bluuebunny/crossref_metadata_2025 huggingface dataset:
Logistics of air medical transport: When and where does helicopter transport reduce prehospital time for trauma?
Authors: Chen Xilin , Gestring Mark L. , Rosengart Matthew R. , Peitzman Andrew B. , Billiar Timothy R. , Sperry Jason L. , Brown Joshua B.
Published: May 2018

BACKGROUND

Trauma is a time sensitive disease. Helicopter emergency medical services (HEMS) have shown benefit over ground emergency medical services (GEMS), which may be related to reduced prehospital time. The distance at which this time benefit emerges depends on many factors that can vary across regions. Our objective was to determine the threshold distance at which HEMS has shorter prehospital time than GEMS under different conditions.

METHODS

Patients in the Pennsylvania trauma registry 2000 to 2013 were included. Distance between zip centroid and trauma center was calculated using straight-line distance for HEMS and driving distance from geographic information systems network analysis for GEMS. Contrast margins from linear regression identified the threshold distance at which HEMS had a significantly lower prehospital time than GEMS, indicated by nonoverlapping 95% confidence intervals. The effect of peak traffic times and adverse weather on the threshold distance was evaluated. Geographic effects across EMS regions were also evaluated.

RESULTS

A total of 144,741 patients were included with 19% transported by HEMS. Overall, HEMS became faster than GEMS at 7.7 miles from the trauma center (p = 0.043). Helicopter emergency medical services became faster at 6.5 miles during peak traffic (p = 0.025) compared with 7.9 miles during off-peak traffic (p = 0.048). Adverse weather increased the distance at which HEMS was faster to 17.1 miles (p = 0.046) from 7.3 miles in clear weather (p = 0.036). Significant variation occurred across EMS regions, with threshold distances ranging from 5.4 to 35.3 miles. There was an inverse but non-significant relationship between urban population and threshold distance across EMS regions (ρ, −0.351, p = 0.28).

CONCLUSION

This is the first study to demonstrate that traffic, weather, and geographic region significantly impact the threshold distance at which HEMS are faster than GEMS. Helicopter emergency medical services was faster at shorter distances during peak traffic while adverse weather increased this distance. The threshold distance varied widely across geographic region. These factors must be considered to guide appropriate HEMS triage protocols.

LEVEL OF EVIDENCE

Therapeutic, level IV.
Hemorrhagic shock alters fracture callus composition and activates the IL6 and RANKL/OPG pathway in mice
Authors: Bundkirchen Katrin , Macke Christian , Angrisani Nina , Schäck Luisa M. , Noack Sandra , Fehr Michael , Krettek Christian , Neunaber Claudia
Published: May 2018

Background

Fracture and hemorrhagic shock often lead to impaired fracture healing. To elucidate underlying pathogenesis, this study aimed to analyze histological properties during fracture healing after hemorrhagic shock and involved signaling pathways in mice.

METHODS

Male C57BL/6NCrl mice were assigned into five groups. Control group underwent no interventions. Sham group had a catheter and external fixator but neither blood loss nor osteotomy. Trauma-hemorrhage (TH) group received a pressure-controlled hemorrhagic shock; osteotomy (Fx) group, an osteotomy and fixator; and combined trauma (THFx) group, both hemorrhagic shock and externally fixed osteotomy. After 1, 2, 3, and 4 weeks, the animals were killed. Undecalcified bones were analyzed histologically and signaling pathways relevant for fracture healing by polymerase chain reaction and Western blot. Statistical significance was set at 0.05 or less. Comparisons were performed using the Mann-Whitney U or Kruskal-Wallis test.

RESULTS

In the THFx group, a decreased bone formation after 3 weeks, a reduction of both bone and cartilage after 2 weeks, and an enhanced activation of the RANKL/OPG and IL6 signaling pathway after 1 week were shown in comparison to Fx.

CONCLUSIONS

Hemorrhagic shock has a retarding effect on fracture healing in the early phase of fracture healing and leads to activation of the IL6 and RANKL/OPG signaling pathways.
Extending the golden hour for Zone 1 resuscitative endovascular balloon occlusion of the aorta: Improved survival and reperfusion injury with intermittent versus continuous resuscitative endovascular balloon occlusion of the aorta in a porcine severe truncal hemorrhage model
Authors: Kuckelman John P. , Barron Morgan , Moe Donald , Derickson Michael , Phillips Cody , Kononchik Joseph , Lallemand Michael , Marko Shannon , Eckert Matthew , Martin Matthew J.
Published: May 2018

BACKGROUND

Noncompressible hemorrhage can be controlled using resuscitative endovascular balloon occlusion of the aorta (REBOA). Prolonged ischemia limits REBOA application during Zone 1 deployment. Intermittent inflation/deflation may effectively mitigate this problem.

METHODS

A lethal abdominal vascular injury was created in 28 swines. Animals were randomized to controls (n = 7), 60 minutes full REBOA (FR, n = 5), time-based intermittent REBOA (iRT, n = 7), and pressure-based REBOA (iRP, n = 9). Intermittent groups had an initial inflation for 15 minutes, followed by 10-minute inflation: 3-minute deflation cycles (iRT), or an inflate/deflate schedule based on mean arterial pressure (MAP) less than 40 mm Hg (iRP). Experiments were concluded after 120 minutes or death (MAP < 20 mm Hg).

