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Objective To date, no biomarkers have been established to predict haematological complications and outcomes of extracorporeal membrane oxygenation (ECMO). The aim of this study was to investigate the expression of a panel of microRNAs (miRNAs), which are promising biomarkers in many clinical fields, in patients before and after initiating ECMO. Methods Serum miRNA levels from 14 patients hospitalized for acute respiratory failure and supported with ECMO in our medical intensive care unit were analysed before and 24 hours after ECMO. In total, 179 serum-enriched miRNAs were profiled by using a real-time PCR panel. For validation, differentially expressed miRNAs were individually quantified with conventional real-time quantitative PCR at 0, 24, and 72 hours. Results Under ECMO support, platelet count significantly decreased by 65 × 103/µL (25th percentile = 154.3 × 103/µL; 75th percentile = 33 × 103/µL). Expression of the 179 miRNAs investigated in this study did not change significantly throughout the observational period. Conclusions According to our data, the expression of serum miRNAs was not altered by ECMO therapy itself. We conclude that ECMO does not limit the application of miRNAs as specific clinical biomarkers for the patients’ underlying disease.

Agricultural producers have many incentives to clear small natural areas from their fields, as this can expand their cultivated land base. However, natural areas can play a role in delivering ecosy...

In the agricultural landscape of south-central Alberta, Canada, producers have many incentives to clear small natural habitats from their fields, as this can expand their cultivated land base and r...

microRNAs (miRNAs) have shown promise as potential new biomarkers for myocardial injury and myocardial ischemia. New digital polymerase chain reaction (PCR) techniques allow for highly precise and reliable absolute direct quantification.In this pilot study we used droplet digital PCR (ddPCR) to assess if miRNAs might be released into circulation in patients with functionally relevant coronary artery disease (CAD). Blood samples for measurement of high-sensitivity cardiac troponin I (hs-cTnI) and miRNAs were obtained before, immediately after peak stress, and 2 h after stress testing in a blinded manner in consecutive patients referred for rest/stress myocardial perfusion single-photon emission tomography/computer tomography (MPI-SPECT/CT). ddPCR was used to directly quantify the serum concentrations of miR-21, miR-208a, and miR-499 as potential markers of myocardial injury/ischemia. Functionally relevant CAD was determined by expert interpretation of MPI-SPECT/CT, coronary angiography and fractional flow reserve, if performed.Overall, 200 patients were included and functionally relevant CAD was detected in 85 of them (42%). Neither miR-21, miR-208a, nor miR-499 concentrations differed at rest, stress, or 2-h after stress when comparing patients with versus without functionally relevant CAD, while hs-cTnI concentrations were significantly higher in patients with functionally relevant CAD (P 0.001). Exercise-induced changes in miRNA or hs-cTnI concentrations did not have diagnostic utility and were similar in patients with versus without functionally relevant CAD.miR-208a, miR-21 and miR-499 concentrations at rest, after exercise and exercise-induced changes do not provide additional clinical value regarding the detection of functionally relevant CAD.

For decades, fetal bovine serum (FBS) has been used routinely for culturing many cell types, based on its empirically demonstrated effects on cell growth, and the lack of suitable non-xenogeneic alternatives. The FBS-based culture media do not represent the human physiological conditions, and can compromise biomimicry of preclinical models. To recapitulate in vitro the features of human bone and bone cancer, we investigated the effects of human serum and human platelet lysate on modeling osteogenesis, osteoclastogenesis, and bone cancer in two-dimensional (2D) and three-dimensional (3D) settings. For monitoring tumor growth within tissue-engineered bone in a non-destructive fashion, we generated cancer cell lines expressing and secreting luciferase. Culture media containing human serum enhanced osteogenesis and osteoclasts differentiation, and provided a more realistic in vitro mimic of human cancer cell proliferation. When human serum was used for building 3D engineered bone, the tissue recapitulated bone homeostasis and response to bisphosphonates observed in native bone. We found disparities in cell behavior and drug responses between the metastatic and primary cancer cells cultured in the bone niche, with the effectiveness of bisphosphonates observed only in metastatic models. Overall, these data support the utility of human serum for bioengineering of bone and bone cancers.

Joint disorders can be detrimental to quality of life. There is an unmet need for precise functional reconstruction of native-like cartilage and bone tissues in the craniofacial space and particularly for the temporomandibular joint (TMJ). Current surgical methods suffer from lack of precision and comorbidities and frequently involve multiple operations. Studies have sought to improve craniofacial bone grafts without addressing the cartilage, which is essential to TMJ function. For the human-sized TMJ in the Yucatan minipig model, we engineered autologous, biologically, and anatomically matched cartilage-bone grafts for repairing the ramus-condyle unit (RCU), a geometrically intricate structure subjected to complex loading forces. Using image-guided micromilling, anatomically precise scaffolds were created from decellularized bone matrix and infused with autologous adipose-derived chondrogenic and osteogenic progenitor cells. The resulting constructs were cultured in a dual perfusion bioreactor for 5 weeks before implantation. Six months after implantation, the bioengineered RCUs maintained their predefined anatomical structure and regenerated full-thickness, stratified, and mechanically robust cartilage over the underlying bone, to a greater extent than either autologous bone-only engineered grafts or acellular scaffolds. Tracking of implanted cells and parallel bioreactor studies enabled additional insights into the progression of cartilage and bone regeneration. This study demonstrates the feasibility of TMJ regeneration using anatomically precise, autologous, living cartilage-bone grafts for functional, personalized total joint replacement. Inclusion of the adjacent tissues such as soft connective tissues and the TMJ disc could further extend the functional integration of engineered RCUs with the host.

