Analgesia use in trauma patients at a university teaching hospital in Kigali, Rwanda.

Introduction: While trends in analgesia have been identified in high-income countries, little research exists regarding analgesia administration in low- and middle-income countries (LMIC). This study evaluates analgesia administration and clinical characteristics among patients seeking emergency injury care at University Teaching Hospital-Kigali in Kigali, Rwanda. Methods: This retrospective, cross-sectional study utilized a random sample of emergency center (EC) cases accrued between July 2015 and June 2016. Data was extracted from the medical record for patients who had an injury and were ≥ 15 years of age. Injury-related EC visits were identified by presenting complaint or final discharge diagnosis. Sociodemographic information, injury mechanism and type, and analgesic medications ordered and administered were analyzed. Results: Of the 3,609 random cases, 1,329 met eligibility and were analyzed. The study population was predominantly male (72%) with a median age of 32 years and range between 15 and 81 years. In the studied sample, 728 (54.8%) were treated with analgesia in the EC. In unadjusted logistic regression, only age was not a significant predictor of receiving pain medication and was excluded from the adjusted analysis. In the adjusted model, all predictors remained significant, with being male, having at least one severe injury, and road traffic accident (RTA) as injury mechanism being significant predictors of analgesia administration. Conclusion: In the study setting of injured patients in Rwanda, being male, involved in RTA or having more than one serious injury was associated with higher odds of receiving pain medication. Approximately half of the patients with traumatic injuries received pain medications, predominantly opioids with no factors predicting whether a patient would receive opioids versus other medications. Further research on implementation of pain guidelines and drug shortages is warranted to improve pain management for injured patients in the LMIC setting.

A peptide immunoaffinity LC-MS/MS strategy for quantifying the GPCR protein, S1PR1 in human colon biopsies.

Background: S1PR1, a G protein-coupled receptor (GPCR) protein, is a therapeutic target for treatment of autoimmune diseases. As a potential biomarker for drug effect and patient stratification, it is of great significance to measure it in biological samples. However, due to the hydrophobic nature of S1PR1 and the difficulties in extraction and solubilization, as well as low expression levels, quantitative determination of S1PR1 remains challenging. Results: In this work, a peptide immunoaffinity LC-MS/MS method was developed to quantify S1PR1 in biopsy-sized colon samples with an LLOQ of 7.81 pM. Conclusion: Peptide immunoaffinity LC-MS/MS based strategy has achieved the desired sensitivity for low abundance S1PR1, and the same strategy could be applied to quantify S1PR1 in multiple species and other GPCR proteins.

LC/MS/MS-based quantitation of pig and human S100A1 protein in cardiac tissues: Application to gene therapy.

The S100A1 protein is a target of interest for the treatment of heart failure as it has been previously reported to be depleted in failing cardiomyocytes. A gene therapy approach leading to increased expression levels of the protein directly in the heart could potentially lead to restoration of contractile function and improve overall cell survival. S100A1 is a relatively small soluble protein that is extremely well conserved across species with only a single amino acid difference between the sequences in human and pig, a commonly used pre-clinical model for evaluation of efficacy, biodistribution and safety for cardiac-directed gene therapy approaches. This high homology presents a bioanalytical challenge for the accurate detection and quantitation of both endogenous (pig) and exogenous (human) transduced S100A1 proteins post treatment using a human S100A1 gene therapy in pigs. Here we present a sensitive and selective LC-MS/MS approach that can easily differentiate and simultaneously quantitate both human and pig S100A1 proteins. Additionally, we report on a detailed profiling of S100A1 protein in various pig tissues, a comprehensive evaluation of S100A1 distribution in pig hearts and a comparison to S100A1 levels in human cardiac samples.

Demolished concretes recycling by the use of pneumatic jigs.