RESULTS

Intermittent REBOA animals all survived to 120 minutes versus 15 minutes for controls and 63 minutes for FR (p < 0.001). After 60 minutes, FR animals were more hypotensive (MAP 20 mm Hg vs. 80 mm Hg [iRP] and 100 mm Hg [iRT]; p < 0.001), had lower cardiac output (1.06 mL/min vs. 5.1 L/min [iRP] and 8.2 L/min [iRT]; p < 0.001), higher lactate (12.5 mg/dL vs. 8.5 mg/dL [iRP], p = 0.02), and decreased clot firmness on rotational thromboelastometry than iRP/T (64 mm vs. 69 mm [iRP] and 69 mm [iRT], p = 0.04). Acidosis was worse in iRT versus iRP at 120 minutes (pH 7.28 vs. pH 7.12; p = 0.02), improved lactate (11.9 mg/dL vs. 16.3 mg/dL; p = 0.04), and decreased whole blood resuscitation (452 mL vs. 646 mL, p = 0.05). Blood loss (clot weight) was higher in controls (2.0 kg) versus iRT and iRP (1.16 kg and 1.23 kg; p < 0.01) and not different from FR (0.87 kg; p = 0.10).

CONCLUSION

Intermittent REBOA can maintain supraceliac hemorrhage control while decreasing distal ischemia in a swine model. Prolonged survival times, decreased acidosis, and lower resuscitation requirements indicate that this technique could potentially extend Zone 1 REBOA deployment times. Schedules based on MAP may be superior to time-based regimens.
A clinical prediction model for raised intracranial pressure in patients with traumatic brain injuries
Authors: Pace Jacob , Parry Neil , Vogt Kelly , Hilsden Rich , Leeper R. Robert , Markova Zarina , Priestap Fran , Younan Julia , Ball Ian
Published: May 2018

BACKGROUND

Intracranial hypertension is believed to contribute to secondary brain insult in traumatically brain injured patients. Currently, the diagnosis of intracranial hypertension requires intracranial monitoring or advanced imaging. Unfortunately, prehospital transport times can be prolonged, delaying time to the initial radiographic assessment. The aim of this study was to identify clinical variables associated with raised intracranial pressure (ICP) prior to the completion of neuroimaging.

METHODS

We performed a retrospective cohort study of head injured patients over a 3-year period. Patients were labeled as having increased ICP if they had a single reading of ICP greater than 20 mm Hg within 1 hour of ICP monitor insertion or computed tomography findings suggestive of raised ICP. Patient and clinical characteristics were analyzed using stepwise multivariable logistic regression with ICP as the dependent variable.

RESULTS

Of 701 head injured patients identified, 580 patients met inclusion criteria. Mean age was 48.65 ± 21 years, 73.3% were male. The mean Injury Severity Score was 22.71 ± 12.38, and the mean Abbreviated Injury Scale for body region head was 3.34 ± 1.06. Overall mortality was 14.7%. Only 46 (7.9%) patients had an ICP monitor inserted; however, a total of 107 (18%) patients met the definition of raised ICP. The mortality rate for patients with raised ICP was 50.4%. Independent predictors of raised ICP were as follows: age, older than 55 years (odds ratio [OR], 2.26; 95% confidence interval [CI], 1.35–3.76), pupillary fixation (OR, 5.76; 95% CI, 3.16–10.50), signs of significant head trauma (OR, 2.431; 95% CI, 1.39–4.26), and need for intubation (OR, 3.589; 95% CI, 2.10–6.14).

CONCLUSION

This study identified four independent variables associated with raised ICP and incorporated these findings into a preliminary risk assessment scale that can be implemented at the bedside to identify patients at significant risk of raised ICP. Future work is needed to prospectively validate these findings prior to clinical implementation.

LEVEL OF EVIDENCE

Prognostic, Epidemiological, level III.
Recent advances in austere combat surgery: Use of aortic balloon occlusion as well as blood challenges by special operations medical forces in recent combat operations
Authors: Northern David Marc , Manley Justin D. , Lyon Regan , Farber Daniel , Mitchell Benjamin J. , Filak Kristopher J. , Lundy Jonathan , DuBose Joe J. , Rasmussen Todd E. , Holcomb John B.
Published: May 2018

BACKGROUND

Resuscitative endovascular balloon occlusion of the aorta (REBOA) for control of noncompressible torso hemorrhage is a technology that is increasingly being utilized in the combat casualty setting. Its use in the resource restricted environment holds potential to improve hemorrhage control, decrease blood product utilization, decrease morbidity, and improve combat mortality. The objective of this report is to present the single largest series of REBOA use on severely injured combat casualties.

METHODS

Over an 18-month period, austere surgical teams comprised of coalition partners provided initial damage control resuscitation (DCR) and surgical stabilization for over 2,300 combat casualties prior to transferring patients to the next level of trauma care.

RESULTS

Twenty patients presented with injuries from explosion and gunshot wounds with mean initial heart rate of 129 bpm and mean initial systolic blood pressure of 71 mm Hg. Femoral cutdowns were used in six patients. Aortic occlusion was achieved with REBOA catheter placement in Zone 1 (n = 17) and Zone 3 (n = 2). Systolic blood pressure increased an average of 56 mm Hg with aortic occlusion. There were no access related site complications. All patients survived transport to the next level of care. The majority of blood products transfused in this cohort were whole blood, largely supported by emergent blood drives.

CONCLUSION

This series demonstrates the potential for REBOA as a lifesaving technique for the patient who presents with hemodynamic instability and noncompressible torso hemorrhage. Resuscitative endovascular balloon occlusion of the aorta allows austere surgical teams to rapidly stabilize severely injured combat casualties, expand capability, and provide enhanced DCR while minimizing personnel, resources, and blood product utilization. The use of “whole blood only” strategy for DCR shows potential to be superior to traditional component therapy, and when combined with “proactive” REBOA utilization, provides significant improvements in hemodynamics and hemorrhage control.

