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Background Inflammatory bowel diseases (IBDs) involve an aberrant host response to intestinal microbiota causing mucosal inflammation and gastrointestinal symptoms. Patient-reported outcomes (PROs) are increasingly important in clinical care and research. Our aim was to examine associations between PROs and fecal microbiota in patients 0 to 22 years of age with IBD. Methods A longitudinal, prospective, single-center study tested for associations between microbial community composition via shotgun metagenomics and PROs including stool frequency and rectal bleeding in ulcerative colitis (UC) and abdominal pain and stool frequency in Crohn’s disease (CD). Mucosal inflammation was assessed with fecal calprotectin. A negative binomial mixed-effects model including clinical characteristics and fecal calprotectin tested for differentially abundant species and metabolic pathways by PROs. Results In 70 CD patients with 244 stool samples, abdominal pain correlated with increased relative abundance of Haemophilus and reduced Clostridium spp. There were no differences relative to calprotectin level. In 23 UC patients with 76 samples, both rectal bleeding and increased stool frequency correlated with increased Klebsiella and reduced Bacteroides spp. Conversely, UC patients with lower calprotectin had reduced Klebsiella. Both UC and CD patients with active symptoms exhibited less longitudinal microbial community stability. No differences in metabolic pathways were observed in CD. Increased sulfoglycolysis and ornithine biosynthesis correlated with symptomatic UC. Conclusions Microbial community composition correlated with PROs in both CD and UC. Metabolic pathways differed relative to PROs in UC, but not CD. Data suggest that microbiota may contribute to patient symptoms in IBD, in addition to effects of mucosal inflammation.

Accurate presumptive and confirmatory test use for forensic body fluid identification is essential for gaining contextual information for crime scene investigators. Loop-mediated isothermal amplification (LAMP) is an ideal method for forensic body fluid identification because it is highly specific and generates multi-sized amplicon DNA, and successful amplification results can be read out colorimetrically. Here, we show preliminary data on a LAMP method that rapidly identifies body fluids including venous blood, semen, and saliva, based on colorimetric response and image analysis. The method is designed for easy implementation into forensic casework protocols with minimal disruption to DNA analysis. LAMP naturally increases target specificity due to the use of multiple primers for one target and mRNA targets were used for tissue and human specificity. With colorimetric detection as an inherent part of LAMP, samples that are positive or negative for any of the body fluids are readily identified by image capture and analysis, thus eliminating subjectivity. Results show by using the 3D-printed imaging system specific color ranges can be set for easy determination of body fluids. The resulting color change can be seen in

Ana Lily Amirpour’s 2017 film The Bad Batch is a nightmare of postmodern abjection. Set in a desert wasteland in Texas, the film depicts a quasi-futuristic society that starkly reveals the dark underside of contemporary society, here portrayed in two realms, both exhibiting the height of abjection: the cannibal town called the Bridge and the shanty town of Comfort, where a lone perverse patriarch impregnates all the women while doling out steady doses of LSD to contain the masses. Borrowing from Julia Kristeva’s description of the ‘deject’ in her work Powers of Horror, this analysis focuses on those characters who ultimately choose neither of these options. Having confronted and internalized the abject, these characters become eternal exiles, achieving a measure of liberation by assuming and embodying their partiality and by embracing ‘a weight of meaninglessness, about which there is nothing insignificant’ (Kristeva, J., 1982: 2).

