Evaluating the Effectiveness of Tethered Bis(urazolyl) Diradicals as Molecular Building Blocks for Dynamic Covalent Chemistry.

Dynamic covalent chemistry (DCvC) is a powerful means by which to rapidly prepare complex structures from simple molecular building blocks. Effective DCvC behavior is contingent upon the reversibility of covalent bond formation. Stabilized radical species, therefore, have been effectively used for these applications. In earlier work we demonstrated that properly substituted 1-arylurazolyl radicals showed promise as oxygen-insensitive heterocyclic N-centered radicals with a propensity for reversible bond formation. In this work we have synthesized several tethered bis(urazolyl) diradicals, varying by the type and length of connectivity between the urazole rings, and tested them for DCvC behavior. We have found that when the two aryl rings to which the urazolyl radical sites are attached are tethered by a chain of five or more carbons, equilibrium mixtures of monomeric and dimeric species are formed by N-N bond formation between two radical sites. DCvC behavior is observed that is sensitive to changes in temperature, concentration, and (to a lesser extent) solvent. In general, the dimer species is favored at lower temperatures and higher concentrations.

Endoplasmic Reticulum Stress-Regulated Chaperones as a Serum Biomarker Panel for Parkinson's Disease.

Examination of post-mortem brain tissues has previously revealed a strong association between Parkinson's disease (PD) pathophysiology and endoplasmic reticulum (ER) stress. Evidence in the literature regarding the circulation of ER stress-regulated factors released from neurons provides a rationale for investigating ER stress biomarkers in the blood to aid diagnosis of PD. The levels of ER stress-regulated proteins in serum collected from 29 PD patients and 24 non-PD controls were measured using enzyme-linked immunosorbent assays. A panel of four biomarkers, protein disulfide-isomerase A1, protein disulfide-isomerase A3, mesencephalic astrocyte-derived neurotrophic factor, and clusterin, together with age and gender had higher ability (area under the curve 0.64, sensitivity 66%, specificity 57%) and net benefit to discriminate PD patients from the non-PD group compared with other analyzed models. Addition of oligomeric and total α-synuclein to the model did not improve the diagnostic power of the biomarker panel. We provide evidence that ER stress-regulated proteins merit further investigation for their potential as diagnostic biomarkers of PD.

International Consensus Statement on the diagnosis, multidisciplinary management and lifelong care of individuals with achondroplasia.

Achondroplasia, the most common skeletal dysplasia, is characterized by a variety of medical, functional and psychosocial challenges across the lifespan. The condition is caused by a common, recurring, gain-of-function mutation in FGFR3, the gene that encodes fibroblast growth factor receptor 3. This mutation leads to impaired endochondral ossification of the human skeleton. The clinical and radiographic hallmarks of achondroplasia make accurate diagnosis possible in most patients. However, marked variability exists in the clinical care pathways and protocols practised by clinicians who manage children and adults with this condition. A group of 55 international experts from 16 countries and 5 continents have developed consensus statements and recommendations that aim to capture the key challenges and optimal management of achondroplasia across each major life stage and sub-specialty area, using a modified Delphi process. The primary purpose of this first International Consensus Statement is to facilitate the improvement and standardization of care for children and adults with achondroplasia worldwide in order to optimize their clinical outcomes and quality of life.

Long-term outcomes of patients with mucopolysaccharidosis VI treated with galsulfase enzyme replacement therapy since infancy.