micro-RNAs have shown promise as potential biomarkers for acute myocardial infarction and ischemia-reperfusion injury (I/R). Most recently droplet digital polymerase chain reaction (ddPCR) has been introduced as a more reliable and reproducible method for detecting micro-RNAs.We aimed to demonstrate the improved technical performance and diagnostic potential of ddPCR by measuring micro-RNAs in ST-elevation myocardial infarction (STEMI).A dilution series was performed in duplicate on synthetic Caenorrhabditis elegans-miR-39, comparing quantitative real-time PCR (qRT-PCR) and ddPCR. We used ddPCR and qRT-PCR to quantify the serum levels of miR-21, miR-208a and miR-499 between STEMI patients (n=24) and stable coronary artery disease (CAD) patients (n=20). In STEMI, I/R injury was assessed via measurement of ST-segment resolution.In the dilution series, ddPCR demonstrated superior coefficient of variation (12.1%vs.32.9%) and limit of detection (0.9325 vs.2.425copies/μl). In the patient cohort, ddPCR demonstrated greater differences in miR-21 levels (2190.5 vs. 484.7copies/μl; p=0.0004 for ddPCR and 136.4 vs. 122.8copies/μl; p=0.2273 for qRT-PCR) and in miR-208a (0 vs. 24.1copies/μl, p=0.0013 for ddPCR and 0 vs. 0copies/μl, p=0.0032 for qRT-PCR), with similar differences observed in miR-499 levels (9.4 vs. 81.5copies/μl, p0.0001 for ddPCR and 0 vs. 19.41copies/μl, p0.0001 for qRT-PCR). ddPCR also more accurately defined STEMI for all miRNAs (area under the curve (AUC) of 0.8021/0.7740/0.9063 for miR-21/208a/499 with ddPCR vs. AUC of 0.6083/0.6917/0.8417 with qRT-PCR). However, there was no association between miR-21/208a/499 levels and ischemia-reperfusion injury.ddPCR demonstrates superiority in both technical performance and diagnostic potential compared to qRT-PCR. Ultimately, this supports its use as a diagnostic method for quantifying micro-RNAs, particularly in large multi-center trials.

Animal venoms are complex chemical arsenals. Most venoms are rich in bioactive peptides with proven potential as research tools, drug leads and drugs. Areas covered: We review recent advances in venom-peptide discovery, particularly the adoption of combined transcriptomic/proteomic approaches for the exploration of venom composition. Expert commentary: Advances in transcriptomics and proteomics have dramatically altered the manner and rate of venom-peptide discovery. The increasing trend towards a toxin-driven approach, as opposed to traditional target-based screening of venoms, is likely to expedite the discovery of venom-peptides with novel structures and new and unanticipated mechanisms of action. At the same time, these advances will drive the development of higher-throughput approaches for target identification. Taken together, these approaches should enhance our understanding of the natural ecological function of venom peptides and increase the rate of identification of novel venom-derived pharmacological tools, drug leads and drugs.

Background Ischemia-reperfusion (I/R) injury in ST-segment elevation myocardial infarction (STEMI) significantly contributes to overall myocardial damage. As a consequence of I/R injury in the heart, the high-temperature requirement protein A2 (HtrA2) is released from the mitochondrial intermembrane space of cardiomyocytes to the cytoplasm, whereupon it induces apoptosis. Methods Serum was obtained from STEMI ( n =37), non–ST-segment elevation myocardial infarction (NSTEMI) ( n =20), stable coronary artery disease (CAD) ( n =17) and patients with CAD excluded ( n =9). In STEMI, I/R injury was assessed via measurement of ST-segment resolution. Results HtrA2 was significantly increased in STEMI compared to NSTEMI, stable CAD and patients with CAD excluded (981.3 (IQR: 543.5–1526.2)pg/mL vs. 494.5 (IQR: 413.8–607)pg/mL vs. 291 (IQR: 239–458.5)pg/mL vs. 692.2 (IQR: 276.6–964.7)pg/mL; p ≤0.0001). STEMI patients with HtrA2 level of at least the median or above had a higher peak creatine kinase (CK) ( p =0.0002) and cardiac troponin T levels (cTnT) ( p =0.0019). Significantly more STEMI patients with HtrA2 levels of at least the median or above were identified as I/R injury (87% vs. 42%; p Conclusion HtrA2 shows promise as a novel potential biomarker for mitochondrial-induced cardiomyocyte apoptosis and may help to identify I/R injury after STEMI.