LEVEL OF EVIDENCE

Case series, level V.
Transfusion of cryopreserved platelets exacerbates inflammatory liver and lung injury in a mice model of hemorrhage
Authors: Zhao Jingxiang , Sun Zhenwei , You Guoxing , Wang Ying , Chen Gan , Zhao Lian , Zhou Hong
Published: May 2018

BACKGROUND

Platelets are essential for primary hemostasis and also play an important role in inflammatory reactions. The hemostatic property of cryopreserved platelets (CPPs) has been confirmed in the treatment of bleeding casualties, but inflammatory injury induced by CPP transfusion is relatively unclear. We aim to investigate the effects of CPP transfusion on inflammatory organ injury in mice after hemorrhage.

METHODS

Mice were subjected to a volume-controlled hemorrhage over 1 hour, and then were transfused with fresh platelets (FPs), Liquid-stored platelets (LPPs), CPPs, or fresh frozen plasma (FFP, control). At 6 hours posttransfusion, mice were sacrificed, and blood and tissues were sampled. Tissue sections were examined histologically and by immunohistochemical staining of neutrophils and macrophages. Plasma alanine aminotransferase, hepatic myeloperoxidase activity and inflammatory cytokine levels were measured.

RESULTS

Transfusion of stored platelets (LPPs and CPPs) caused more serious histological injury in liver and lung compared with FPs and FFP (p < 0.05). However, kidney histological injury was similar among groups. Significantly higher numbers of Ly-6G–positive neutrophils were detected in liver and of F4/80-positive macrophages in liver and lung of mice transfused with LPPs or CPPs compared with FPs or FFP (p < 0.05). Transfusion of CPPs caused the most severe inflammatory liver injury, as reflected by alanine aminotransferase levels, hepatic macrophage infiltration, and hepatic myeloperoxidase activity and inflammatory cytokine levels (macrophage inflammatory protein-2, tumor necrosis factor-α, and interleukin-1β).

CONCLUSION

Cryopreserved platelet transfusion is more likely to aggravate hemorrhage-induced liver and lung injury by activating macrophage and facilitating neutrophil infiltration into hepatic tissues.
Unrealized potential of the US military battlefield trauma system: DOW rate is higher in Iraq and Afghanistan than in Vietnam, but CFR and KIA rate are lower
Authors: Nessen Shawn C. , Gurney Jennifer , Rasmussen Todd E. , Cap Andrew P. , Mann-Salinas Elizabeth , Le Tuan D. , Shackelford Stacy , Remick Kyle N. , Akers Kevin S. , Eastridge Brian J. , Jenkins Donald , Stockinger Zsolt T. , Murray Clinton K. , Gross Kirby R. , Seery Jason , Mabry Robert , Holcomb John B.
Published: May 2018

Level of Evidence

Observational/retrospective/historic controls, level IV.
A metal organic framework reduces thrombus formation and platelet aggregation ex vivo
Authors: Roberts Teryn R. , Neufeld Megan J. , Meledeo Michael A. , Cap Andrew P. , Cancio Leopoldo C. , Reynolds Melissa M. , Batchinsky Andriy I.
Published: May 2018

BACKGROUND

Management of hemostasis is a key challenge during extracorporeal life support (ECLS). Metal organic frameworks are being investigated for use as nitric oxide (NO) catalysts for incorporation into ECLS circuitry to prevent thrombosis at the blood–biomaterial interface. A specific metal organic framework, CuBTTri, has been shown to accelerate NO release from bioavailable donors like S-nitrosoglutathione (GSNO). We hypothesized that CuBTTri would reduce thrombus formation in whole blood (WB) and inhibit platelet aggregation.

METHODS

CuBTTri particles were added to WB and analyzed by thromboelastography. Biostable metal-based frameworks (MIL-100, Zeolite USY) were added to blood as controls, in addition to a saline vehicle control. Reaction time (R), clot formation time (K), alpha-angle, clot strength (MA), and percent fibrinolysis (LY30/LY60) were recorded. The effect of CuBTTri on platelet aggregation was assessed in WB and platelet-rich plasma (PRP), both with and without addition of GSNO.

RESULTS

CuBTTri significantly prolonged R and K and decreased alpha-angle and MA relative to the metal framework controls. Dose escalation results suggest that the control metal-based particles induce thrombus formation, as R and K were significantly reduced compared with the saline control; however, this did not occur in the CuBTTri group. LY30/LY60 were elevated in the CuBTTri group versus saline (p = 0.014) but were not different from metal framework controls. CuBTTri alone and with GSNO reduced platelet aggregation in WB (p < 0.0001), whereas GSNO alone had no effect. In PRP, GSNO and CuBTTri inhibited platelet aggregation separately, and together decreased aggregation by 35% relative to GSNO alone (p = 0.004).

CONCLUSIONS

CuBTTri reduced thrombus formation and inhibited platelet aggregation. CuBTTri enhanced platelet inhibition with GSNO, which was consistent with reports that CuBTTri accelerates NO release from endogenous NO donors. This initial characterization of CuBTTri demonstrated its potential as an antithrombogenic agent to be further evaluated with incorporation into ECLS circuitry.
Motor vehicle crash fatalaties and undercompensated care associated with legalization of marijuana
Authors: Steinemann Susan , Galanis Daniel , Nguyen Tiffany , Biffl Walter
Published: May 2018

BACKGROUND

Half of the US states have legalized medical cannabis (marijuana), some allow recreational use. The economic and public health effects of these policies are still being evaluated. We hypothesized that cannabis legalization was associated with an increase in the proportion of motor vehicle crash fatalities involving cannabis-positive drivers, and that cannabis use is associated with high-risk behavior and poor insurance status.