OBJECTIVE: Long-term outcomes of patients with mucopolysaccharidosis (MPS) VI treated with galsulfase enzyme replacement therapy (ERT) since infancy were evaluated. METHODS: The study was a multicenter, prospective evaluation using data from infants with MPS VI generated during a phase 4 study (ASB-008; Clinicaltrials.govNCT00299000) and clinical data collected ≥5 years after completion of the study. RESULTS: Parents of three subjects from ASB-008 (subjects 1, 2, and 4) provided written informed consent to participate in the follow-up study. One subject was excluded as consent was not provided. Subjects 1, 2, and 4 were aged 0.7, 0.3, and 1.1 years, respectively, at initiation of galsulfase and 10.5, 7.9, and 10.5 years, respectively, at follow-up. All subjects had classical MPS VI based on pre-treatment urinary glycosaminoglycans and the early onset of clinical manifestations. At follow-up, subject 4 had normal stature for age; subjects 1 and 2 had short stature, but height remained around the 90th percentile of growth curves for untreated classical MPS VI. Six-minute walk distance was normal for age/height in subjects 1 (550 m) and 4 (506 m), and reduced for subject 2 (340 m). Subject 2 preserved normal respiratory function, while percent predicted forced vital capacity and forced expiratory volume in 1 s decreased over time in the other subjects. Skeletal dysplasia was already apparent in all subjects at baseline and continued to progress. Cardiac valve disease showed mild progression in subject 1, mild improvement in subject 4, and remained trivial in subject 2. All subjects had considerably reduced pinch and grip strength at follow-up, but functional dexterity was relatively normal for age and there was limited impact on activities of daily living. Bruininks-Oseretsky Test of Motor Proficiency (BOT-2) results showed that subjects 2 and 4 had numerous fine and gross motor competencies. Corneal clouding progressed in all subjects, while progression of hearing impairment was variable. Liver size normalized from baseline in subjects 1 and 4, and remained normal in subject 2. CONCLUSION: Very early and continuous ERT appears to slow down the clinical course of MPS VI, as shown by preservation of endurance, functional dexterity, and several fine and gross motor competencies after 7.7-9.8 years of treatment, and less growth impairment or progression of cardiac disease than could be expected based on the patients' classical phenotype. ERT does not seem to prevent progression of skeletal or eye disease in the long term.

Integrating inflammatory serum biomarkers into a risk calculator for prostate cancer detection.

Improved prostate cancer detection methods would avoid over-diagnosis of clinically indolent disease informing appropriate treatment decisions. The aims of this study were to investigate the role of a panel of Inflammation biomarkers to inform the need for a biopsy to diagnose prostate cancer. Peripheral blood serum obtained from 436 men undergoing transrectal ultrasound guided biopsy were assessed for a panel of 18 inflammatory serum biomarkers in addition to Total and Free Prostate Specific Antigen (PSA). This panel was integrated into a previously developed Irish clinical risk calculator (IPRC) for the detection of prostate cancer and high-grade prostate cancer (Gleason Score ≥ 7). Using logistic regression and multinomial regression methods, two models (Logst-RC and Multi-RC) were developed considering linear and nonlinear effects of the panel in conjunction with clinical and demographic parameters for determination of the two endpoints. Both models significantly improved the predictive ability of the clinical model for detection of prostate cancer (from 0.656 to 0.731 for Logst-RC and 0.713 for Multi-RC) and high-grade prostate cancer (from 0.716 to 0.785 for Logst-RC and 0.767 for Multi-RC) and demonstrated higher clinical net benefit. This improved discriminatory power and clinical utility may allow for individualised risk stratification improving clinical decision making.

Probing the Dynamic Covalent Chemistry Behavior of Nitrogen-Centered Di- and Triurazole Radicals.

Dynamic covalent chemistry (DCvC) describes systems in which readily reversible bond formation allows for control of product distributions by straightforward manipulation of reaction conditions (e.g., changes in temperature, solvent, concentration, etc). Nitrogen-centered 1-aryl urazole radicals reversibly form tetrazane dimers in solution via N-N bond formation. When two such urazole units are attached to a single, appropriately substituted benzene ring, the resulting diradical system engages in DCvC. At room temperature, a polymeric network of units is created that exhibits gel-like properties, while at higher temperatures, near quantitative dimerization to form a molecular cage is observed.However, attaching three such urazole units to a single appropriately substituted benzene ring inhibits DCvC behavior.

Structural Consequences of the 1,2,3-Triazole as an Amide Bioisostere in Analogues of the Cystic Fibrosis Drugs VX-809 and VX-770.

Although the 1,2,3-triazole is a commonly used amide bioisostere in medicinal chemistry, the structural implications of this replacement have not been fully studied. Employing X-ray crystallography and computational studies, we report the spatial and electronic consequences of replacing an amide with the triazole in analogues of cystic fibrosis drugs in the VX-770 and VX-809 series. Crystallographic analyses quantify subtle differences in the relative positions and conformational preferences of the R1 and R2 substituents attached to the amide and triazole bioisosteres. Computational studies derived from the X-ray data highlight the improved hydrogen bonding donor and acceptor capabilities of the amide in comparison to the triazole. This analysis of the spatial and electronic differences between the amide and 1,2,3-triazole will inform medicinal chemists as they consider using the triazole as an amide bioisostere.

Application of a New Multiplexed Array for Rapid, Sensitive, Simultaneous and Quantitative Assessment of Spliced and Unspliced XBP1.

