Hydrogen-Deuterium-Exchange-Based Mass Distribution Shifts in High-Resolution Cyclic Ion Mobility Separations.

The mass distribution of ions influences separations in ion mobility spectrometry-mass spectrometry (IMS-MS). Herein, we introduce a method to induce mass distribution shifts for various analytes using hydrogen-deuterium exchange (HDX) immediately prior to ionization using a dual syringe approach. By replacing labile hydrogens on analytes with deuteriums, we were able to differentiate isomers using separations of isotopologues. For each analyte studied, every possible level of deuteration (from undeuterated to fully deuterated) was generated and then separated using cyclic ion mobility spectrometry-mass spectrometry (cIMS-MS). The information gained from such separations (relative arrival times; tRel. values) was found to be orthogonal to conventional IMS-MS separations. Additionally, the observed shifts were linearly additive with increasing deuteration, suggesting that this methodology could be extended to analytes with a larger number of labile hydrogens. For one isomer pair, as few as two deuteriums were able to produce a large enough mass distribution shift to differentiate isomers. In another experiment, we found that the mass distribution shift was large enough to overcome the reduced mass contribution, resulting in a "flipped" arrival time where the heavier deuterated isotopologue arrived before the lighter one. In this work, we present a proof-of-concept demonstration that mass-distribution-based shifts, tRel. values, could potentially act as an added dimension to characterize molecules in IMS-MS. We anticipate, along with future work in this area, that mass-distribution-based shifts could enable the identification of unknown molecules through a database-driven approach in an analogous fashion to collision cross section (CCS) measurements.

Endoscopic Fascia Release for Forearm Chronic Exertional Compartment Syndrome: Case Report and Surgical Technique.

BACKGROUND: Chronic exertional compartment syndrome (CECS) of the forearm is traditionally treated with open compartment release requiring large incisions that can result in less than optimal esthetic results. The purpose of this study is to describe a case report of 2 professional motocross patients with forearm CECS treated endoscopically using a minimally invasive technique. METHODS: Two professional motocross racers presented with a history of chronic proximal volar forearm pain when motocross riding. Other symptoms included paresthesia and weakness, which, at times, led to an inability to continue riding. Both failed conservative management. Compartment pressure measurements were performed before and after provocative exercises to confirm diagnosis of CECS. Release of both the volar and dorsal compartments was performed endoscopically through a single incision. RESULTS: Symptoms resolved after surgery. The first patient resumed riding at 1 week, competing at 3 weeks, and continues to ride competitively without symptoms at 3 years postoperative. The second patient began riding at 1 week and won second place in the National Supercross finals 5 weeks after simultaneous bilateral release. CONCLUSIONS: This technique is simple and effective. The cannula used protects the superficial nerves while allowing release through a small, cosmetically pleasing incision.

The Critical Importance of Retrieval--and Spacing--for Learning.

We examined the impact of repeated testing and repeated studying on long-term learning. In Experiment 1, we replicated Karpicke and Roediger's (2008) influential results showing that once information can be recalled, repeated testing on that information enhances learning, whereas restudying that information does not. We then examined whether the apparent ineffectiveness of restudying might be attributable to the spacing differences between items that were inherent in the between-subjects design employed by Karpicke and Roediger. When we controlled for these spacing differences by manipulating the various learning conditions within subjects in Experiment 2, we found that both repeated testing and restudying improved learning, and that learners' awareness of the relative mnemonic benefits of these strategies was enhanced. These findings contribute to understanding how two important factors in learning-test-induced retrieval processes and spacing-can interact, and they illustrate that such interactions can play out differently in between-subjects and within-subjects experimental designs.

Distress in patients with bleeding disorders: a single institutional cross-sectional study.