Subtherapeutic drug concentrations contribute to both primary and secondary nonresponse to infliximab in children with Crohn disease (CD). The aim of this study was to evaluate treatment outcomes and infliximab concentrations at infusions 2 and 3 with an objective to establish infliximab targets during induction for primary responders.Single-center, prospective cohort of anti- tumor necrosis factor-alpha naïve CD patients younger than 22 years starting infliximab. Clinical response was defined with the weighted pediatric CD activity index at the fourth infusion. Rates of biological response (50% improvement in fecal calprotectin) and maintenance concentrations ≥5 μg/mL were secondary outcomes.We enrolled 72 patients with CD with 70 of 72 receiving infliximab monotherapy. Clinical response, biological response, and start of maintenance concentrations ≥5 μg/mL were achieved in 64%, 54%, and 22%, respectively. The median (interquartile range) infliximab concentrations at infusion 2 and 3 in clinical responders were 27.8 μg/mL (19.5-40) and 14 μg/mL (8.3-24) compared to 18.8 μg/mL (9.1-23, P 0.001) and 7.8 μg/mL (4-13.2, P 0.01) in nonresponders. Receiver operating characteristic analysis determined that an infliximab concentration ≥15.9 μg/mL at infusion 3 was associated with clinical response (area under the curve [AUC] 0.73), whereas an infusion 3 level ≥18 μg/mL was associated with a start of maintenance concentration5 μg/mL (AUC 0.85). Independent predictors for infusion 3 levels18 μg/mL included pretreatment prednisone, low body mass index, elevated erythrocyte sedimentation rate and C-reactive protein, hypoalbuminemia, and an infusion 2 infliximab level29 μg/mL.We found that infusion 2 (≥29 μg/mL) and infusion 3 (≥18 μg/mL) infliximab concentrations were strongly associated with improved early outcomes and higher first maintenance dose levels.

We report the successful separation of sperm cells from a relevant composition of mock sexual assault samples using a novel acoustic differential extraction (ADE) technology. A multi-layer microfluidic device fabricated in a non-photolithographic process from glass and polydimethylsiloxane (PDMS) was capable of interfacing with custom-built instrumentation to exploit a standing acoustic wave for the trapping of individual sperm cells in a sample containing an abundance of epithelial cells. Samples were generated from buccal and vaginal swabs to mimic post-coital vaginal swabs, and processed through the ADE system followed by DNA extraction of the captured cells with amplification of DNA using a custom short tandem repeat (STR) chemistry. The prototype acoustic trapping technology was fully capable of isolating intact sperm cells from mock samples with disparate masses of male and female DNA. Other biological components were evaluated for adverse effects on sperm cell trapping, including blood, yeast, and bacteria (E. coli), and these had negligible effects on observed sperm cell trapping. Finally, we demonstrate the successful capture of sperm cells from mock samples containing a 40-fold excess in female epithelial cells over sperm cells. The effectiveness of sperm cell purification was ascertained with polymerase chain reaction (PCR) amplification of STR loci from the male fraction post separation with an 18-plex amplification kit, which resulted in male-only profiles.

Standard-of-care infliximab dosing regimens were developed prior to the routine use of therapeutic drug monitoring and identification of target concentrations. Not surprisingly, subtherapeutic infliximab concentrations in pediatric Crohn’s disease (CD) are common. The primary aim was to conduct a real-world pharmacokinetic (PK) evaluation to discover blood biomarkers of rapid clearance, identify exposure targets, and a secondary aim to translate PK modeling to the clinic. In a multicenter observational study, 671 peak and trough infliximab concentrations from 78 patients with CD were analyzed with a drug-tolerant assay (Esoterix; LabCorp, Calabasas, CA). Individual area under the curve (AUC) estimates were generated as a measure of drug exposure over time. Population PK modeling (nonlinear mixed-effect modeling) identified serum albumin, antibody to infliximab, erythrocyte sedimentation rate (ESR), and neutrophil CD64 as biomarkers for drug clearance. Week 14 and week 52 biochemical remitters (fecal calprotectin < 250 μg/g) had higher infliximab exposure (AUC) throughout induction. The optimal infliximab AUC target during induction for week 14 biochemical remission was 79,348 μg*h/mL (area under the receiver operating characteristic curve (AUROC) 0.77, [0.63–0.90], 85.7% sensitive, and 64.3% specific) with those exceeding the AUC target more likely to achieve a surgery-free week 52 biochemical remission (OR 4.3, [1.2–14.6]). Pretreatment predictors for subtherapeutic week 14 AUC included neutrophil CD64 > 6 (OR 4.5, [1.4–17.8]), ESR > 30 mm/h (OR 3.8, [1.4–11]), age < 10 years old (OR 4.2, [1.2–20]), and weight < 30 kg (OR 6.6, [2.1–25]). We created a decision-support PK dashboard with an iterative process and embedded the modeling program within the electronic health record. Model-informed precision dosing guided by real-world PKs is now available at the bedside in real-time.