Large quantities of construction and demolition waste is generated annually around the world. Part of this material is processed in recycling plants. After removing metals, fines and lights, the construction and demolition waste is crushed and sized and can be used as aggregates for low resistance concrete, for road sub-base, city landfill and other low value-added applications. For their use as coarse aggregate in structural concretes, construction and demolition waste must exhibit high densities and regularity of the material. This material usually is presented in demolished concretes. About 20% of the particles from demolished concretes can be used as coarse aggregates substituting part of natural aggregates in structural concretes. This article presents studies of demolished concretes recycling by the use of a pneumatic jig. All jigging tests were carried out with three different concretes produced in three strength classes: C16/20, ordinary concrete; C50/60, high strength concrete; and C70/85, very high strength concrete. Based on density distribution of the three concretes, there are reasonable masses with densities over 2.7 g cm-3, particle density considered appropriate to the used as coarse aggregate for structural concretes. The concretes present different mass recoveries of the denser particles (different liberation). Coarse aggregates can be recovered with reasonable masses by the use of air jigs: About 65% for high strength concretes and about 75% for the low strength concrete. The jigging concentration efficiency depends on the concrete liberation, density and size distribution.

Cerebral microbleeds in adult survivors of childhood acute lymphoblastic leukemia treated with cranial radiation.

Cranial radiation therapy is associated with white matter-specific brain injury, cortical volume loss, mineralization, microangiopathy and neurocognitive impairment in survivors of childhood acute lymphoblastic leukemia. In this retrospective cross-sectional analysis, neurocognitive testing and 3 T brain MRI's were obtained in 101 survivors treated with cranial radiation. Small focal intracerebral hemorrhages only visible on exquisitely sensitive MRI sequences were identified and localized using susceptibility weighted imaging. Modified Poisson regression was used to assess the effect of cranial radiation on cumulative number and location of microbleeds in each brain region, and multiple linear regression was used to evaluate microbleeds on neurocognitive outcomes, adjusting for age at diagnosis and sex. At least one microbleed was present in 85% of survivors, occurring more frequently in frontal lobes. Radiation dose of 24 Gy conveyed a 5-fold greater risk (95% CI 2.57-10.32) of having multiple microbleeds compared to a dose of 18 Gy. No significant difference was found in neurocognitive scores with either the absence or presence of microbleeds or their location. Greater prevalence of microbleeds in our study compared to prior reports is likely related to longer time since treatment, better sensitivity of SWI for detection of microbleeds and the use of a 3 T MRI platform.

In Vitro Assembly of Virus-Derived Designer Shells Around Inorganic Nanoparticles.

Nanoparticle-templated assembly of virus shells provides a promising approach to the production of hybrid nanomaterials and a potential avenue toward new mechanistic insights in virus phenomena originating in many-body effects, which cannot be understood from examining the properties of molecular subunits alone. This approach complements the successful molecular biology perspective traditionally used in virology, and promises a deeper understanding of viruses and virus-like particles through an expanded methodological toolbox. Here we present protocols for forming a virus coat protein shell around functionalized inorganic nanoparticles.

Successful re-ablation of a permanent junctional reciprocating tachycardia with cryoenergy : Case report of a 19-year-old patient.

We report the case of a 19-year-old male patient who presented with a permanent junctional reciprocating tachycardia (PJRT). After a primarily successful radiofrequency ablation of a para-Hisian, midseptal, accessory pathway, recurrence of tachycardia was documented. Thereafter, successful ablation using cryoenergy was performed. Since this second ablation the patient has been free of tachycardia. Our case study shows that the treatment of PJRT in young adults using cryoenergy can be successfully and safely conducted, especially after tachycardia recurrence following an initial radiofrequency ablation.

Discovery of Clinical Candidate 2-((2S,6S)-2-Phenyl-6-hydroxyadamantan-2-yl)-1-(3'-hydroxyazetidin-1-yl)ethanone [BMS-816336], an Orally Active Novel Selective 11ß-Hydroxysteroid Dehydrogenase Type 1 Inhibitor.

