaries, although it remains unclear if one tilt angle is dominant through most of the nematic sheet (with abrupt jumps near the boundaries) or if the tilt angle varies smoothly between two extreme values (∼90 and 0°). The technique presented here could be applied to perform high-throughput measurements for assessing the influence of different surfactants on the orientation of nematic phases and may lead to further improvements in areas such as boundary lubrication and clarifying the nature of defect structures in LC displays.

Altered active zones, vesicle pools, nerve terminal conductivity, and morphology during experimental MuSK myasthenia gravis.

Recent studies demonstrate reduced motor-nerve function during autoimmune muscle-specific tyrosine kinase (MuSK) myasthenia gravis (MG). To further understand the basis of motor-nerve dysfunction during MuSK-MG, we immunized female C57/B6 mice with purified rat MuSK ectodomain. Nerve-muscle preparations were dissected and neuromuscular junctions (NMJs) studied electrophysiologically, morphologically, and biochemically. While all mice produced antibodies to MuSK, only 40% developed respiratory muscle weakness. In vitro study of respiratory nerve-muscle preparations isolated from these affected mice revealed that 78% of NMJs produced endplate currents (EPCs) with significantly reduced quantal content, although potentiation and depression at 50 Hz remained qualitatively normal. EPC and mEPC amplitude variability indicated significantly reduced number of vesicle-release sites (active zones) and reduced probability of vesicle release. The readily releasable vesicle pool size and the frequency of large amplitude mEPCs also declined. The remaining NMJs had intermittent (4%) or complete (18%) failure of neurotransmitter release in response to 50 Hz nerve stimulation, presumably due to blocked action potential entry into the nerve terminal, which may arise from nerve terminal swelling and thinning. Since MuSK-MG-affected muscles do not express the AChR γ subunit, the observed prolongation of EPC decay time was not due to inactivity-induced expression of embryonic acetylcholine receptor, but rather to reduced catalytic activity of acetylcholinesterase. Muscle protein levels of MuSK did not change. These findings provide novel insight into the pathophysiology of autoimmune MuSK-MG.

Structural basis of the pH-dependent assembly of a botulinum neurotoxin complex.

Botulinum neurotoxins (BoNTs) are among the most poisonous biological substances known. They assemble with non-toxic non-hemagglutinin (NTNHA) protein to form the minimally functional progenitor toxin complexes (M-PTC), which protects BoNT in the gastrointestinal tract and releases it upon entry into the circulation. Here we provide molecular insight into the assembly between BoNT/A and NTNHA-A using small-angle X-ray scattering. We found that the free form BoNT/A maintains a pH-independent conformation with limited domain flexibility. Intriguingly, the free form NTNHA-A adopts pH-dependent conformational changes due to a torsional motion of its C-terminal domain. Once forming a complex at acidic pH, they each adopt a stable conformation that is similar to that observed in the crystal structure of the M-PTC. Our results suggest that assembly of the M-PTC depends on the environmental pH and that the complex form of BoNT/A is induced by interacting with NTNHA-A at acidic pH.