Background The neutrophil fecal biomarkers, calprotectin (FCP) and lactoferrin (LCT), and peripheral blood neutrophil CD64 surface receptor (nCD64) are biomarkers for mucosal inflammation in inflammatory bowel disease (IBD). Although FCP has been evaluated as a biomarker for mucosal healing, cut points for LCT and nCD64 are less known. We aimed to identify the cut points for LCT and nCD64 that were associated with FCP remission, with a secondary aim to evaluate the relationship between biochemical outcomes and infliximab (IFX) trough concentrations. Methods We analyzed FCP, LCT, and nCD64 before and after IFX induction in a pediatric Crohn’s disease (CD) cohort study. Week-14 FCP biomarker remission was defined as FCP 17.5 improvement. Predictive outcomes were calculated by receiver operating characteristics (ROCs). Results Among 56 CD patients, ROC analysis identified an infusion 4 LCT 9.4 µg/mL (AUROC, 0.799, P = 0.002) and >11.5 µg/mL (AUROC, 0.835, P = 0.003) were associated with a FCP 5 µg/mL had a median FCP improvement (dose 1 to dose 4) of 90% compared with a median of 35% with levels Conclusions This study establishes cut points in neutrophil stool and blood biomarkers for both biochemical remission and therapeutic trough levels following induction therapy. Further studies that evaluate pharmacodynamic biomarker targets for endoscopic and histologic healing are warranted.

Author(s): Denson, Lee A; Jurickova, Ingrid; Karns, Rebekah; Shaw, Kelly A; Cutler, David J; Okou, David; Valencia, C Alexander; Dodd, Anne; Mondal, Kajari; Aronow, Bruce J; Haberman, Yael; Linn, Aaron; Price, Adam; Bezold, Ramona; Lake, Kathleen; Jackson, Kimberly; Walters, Thomas D; Griffiths, Anne; Baldassano, Robert N; Noe, Joshua D; Hyams, Jeffrey S; Crandall, Wallace V; Kirschner, Barbara S; Heyman, Melvin B; Snapper, Scott; Guthery, Stephen L; Dubinsky, Marla C; Leleiko, Neal S; Otley, Anthony R; Xavier, Ramnik J; Stevens, Christine; Daly, Mark J; Zwick, Michael E; Kugathasan, Subra | Abstract: BACKGROUND:Granulocyte-macrophage colony-stimulating factor auto-antibodies (GMAbs) suppress neutrophil-extrinsic GM-CSF signaling and increase risk for stricturing behavior in Crohn's disease (CD). We aimed to define clinical, genomic, and functional associations with neutrophil-intrinsic GM-CSF signaling. METHODS:Missense mutations in CSF2RA, CSF2RB, JAK2, STAT5A, and STAT5B were identified using whole-exome sequencing in 543 pediatric inflammatory bowel disease (IBD) patients. Neutrophil-intrinsic GM-CSF signaling was defined using the GM-CSF-induced STAT5 stimulation index (GMSI) in 180 pediatric IBD patients and 26 non-IBD controls. Reduced GM-CSF signaling (GMSI-Lo) was defined as the 20th percentile within the control group. Variation in neutrophil phospho-protein abundance, bacterial killing, and the global pattern of gene expression with the GMSI was determined. RESULTS:We validated 18 potentially damaging missense mutations in CSF2RA and CSF2RB. CSF2RA A17G carriage increased from 10% in those with intact neutrophil GMSI to 32% in those with low GMSI (P = 0.02). The frequency of reduced Staphylococcus aureus killing increased from 17% in those with intact neutrophil GMSI to 35% in GMSI-Lo neutrophils (P = 0.043). Crohn's disease neutrophils with low GMSI exhibited specific alterations in phospho-protein networks and genes regulating cytokine production, wound healing, and cell survival and proliferation. Stricturing behavior increased from 7% in patients with both low GMAb and intact GMSI to 64% in patients with both elevated GMAb and low GMSI (P l 0.0001). CONCLUSIONS:Low/normal neutrophil-intrinsic GM-CSF signaling is associated with CSF2RA missense mutations, alterations in gene expression networks, and higher rates of disease complications in pediatric CD.