Background: The extent of myocardial damage in patients with ST-segment elevation myocardial infarction (STEMI) depends on both the time to reperfusion as well as injury induced by ischaemia–reperfusion resulting in a cascade of cellular and humoral reactions. As a consequence of ischaemia–reperfusion in the heart, the high-temperature requirement serine peptidase 2 (HtrA2) is translocated from the mitochondria to the cytosol, whereupon it induces protease activity-dependent apoptosis mediated via caspases. Myocardial damage induced by reperfusion cannot be monitored due to a current lack in specific biomarkers. We examined the serum level of HtrA2 as a potentially novel biomarker for mitochondrial-induced cardiomyocyte apoptosis. Methods: After informed consent, peripheral blood was obtained from patients ( n=19) with first-time acute anterior STEMI after percutaneous coronary intervention. Within this group, 10 of the patients received the mitochondria-targeting peptide elamipretide (phase 2a clinical study EMBRACE (NCT01572909)). Blood was also obtained from a control group of healthy donors ( n=16). The serum level of HtrA2 was measured by an enzyme-linked immunosorbent assay (ELISA). In a murine model of myocardial ischaemia–reperfusion injury, HtrA2 was determined in plasma by ELISA after left anterior descending artery occlusion. Results: HtrA2 median was significantly increased in patients with STEMI compared to healthy controls 392.4 (240.7–502.8) pg/mL vs. 1805.5 (981.3–2220.1) pg/mL ( P⩽0.05). Elamipretide significantly reduced the HtrA2 median serum level after myocardial infarction 1805.5 (981.3–2220.1) pg/mL vs. 496.5 (379.4–703.8) pg/mL ( P⩽0.05). Left anterior descending artery occlusion in mice significantly increased HtrA2 mean in plasma (117.4 fg/ml±SEM 28.1 vs. 525.2 fg/ml±SEM 96; P⩽0.05). Conclusion: Compared to healthy controls, we found significantly increased serum levels of HtrA2 in patients with STEMI. The result was validated in a murine model of myocardial ischaemia–reperfusion injury. In humans the increased serum level was significantly reduced by the mitochondria-targeting peptide elamipretide. In conclusion, HtrA2 is detectable in serum of patients with STEMI and might present a novel biomarker for mitochondrial-induced cardiomyocyte apoptosis. Consequently, HtrA2 may also show promise as a biomarker for the identification of ischaemia–reperfusion injury. However, this must be validated in a lager clinical trial.

INTRODUCTION Decreased expression of highly immunogenic cancer-testis antigens (CTA) might help tumor to achieve low immunogenicity, escape immune surveillance and grow unimpeded. Our aim was to evaluate CTA expression in tumor and normal tissues and to investigate possible means of improving the immune response in a murine prostate cancer (CaP) model by using the combination of epigenetic modifier 5-azacitidine (5-AzaC) and immunomodulator lenalidomide. No study to date has examined the effect of this combination on the prostate cancer or its impact on antigen-presenting cells (APC). MATERIALS AND METHODS Gene microarrays were performed to compare expression of several CTA in murine prostate cancer (RM-1 cells) and normal prostate. RM-1 cells were treated with 5-AzaC and real-time PCR was performed to investigate the expression of several CTA. Western blotting was used to determine whether expression of CTA-specific mRNA induced by 5-AzaC resulted in increase in the corresponding protein. Effect of the epigenetic agents and immunomodulators was assessed on dendritic cells (DC) using flow cytometry, ELISA and T-cell proliferation assay. RESULTS Gene arrays demonstrated decreased expression of 35 CTA in CaP tissue compared to normal prostate. 5-AzaC treatment of RM-1 prostate cancer cells upregulated the expression of all 13 CTA tested in a dose-dependent fashion. DC were treated with 5-AzaC and lenalidomide and the expression of surface markers MHC Class I, MHC Class II, CD80, CD86, CD 205, and CD40 was increased. Combination of 5-AzaC and lenalidomide enhances the ability of DC to stimulate T-cell proliferation in mixed leukocyte reaction. Secretion of IL-12 and IL-15 by DC increased significantly with addition of 5-AzaC or 5-AzaC and lenalidomide. CONCLUSIONS Decreased expression of CTA by prostate cancer may be a means of escaping immune monitoring. Combination of epigenetic modifications and immunomodulation by 5-AzaC and lenalidomide increased tumor immunogenicity and enhanced DC function and may be used in the treatment of advanced prostate cancer. Prostate 77: 361-373, 2017. © 2016 Wiley Periodicals, Inc.

AIMS The purpose of this study was to evaluate lower urinary tract symptoms (LUTS) in idiopathic normal pressure hydrocephalus (iNPH). METHODS Patients with new-onset iNPH were prospectively evaluated for LUTS via detailed history and physical, and administration of questionnaires from the International Consultation on Incontinence to assess incontinence (ICIq-UI), overactive bladder (ICIq-OAB), and quality of life (ICIq-LUTqol), as well as the American Urological Association Symptom Score bother scale. All patients with moderate-to-severe LUTS were offered urodynamic testing. Sub-analysis was performed based on gender, medical comorbidities, and age. RESULTS Fifty-five consecutive patients with iNPH completed the initial evaluation and surveys. Total urinary incontinence score was mild to moderate (8.71 ± 0.64: 0–21 scale) with 90.9% experiencing leakage and 74.5% reporting urge incontinence. The most common OAB symptom was nocturia (2.2 ± 0.14: 0–4 scale) with urge incontinence the most bothersome (3.71 ± 0.44: 0–10 scale). Quality-of-life impact was moderate (4.47 ± 0.4: 0–10 scale) and American Urological Association Symptom Score bother scale was 2.89 ± 0.22 (0–6 scale). Urodynamics testing revealed 100% detrusor overactivity and mean bladder capacity of 200 mL. Several differences were identified based on gender, medical comorbidities, and age. CONCLUSIONS Patients with iNPH present with mild-moderate incontinence of which nocturia is the most common symptom, urge incontinence the most bothersome, with 100% of patients having detrusor overactivity. Younger patients experienced greater bother related to LUTS. To our knowledge, this is the only prospective evaluation of urinary symptoms in patients with new-onset iNPH.