BMS-816336 (6n-2), a hydroxy-substituted adamantyl acetamide, has been identified as a novel, potent inhibitor against human 11ß-hydroxysteroid dehydrogenase type 1 (11ß-HSD1) enzyme (IC50 3.0 nM) with >10000-fold selectivity over human 11ß-hydroxysteroid dehydrogenase type 2 (11ß-HSD2). 6n-2 exhibits a robust acute pharmacodynamic effect in cynomolgus monkeys (ED50 0.12 mg/kg) and in DIO mice. It is orally bioavailable (%F ranges from 20 to 72% in preclinical species) and has a predicted pharmacokinetic profile of a high peak to trough ratio and short half-life in humans. This ADME profile met our selection criteria for once daily administration, targeting robust inhibition of 11ß-HSD1 enzyme for the first 12 h period after dosing followed by an "inhibition holiday" so that the potential for hypothalamic-pituitary-adrenal (HPA) axis activation might be mitigated. 6n-2 was found to be well-tolerated in phase 1 clinical studies and represents a potential new treatment for type 2 diabetes, metabolic syndrome, and other human diseases modulated by glucocorticoid control.

Late deterioration of left ventricular function after right ventricular pacemaker implantation.

OBJECTIVES: Right ventricular (RV) pacing induces a left bundle branch block pattern on ECG and may promote heart failure. Patients with dual chamber pacemakers (DCPs) who present with progressive reduction in left ventricular ejection fraction (LVEF) secondary to RV pacing are candidates for cardiac resynchronization therapy (CRT). This study analyzes whether upgrading DCP to CRT with the additional implantation of a left ventricular (LV) lead improves LV function in patients with reduced LVEF following DCP implantation. METHODS: Twenty-two patients (13 males) implanted with DCPs and a high RV pacing percentage (>90%) were evaluated in term of new-onset heart failure symptoms. The patients were enrolled in this retrospective single-center study after obvious causes for a reduced LVEF were excluded with echocardiography and coronary angiography. In all patients, DCPs were then upgraded to biventricular devices. LVEF was analyzed with a two-sided t-test. QRS duration and brain natriuretic peptide (BNP) levels were analyzed with the unpaired t-test. RESULTS: LVEF declined after DCP implantation from 54±10% to 31±7%, and the mean QRS duration was 161±20 ms during RV pacing. NT-pro BNP levels were elevated (3365±11436 pmol/L). After upgrading to a biventricular device, a biventricular pacing percentage of 98.1±2% was achieved. QRS duration decreased to 108±16 ms and 106±20 ms after 1 and 6 months, respectively. There was a significant increase in LVEF to 38±8% and 41±11% and a decrease in NT-pro BNP levels to 3088±2326 pmol/L and 1860±1838 pmol/L at 1 and 6 months, respectively. CONCLUSION: Upgrading to CRT may be beneficial in patients with DCPs and heart failure induced by a high RV pacing percentage.

Quantification of in vivo site-specific Asp isomerization and Asn deamidation of mAbs in animal serum using IP-LC-MS.

BACKGROUND: Isomerization of aspartic acid and deamidation of asparagine are two common amino acid modifications that are of particular concern if located within the complementarity-determining region of therapeutic antibodies. Questions arise as to the extent of modification occurring in circulation due to potential exposure of the therapeutic antibody to different pH regimes. RESULTS: To enable evaluation of site-specific isomerization and deamidation of human mAbs in vivo, immunoprecipitation (IP) has been combined with LC-MS providing selective enrichment, separation and detection of naive and modified forms of tryptic peptides comprising complementarity-determining region sequences. CONCLUSION: IP-LC-MS can be applied to simultaneously quantify in vivo drug concentrations and measure the extent of isomerization or deamidation in PK studies conducted during the drug discovery stage.

Paired self-compensated spin-lock preparation for improved T1ρ quantification.