METHODS

Hawaii legalized cannabis in 2000. Fatality Analysis Reporting System data were analyzed before (1993–2000) and after (2001–2015) legalization. The presence of cannabis (THC), methamphetamine, and alcohol in fatally injured drivers was compared. Data from the state's highest level trauma center were reviewed for THC status from 1997 to 2013. State Trauma Registry data from 2011 to 2015 were reviewed to evaluate association between cannabis, helmet/seatbelt use, and payor mix.

RESULTS

THC positivity among driver fatalities increased since legalization, with a threefold increase from 1993–2000 to 2001–2015. Methamphetamine, which has remained illegal, and alcohol positivity were not significantly different before versus after 2000. THC-positive fatalities were younger, and more likely, single-vehicle accidents, nighttime crashes, and speeding. They were less likely to have used a seatbelt or helmet. THC positivity among all injured patients tested at our highest level trauma center increased from 11% before to 20% after legalization. From 2011 to 2015, THC-positive patients were significantly less likely to wear a seatbelt or helmet (33% vs 56%). They were twice as likely to have Medicaid insurance (28% vs 14%).

CONCLUSION

Since the legalization of cannabis, THC positivity among MVC fatalities has tripled statewide, and THC positivity among patients presenting to the highest level trauma center has doubled. THC-positive patients are less likely to use protective devices and more likely to rely on publically funded medical insurance. These findings have implications nationally and underscore the need for further research and policy development to address the public health effects and the costs of cannabis-related trauma.

LEVEL OF EVIDENCE

Prognostic, level III.
Crisis under the radar: Illicit amphetamine use is reaching epidemic proportions and contributing to resource overutilization at a Level I trauma center
Authors: Gemma Vincent A. , Chapple Kristina M. , Goslar Pamela W. , Israr Sharjeel , Petersen Scott R. , Weinberg Jordan A.
Published: May 2018

INTRODUCTION

Trauma centers reported illicit amphetamine use in approximately 10% of trauma admissions in the previous decade. From experience at a trauma center located in a southwestern metropolis, our perception is that illicit amphetamine use is on the rise and that these patients utilize in-hospital resources beyond what would be expected for their injuries. The purposes of this study were to document the incidence of illicit amphetamine use among our trauma patients and to evaluate its impact on resource utilization.

METHODS

We conducted a retrospective cohort study using 7 consecutive years of data (starting July 2010) from our institution's trauma registry. Toxicology screenings were used to categorize patients into one of three groups: illicit amphetamine, other drugs, or drug-free. Adjusted linear and logistic regression models were used to predict hospital cost, length of stay, intensive care unit admission, and ventilation between drug groups. Models were conducted with combined injury severity (Injury Severity Score [ISS]) and then repeated for ISS of less than 9, ISS 9 to 15, and ISS 16 or greater.

RESULTS

Eight thousand five hundred eighty-nine patients were categorized into the following three toxicology groups: 1,255 (14.6%) illicit amphetamine, 2,214 (25.8%) other drugs, and 5,120 (59.6%) drug-free. Illicit amphetamine use increased threefold over the course of the study (from 7.85% to 25.0% of annual trauma admissions). Adjusted linear models demonstrated that illicit amphetamine among patients with ISS of less than 9 was associated with 4.6% increase in hospital cost (p = 0.019) and 7.4% increase in length of stay (p = 0.043). Logistic models revealed significantly increased odds of ventilation across all ISS groups and increased odds of intensive care unit admission when all ISS groups were combined (p = 0.001) and within the group with ISS of less than 9 (p = 0.002).

CONCLUSIONS

Hospital resource utilization of amphetamine patients with minor injuries is significant. Trauma centers with similar epidemic growth in proportion of amphetamine patients face a potentially significant resource strain relative to other centers.

LEVEL OF EVIDENCE

Prognostic/Epidemiological, level II; Therapeutic, level III.
Increased trauma activation is not equally beneficial for all elderly trauma patients
Authors: Carr Bryan W. , Hammer Peter M. , Timsina Lava , Rozycki Grace , Feliciano David V. , Coleman Jamie J.
Published: May 2018

BACKGROUND

Physiologic changes in the elderly lead to higher morbidity and mortality after injury. Increasing level of trauma activation has been proposed to improve geriatric outcomes, but the increased cost to the patient and stress to the hospital system are significant downsides. The purpose of this study was to identify the age at which an increase in activation status is beneficial.

METHODS

A retrospective review of trauma patients 70 years or older from October 1, 2011, to October 1, 2016, was performed. On October 1, 2013, a policy change increased the activation criteria to the highest level for patients 70 years or older with a significant mechanism of injury. Patients who presented prior to (PRE) were compared with those after the change (POST). Data collected included age, Injury Severity Score (ISS), length of stay (LOS), complications, and mortality. Primary outcome was mortality, and secondary outcome was LOS. Multivariable regressions controlled for age, ISS, injury mechanism, and number of complications.

RESULTS

A total of 4,341 patients met the inclusion criteria, 1,919 in PRE and 2,422 in POST. Mean age was 80.4 and 81 years in PRE and POST groups, respectively (p = 0.0155). Mean ISS values were 11.6 and 12.4 (p < 0.0001) for the PRE and POST groups. POST had more Level 1 activations (696 vs. 220, p < 0.0001). After controlling for age, ISS, mechanism of injury, and number of complications, mortality was significantly reduced in the POST group 77 years or older (odds ratio, 0.53; 95% confidence interval, 0.3–0.87) (Fig. 1). Hospital LOS was significantly reduced in the POST group age 78 years or older (regression coefficient, −0.55; 95% confidence interval, −1.09 to −0.01) (Fig. 2).