Anti-ischemic therapy remains a challenge due to the complexity of hypoxia response pathways. Hypoxia-inducible factor (HIF)-1 is a heterodimer transcription factor consisting of 2 subunits, HIF-1α and HIF-1β. Hypoxia-dependent activation of HIF-1α regulates cellular O(2) homeostasis. Raynaud syndrome (RS), as a comorbidity of the autoimmune disease systemic sclerosis (SS), is characterized by vasospasms that limit blood flow to the limbs, resulting in hypoxia. A single-center randomized study was conducted to compare prostaglandin E1 (PgE1) therapy with a treatment combining PgE1 and an endothelin-1 blocker, bosentan. A total of 30 patients suffering from SS with RS were enrolled. We examined the regulation of HIF-1α, its target heme oxygenase-1 (HMOX-1), and the serum levels of the HIF-1α protein in a subset of patients as well as in ten healthy individuals. The expression of HIF-1α and HMOX-1 in monocytes was measured using absolute plasmid-based quantitative real-time PCR, whereas serum HIF-1α levels were measured with ELISA. Samples were taken at the time of randomization and after 24 weeks. We found that HIF-1α and HMOX-1 mRNA expression in monocytes and serum HIF-1α protein levels were significantly higher in the SS/RS patients compared to the healthy control group. Single-drug therapy significantly increased HIF-1α and HMOX-1 mRNA expression in monocytes and serum HIF-1α protein levels in the SS/RS patients compared to those at the time of randomization, whereas combining PgE1 with an endothelin-1 blocker prevented the further increases in HIF-1α and HMOX-1 expression. We propose HIF-1α and HMOX-1 as novel markers for anti-ischemic therapy in RS.

In the immediate post-war years Britain needed to decide which of the sciences that had emerged during the war as ‘militarily relevant’, the “nation” needed to continue investing in to ensure national security in peacetime. Into this melee of political policy making George Deacon emerged as a go-between for the oceanographic community, Whitehall (the British Civil Service) officials, and the military establishment. Charting these interactions reveals the second major argument of the book—that the day-to-day organisation for science was formulated and organised within the sub-government below the direct political oversight of ministers. Utilising minute sheets and internal government letters the chapter burrows into committee-work to reveal the inner-workings of the sub-government, arguing that the relationships forged during the war were essential to gaining financial patronage for oceanography in its aftermath.

miRNAs have shown promise as potential biomarkers for acute myocardial infarction (AMI). However, the current used quantitative real-time PCR (qRT-PCR) allows solely for relative expression of nucleic acids and it is susceptible to day-to-day variability, which has limited the validity of using the miRNAs as biomarkers. In this study we explored the technical qualities and diagnostic potential of a new technique, chip-based digital PCR, in quantifying the miRNAs in patients with AMI and ischaemia-reperfusion injury (I/R). In a dilution series of synthetic C.elegans-miR-39, chip-based digital PCR displayed a lower coefficient of variation (8.9% vs 46.3%) and a lower limit of detection (0.2 copies/μL vs 1.1 copies/μL) compared with qRT-PCR. In the serum collected from 24 patients with ST-elevation myocardial infarction (STEMI) and 20 patients with stable coronary artery disease (CAD) patients after percutaneous coronary intervention (PCI), we used qRT-PCR and multiplexed chip-based digital PCR to quantify the serum levels of miRNA-21 and miRNA-499 as they have been validated in AMI in prior studies. In STEMI, I/R injury was assessed via measurement of ST-segment resolution (ST-R). Chip-based digital PCR revealed a statistical significance in the difference of miR-21 levels between stable CAD and STEMI groups (118.8 copies/μL vs 59 copies/μL; P=0.0300), whereas qRT-PCR was unable to reach significance (136.4 copies/μL vs 122.8 copies/μL; P=0.2273). For miR-499 levels, both chip-based digital PCR and qRT-PCR revealed statistically significant differences between stable CAD and STEMI groups (2 copies/μL vs 8.5 copies/μL, P=0.0011; 0 copies/μL vs 19.4 copies/μL; P

Study Design Biomechanical cadaveric study. Objective Clinical studies indicate that using less-rigid fixation techniques in place of the standard all-pedicle screw construct when correcting for scoliosis may reduce the incidence of proximal junctional kyphosis and improve patient outcomes. The purpose of this study is to investigate whether there is a biomechanical advantage to using supralaminar hooks in place of pedicle screws at the upper-instrumented vertebrae in a multilevel thoracic construct. Methods T7–T12 spines were biomechanically tested: (1) intact; (2) following a two-level pedicles screw fusion from T9 to T11; and after proximal extension of the fusion to T8–T9 with (3) bilateral supra-laminar hooks, (4) a unilateral hook + unilateral screw hybrid, or (5) bilateral pedicle screws. Specimens were nondestructively loaded while three-dimensional kinematics and intradiscal pressure at the supra-adjacent level were recorded. Results Supra-adjacent hypermobility was reduced when bilateral hooks were used in place of pedicle screws at the upper-instrumented level, with statistically significant differences in lateral bending and torsion ( p < 0.05 and p < 0.001, respectively). Disk pressures in the supra-adjacent segment were not statistically different among top-off techniques. Conclusions The use of supralaminar hooks at the top of a multilevel posterior fusion construct reduces the stress at the proximal uninstrumented motion segment. Although further data is needed to provide a definitive link to the clinical occurrence of PJK, this in vitro study demonstrates the potential benefit of “easing” the transition between the stiff instrumented spine and the flexible native spine and is the first to demonstrate these results with laminar hooks.