INTRODUCTION: Distress may affect a patient's ability to cope with and manage disease. AIM: To report distress prevalence in adult patients with bleeding disorders and determine whether specific clinical and health characteristics, including disease severity and employment status, are associated with distress. METHODS: Patients who visited a Haemophilia Treatment Centre (HTC) between January 1st, 2012 through February 28th, 2014 and who completed a distress screen, pain screen and questionnaire were evaluated cross sectionally. Distress was measured by the National Comprehensive Cancer Network Distress Management Tool, which allowed patients to rate recent distress on a 0-10 point scale. A rating of five or more was categorized as high distress. Pain was measured by the Brief Pain Inventory Short Form, which asked patients to rate pain types on 0-10 point scales. Patients reported employment and other demographic and behavioural information on the questionnaire. Primary diagnosis, age, HIV and HCV status were abstracted from medical records. Adjusted logistic regression was used to identify distress associations. RESULTS: High distress prevalence among 152 patients with bleeding disorders was 31.6%. Unemployment, disability, greater depressive symptoms and higher pain were associated with high distress in multivariable models. Bleeding disorder diagnosis, race/ethnicity, HIV/HCV status and on-demand treatment regimen were not associated with high distress. CONCLUSION: Distress among patients with congenital bleeding disorders followed at a comprehensive HTC was high and similar to that reported among patients with cancer. Future research should determine whether distress impacts clinical outcomes in patients with bleeding disorders as demonstrated in other chronic disorders.

Long-Term Outcome of Arthroscopic Resection Arthroplasty With or Without Interposition for Thumb Basal Joint Arthritis.

PURPOSE: To report results on 144 cases following arthroscopic resection arthroplasty (ARA) with or without interposition for basal joint arthritis. METHODS: Cases undergoing ARA for thumb carpometacarpal osteoarthritis between 2004 and 2011 were prospectively enrolled (n = 178). Data were collected before surgery and after surgery at 1, 3, 6, and 12 months and annually thereafter. Patients were excluded for less than 1-year follow-up or concomitant procedures that interfered with evaluation of the variable of interest (interposition). Human acellular dermal matrix (GRAFTJACKET) was the most commonly used interposition. Outcomes on 19 cases of interposition using collagen bioimplant (OrthADAPT) and porous polyurethaneurea (Artelon) scaffolds were also reported. Comparative analyses were performed on 52 patients with GRAFTJACKET interposition and on 73 without. Mean follow-up was 7.4 and 5.6 years with and without interposition, respectively. Descriptive statistics were evaluated on all baseline variables. Raw change scores of grip, pinch, and pain outcomes were evaluated. Confounding variables at a significance level of P less than .05 were adjusted for in linear mixed models, and an analysis of covariance was employed through an unstructured type of variance-covariance matrix. RESULTS: Change scores from baseline to 1 year for the interposition group for pain (numerical rating scale, 0-10), pinch, and grip was -5.8, 3.3, and 7, respectively, and -5.1, 2.1, and 9 for the noninterposition group. Postoperative mean satisfaction was 4.7 and 4.4 for the with- and without-interposition groups, respectively. There were 4 failures with and 2 without interposition. Artelon and OrthADAPT did poorly with unacceptably high failure rates. CONCLUSIONS: This study suggested that interposition is not necessary following ARA for thumb basal joint arthritis. Because arthroscopic interposition of material contributes to health care costs in terms of patient and facility costs without clear benefit to the patient, routine use of expensive interposition products should be abandoned or carefully evaluated with a prospective randomized controlled trial. TYPE OF STUDY/LEVEL OF EVIDENCE: Therapeutic IV.

Setting expectations following endoscopic cubital tunnel release.

BACKGROUND: The objective was to evaluate recovery characteristics of patients undergoing endoscopic cubital tunnel release (ECuTR) by determining the following: (1) return to work (RTW) times following ECuTR compared with RTW times of patients that underwent anterior transposition of the ulnar nerve (ATUN), (2) satisfaction rates and factors affecting satisfaction, (3) resolution rates of common preoperative complaints and findings, and (4) effect of preoperative ulnar nerve subluxation on postoperative outcomes. METHODS: A total of 172 cases in 148 patients undergoing ECuTR were prospectively enrolled including 56 women and 92 men. Kaplan-Meier analyses were performed to determine RTW time for ECuTR patients and for a cohort of 15 patients that underwent ATUN. Patients were evaluated for subjective and objective complaints preoperatively and postoperatively. Cases were grouped by Dellon's classification preoperatively and modified by Bishop's postoperatively. RESULTS: Half of ECuTR patients returned to normal work within 8 days postoperatively versus 71 days following ATUN. Variables significantly negatively affecting RTW were male sex, manual labor, and worker's compensation status. Dellon's was the best predictor of postoperative satisfaction. Complete resolution of symptoms occurred in 86 % of patients for weakness, 81 % for pain, 79 % for numbness and tingling (N/T), 78 % for atrophy, 76 % for abnormal two-point discrimination, and 65 % for Wartenberg's. Preoperative ulnar nerve subluxation had no effect on outcome. CONCLUSIONS: Improved RTW time following ECuTR versus ATUN indicates potential and substantial cost-saving implications with respect to reduced worker productivity loss. Patients with more severe preoperative Dellon's classification can expect less optimal results regarding postoperative satisfaction and resolution rates of N/T and pain.