Background & aimsIndividuals with monogenic disorders of phagocyte function develop chronic colitis that resembles Crohn's disease (CD). We tested for associations between mutations in genes encoding reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidases, neutrophil function, and phenotypes of CD in pediatric patients.MethodsWe performed whole-exome sequence analysis to identify mutations in genes encoding NADPH oxidases (such as CYBA, CYBB, NCF1, NCF2, NCF4, RAC1, and RAC2) using DNA from 543 pediatric patients with inflammatory bowel diseases. Blood samples were collected from an additional 129 pediatric patients with CD and 26 children without IBD (controls); we performed assays for neutrophil activation, reactive oxygen species (ROS) production, and bacteria uptake and killing. Whole-exome sequence analysis was performed using DNA from 46 of the children with CD to examine associations with NADPH gene mutations; RNA sequence analyses were performed using blood cells from 46 children with CD to test for variations in neutrophil gene expression associated with ROS production.ResultsWe identified 26 missense mutations in CYBA, CYBB, NCF1, NCF2, and NCF4. Patients with CD who carried mutations in these genes were 3-fold more likely to have perianal disease (P= .0008) and stricturing complications (P= .002) than children with CD without these mutations. Among patients withCD with none of these mutations, 9% had undergone abdominal surgery; among patients with mutations in these NADPH oxidase genes, 31% had undergone abdominal surgery (P=.0004). A higher proportion of neutrophils from children with CD had low ROS production (47%) than from controls (15%) among the 129 patients tested for ROS (P= .002). Minor alleles of the NADPH genes were detected in 7% of children with CD whose neutrophils produced normal levels of ROS vs 38% of children whose neutrophils produced low levels of ROS (P= .009). Neutrophils that produced low levels of ROS had specific alterations in genes that regulate glucose metabolism and antimicrobial responses.ConclusionsWe identified missense mutations in genes that encode NADPH oxidases in children with CD; these were associated with a more aggressive disease course and reduced ROS production by neutrophils from the patients.

Objectives Inflammatory bowel diseases (IBD), including Crohn’s disease (CD) and ulcerative colitis (UC), are characterized by an aberrant host response to intestinal microbiota causing mucosal inflammation and gastrointestinal symptoms. Patient reported outcome measures (PROs) are increasingly utilized in clinical care and research. Our study aim was to determine if fecal microbial shifts were associated with PROs in children and young adults with IBD. Methods A longitudinal prospective single center study of 93 patients tested for association between fecal microbial shifts, mucosal inflammation as measured by fecal calprotectin, and self-reported symptoms including diarrhea, rectal bleeding, and abdominal pain. For CD, abdominal pain and diarrhea determined disease activity or overall PRO. For UC, diarrhea and rectal bleeding were used. Fecal calprotectin and shotgun metagenomic sequencing were performed on all samples. Demographic and clinical characteristics (Table 1) were incorporated into a negative binomial mixed-effects model in “R” to identify differentially abundant species by PRO and fecal calprotectin. Metabolic pathways were mapped using the HUMAnN2 pipeline and compared to overall PRO, individual symptoms and fecal calprotectin level. Results In 70 CD patients with 244 stool samples, there was no association between symptoms and fecal calprotectin. In 23 UC patients with 76 stool samples, increased fecal calprotectin was associated with rectal bleeding (OR 4.93 [1.18, 20.64], p=0.03). Examination of differentially abundant species in those with self-reported active UC showed increased Klebsiella species and reduced Bacteroides. Conversely, UC patients with fecal calprotectin < 100 µg/gm had reduction in Klebsiella and increase in Bifidobacteria and Bacteroides (Figure 1). Analysis of differentially abundant species in those with abdominal pain in CD showed increase in Haemophilus and reduction in Bacteroides. No microbial shifts were identified in CD patients in association with overall PRO, diarrhea, nor with fecal calprotectin < 250 µg/gm. Metabolic pathway analysis showed no differences in those with CD. In UC patients, increases in sulfoglycolysis and ornithine biosynthesis were associated with overall PROs. Conclusions Fecal microbial shifts including decreased commensals such as Bacteroides correlate with UC patient symptoms. Increased fecal calprotectin level was associated with rectal bleeding in these patients, but not diarrhea. In CD, there was no association with fecal calprotectin and symptoms, and microbial shifts were detected in association with abdominal pain. Similarly, metabolic pathways differed relative to patient-reported symptoms in UC, but not in CD. Data suggests that microbial shifts may directly contribute to symptoms in children and young adults with IBD.