CONCLUSIONS

This study suggests geriatric trauma patients 77 years or older benefit from the highest level of trauma activation with shorter LOS and lower mortality. A focused approach to increasing activation level for elderly patients may decrease patient cost.

LEVEL OF EVIDENCE

Therapeutic/Care Management, level IV.
The Impact of Implantation Time During Liver Transplantation on Outcome: A Eurotransplant Cohort Study
Authors: Jochmans Ina , Fieuws Steffen , Tieken Ineke , Samuel Undine , Pirenne Jacques
Published: May 2018

Background

The liver graft quickly rewarms during transplantation when the vascular anastomoses are being performed, potentially impacting on outcomes.

Methods

We investigated the relationship between implantation time and outcome in 5223 recipients of deceased-donor livers transplanted in Eurotransplant (2004-2013). Cox regression analyses were corrected for donor, preservation, and recipient variables. Transplant loss represents all-cause graft failure.

Results

Median implantation time was 41 minutes (interquartile range, 34-51). Implantation time independently associated with transplant loss (adjusted hazard ratio, 1.04 for every 10-minute increase; 95% confidence interval, 1.01-1.07; P = 0.007). The magnitude of the implantation time effect was comparable to the effect of each additional hour of cold ischemia (adjusted hazard ratio, 1.03; 95% confidence interval, 1.02-1.05; P < 0.001). The effect was most pronounced early posttransplant with no evidence of a significant effect beyond 3 months. A similar detrimental effect of implantation time was seen for graft and patient survivals. The increased risk for transplant loss in livers donated after circulatory determination of death could be attributed to donor warm ischemia time.

Conclusions

Implantation time associates with inferior liver transplant outcome in a continuous way. These findings need confirmation and further study of confounding factors is needed so steps toward improving outcomes can be made.
Microrna-184 is Induced by Store-Operated Calcium Entry and Regulates Early Keratinocyte Differentiation
Authors: Richardson Adam , Powell Andrew , Sexton Darren W. , Parsons Jason L. , Reynolds Nicholas J. , Ross Kehinde
Published: May 2018

Extracellular calcium (Ca2+) and store-operated Ca2+ entry (SOCE) govern homeostasis in the mammalian epidermis. Multiple microRNAs (miRNA) also regulate epidermal differentiation, and raised external Ca2+ modulates the expression of several such miRNAs in keratinocytes. However, little is known about the regulation of miR-184 in keratinocytes or the roles of miR-184 in keratinocyte differentiation. Here we report exogenous Ca2+ stimulates miR-184 expression in primary epidermal keratinocytes and that this occurs in a SOCE-dependent manner. Levels of miR-184 were raised by about 30-fold after exposure to 1.5 mM Ca2+ for 5 days. In contrast, neither phorbol ester nor 1, 25-dihydroxyvitamin D3 had any effect on miR-184 levels. Pharmacologic and genetic inhibitors of SOCE abrogated Ca2+-dependent miR-184 induction by 70% or more. Ectopic miR-184 inhibited keratinocyte proliferation and led to a 4-fold increase in the expression of involucrin, a marker of early keratinocyte differentiation. Exogenous miR-184 also triggered a 3-fold rise in levels of cyclin E and doubled the levels of γH2AX, a marker of DNA double strand breaks. The p21 cyclin-dependent kinase (CDK) inhibitor, which supports keratinocyte growth arrest, was also induced by miR-184. Together our findings point to a SOCE:miR-184 pathway that targets a cyclin E/DNA damage regulatory node to facilitate keratinocyte differentiation.
Supplemental vibrotactile feedback of real-time limb position enhances precision of goal-directed reaching
Authors: Risi Nicoletta , Shah Valay , Mrotek Leigh A. , Casadio Maura , Scheidt Robert A.
Published: May 2018

We examined vibrotactile stimulation as a form of supplemental limb state feedback to enhance on-going control goal-directed movements. Subjects wore a two-dimensional vibrotactile display on their non-dominant arm while performing horizontal planar reaching movements with their dominant arm. The vibrotactile display provided feedback of hand position such that small hand displacements were more easily discriminable using vibrotactile feedback than with intrinsic proprioceptive feedback. When subjects relied solely on proprioceptive feedback to capture visuospatial targets, target capture performance was degraded by proprioceptive drift and an expansion of task space. By contrast, reach accuracy was enhanced immediately when subjects were provided vibrotactile feedback, and further improved over two days of training. Improvements reflected a resolution of proprioceptive drift which occurred only when vibrotactile feedback was active, demonstrating that the benefits of vibrotactile feedback are due in part to its integration into the ongoing control of movement. A partial resolution of task space expansion that persisted even when the vibrotactile feedback was inactive demonstrated that training with vibrotactile feedback also induced changes in movement planning. However, the benefits of vibrotactile feedback come at a cognitive cost. All subjects adopted a stereotyped, movement decomposition strategy wherein they attempted to capture targets by moving first along one axis of the vibrotactile display and then the other. For most subjects, this inefficient movement approach did not resolve over two bouts of training performed on separate days, suggesting that additional training is needed to fully integrate vibrotactile feedback into the planning and online control of goal-directed reaching.
Single-cell mRNA profiling reveals heterogeneous combinatorial expression of Hoxd genes during limb development
Authors: Fabre Pierre J , Leleu Marion , Mascrez Benedicte , Lo Giudice Quentin , Cobb John , Duboule Denis
Published: May 2018