BACKGROUND: IRE1α-mediated unconventional splicing of XBP1 is emerging as a biomarker in several disease states and is indicative of activation of the unfolded protein response sensor IRE1. Splicing of XBP1 mRNA results in the translation of two distinct XBP1 protein isoforms (XBP1s and XBP1u) which, due to post-translational regulation, do not correlate with mRNA levels. As both XBP1 isoforms are implicated in pathogenic or disease progression mechanisms there is a need for a reliable, clinically applicable method to detect them. METHODS: A multiplexed isoform-specific XBP1 array utilising Biochip array technology (BAT™) was assessed for specificity and suitability when using cell protein lysates. The array was applied to RIPA protein lysates from several relevant pre-clinical models with an aim to quantify XBP1 isoforms in comparison with RT-PCR or immunoblot reference methods. RESULTS: A novel reliable, specific and sensitive XBP1 biochip was successfully utilised in pre-clinical research. Application of this biochip to detect XBP1 splicing at the protein level in relevant breast cancer models, under basal conditions as well as pharmacological inhibition and paclitaxel induction, confirmed the findings of previous studies. The biochip was also applied to non-adherent cells and used to quantify changes in the XBP1 isoforms upon activation of the NLRP3 inflammasome. CONCLUSIONS: The XBP1 biochip enables isoform specific quantification of protein level changes upon activation and inhibition of IRE1α RNase activity, using a routine clinical methodology. As such it provides a research tool and potential clinical tool with a quantified, simultaneous, rapid output that is not available from any other published method.

Structural verification of a tetrahydrotetrazole compound.

Tetrahydrotetrazoles are five-membered-ring heterocycles containing four contiguous saturated nitrogen atoms. Very few examples of such compounds have been reported in the literature. Our previous attempt at the synthesis of a member of this class of compound suggested that the N-N bonds may be more labile than expected. This finding raised the question as to whether the structures of any of the previously reported tetrahydrotetrazoles had been properly assigned. We have reproduced the synthesis of a reported tetrahydrotetrazole, namely 1,2-di-tert-butyl 3-phenyl-1H,2H,3H,10bH-[1,2,3,4]tetrazolo[5,1-a]isoquinoline-1,2-dicarboxylate, C25H30N4O4, and have now confidently confirmed its structure via X-ray crystallography. However, while sufficiently stable in the crystal phase, we discovered that it remains very labile in solution (having a half-life of only 15 min at 20 °C in CDCl3). A tentative reaction pathway for its dissociation based on 1H NMR spectral evidence is provided.

Cross-Platform Ubiquitous Volume Rendering Using Programmable Shaders in VTK for Scientific and Medical Visualization.

The visualization toolkit (VTK) is a popular cross-platform, open source toolkit for scientific and medical data visualization, processing, and analysis. It supports a wide variety of data formats, algorithms, and rendering techniques for both polygonal and volumetric data. In particular, VTK's volume rendering module has long provided a comprehensive set of features such as plane clipping, color and opacity transfer functions, lighting, and other controls needed for visualization. However, due to VTK's legacy OpenGL backend and its reliance on a deprecated API, the system did not take advantage of the latest improvements in graphics hardware or the flexibility of a programmable pipeline. Additionally, this dependence on an antiquated pipeline posed restrictions when running on emerging computing platforms, thereby limiting its overall applicability. In response to these shortcomings, the VTK community developed a new and improved volume rendering module, which not only provides a modern graphics processing unit-based implementation, but also augments its capabilities with new features such as fast volume clipping, gradient-magnitude-based opacity modulation, render to texture, and hardware-based volume picking.

Surface coatings including fingerprint residues can significantly alter the size and shape of bloodstains.

When conducting a blood pattern analysis (BPA) the size, shape, distribution, and location of bloodstains found at a crime scene may be critical in forming a hypothesis as to what transpired during a bloody event. Prior studies have demonstrated that the size and shape of a bloodstain on a smooth surface are determined from impact dynamics and to a lesser degree by the target material itself. Yet, these studies have relied on clean surfaces, and it is unclear whether the presence of microscopic coatings and residues could significantly alter the size or shape of the dried stain. Here, in the present work, experiments are conducted to demonstrate that various coatings, such as the sebaceous residue from a latent fingerprint, can dramatically alter the size and shape of the stain from the moment of impact through the drying process. These experiments also highlight that a drop impacting a tilted superhydrophobic-coating glass substrate can cause the blood drop to completely recoil without leaving a stain. Relying on a combination of high-speed and time-lapse photography, the specific stages in the stain evolution responsible for the deviations from the current models are identified. At a relatively low impact velocity, the stain sizes on the coated glass surfaces were 35-72% smaller than on the clean glass surface. At a higher impact velocity, the stains on the coated surfaces were not only smaller, but also contained drop spatter around the primary stain that was not observable in the absence of the microscopic coatings. The reduction in bloodstain size did not appreciably change when a chemical was added to deactivate the anticoagulant and allow the blood to clot.