AIM To determine the vaccination rates in pediatric immunosuppression-dependent inflammatory bowel disease (IBD) and review the safety and efficacy of vaccinations in this population. METHODS The electronic medical records from October 2009 to December 2015 of patients diagnosed with IBD at 10 years of age or younger and prescribed anti-tumor necrosis factor alpha (anti-TNF-α) therapy were reviewed for clinical history, medication history, vaccination history, and hepatitis B and varicella titers. Literature discussing vaccination response in IBD patients were identified through search of the MEDLINE database and reviewed using the key words “inflammatory bowel disease”, “immunization”, “vaccination”, “pneumococcal”, “varicella”, and “hepatitis B”. Non-human and non-English language studies were excluded. Search results were reviewed by authors to select articles that addressed safety and efficacy of immunizations in inflammatory bowel disease. RESULTS A total of 51 patients diagnosed with IBD prior to the age of 10 and receiving anti-TNF-α therapy were identified. Thirty-three percent of patients (17/51) had incomplete or no documentation of vaccinations. Sixteen case reports, cohort studies, cross-sectional studies, and randomized trials were determined through review of the literature to describe the safety and efficacy of hepatitis B, pneumococcal, and varicella immunizations in adult and pediatric patients with IBD. These studies showed that patients safely tolerated the vaccines without significant adverse effects. Importantly, IBD patients receiving immunosuppressive medications, particularly anti-TNF-α treatment, have decreased vaccine response compared to controls. However, the majority of patients are still able to achieve protective levels of specific antibodies. CONCLUSION Immunizations have been shown to be well-tolerated and protective immunity can be achieved in patients with IBD requiring immunosuppressive therapy.

Lab Meeting: How do Black Lives Matter in Teaching, Lab Practices and Research?Curated by Anne Pollock and Deboleena Roy

OBJECTIVES Exocrine pancreatic insufficiency (EPI) can have a significant impact on a child's growth and nutrition. Our aim was to evaluate the utility of direct endoscopic pancreatic function testing (ePFT) in pediatrics. METHODS A single-center retrospective chart review was performed of children who underwent ePFT from December 2007 through February 2015. Endoscopic pancreatic function testings were performed by 1 of 2 methods: (1) intravenous cholecystokinin, followed by the collection of a single duodenal aspirate at 10 minutes, or (2) intravenous cholecystokinin or secretin, followed by the collection of 3 duodenal aspirates at a 5, 10, and 15 minutes. Samples were tested for pH and enzyme activities. RESULTS A total of 508 ePFTs were performed (481 single-sample tests, 27 multiple-sample tests). Based on the multiple-sample group, enzyme levels for chymotrypsin, amylase, and lipase peaked at 5 minutes, followed by a decrease in activity over time. Exocrine pancreatic sufficiency was identified in 373 (73.4%) and EPI in 93 (18.3%). Exocrine pancreatic sufficiency analysis found all pancreatic enzyme activities significantly increase with age: trypsin, chymotrypsin, amylase, and lipase, (P < 0.05). CONCLUSIONS Endoscopic pancreatic function testing can be used in the evaluation of EPI in children. Normative data suggest that pancreatic enzyme activities mature with age.