PURPOSE: Spin-lock (SL) imaging allows quantification of the spin-lattice relaxation time in the rotating frame (T1ρ). B0 and B1 inhomogeneities impact T1ρ quantification because the preparatory block in SL imaging is sensitive to the field heterogeneities. Here, a modified preparatory block (PSC-SL) is proposed that attempts to alleviate SL sensitivity to field inhomogeneities in scenarios where existing approaches fail, i.e. high SL frequencies. METHODS: Computer simulations, phantom and in vivo experiments were used to determine the effect of field inhomogeneities on T1ρ quantification. Existing SL preparations were compared with PSC-SL in different conditions to assess the advantages and disadvantages of each method. RESULTS: Phantom experiments and computer modeling demonstrate that PSC-SL provides superior T1ρ quantification at high SL frequencies in situations where the existing SL preparation methods fail. This result has been confirmed in pre-clinical neuro and body imaging at 7T. CONCLUSION: PSC-SL complements existing methods by increasing the accuracy of T1ρ quantification at high spin-lock frequencies when large field inhomogeneities are present. A-priory information about the experimental conditions such, as field distribution and spinlock frequency are useful for selecting an appropriate spin-lock preparation for specific applications.

Successful intraprocedural anticoagulation with bivalirudin during pulmonary vein isolation in a patient with known heparin-induced thrombocytopenia type II.

We report the case of a 56-year-old female who presented with symptomatic paroxysmal atrial fibrillation. Anamnestic heparin-induced thrombocytopenia (HIT) type II was suspected, and a rapid diagnostic test showed antibodies against platelet factor 4. The heparin-induced platelet activation-assay was negative. Radiofrequency pulmonary vein isolation with intraprocedural anticoagulation using bivalirudin was ultimately performed. Dosing was controlled by monitoring the activated clotting time. Post-procedural blood tests were normal. There were no thromboembolic or bleeding events. Bivalirudin is a therapeutic option for anticoagulation during pulmonary vein isolation procedures in patients with a history of HIT type II.

Examining the Heterogeneous Genome Content of Multipartite Viruses BMV and CCMV by Native Mass Spectrometry.

Since the concept was first introduced by Brian Chait and co-workers in 1991, mass spectrometry of proteins and protein complexes under non-denaturing conditions (native MS) has strongly developed, through parallel advances in instrumentation, sample preparation, and data analysis tools. However, the success rate of native MS analysis, particularly in heterogeneous mega-Dalton (MDa) protein complexes, still strongly depends on careful instrument modification. Here, we further explore these boundaries in native mass spectrometry, analyzing two related endogenous multipartite viruses: the Brome Mosaic Virus (BMV) and the Cowpea Chlorotic Mottle Virus (CCMV). Both CCMV and BMV are approximately 4.6 megadalton (MDa) in mass, of which approximately 1 MDA originates from the genomic content of the virion. Both viruses are produced as mixtures of three particles carrying different segments of the genome, varying by approximately 0.1 MDA in mass (~2%). This mixture of particles poses a challenging analytical problem for high-resolution native MS analysis, given the large mass scales involved. We attempt to unravel the particle heterogeneity using both Q-TOF and Orbitrap mass spectrometers extensively modified for analysis of very large assemblies. We show that manipulation of the charging behavior can provide assistance in assigning the correct charge states. Despite their challenging size and heterogeneity, we obtained native mass spectra with resolved series of charge states for both BMV and CCMV, demonstrating that native MS of endogenous multipartite virions is feasible. Graphical Abstract ᅟ.

[Transseptal puncture after device closure of patent foramen ovale: Is it feasible?]. / Transseptale Punktion nach PFO-Verschluss: Geht das?

CASE REPORT: The case of a 40-year-old woman with paroxysmal symptomatic atrial fibrillation and implanted occluder of a patent foramen ovale (PFO; AMPLATZER™ Septal Occluder, St. Jude Medical) is reported. Due to the symptomic atrial fibrillation, pulmonary vein isolation was planned. METHODS: Under transesophageal, echocardiographic control the transseptal puncture was performed posterior inferior of the occluder without any complications. The pulmonary vein was successfully isolatedusing radiofrequency energy. CONCLUSION: This case demonstrates that transseptal puncture in pulmonary vein isolation with an inserted PFO occluder under additional transesophageal, echocardiographic monitoring is safe and feasible.

[Recurrent microdislocation of pacemaker leads due to an aortic aneurysm of the aorta ascendens]. / Rezidivierende Mikrodislokationen von Schrittmachersonden aufgrund eines Aortenaneurysmas der Aorta ascendens.