Surgical management of congenital thoracic kyphosis and multilevel bilateral thoracic pedicle aplasia: case report.

Pedicle aplasia is an uncommon congenital anomaly most frequently involving the absence of a single pedicle at a single vertebral level. Bilateral pedicle aplasia at multiple levels is exceedingly rare and has only been described once previously in the literature. While single-level pedicle aplasia is often asymptomatic and discovered incidentally, pedicle aplasia of multiple levels may produce severe spinal deformities and neurological deficits. Due to the rarity of this condition, optimal management remains uncertain. In this case report, the authors describe the surgical management of a healthy 9-year-old boy who presented with frequent falls, difficulty running, and severe thoracic kyphotic deformity and was found to have bilateral pedicle aplasia from T3 to T9. A review of the literature regarding pedicle aplasia is also presented.

Sleep disorders in children with incidental pineal cyst on MRI: a pilot study.

INTRODUCTION: For several years, pineal cysts have been considered an incidental finding on brain MRI's even though research and case reports have shown a possible association of pineal cysts with headaches and sleep disturbances. This pilot study aims to evaluate sleep disorders in school-age children with an incidental pineal cyst in an otherwise normal brain MRI. METHOD: Children aged 6-12 years, who were referred for evaluation of headaches, tics, or syncope, and had an incidental pineal cyst on an otherwise normal brain MRI were included and compared to a control group of children with the same referral reasons but with a normal MRI and to a cohort of normal controls. The Sleep Disturbance Scale for Children (SDSC) was administered to the parents. Exclusion criteria included use of medications that affect sleep, seizures, brain abnormalities, tumors, or comorbid medical conditions that affect sleep. RESULTS: Eighteen children (11 females) with pineal cysts, 19 children with normal MRI, and 100 age- and sex-matched controls were included in our study. There were statistically significant differences in the total SDSC score (with a difference of 10 between the median scores) and in two of the six domains of this scale. Children with pineal cysts scored significantly higher in the domains of disorders of excessive sleepiness and disorders of initiating and maintaining sleep than the two control groups. The scores in these two domains correlated significantly with the size of the cyst. CONCLUSION: School-age children with pineal cysts have significantly increased levels of sleepiness and difficulty with sleep initiation and maintenance.

Transient Central Sleep Apnea Runs Triggered by Disorder of Arousal in a Child.

ABSTRACT: We report the case of a 5-year-old girl with frequent nocturnal episodes of disorder of arousal (confusional arousals, sleep terrors, and sleep walking), occurring at the end of periods of slow wave sleep, followed by return to sleep accompanied by the occurrence of periodic breathing with a run of approximately 10 to 20 central events. The duration of the central events and oxyhemoglobin desaturation were both maximum at the beginning of each run and became progressively less prominent with the development of the sequences. Night episodes disappeared with bedtime clonazepam but behavioral problems occurred as a paradoxical response; thus, clonazepam was stopped. Sleep extension and melatonin were then started, which were followed by a reduction of night episode frequency and intensity. This observation appears to be the first report of central sleep apnea sequences triggered by parasomnia and, if confirmed by additional reports, it might be considered to be a possible new classification of "complex parasomnia."

Unanticipated formation of a novel octaazacyclodecane ring upon oxidation of a 1,1-bis-urazole.

Tetrahydrotetrazoles are a little-explored class of five-membered heterocycles with four contiguous singly-bonded N atoms. Recent work in our labs has demonstrated that urazole radicals are amenable to N-N bond formation via radical combination to form such a chain of four N atoms. Previously described 1,1-bis-urazole compounds appeared to be convenient precursors to the target tetrazoles via their oxidation to intermediate urazole diradicals, which upon N-N bond formation would complete the tetrazole framework. While oxidation proceeded smoothly, the novel 10-membered octaaza heterocycle 7,7,18,18-tetraacetyl-4,10,15,21-tetraphenyl-1,2,4,6,8,10,12,13,15,17,19,21-dodecaazapentacyclo[17.3.0.02,6.08,12.013,17]docosan-3,5,9,11,14,16,20,22-octone, C42H32N12O12, was obtained (36% yield) instead of the expected tetrazole product, as confirmed by X-ray crystallography. Calculations at the (U)B3LYP/6-311G(d,p) level of theory suggest that the desired tetrazoles have weak N-N bonds connecting the two urazole units.