This report describes what professional experiences and other characteristics the general and flag officers in each of the military services tend to share as a result of each service's approach to personnel management and other related factors, such as service culture, and how these approaches might influence the ways in which general and flag officers lead, manage, and advise.

Background Clostridioides difficile infection and colonization are common in pediatric Crohn’s disease (CD). Our aims were to test the relationship between C. difficile positivity and bowel resection surgery and to characterize microbial shifts associated with C. difficile carriage and surgery. Methods A retrospective single-center study of 75 pediatric CD patients tested for association between C. difficile carriage and bowel resection surgery. A prospective single-center study of 70 CD patients utilized C. difficile testing and shotgun metagenomic sequencing of fecal samples to define microbiota variation stratified by C. difficile carriage or history of surgery. Results The rate of bowel resection surgery increased from 21% in those without C. difficile to 67% in those with (P = 0.003). From a Kaplan-Meier survival model, the hazard ratio for time to first surgery was 4.4 (95% CI, 1.2–16.2; P = 0.00) in patients with positive C. difficile testing in the first year after diagnosis. Multivariable logistic regression analysis confirmed this association (odds ratio 16.2; 95% CI, 2.2–120; P = 0.006). Larger differences in microbial abundance and metabolic pathways were observed in patients with prior surgery than in those with C. difficile carriage. Depletion of Alistipes and Ruminococcus species and reduction in methionine biosynthesis were noted in patients with both C. difficile carriage and past surgery. Conclusions A positive C. difficile test during the first year after diagnosis is associated with decreased time to first bowel resection surgery in pediatric Crohn’s disease. Depletion of beneficial commensals and methionine biosynthesis in patients with C. difficile carriage may contribute to increased risk for surgery.

Background We hypothesized that elevations of plasma Oncostatin M (OSM) would be associated with infliximab nonresponse. Methods Plasma OSM was measured in Crohn disease patients pre-infliximab with biochemical response (>50% reduction in fecal calprotectin) as the primary outcome. Results The median OSM in biochemical responders was 86 (69–148) pg/mL compared with 166 (74–1766) pg/mL in nonresponders (P = 0.03). Plasma OSM > 143.5 pg/mL was 71% sensitive and 78% specific for biochemical nonresponse (area under the curve 0.71). Early biochemical nonremission was also associated with an elevated neutrophil CD64 expression (odds ratio 8.9, P = 0.011). Conclusions Elevated preinfliximab plasma OSM and nCD64 surface expression were both associated with poor biochemical outcomes., Pretreatment elevations in plasma Oncostatin M and whole blood neutrophil CD64 are associated with unfavorable early and late treatment outcomes to infliximab in patients with Crohn disease.

Purpose Diffusion-weighted imaging (DWI) and 18F-fluorodeoxyglucose–positron emission tomography (18F-FDG–PET) independently correlate with malignancy in breast cancer, but the relationship between their structural and metabolic metrics is not completely understood. This study spatially correlates diffusion, perfusion, and glucose avidity in breast cancer with simultaneous PET/MR imaging and compares correlations with clinical prognostics. Methods In this Health Insurance Portability and Accountability Act-compliant prospective study, with written informed consent and approval of the institutional review board and using simultaneously acquired FDG-PET and DWI, tissue diffusion (Dt), and perfusion fraction (fp) from intravoxel incoherent motion (IVIM) analysis were registered to FDG-PET within 14 locally advanced breast cancers. Lesions were analyzed using 2D histograms and correlation coefficients between Dt, fp, and standardized uptake value (SUV). Correlations were compared with prognostics from biopsy, metastatic burden from whole-body PET, and treatment history. Results SUV||Dt correlation coefficient significantly distinguished treated (0.11 ± 0.24) from nontreated (−0.33 ± 0.26) patients (P = 0.005). SUV||fp correlations were on average negative for the whole cohort (−0.17 ± 0.13). Conclusion Simultaneously acquired and registered FDG-PET/DWI allowed quantifiable descriptions of breast cancer microenvironments that may provide a framework for monitoring and predicting response to treatment. Magn Reson Med, 2016. © 2016 International Society for Magnetic Resonance in Medicine