Historians of science and technology have recently been grappling with the thorny issue of expertise, questions around what constitutes an expert, how such a status is acquired and maintained, and,...

Menopause is associated with bone loss and enhanced visceral adiposity. A polyclonal antibody that targets the β-subunit of the pituitary hormone follicle-stimulating hormone (Fsh) increases bone mass in mice. Here, we report that this antibody sharply reduces adipose tissue in wild-type mice, phenocopying genetic haploinsufficiency for the Fsh receptor gene Fshr. The antibody also causes profound beiging, increases cellular mitochondrial density, activates brown adipose tissue and enhances thermogenesis. These actions result from the specific binding of the antibody to the β-subunit of Fsh to block its action. Our studies uncover opportunities for simultaneously treating obesity and osteoporosis.

AIMS The purpose of this study was to evaluate the change in lower urinary tract symptoms following ventriculoperitoneal shunting in patients with idiopathic normal pressure hydrocephalus (iNPH). METHODS Lower urinary tract symptoms in patients with new-onset iNPH were prospectively evaluated using validated questionnaires from the International Consultation on Incontinence to assess overactive bladder (ICIq-OAB), incontinence (ICIq-UI), and quality of life (ICIq-LUTqol), as well as the American Urological Association Symptom Score bother scale, prior to and following ventriculoperitoneal shunting for iNPH. Sub-analysis was performed based on gender, age, and medical comorbidities. RESULTS Twenty-three consecutive patients with new-onset iNPH were evaluated prior to, and following, surgical intervention for iNPH via ventriculoperitoneal shunting. Shunting resulted in a significant improvement in urinary urgency, urge incontinence, ability to perform physical activities, and overall quality of life. Women had improvement across more domains than men following shunting, particularly in terms of urinary urgency and overall quality of life. Younger patients experienced significant improvement in scores following shunting as compared to older patients. Patients with two or more medical comorbidities, as well as those with fewer than two comorbidities, reported a significant improvement in overall quality of life. CONCLUSIONS Surgical intervention for iNPH results in significant improvement in urinary symptoms, specifically in terms of urinary urgency and urge incontinence as well as overall quality of life, particularly in women and younger patients.

Cone snails (genus Conus) are venomous marine snails that inject prey with a lethal cocktail of conotoxins, small, secreted, and cysteine-rich peptides. Given the diversity and often high affinity for their molecular targets, consisting of ion channels, receptors or transporters, many conotoxins have become invaluable pharmacological probes, drug leads, and therapeutics. Transcriptome sequencing of Conus venom glands followed by de novo assembly and homology-based toxin identification and annotation is currently the state-of-the-art for discovery of new conotoxins. However, homology-based search techniques, by definition, can only detect novel toxins that are homologous to previously reported conotoxins. To overcome these obstacles for discovery, we have created ConusPipe, a machine learning tool that utilizes prominent chemical characters of conotoxins to predict whether a certain transcript in a Conus transcriptome, which has no otherwise detectable homologs in current reference databases, is a putative conotoxin. By using ConusPipe on RNASeq data of 10 species, we report 5148 new putative conotoxin transcripts that have no homologues in current reference databases. 896 of these were identified by at least three out of four models used. These data significantly expand current publicly available conotoxin datasets and our approach provides a new computational avenue for the discovery of novel toxin families.

Surgical treatment of symptomatic adjacent segment disease (ASD) typically involves extension of previous instrumentation to include the newly affected level(s). Disruption of the incision site can present challenges and increases the risk of complication. Lateral-based interbody fusion techniques may provide a viable surgical alternative that avoids these risks. This study is the first to analyze the biomechanical effect of adding a lateral-based construct to an existing fusion.The study aimed to determine whether a minimally invasive lateral interbody device, with and without supplemental instrumentation, can effectively stabilize the rostral segment adjacent to a two-level fusion when compared with a traditional posterior revision approach.This is a cadaveric biomechanical study of lateral-based interbody strategies as add-on techniques to an existing fusion for the treatment of ASD.Twelve lumbosacral specimens were non-destructively loaded in flexion, extension, lateral bending, and torsion. Sequentially, the tested conditions were intact, two-level transforaminal lumbar interbody fusion (TLIF) (L3-L5), followed by lateral lumbar interbody fusion procedures at L2-L3 including interbody alone, a supplemental lateral plate, a supplemental spinous process plate, and then either cortical screw or pedicle screw fixation. A three-level TLIF was the final instrumented condition. In all conditions, three-dimensional kinematics were tracked and range of motion (ROM) was calculated for comparisons. Institutional funds ($50,000) in support of this work were provided by Medtronic Spine.The addition of a lateral interbody device superadjacent to a two-level fusion significantly reduced motion in flexion, extension, and lateral bending (p.05). Supplementing with a lateral plate further reduced ROM during lateral bending and torsion, whereas a spinous process plate further reduced ROM during flexion and extension. The addition of posterior cortical screws provided the most stable lateral lumbar interbody fusion construct, demonstrating ROM comparable with a traditional three-level TLIF.The data presented suggest that a lateral-based interbody fusion supplemented with additional minimally invasive instrumentation may provide comparable stability with a traditional posterior revision approach without removal of the existing two-level rod in an ASD revision scenario.