Arthroscopic Resection Arthroplasty of the Radial Column for SLAC Wrist.

Background Symptomatic advanced scapholunate advanced collapse (SLAC) wrists are typically treated with extensive open procedures, including but not limited to scaphoidectomy plus four-corner fusion (4CF) and proximal row carpectomy (PRC). Although a minimally invasive arthroscopic option would be desirable, no convincing reports exist in the literature. The purpose of this paper is to describe a new surgical technique and outcomes on 14 patients who underwent arthroscopic resection arthroplasty of the radial column (ARARC) for arthroscopic stage II through stage IIIB SLAC wrists and to describe an arthroscopic staging classification of the radiocarpal joint for patients with SLAC wrist. Patients and Methods Data were collected prospectively on 17 patients presenting with radiographic stage I through III SLAC wrist who underwent ARARC in lieu of scaphoidectomy and 4CF or PRC. Fourteen patients (12 men and 2 women) subject to 1-year follow-up were included. The average age was 57 years (range 41 to 78). The mean follow-up was 24 months (range 12 to 61). Arthroscopic resection arthroplasty of the radial column is described for varying stages of arthritic changes of the radioscaphoid joint. Midcarpal resection was not performed. Results The mean Disabilities of the Arm, Shoulder, and Hand (DASH) score was 66 preoperatively and 28 at final follow-up. The mean satisfaction (0 = not satisfied, 5 = completely satisfied) at final follow-up was 4.5 (range 3 to 5). The pain level (on 0-10 scale) improved from 6.6 to 1.3. The total arc of motion changed from 124° preoperatively to 142° postoperatively following an ARARC. Grip was 16 kg preoperatively and 18 kg postoperatively. Radiographic stages typically underestimated arthroscopic staging. Although four of our patients appeared to be radiographic stage I, all were found to have arthritis involving some or all of the radioscaphoid articulation at the time of arthroscopy. Clinical Relevance Pain relief is rapid and remains consistent over time following ARARC. ARARC may be a viable surgical option for patients with SLAC wrist who desire a minimally invasive procedure. Radiographic stages underestimate the degree of arthritic change. Accurate staging requires arthroscopy. The indications and long-term outcome are not well defined; continued surveillance is warranted. Level of Evidence Level IV, Therapeutic study.

Wrong site surgery-where are we and what is the next step?

BACKGROUND: Wrong site surgery is estimated to occur 40 times per week in hospitals and clinics in USA. The universal protocol was implemented by the joint commission board of commissioners to address wrong site, wrong procedure, and wrong person surgery. DISCUSSION: The universal protocol has three principal components: preoperative verification, marking of the operative site, and a time-out. Despite this organized approach to this problem, current data do not demonstrate any progress. In fact some data suggest that the problem may be getting worse. It is apparent that a process relying on surgeon and surgical team memory is doomed to ultimate failure. Recommendations are made for a more in depth checklist process based on the recommendations of the World Health Organization, reports in the literature of known areas of weakness in the current process, and personal experience in hopes of establishing a more bullet proof system to avoid wrong site procedure.

New frontiers in hand arthroscopy.

This article covers new and emerging techniques in small joint arthroscopy in the hand. Recent improvement in the quality of small joint scopes and advancement in techniques have allowed for many new small joint arthroscopic procedures in the hand. The arthroscopic classification for thumb carpometacarpal (CMC) arthritis as well as treatment of each stage are described. Arthroscopic treatment of pantrapezial arthrosis is reviewed. Metacarpophalangeal arthroscopy for the treatment of synovitis, arthritis, fractures, and gamekeeper injuries are discussed, as is arthroscopy of the proximal interphalangeal, pisotriquetral, fourth and fifth CMC, and distal interphalangeal joints.