A global analysis of gene expression during development reveals specific transcription patterns associated with the emergence of various cell types, tissues and organs. These heterogeneous patterns are instrumental to ensure the proper formation of the different parts of our body, as shown by the phenotypic effects generated by functional genetic approaches. However, variations at the cellular level can be observed within each structure or organ. In the developing mammalian limbs, expression of Hoxd genes is differentially controlled in space and time in cells that will pattern the digits and the arms. Here we analyze single-cell transcriptomes of limb bud cells and show that Hox genes are expressed in specific combinations that match particular cell types. In the presumptive digits, we find that the expression of Hoxd gene is unbalanced, despite their common genomic proximity to known global enhancers, often expressing only a subset of the five genes transcribed in these cells. We also report that combinatorial expression follows a pseudo-time sequence, suggesting that a progression in combinatorial expression may be associated with cellular diversity in developing digits.
Some generic measures of the extent of chemical disequilibrium applied to living and abiotic systems
Authors: Intoy B. F. , Halley J. W.
Published: May 2018

We report results of evaluation of several measures of chemical disequilibrium in living and abiotic systems. The previously defined measures includeRTandRLwhich are Euclidean distances of a coarse grained polymer length distribution from two different chemical equilibrium states associated with equilibration to an external temperature bath and with isolated equilibration to a distribution determined by the bond energy of the system, respectively. The determination uses a simplified model of the energetics of the constituent molecules introduced earlier. We evaluated the measures for data from the ribosome of E. Coli, a variety of yeast, the proteomes (with certain assumptions) of a large family of prokaryotes, for mass spectrometric data from the atmosphere the Saturn satellite Titan and for commercial copolymers. We find with surprising consistency thatRLis much smaller thanRTfor all these systems. SmallRLmay be characteristic of systems in the biosphere.
Evaluation of whole genome sequencing for the identification and typing of Vibrio cholerae
Authors: Greig David R. , Schafer Ulf , Octavia Sophie , Hunter Ebony , Chattaway Marie A. , Dallman Timothy J. , Jenkins Claire
Published: May 2018

Epidemiological and microbiological data on Vibrio cholerae isolated between 2004 and 2017 (n=836) and held in the Public Health England culture archive were reviewed. The traditional biochemical species identification and serological typing results were compared with the genome derived species identification and serotype for a sub-set of isolates (n=152). Of the 836 isolates, 750 (89.7%) were from faecal specimens, 206 (24.6%) belonged to serogroup O1 and seven (0.8%) were serogroup O139, and 792 (94.7%) isolates from patients reporting recent travel abroad, most commonly to India (n=209) and Pakistan (n=104). Of the 152 isolates of V. cholerae speciated by kmer identification, 149 (98.1%) were concordant with the traditional biochemical approach. Traditional serotyping results were 100% concordant with the whole genome sequencing (WGS) analysis for identification of serogroups O1 and O139 and Classical and El Tor biotypes. ctxA was detected in all isolates of V. cholerae O1 El Tor and O139 belonging to sequence type (ST) 69, and in V. cholerae O1 Classical variants belonging to ST73. A phylogeny of isolates belonging to ST69 from UK travellers clustered geographically, with isolates from India and Pakistan located on separate branches. Moving forward, WGS data from UK travellers will contribute to global surveillance programs, and the monitoring of emerging threats to public health and the global dissemination of pathogenic lineages. At the national level, these WGS data will inform the timely reinforcement of direct public health messaging to travellers and mitigate the impact of imported infections and the associated risks to public health.
Using Core Genome Alignments to Assign Bacterial Species
Authors: Chung Matthew , Munro James B. , Dunning Hotopp Julie C.
Published: May 2018

With the exponential increase in the number of bacterial taxa with genome sequence data, a new standardized method is needed to assign bacterial species designations using genomic data that is consistent with the classically-obtained taxonomy. This is particularly acute for unculturable obligate intracellular bacteria like those in the Rickettsiales, where classical methods like DNA-DNA hybridization cannot be used to define species. Within the Rickettsiales, species designations have been applied inconsistently, often obfuscating the relationship between organisms and the context for experimental results. In this study, we generated core genome alignments for a wide range of genera with classically defined species, includingArcobacter,Caulobacter,Erwinia,Neisseria,Polaribacter,Ralstonia,Thermus, as well as genera within the Rickettsiales includingRickettsia,Orientia,Ehrlichia,Neoehrlichia,Anaplasma,eorickettsia, andWolbachia. A core genome alignment sequence identity (CGASI) threshold of 96.8% was found to maximize the prediction of classically-defined species. Using the CGASI cutoff, theWolbachiagenus can be delineated into species that differ from the currently used supergroup designations, while theRickettsiagenus is delineated into nine species, as opposed to the current 27 species. Additionally, we find that core genome alignments cannot be constructed between genomes belonging to different genera, establishing a bacterial genus cutoff that suggests the need to create new genera from theAnaplasmaandNeorickettsia. By using core genome alignments to assign taxonomic designations, we aim to provide a high-resolution, robust method for bacterial nomenclature that is aligned with classically-obtained results.
Abnormal microtubule dynamics disrupt nucleocytoplasmic transport in tau-mediated frontotemporal dementia
Authors: Paonessa F. , Evans L. , Solanki R. , Larrieu D. , Wray S. , Hardy J. , Jackson S.P. , Livesey F.J.
Published: May 2018