Multiple myeloma (MM) cells secrete osteoclastogenic factors that promote osteolytic lesions; however, the identity of these factors is largely unknown. Here, we performed a screen of human myeloma cells to identify pro-osteoclastogenic agents that could potentially serve as therapeutic targets for ameliorating MM-associated bone disease. We found that myeloma cells express high levels of the matrix metalloproteinase MMP-13 and determined that MMP-13 directly enhances osteoclast multinucleation and bone-resorptive activity by triggering upregulation of the cell fusogen DC-STAMP. Moreover, this effect was independent of the proteolytic activity of the enzyme. Further, in mouse xenograft models, silencing MMP-13 expression in myeloma cells inhibited the development of osteolytic lesions. In patient cohorts, MMP-13 expression was localized to BM-associated myeloma cells, while elevated MMP-13 serum levels were able to correctly predict the presence of active bone disease. Together, these data demonstrate that MMP-13 is critical for the development of osteolytic lesions in MM and that targeting the MMP-13 protein - rather than its catalytic activity - constitutes a potential approach to mitigating bone disease in affected patients.

Background: Reduced upper airway muscle activity during sleep is fundamental to obstructive sleep apnea (OSA) pathogenesis. Hypoglossal nerve stimulation (HGNS) counteracts this problem, with potential to reduce OSA severity. Study objectives: To examine safety and efficacy of a novel HGNS system (HGNS, Apnex Medical, Inc.) in treating OSA. Participants: Twenty-one patients, 67% male, age (mean ± SD) 53.6 ± 9.2 years, with moderate to severe OSA and unable to tolerate continuous positive airway pressure (CPAP). Design: Each participant underwent surgical implantation of the HGNS system in a prospective single-arm interventional trial. OSA severity was defined by apnea-hypopnea index (AHI) during in-laboratory polysomnography (PSG) at baseline and 3 and 6 months post-implant. Therapy compliance was assessed by nightly hours of use. Symptoms were assessed using the Epworth Sleepiness Scale (ESS), Functional Outcomes of Sleep Questionnaire (FOSQ), Calgary Sleep Apnea Quality of Life Index (SAQLI), and the Beck Depression Inventory (BDI). Results: HGNS was used on 89% ± 15% of nights (n = 21). On these nights, it was used for 5.8 ± 1.6 h per night. Nineteen of 21 participants had baseline and 6-month PSGs. There was a significant improvement (all P < 0.05) from baseline to 6 months in: AHI (43.1 ± 17.5 to 19.5 ± 16.7), ESS (12.1 ± 4.7 to 8.1 ± 4.4), FOSQ (14.4 ± 2.0 to 16.7 ± 2.2), SAQLI (3.2 ± 1.0 to 4.9 ± 1.3), and BDI (15.8 ± 9.0 to 9.7 ± 7.6). Two serious device-related adverse events occurred: an infection requiring device removal and a stimulation lead cuff dislodgement requiring replacement. Conclusions: HGNS demonstrated favorable safety, efficacy, and compliance. Participants experienced a significant decrease in OSA severity and OSA-associated symptoms. Clinical trial information: NAME: Australian Clinical Study of the Apnex Medical HGNS System to Treat Obstructive Sleep Apnea. Registration number: NCT01186926. URL: http://clinicaltrials.gov/ct2/show/NCT01186926.

An in vivo dosing study of vitamin D in a rat posterolateral spinal fusion model with autogenous bone grafting. Rats randomized to 4 levels of vitamin D-adjusted rat chow, longitudinal serum validation, surgeons/observers blinded to dietary conditions, and rats followed prospectively for fusion endpoint.To assess the impact of dietary and serum levels of vitamin D on fusion success, consolidation of fusion mass, and biomechanical stiffness after posterolateral spinal fusion procedure.Metabolic risk factors, including vitamin D insufficiency, are often overlooked by spine surgeons. Currently, there are no published data on the causal effect of insufficient or deficient vitamin D levels on the success of establishing solid bony union after a spinal fusion procedure.Fifty rats were randomized to 4 experimentally controlled rat chow diets: normal control, vitamin D-deficient, vitamin D-insufficient, and a nontoxic high dose of vitamin D, 4 weeks prior to surgery and maintained postsurgery until sacrifice. Serum levels of 25(OH)D were determined at surgery and sacrifice using radioimmunoassay. Posterolateral fusion surgery with tail autograft was performed. Rats were sacrificed 12 weeks postoperatively, and fusion was evaluated via manual palpation, high-resolution radiographs, micro-computed tomographic scans, and biomechanical testing.Serum 25(OH)D and calcium levels were significantly correlated with vitamin D-adjusted chow (P0.001). There was a dose-dependent relationship between vitamin D-adjusted chow and manual palpation fusion, with greatest differences found in measures of radiographical density between high and deficient vitamin D (P0.05). Adequate levels of vitamin D (high and normal control) yielded stiffer fusion than inadequate levels (insufficient and deficient) (P0.05).Manual palpation fusion rates increased with supplementation of dietary vitamin D. Biomechanical stiffness, bone volume, and density were also positively related to vitamin D and calcium.N/A.