Endoscopic cubital tunnel release.

A minimally invasive endoscopic approach has been successfully applied to surgical treatment of cubital tunnel syndrome. This procedure allows for smaller incisions with faster recovery time. This article details relevant surgical anatomy, indications, contraindications, surgical technique, complications, and postoperative management.

Endoscopic cubital tunnel recurrence rates.

Cubital tunnel syndrome is the second most common nerve entrapment in the upper extremity. There are no current publications concerning the recurrence rates after endoscopic cubital tunnel release. The purpose of this study is to evaluate the recurrence rate of endoscopic cubital tunnel release compared to published reports of recurrence following open cubital tunnel procedures. We reviewed 134 consecutive cases of endoscopic cubital tunnel release in 117 patients. There were 104 cases in 94 patients with greater than 3 months follow-up. The mean follow-up time was 736 days. They were grouped using Dellon's classification. Two literature control groups were used from published reports of recurrence rate following open cubital tunnel release. A recurrence was identified if the patient was symptom-free following surgery but had symptoms reappear 3 months or more after surgery as defined in the literature. Of the 104 cases, 92.31% had more than a 4-month follow-up. One case (0.96%) met the criteria for recurrence at 4 months postprocedure. Data were then compared to the literature control groups used from published reports of recurrence rates following open cubital tunnel release. Pooled, the combined controls had 22 of 180 cases (12.22%) with recurrences. The percentage of procedure recurrence varied significantly with p value equal to 0.0004. It is recognized that there is a lack of common classification and comparative analysis of these studies, but they do classify preoperative grading and recurrence similarly. We are 95% confident that our true recurrence rate is between 0.02% and 5.24% and that endoscopic cubital tunnel release has a recurrence rate, which is not higher than open cubital tunnel release literature controls.

Arthroscopic distal interphalangeal joint arthrodesis.

Arthrodesis using a headless screw has been shown to be an effective surgical treatment for pain, deformity, or instability arising at the distal interphalangeal joint of the fingers or interphalangeal joint of the thumb. Previous publications have documented the utility of this treatment using open techniques. Common indications include degenerative, inflammatory, or posttraumatic arthritis and chronic mallet finger or chronic flexor digitorum profundus injury. The purpose of this study is to present an arthroscopic technique for arthrodesis of the distal interphalangeal joint of the fingers or interphalangeal joint of the thumb using the headless screw. The surgical technique is carefully outlined including surgical setup and necessary equipment. Potential technical difficulties and complications are also reviewed.

Biochemical and functional characterization of Salp20, an Ixodes scapularis tick salivary protein that inhibits the complement pathway.

Ixodes ticks are vectors of several pathogens including Borrelia burgdorferi. Tick saliva contains numerous molecules that facilitate blood feeding without host immune recognition and rejection. We have expressed, purified, and characterized Ixodes scapularis salivary protein 20 (Salp20), a potential inhibitor of the alternative complement pathway that shares homology with the Isac protein family. When analysed by SDS-PAGE and size exclusion chromatography, Salp20 was approximately 48 kDa, more than double its predicted mass, primarily due N- and O-linked glycosylations. Recombinant Salp20 inhibited the alternative complement pathway by dissociating the C3 convertase, and partially protected a serum sensitive species of Borrelia from lysis by normal human serum. We propose that Salp20 facilitates tick feeding and possibly protects tick-borne pathogens from complement components.

hAG-2 and hAG-3, human homologues of genes involved in differentiation, are associated with oestrogen receptor-positive breast tumours and interact with metastasis gene C4.4a and dystroglycan.