The neuronal microtubule-associated protein tau (MAPT) is central to the pathogenesis of many dementias, including Alzheimer’s disease. Autosomal dominant mutations inMAPTcause inherited frontotemporal dementia (FTD), but the underlying pathogenic mechanisms are unclear. Using human stem cell models of FTD due toMAPTmutations, we find that tau becomes hyperphosphorylated and mislocalises to neuronal cell bodies and dendrites in cortical neurons, recapitulating a key early event in FTD. Mislocalised tau in the cell body leads to abnormal microtubule dynamics in FTD-MAPT neurons that grossly deform the nuclear membrane, resulting in defective nucleocytoplasmic transport. Neurons in the post-mortem human FTD-MAPT cortex have a high incidence of nuclear deformation, indicating that tau-mediated nuclear membrane dysfunction is an important pathogenic process in FTD. Defects in nucleocytoplasmic transport in FTD point to important commonalities in the pathogenic mechanisms of both tau-mediated dementias and ALS-FTD due to TDP-43 and C9orf72 mutations.
Practical bioinstrumentation developments for AC magnetic field-mediated magnetic nanoparticle heating applications
Authors: Subramanian Mahendran , Miaskowski Arkadiusz , Mahapatro Ajit K. , Hovorka Ondrej , Dobson Jon
Published: May 2018

Heat dissipation during magnetization reversal processes in magnetic nanoparticles (MNP), upon exposure to alternating magnetic fields (AMF), has been extensively studied in relation to applications in magnetic fluid hyperthermia (MFH). This current paper demonstrates the design, fabrication, and evaluation of an efficient instrument, operating on this principle, for use as (i) a non-contact, in vitro, real-time temperature monitor; (ii) a drug release analysis system (DRAS); (iii) a high flux density module for AMF-mediated MNP studies; and (iv) an in vivo coil setup for real-time, whole body thermal imaging. The proposed DRAS is demonstrated by an AMF-mediated drug release proof-of-principle experiment. Also, the technique described facilitates non-contact temperature measurements of specific absorption rate (SAR) as accurately as temperature measurements using a probe in contact with the sample. Numerical calculations estimating the absolute and root mean squared flux densities, and other MNP – AMF studies suggest that the proposed stacked planar coil module could be employed for calorimetry. Even though the proposed in vivo coil setup could be used for real-time, whole body thermal imaging (within the limitations due to issues of penetration depth), further design effort is required in order to enhance the energy transfer efficiency.
A survey of public attitudes towards third-party reproduction in Japan in 2014
Authors: Yamamoto Naoko , Hirata Tetsuya , Izumi Gentaro , Nakazawa Akari , Fukuda Shinya , Neriishi Kazuaki , Arakawa Tomoko , Takamura Masashi , Harada Miyuki , Hirota Yasushi , Koga Kaori , Wada-Hiraike Osamu , Fujii Tomoyuki , Irahara Minoru , Osuga Yutaka
Published: May 2018

Objective: The objective of this study was to examine public attitudes towards third-party reproduction and the disclosure of conception through third-party reproduction.

Methods: We conducted the web-based survey for the public attitude towards third-party reproduction in February 2014. Twenty-five hundred people were recruited with equal segregation of age (20s, 30s, 40s, and 50s) and gender. We analyzed the association between gender, age, infertility, and ethical view using a questionnaire regarding donor sperm, donor oocyte, donor embryo, gestational surrogacy, and disclosure to offspring.

Results: Of the respondents, 36.2% approved and 26.6% disapproved with gamete or embryo donation. The frequency of those who approved was the lowest in females in the 50-59 years age group, and was significantly higher in males or females with infertility. Secondly, 40.9% approved and 21.8% disapproved with gestational surrogacy. The frequency of those who approved gestational surrogacy was higher in males or females with infertility. Thirdly, 46.3% of respondents agreed and 20.4% disagreed with “offspring have the right to know their origin”. Those who disagreed were primarily in the 50-59 age group of both genders, and disagreement was significantly higher in the infertility group compared with non-infertility group.

Conclusion: In this study, public attitudes were affected by gender, age, and experience of infertility. These study findings are important in understanding the attitude towards third-party reproduction and disclosure to the offspring. Respondents having indecisive attitudes were >30%, which might indicate an increased requirement for information and education to enhance the discussion on the ethical consensus on third-party reproduction in Japan.
Single-fluorescent protein reporters allow parallel quantification of NK cell-mediated granzyme and caspase activities in single target cells
Authors: Liesche Clarissa , Sauer Patricia , Claus Maren , Eils Roland , Beaudouin Joël , Watzl Carsten
Published: May 2018

1.

Natural killer (NK) cells eliminate infected and tumorigenic cells through delivery of granzymes via perforin pores or by activation of caspases via death receptors. In order to understand how NK cells combine different cell death mechanisms it is important to quantify target cell responses on a single cell level. However, currently existing reporters do not allow the measurement of several protease activities inside the same cell. Here we present a strategy for the comparison of two different proteases at a time inside individual target cells upon engagement by NK cells. We developed single-fluorescent protein reporters containing the RIEAD or the VGPD cleavage site for the measurement of granzyme B activity. We show that these two granzyme B reporters can be applied in combination with caspase-8 or caspase-3 reporters. While we did not find that caspase-8 was activated by granzyme B, our method revealed that caspase-3 activity follows granzyme B activity with a delay of about 6 minutes. Finally, we illustrate the comparison of several different reporters for granzyme A, M, K and H. The here presented approach is a valuable means for the investigation of the temporal evolution of cell death mediated by cytotoxic lymphocytes.
Antimicrobial peptides modulate long-term memory
Authors: Barajas Azpeleta Raquel , Wu Jianping , Gill Jason , Welte Ryan , Seidel Chris , Si Kausik
Published: May 2018