We report about a 79 years old female patient which was admitted due to a symptomatic AV block 3rd degree. The coronary angiography excluded a coronary artery disease and the echocardiography revealed a normal left ventricular systolic function. Therefore a dual-chamber pacemaker was implanted. Following two micro-dislocations of the right ventricular lead, which required operative revisions, a computed tomography of the heart was performed. This detected an aneurysm of the ascending aorta (5 cm maximum diameter) with compression of the superior caval vein. This case shows that a possible cause of recurrent micro-dislocations could be a pathological anatomy of the heart.

[Wide QRS complex tachycardia in a patient with ischemic cardiomyopathy and prior ventricular tachycardia ablation]. / Breitkomplex-Tachykardie bei einem Patienten mit ischämischer Kardiomyopathie und stattgehabter Ablation einer ventrikulären Tachykardie.

CASE REPORT: This article reports the case of a 76-year-old patient with multiple implantable cardioverter defibrillator (ICD) interventions of the single chamber ICD. On admission the 12-lead electrocardiogram (ECG) documented a wide QRS complex tachycardia without clearly identifiable P waves. The patient had previously had two ventricular tachycardia (VT) ablations due to VT storms. The resting ECG revealed a sinus rhythm with a wide QRS complex. During the electrophysiological investigation typical atrial flutter could be detected and an ablation of the cavotricuspid isthmus was performed. During the follow-up period, the patient has been free from tachycardia. CONCLUSION: The case described emphasizes that a differentiation between supraventricular tachycardia (SVT) and VT based on a stored ECG without an atrial channel can be challenging.

Segmented GFP-like aptamer probes for functional imaging of viral genome trafficking.

Recently developed GFP-like RNA aptamers harbor a few unique potential benefits for in vivo RNA imaging applications, including co-packaging of viral genomes. Here we examine them in the context of co-packaging of RNA strands during virion assembly and trafficking. The approach is applicable both in vitro and in vivo, thus bridging an existing methodological gap. We have found that splitting the aptamer sequence in the loop region into two separate parts allows for subsequent self-assembly into a functional unit, which preserves the dye-binding pocket. In presence of the dye, virus-like particles encapsulating segmented GFP-like aptamers provided bright fluorescence emission and showed negligible bleaching due to continuous chromophore exchange: two desirable characteristics for real-time in vivo single particle studies requiring a broader dynamic range than currently available. Proof-of-principle in vivo imaging experiments confirmed detectability of aptamer-loaded virus-like particles in barley root cells even in presence of significant autofluorescence background.

Encapsulation of nanoparticles in virus protein shells.

The self-assembly of virus-like particles may lead to materials which combine the unique characteristics of viruses, such as precise size control and responsivity to environmental cues, with the properties of abiotic cargo. For a few different viruses, shell proteins are amenable to the in vitro encapsulation of non-genomic cargo in a regular protein cage. In this chapter we describe protocols of high-efficiency in vitro self-assembly around functionalized gold nanoparticles for three examples of icosahedral and non-icosahedral viral protein cages derived from a plant virus, an animal virus, and a human retrovirus. These protocols can be readily adapted with small modifications to work for a broad variety of inorganic and organic nanoparticles.

Quantification of human mAbs in mouse tissues using generic affinity enrichment procedures and LC-MS detection.

BACKGROUND: The disease state can modulate the penetration of large antibody-sized therapeutic molecules into affected tissues. Suitable bioanalytical methods are required for the quantitative analysis of drug tissue levels to enable a better understanding of the parameters influencing drug penetration and target engagement. RESULTS: Described is a sensitive and selective LC-MS/MS assay for the quantification of human mAb molecules in mouse tissues. By homogenizing tissues directly into serum, a common serum calibration curve can be used for multiple tissues. A generic procedure was used for affinity enrichment. An analytical range of 20 - 20,000 ng/ml was achieved in serum. CONCLUSION: The method described here can be applied for the quantitative analysis of mAb and Fc-fusion therapeutic molecules in a variety of animal tissue matrices.