This work describes the development of a novel microdevice for forensic DNA processing of reference swabs. This microdevice incorporates an enzyme-based assay for DNA preparation, which allows for faster processing times and reduced sample handling. Infrared-mediated PCR (IR-PCR) is used for STR amplification using a custom reaction mixture, allowing for amplification of STR loci in 45 min while circumventing the limitations of traditional block thermocyclers. Uniquely positioned valves coupled with a simple rotational platform are used to exert fluidic control, eliminating the need for bulky external equipment. All microdevices were fabricated using laser ablation and thermal bonding of PMMA layers. Using this microdevice, the enzyme-mediated DNA liberation module produced DNA yields similar to or higher than those produced using the traditional (tube-based) protocol. Initial microdevice IR-PCR experiments to test the amplification module and reaction (using Phusion Flash/SpeedSTAR) generated near-full profiles that suffered from interlocus peak imbalance and poor adenylation (significant -A). However, subsequent attempts using KAPA 2G and Pfu Ultra polymerases generated full STR profiles with improved interlocus balance and the expected adenylated product. A fully integrated run designed to test microfluidic control successfully generated CE-ready STR amplicons in less than 2 h (

We report the development of a disposable polyester toner centrifugal device for semi-automated, dynamic solid phase DNA extraction (dSPE) from whole blood samples. The integration of a novel adhesive and hydrophobic valving with a simple and low cost microfabrication method allowed for sequential addition of reagents without the need for external equipment for fluid flow control. The spin-dSPE method yielded an average extraction efficiency of ∼45% from 0.6 μL of whole blood. The device performed single sample extractions or accommodate up to four samples for simultaneous DNA extraction, with PCR-readiness DNA confirmed by effective amplification of a β-globin gene. The purity of the DNA was challenged by a multiplex amplification with 16 targeted amplification sites. Successful multiplexed amplification could routinely be obtained using the purified DNA collected post an on-chip extraction, with the results comparable to those obtained with commercial DNA extraction methods. This proof-of-principle work represents a significant step towards a fully-automated low cost DNA extraction device.

This report analyzes the cultural characteristics and competitive strategies and tactics exhibited by the Army, Navy, Air Force, Marine Corps, and U.S. Special Operations Command (USSOCOM). Based on this analysis, the authors assess how each of the services and USSOCOM might adapt and respond if facing major policy shifts in the future, focusing specifically on contingencies in the Asia-Pacific.

Messenger RNA profiling for body fluid identification (bfID) is a useful approach to collect contextual information associated with a crime. Current methods require costly fluorescent probes, lengthy amplification protocols and/or time-consuming sample preparation. To simplify this process, we developed a bfID method that has the potential to be rapid in analysis time, inexpensive and fluorescence-free, combining a universal operating procedure with a high-throughout (microwell plate) platform for simultaneous detection of mRNA markers from whole blood, semen, saliva, and vaginal fluid. Full bfID sample preparation and analysis of 23 samples was completed in under 3 h using smart phone optical detection and analysis and show efficacy of the method in a validated blind study. The results provide an efficient, sensitive and specific approach to supplement the current biochemical tests in a forensic laboratory.

U.S. security-sector assistance has a mixed record in Africa. RAND experts offer recommendations to promote the success of such assistance in Africa. The recommendations focus on strategies, programs, and their assessment, monitoring, and evaluation.

The authors developed and employed a methodology to review general and flag officer (G/FO) requirements in the U.S. military. The methodology examines G/FO positions within and across organizations, reviews positions against criteria, evaluates how positions affect development of G/FOs within functional areas, and explores implications of reductions. Results were used to identify opportunities to eliminate or downgrade positions.