Antimicrobial peptides act as a host defense mechanism and regulate the commensal microbiome. To obtain a comprehensive view of genes contributing to long-term memory we performed mRNA sequencing from singleDrosophilaheads following behavioral training that produces long-lasting memory. Surprisingly, we find that two immune peptides with antimicrobial activity, Diptericin B and Gram-Negative Bacteria Binding Protein like 3, regulate long-term but not short-term memory or instinctive behavior inDrosophila. The cellular requirement of these two peptides is distinct: head fat body for DptB, and neurons for GNBP-like3. That antimicrobial peptides influence memory provides a novel example of the emerging link between the immune and nervous systems and reveals that some immune peptides may have been repurposed in the nervous system.
An intermittent hypercaloric diet alters gut microbiota, prefrontal cortical gene expression and social behaviours in rats
Authors: Reichelt Amy C , Loughman Amy , Bernard Ashton , Raipuria Mukesh , Abott Kirsten N , Dachtler James , Van Thi Thu Hao , Moore Robert J
Published: May 2018

Excessive consumption of high fat and high sugar (HFHS) diets are known to alter reward processing and aspects of behaviour, and change microbiota profiles. Studies in gnotobiotic mice also provide evidence that gut microorganisms influence social behaviour. To further investigate these interactions, the impact of intermittent access to a HFHS diet on social behaviour, gene expression and microbiota composition was examined. Rats were permitted intermittent daily access (2h / day) to a palatable HFHS diet for 28 days across the adolescent period. Social interaction, social memory and novel object recognition were assessed during this period. Following testing, RT-PCR was conducted on hippocampal and prefrontal cortex (PFC) samples. 16S ribosomal RNA amplicon sequencing was used for identification and relative quantification of bacterial taxa. Reduced social interaction behaviours, and impaired social memory and novel object recognition were observed in HFHS diet rats. Reduced levels of monoamine oxidase A (Maoa), catechol-O-methyltransferase (Comt) and brain derived neurotrophic factor (Bdnf) mRNA were observed in the PFC of HFHS diet rats. The relative abundance of a number of specific taxa differed significantly between the two diet groups, in particular,LachnospiraceaeandRuminoccoceaebacteria, which also predicted social behaviours, novel object recognition performance and Maoa expression. This is the first study to show that limited daily access to HFHS diet alters social behaviour and cognition in rats. Furthermore, behavioural changes are associated with alterations to cortical gene expression of enzymes involved in monoamine synthesis and neuroplasticity, and microbiota profiles predicted diet-induced changes to behaviour and gene expression.
Simultaneous multiplexed amplicon sequencing and transcriptome profiling in single cells
Authors: Saikia Mridusmita , Burnham Philip , Keshavjee Sara H. , Wang Michael F. Z. , Heyang Michael , Moral-Lopez Pablo , Hinchman Meleana M. , Danko Charles G. , Parker John S. L. , Vlaminck Iwijn De
Published: May 2018

We describe Droplet Assisted RNA Targeting by single cell sequencing (DART-seq), a versatile technology that enables multiplexed amplicon sequencing and transcriptome profiling in single cells. We applied DART-seq to simultaneously characterize the non-A-tailed transcripts of a segmented dsRNA virus and the transcriptome of the infected cell. In addition, we used DART-seq to simultaneously determine the natively paired, variable region heavy and light chain amplicons and the transcriptome of B lymphocytes.
Description and charactrization of the Amazonian entomopathogenic bacteriumPhotorhabdus luminescensMN7
Authors: Kamitani Fernando L. , Almenara Daniela P. , Rossi Carolina , Camargo Neves Maira R. , Müller Lissandra M. A. , Gruber Arthur , Alves João M. P. , Yamaguchi Lydia F. , Yoshida Nídia C. , Kato Massuo J. , Winter Carlos E.
Published: May 2018

Many isolates of the genusPhotorhabdushave been reported around the world. Here we describe the first BrazilianPhotorhabdusisolate, found in association with the entomopathogenic nematodeHeterorhabditis baujardiLPP7, from the Amazonian forest in Monte Negro (RO, Brazil). The new isolate can be grouped with the Hb-Hm clade ofP. luminescenssubsp.luminescens, close to the new subspeciesP. luminescenssubsp.sonorensis. P. luminescensMN7 has several characteristics expected of variant form I cells, such as the presence of intracellular crystals, secretion of hydrolytic enzymes (lipases and proteases) and bioluminescence. AlthoughH. baujardiLPP7 is not prolific when compared toH. bacteriophoraHP88,P. luminescensMN7 is clearly pathogenic and probably secretes the same toxins asP. luminescenssubsp.luminescensW14, when fed to larvae of the greater wax mothGalleria mellonella. This behavior is different from what is found inPhotorhabdus luminescenssubsp.laumondiiHP88, which was used as a control in our experiments, andP. l. subsp.laumondiiTT01. Besides the toxin secretion,P. luminescensMN7 secretes proteolytic polypeptides that have molecular masses different from those found inP. l. subsp.laumondiiTT01. Finally, the crude extract from spent culture medium was shown to contain 3,5-dihydroxy-4-isopropyl-cis-stilbene and 1,3,8-trihydroxy-9,10-anthraquinone as the major compounds, similarly to otherPhotorhabdus luminescensstrains.
Capturing single-cell heterogeneity via data fusion improves image-based profiling
Authors: Rohban Mohammad H. , Singh Shantanu , Carpenter Anne E.
Published: May 2018

Single-cell resolution technologies warrant computational methods that capture cell heterogeneity while allowing efficient comparisons of populations. Here, we summarize cell populations by adding features’ dispersion and covariances to population averages, in the context of image-based profiling. We find that data fusion is critical for these metrics to improve results over the prior state-of-the-art, providing at least ~20% better performance in predicting a compound’s mechanism of action (MoA) and a gene’s pathway.