common precursor of testosterone (T) and dihydrotestosterone (DHT). We reported 5a-androstane-3b,17b-diol (3b-diol), which is inactive androgen metabolized from DHT, can be converted back to DHEA and finally to DHT. In this pathway, 3b-hydroxysteroid dehydrogenase (3bHSD) is the key enzyme for the conversion from DHEA and epiandrosterone (epiAND) to DHT, and Rui et al. reported abiraterone inhibited 3b-HSD in CRPC. We examined the effect of CYP17 inhibitors on the intracrine back-conversion pathway induced by 3b-diol in prostate cancer cells. METHODS: LNCaP cells were incubated in the presence of 3bdiol or DHEA 10nmol/L with or without CYP17 inhibitors (abiraterone, ketoconazole, or orteronel) 10nmol/L. In all experiments, LNCaP cells were incubated without any androgens and CYP17 inhibitors as a control (CTRL). After incubation, PSA levels in the media were determined by enzyme immunoassay, and androgen levels in media were measured by LC-MS/MS. RESULTS: By adding DHEA or 3b-diol in media, PSA levels were increased (p 1⁄4 0.013 and p < 0.001, respectively). 3b-diol highly activated AR about five-times compared to DHEA. Inhibition of PSA secretion by abiraterone, ketoconazole and orteronel was found in the presence of DHEA (p < 0.001, p 1⁄4 0.002, and p 1⁄4 0.017, respectively). 3b-diol-induced PSA accumulation in media was inhibited by abiraterone, not ketoconazole or orteronel. By adding 3b-diol in medium, DHEA, androstenediol (A-diol), androstendione (A-dione), T, and DHT levels were increased compared to CTRL. In the media with 3b-diol and abiraterone, A-dione, T, DHT, and 5a-androstanedione (5a-A-dione) levels were decreased, DHEA, 5-diol, and epiAND levels increased compared to 3b-diol only. CONCLUSIONS: AR was highly activated by 3b-diol compared to DHEA through the conversion of 3b-diol to DHT by way of epiAND and 5a-A-dione, which abiraterone blocked by inhibiting 3b-HSD. Abiraterone has a potential to inhibit the intracrine back-conversion pathway.

OBJECTIVE A case is presented in which a 58-year-old man developed a potential unintended and durable treatment of pain at remote sites (cervical region and low back) after sacral nerve stimulation for chronic urinary retention. METHODS Proper placement of the electrodes in the S3 foramen was confirmed by physiological response and fluoroscopy. RESULTS Potential causality was shown by recurrent pain with the stimulators turned off and abatement of pain with the stimulators turned on. DISCUSSION The current case presents a potential example of neurological crosstalk and highlights the inherent complexity in human neural physiology. Further research may reveal novel treatment strategies for patients with voiding dysfunction and chronic pain syndromes.

Purpose The purpose of this study was to compare the tensile strength of hamstring grafts of varying combined pull-through diameters within the clinically relevant range of 6 to 9 mm. Methods We tested 44 non-irradiated allograft hamstring grafts (11 per group). Combined looped semitendinosus and gracilis grafts were allocated to the 6-, 7-, 8-, or 9-mm group based on the smallest-diameter lumen that the graft could be “pulled through” using a surgical sizing instrument. Testing was performed on an Instron materials testing machine (Instron, Norwood, MA). Samples were secured with cryoclamps, prestressed, and pulled to failure at a rate of 10% gauge length per second. Results The mean load to failure was 2,359 ± 474 N, 3,263 ± 677 N, 3,908 ± 556 N, and 4,360 ± 606 N for the 6-, 7-, 8-, and 9-mm grafts, respectively. Minimum failure loads were as low as 1,567 N, 2,288 N, 2,874 N, and 3,720 N for each group, respectively. There were statistically significant differences between the 6- and 7-mm, 6- and 8-mm, 6- and 9-mm, and 7- and 9-mm groups ( P = .01). Conclusions Statistically different increasing tensile strength was seen as graft diameter increased. Significant variability exists in the strength of multi-stranded hamstring allografts within the diameter range of 6 to 9 mm that often falls well below the commonly accepted value of 4,000 N for a hamstring graft. Clinical Relevance Recent evidence suggests a higher early failure rate of hamstring autografts in subsets of patients with graft diameters of 8 mm or less. This study may increase awareness that hamstring grafts may not be nearly as strong as previously appreciated and that increasing tendon diameters by 1 to 2 mm may dramatically affect graft strength. These data may be helpful in preoperative discussions regarding variable hamstring size, strength, and potential intraoperative augmentation options.

Two important human depth cues are accommodation and vergence. Normally, the eyes accommodate and converge or diverge in tandem; changes in viewing distance cause the eyes to simultaneously adjust both focus and orientation. However, ambiguity between accommodation and vergence cues is a well-known limitation in many stereoscopic display technologies. This limitation also arises in state-of-the-art full-flight simulator displays. In current full-flight simulators, the out-the-window (OTW) display (i.e., the front cockpit window display) employs a fixed collimated display technology which allows the pilot and copilot to perceive the OTW training scene without angular errors or distortions; however, accommodation and vergence cues are limited to fixed ranges (e.g., ~ 20 m). While this approach works well for long-range, the ambiguity of depth cues at shorter range hinders the pilot’s ability to gauge distances in critical maneuvers such as vertical take-off and landing (VTOL). This is the first in a series of papers on a novel, variable-collimation display (VCD) technology that is being developed under NAVY SBIR Topic N121-041 funding. The proposed VCD will integrate with rotary-wing and vertical take-off and landing simulators and provide accurate accommodation and vergence cues for distances ranging from approximately 3 m outside the chin window to ~ 20 m. A display that offers dynamic accommodation and vergence could improve pilot safety and training, and impact other applications presently limited by lack of these depth cues.