Non-Invasive Prenatal Testing (NIPT) in pregnancies with trisomy 21, 18 and 13 performed in a public setting - factors of importance for correct interpretation of results.

OBJECTIVES: We have established an open source platform for non-invasive prenatal testing (NIPT) based on massively parallel whole-genome sequencing in a public setting. The objective of this study was to investigate factors of importance for correct interpretation of NIPT results to ensure a high sensitivity and specificity. STUDY DESIGN: This investigation is a retrospective case-control study performed in a public NIPT center. The study included 108 aneuploid cases and 165 euploid controls. MPS was performed on circulating cell-free DNA in maternal blood. The pipeline included automated library preparation and sequencing on a HiSeq1500 (Illumina). The software programmes WISECONDOR and SeqFF were used for data analysis of aneuploidy status and fetal fraction of cell-free DNA, respectively. Lower limit of fetal fraction for aneuploidy testing was 0.02. RESULTS: We identified four false negative aneuploidy cases of which two were explained by a vanishing twin. The number of no-call cases due to low fetal fraction was 8 out of 273 (2.9%). The sensitivity and specificity, when no-calls and vanished twins were excluded, were 100% and 99.5% for T21, 91% and 99.2% for T18, and 100% and 99.6% for T13. By multiple regression analysis we found a significant association between fetal fraction and gestational age, maternal BMI and ART treatment. CONCLUSION: With a non-commercial open source NIPT set-up having the same high test-performance as reported by large private laboratories, we show that fetal fraction, a vanishing twin, BMI, gestational age and ART treatment are important factors in the interpretation of NIPT results.

Compatibility of ZOLL Defibrillators in Simulation-Based Training.

INTRODUCTION: In response to the need for high-quality cardiopulmonary resuscitation (CPR) during cardiac arrest, our institution recently purchased ZOLL R Series monitor/defibrillators. This defibrillator provides CPR quality metrics and displays a filtered rhythm through compressions. Purchase of this defibrillator resulted in a practice change and heavily impacted our simulation-based training courses by requiring providers to practice CPR and defibrillation in as close to the real environment as possible. Thus, our objective was to determine which commercial simulators would be compatible with the ZOLL R Series defibrillator system and its CPR feedback functionality in a simulation-based training setting. METHODS: Our simulation center uses primarily Gaumard Scientific and Laerdal Medical simulators ranging in size from neonate to adult. Through an iterative process in the laboratory, we evaluated if, and to what level, the CPR display metrics, filtered rhythm, and idle time display could be demonstrated with CPR on the different simulators using infant, pediatric, and adult pads. RESULTS: Certain simulators allow demonstration and real-time practice of defibrillator functions better than others with the ZOLL R Series system when used in the context of CPR training. We have no high-fidelity infant-sized simulators that can meet the depth recommendation for chest compressions given by the American Heart Association. Ventricular fibrillation is the only rhythm that offers a filtered option. Idle time can be reliably displayed for simulators where CPR is detected. CONCLUSIONS: When a primary learning objective for simulation-based training involves training on the ZOLL R Series defibrillator, there are a limited number of simulators and rhythms that can accurately represent its features.

Computational, 1H NMR, and X-ray structural studies on 1-arylurazole tetrazane dimers.

Nitrogen-centered urazole radicals exist in equilibrium with tetrazane dimers in solution. The equilibrium established typically favors the free-radical form. However, 1-arylurazole radicals bearing substituents at the ortho position favor the dimeric form. We were able to determine the structure of one of the dimers (substituted at both ortho positions with methyl groups), namely 1,2-(2,4-dimethylphenyl)-2-[2-(2,4-dimethylphenyl)-4-methyl-3,5-dioxo-1,2,4-triazolidin-1-yl]-4-methyl-1,2,4-triazolidine-3,5-dione, C24H28N6O4, via X-ray crystallography. The experimentally determined structure agreed well with the computationally obtained geometry at the B3LYP/6-311G(d,p) level of theory. The preferred syn conformation of these 1-arylurazole dimers results in the two aromatic rings being proximate and nearly parallel, which leads to some interesting shielding effects of certain signals in the 1H NMR spectrum. Armed with this information, we were able to decipher the more complicated 1H NMR spectrum obtained from a dimer that was monosubstituted at the ortho position with a methyl group.