hAG-2 and hAG-3 are recently discovered human homologues of the secreted Xenopus laevis proteins XAG-1/2 (AGR-1/2) that are expressed in the cement gland, an ectodermal organ in the head associated with anteroposterior fate determination during early development. Although the roles of hAG-2 and hAG-3 in mammalian cells are unknown, both proteins share a high degree of protein sequence homology and lie adjacent to one another on chromosome 7p21. hAG-2 mRNA expression has previously been demonstrated in oestrogen receptor (ER)-positive cell lines. In this study, we have used real-time quantitative RT - PCR analysis and immunohistochemistry on tissue microarrays to demonstrate concordant expression of hAG-2 and hAG-3 mRNA and protein in breast tumour tissues. Tumour expression of both genes correlated with OR (hAG2, P=0.0002; hAG-3, P=0.0012), and inversely correlated with epidermal growth factor receptor (EGFR) (P=0.003). Yeast two-hybrid cloning identified metastasis-associated GPI-anchored C4.4a protein and extracellular alpha-dystroglycan (DAG-1) as binding partners for both hAG-2 and hAG-3, which if replicated in clinical oncology would demonstrate a potential role in tumour metastasis through the regulation of receptor adhesion and functioning. hAG-2 and hAG-3 may therefore serve as useful molecular markers and/or potential therapeutic targets for hormone-responsive breast tumours.

Gene expression profiles of pancreatic cancer and stromal desmoplasia.

Gene expression studies were undertaken in normal pancreas and pancreatic adenocarcinomas to determine new candidate genes that can potentially be used as markers of the disease. The characteristic desmoplastic stromal reaction of pancreatic adenocarcinoma greatly hampers expression studies in this tumour type, and usually necessitates time-consuming tissue microdissection for enrichment of the tumour cell population. We show that fine needle aspiration of cancer provides a fast and efficient way of obtaining samples highly enriched in tumour cells with sufficient yields of RNA. Using Atlas cancer cDNA arrays with 588 cancer-related genes, we describe gene expression profiles of normal pancreas, bulk pancreatic tumour tissues and pancreatic tumour aspirates containing more than 95% tumour cells. Analysis of bulk tissue specimens revealed differentially expressed genes belonging predominantly to the stromal component of the tumour. This contrasted with the results obtained from tumour-cell enriched samples. Several genes already described in pancreatic cancer (caspase 8, TIMP1, CD9, IL-13) were also differentially expressed in our study. Furthermore, we found dysregulated expression of genes not previously associated with pancreatic adenocarcinoma, such as Rac 1, GLG1, NEDD5, RPL-13a, RPS9 and members of the Wnt5A gene family. In summary, we present a panel of genes newly identified in the pathogenesis of pancreatic adenocarcinoma and demonstrate that fine needle aspirates of the tumour mass are a convenient source of material for gene expression studies in tumours accompanied by desmoplastic reactions.

Linked chromosome 16q13 chemokines, macrophage-derived chemokine, fractalkine, and thymus- and activation-regulated chemokine, are expressed in human atherosclerotic lesions.

Chemokines are important mediators of macrophage and T-cell recruitment in a number of inflammatory pathologies, and chemokines expressed in atherosclerotic lesions may play an important role in mononuclear cell recruitment and macrophage differentiation. We have analyzed the expression of the linked chromosome 16q13 genes that encode macrophage-derived chemokine (MDC/CCL22), thymus- and activation-regulated chemokine (TARC/CCL17), and the CX(3)C chemokine fractalkine (CX(3)CL1) in primary macrophages and human atherosclerotic lesions by reverse transcription-polymerase chain reaction and immunohistochemistry. We show that macrophage expression of the chemokines MDC, fractalkine, and TARC is upregulated by treatment with the Th2-type cytokines interleukin-4 and interleukin-13. High levels of MDC, TARC, and fractalkine mRNA expression are seen in some, but not all, human arteries with advanced atherosclerotic lesions. Immunohistochemistry shows that MDC, fractalkine, and TARC are expressed by a subset of macrophages within regions of plaques that contain plaque microvessels. We conclude that MDC, fractalkine, and TARC, which are chromosome 16q13 chemokines, could play a role in mononuclear cell recruitment into atherosclerotic lesions and influence the subsequent inflammatory response. Macrophage-expressed chemokines upregulated by interleukin-4 may be useful surrogate markers for the presence of Th2-type immune responses in human atherosclerotic lesions.

Impact of biodiesel source material and chemical structure on emissions of criteria pollutants from a heavy-duty engine.