This chapter focuses on aspects of hydrocephalus that are common to all ages with a particular emphasis on the aging brain and the so called normal pressure hydrocephalus (NPH) syndrome, which is a chronic disease that likely evolved over a period of years. It has been clear for decades that the rate of ventricular dilatation and the state of brain maturation have a significant impact on the pathology, and possibly the pathogenesis of brain damage. Idiopathic NPH has been distinguished from chronic adult hydrocephalus associated with prior meningitis, brain trauma, or subarachnoid hemorrhage. The initial displacement may be at the expense of the subarachnoid compartment, the venous compartment, and the extracellular compartment with negligible damage to brain cells. Blood flow hypoperfusion leads to hypoxic-ischemic changes in the white matter, physical stretching compromises axon integrity, and retarded turnover of the cerebrospinal fluid (CSF) alters the extracellular microenvironment.

We present optical analysis of electrically-actuated adaptive fluidic lenses having apertures of 10 mm and 40 mm. Zernike decomposition and resolution testing is presented for the 10-mm aperture lens. Focal power and imaging resolution characteristics of the 40-mm aperture lens are provided. The data for the lenses show both formats are suitable for numerous adaptive lens applications demanding high optical quality.

A varicocele is a dilatation of the testicular vein and the pampiniform venous plexus within the spermatic cord. Although rare in pediatric populations, the prevalence of varicoceles markedly increases with pubertal development. Varicoceles are progressive lesions that may hinder testicular growth and function over time and are the most common and correctable cause of male infertility. Approximately 40% of men with primary infertility have a varicocele, and more than half of them experience improvements in semen parameters after varicocelectomy. The decision to treat adolescents with varicocele is a controversial one. The task for pediatricians and urologists is to identify those adolescents who are at greatest risk for infertility in adulthood, in an effort to offer early surgical intervention to those most likely to benefit.

P.R. Eastwood, PhD, M. Barnes, MBBS, R.D. McEvoy, MBBS, J. Wheatley, J.H. Walsh, PhD, F.J. O'Donoghue, MB BCh FRACP MRCP PhD, P. Catcheside, PhD, L. Tyler, BN, K.J. Maddison, BSc, P.D. Rochford, BApSc, N. Antic, C. Worsnop, MD, PhD, T. Churchward, B Ap Sc, D.J. Eckert, PhD, M.C. Campbell, MB, BS, FRACS, G. Hee, MB, BS, FRACS, C.E. Palme, MB, BS, FRACS, S.H. Robinson, MB, BS, FRACS, E. Kezirian, MD, G. Goding, MD, A.S. Jordan, PhD, P.L. Smith, MD, A.R. Schwartz, MD, A. Malhotra, MD, D.R. Hillman, MD Sir Charles Gairdner Hospital Perth/AU, Austin Hospital Melbourne/AU, Repatriation General Hospital Adelaide/AU, Westmead Hospital Sydney/AU, Institute for Breathing and Sleep Heidelberg/AU, Ludwig Engel Centre for Respiratory Research, Westmead Millennium Institute, Westmead NSW 2145 Westmead/AU, Austin Health Melbourne/AU, Adelaide Institute for Sleep Heath Adelaide/AU, University of Melbourne, Melbourne/AU, Brigham and Women's Hospital Boston, MA/US, Westmead Private Hospital Sydney/AU, Flinders University Adelaide/AU, University of California San Francisco San Francisco, CA/US, University of Minnesota Minneapolis, MN/US, Johns Hopkins School of Medicine Baltimore, MD/US, Johns Hopkins University Baltimore, MD/US, Brigham and Women's Hospital / Boston, MA/US

Acetabular insufficiency fractures are much less common than acetabular fractures associated with trauma. They most commonly occur in postmenopausal women with a history of rheumatoid arthritis or pelvic irradiation. We present the case of a 93-year-old man with an atraumatic pelvic insufficiency fracture of the fossa acetabuli. The patient had 2 predisposing risk factors: osteoporosis and lower-extremity reconstructive surgery.

Any surgeon experienced with a laparoscopic approach to intra-abdominal surgery on morbidly obese patients is aware of the increased difficulty associated with the patients' obesity. We present a study of the mechanics of laparoscopic surgery that explains the difficulty subjectively experienced in terms of the decreased sensitivity felt by the surgeon as a result of the increased thickness of the abdominal wall and increasing force required for the repositioning of the tip of the operating instrument. We propose that the placement and angling of the trocar in the abdominal wall are of paramount importance in the successful and safe completion of laparoscopic procedures in obese patients.

1. A classification of gastro-duodenal ulcers on an etiological and clinical basis is presented. 2. There is discussion of the limitations of histopathology in the elucidation of the chronic ulcer and other psychogenic diseases. For that reason the pathological picture of the ulcer itself plays a minor role in this classification. 3. Evidence is advanced to show the inadequacy of the term “peptic” as descriptive of gastro-duodenal ulcers. The term “chronic” is shown to be loose and without any specific connotation. 4. The term “psychogenic” is suggested as a substitute because it is descriptive of the causal relationship of the disease. 5. Malignant transformation does not occur in a benign psychogenic ulcer. 6. Traumatic ulcer is classified as brought about by direct causes and therefore could not result in a psychogenic (chronic) ulcer. 7. The subjective, physical and laboratory findings of hemorrhagic erosions should not be included in statistical studies of the psychogenic type of ulcer.

This paper advances the psychogenic theory of “peptic” ulcer. It claims that this disease is found only among susceptible individuals of the white race — usually the long thin type who are given to worry and nervous instability. These individuals with their fears and anxieties develop an imbalance of the vegetative nervous system. This results in a bombardment of stimuli to the lesser curvature of the stomach and duodenum producing vascular spasm, thrombosis, induration, ischemia, and finally necrosis and ulceration.