Biodiesel is an oxygenated diesel fuel made from vegetable oils and animal fats by conversion of the triglyceride fats to esters via transesterification. In this study we examined biodiesels produced from a variety of real-world feedstocks as well as pure (technical grade) fatty acid methyl and ethyl esters for emissions performance in a heavy-duty truck engine. The objective was to understand the impact of biodiesel chemical structure, specifically fatty acid chain length and number of double bonds, on emissions of NOx and particulate matter (PM). A group of seven biodiesels produced from real-world feedstocks and 14 produced from pure fatty acids were tested in a heavy-duty truck engine using the U.S. heavy-duty federal test procedure (transient test). It was found that the molecular structure of biodiesel can have a substantial impact on emissions. The properties of density, cetane number, and iodine number were found to be highly correlated with one another. For neat biodiesels, PM emissions were essentially constant at about 0.07 g/bhp-h for all biodiesels as long as density was less than 0.89 g/cm3 or cetane number was greater than about 45. NOx emissions increased with increasing fuel density or decreasing fuel cetane number. Increasing the number of double bonds, quantified as iodine number, correlated with increasing emissions of NOx. Thus the increased NOx observed for some fuels cannot be explained by the NOx/PM tradeoff and is therefore not driven by thermal NO formation. For fully saturated fatty acid chains the NOx emission increased with decreasing chain length for tests using 18, 16, and 12 carbon chain molecules. Additionally, there was no significant difference in NOx or PM emissions for the methyl and ethyl esters of identical fatty acids.

Cloning and expression profiling of Hpa2, a novel mammalian heparanase family member.

Heparan sulfate proteoglycans are important constituents of the extracellular matrix and basement membrane. Cleavage of heparan sulfate by heparanase, an endoglycosidase, is implicated in the extravasation of leukocytes and metastatic tumour cells, identifying this enzyme(s) as a target for anti-inflammatory and anti-metastatic therapies. The cloning of a cDNA encoding human heparanase (Hpa1) was reported recently, together with evidence indicating that the hpa1 gene is unique and unlikely to belong to a family of related genes. Here we report the cloning of a cDNA encoding a novel human protein, HPA2, with significant homology to Hpa1. Alternative splicing of the hpa2 transcript yields three different mRNAs, encoding putative proteins of 480, 534, and 592 amino acids. Sequence analyses predict that all three Hpa2 proteins are intracellular, membrane-bound enzymes. Hpa2 also shows a markedly different mRNA distribution to Hpa1 in both normal and cancer tissues. The difference in expression profiles and predicted cellular locations suggests that Hpa2 and Hpa1 proteins have distinct biological functions.

Expression of mineralisation-regulating proteins in association with human vascular calcification.

OBJECTIVES: These studies aim to investigate the expression and function of mineralisation-regulating proteins in association with human vascular calcification focussing on the similarities and differences between the two major calcification pathologies in man: atherosclerotic, intimal calcification and Monckeberg's sclerotic medial calcification. BACKGROUND: A number of studies have documented expression of mineralisation-regulating proteins in association with human atherosclerotic calcification leading to the suggestion that human vascular calcification may be a regulated process with similarities to developmental osteogenesis. METHODS: In situ hybridisation, immunohistochemistry and semi-quantitative RT-PCR analysis were used to determine the temporal and spatial expression patterns of mineralisation-regulating proteins within human calcified vascular lesions. Additionally, the expression and regulation of bone-associated proteins was analysed during spontaneous calcification of human VSMCs in vitro. RESULTS: In association with both medial and intimal calcification, the temporal changes in expression of mineralisation-regulating proteins are similar. Some constitutively expressed bone-associated proteins, including matrix Gla protein (MGP), are down-regulated in association with calcification while expression of a number of bone-associated proteins, not normally expressed in the vessel wall, are induced including alkaline phosphatase (ALK), bone sialoprotein (BSP) and bone Gla protein (BGP). In medial calcification the source of expression of these mineralisation-regulating proteins is VSMCs while in intimal lesions both VSMCs and macrophages express them. Furthermore, these bone-associated proteins are spontaneously expressed by VSMCs in vitro suggesting that human VSMCs are capable of simultaneously exhibiting smooth muscle and osteogenic-like properties. CONCLUSIONS: These studies imply that both medial and intimal vascular calcification are regulated processes; however the aetiology of each pathology differs.