Showtime's hit series Dexter, which aired for eight seasons from 2006 to 2013, chronicles the adventures of a serial killer with "standards" who works for Miami Metro Homicide by day and prowls the streets of Miami by night in search of criminals who slipped through the system. As Karin Hoepker contends in her contribution to Transgressive Television, Dexter "set the standards for many shows to follow". The links between Dexter and Wuthering Heights are not immediately apparent, particularly because Dexter disavows its Gothic elements from the start, even and especially with regard to Dexter's murders. Dex and Deb's story resembles that of Catherine and Heathcliff of Wuthering Heights from the very beginning. In Wuthering Heights, Catherine's position in the family is usurped by Heathcliff, who becomes the father's favorite. Like Heathcliff, Dexter is an impostor, using the law and the patriarchal order to exact his own revenge.

Correct identification of probative samples is the first crucial step in the analysis of sexual assault kits (SAKs). We report a nucleic acid-based approach, as an alternative to the widely utilized p30 assay, to screening male DNA from SAKs collected from female victims by combining a rapid lysis protocol with an isothermal amplification method. The enzymatic lysis protocol efficiently digests biological material to release nuclear DNA in 10 min in a single closed tube, including resilient cell types such as sperm cells. The amplification and detection of human male specific DNA is achieved through loop-mediated isothermal amplification (LAMP) accompanied with hydroxynaphthol blue, a colorimetric indicator, producing a visually-distinctive color change in the presence of male DNA. The Y-screen approach demonstrated high specificity to human male DNA, can reliably detect target DNA as low as 50 pg, and correctly identified all probative samples from 14 single-blind mock sexual assault samples. In contrast with the widely used p30 assay which requires at least 2 h incubation time and manual application to a lateral flow pad, this Y-screen assay can be completed in half the time, and can be performed in a 96-well format without the need for a fluorescence detector, making facile high-throughput sample screening possible.

From September 2011 through May 2012, icarusLabs competed in the UAVForge Competition, sponsored by the United States Defense Advanced Research Projects Agency (DARPA). The objective of the competition was to design and build a small unmanned aerial vehicle (UAV) that could complete a local surveillance mission. A unique feature of the competition was an experimental crowdsourcing model, in which competing teams were able to provide technical feedback to each other as part of the design process. The icarusLabs team's solution to the challenge combined a tricopter and blended wing body aircraft into a single hybrid airframe. This paper presents the rationale and theory behind the design as well as the lessons learned from the technical and logistical challenges faced. In addition, the authors reflect on the crowdsourcing aspect of the challenge from a competitor's perspective. Overall, incentives for individual teams' success directly conflicted with the essence of crowdsourcing, but the competition was successful at stimulating interest to fill a void in current UAV solutions, bringing together passionate individuals from the global unmanned systems community, and generating a broad range of novel approaches to address the presented challenges.

Welsh author Arthur Machen (1863–1947) wrote his most popular supernatural tales between 1890 and 1900, a period in which European culture felt itself to be on the decline and in which “decadent” art and literature rose up both as a reflection of and a contribution to this perceived cultural deterioration. While Machen's works have received little critical attention, a recent revival of interest in fin-de-siècle decadence has brought his supernatural tales into the literary limelight. Noteworthy examples of this interest include Julian North's treatment of The Great God Pan in Michael St. John's Romancing Decay: Ideas of Decadence in European Culture and Christine Ferguson's analysis of the same work in her PMLA article “Decadence as Scientific Fulfillment.” Indeed, Machen's supernatural tales could enhance and complicate any exposition of decadent literature and culture; they offer a unique vision of descent into the primordial that differs from the moral and psychological treatment of decadence in other popular works of the time, such as Wilde's The Picture of Dorian Gray and Stevenson's Strange Case of Dr. Jekyll and Mr. Hyde. Like Stevenson and Wilde, Machen employs themes of transgression and metamorphosis to illustrate his characters’ deviations from human nature. However, the forces at work in Machen's tales do not arise from the recesses of the human mind in its modern conception, nor do his protagonists sin primarily against society and the arbitrary nature of its morals and values. Instead, Machen locates mythic forces at work within his contemporary society to highlight a much older form of transgression and to challenge notions of degeneration that held currency at the end of the nineteenth century.

The U.S.-led strategy to defeat the Islamic State has been criticized for a lack of clarity, overemphasis on tactical objectives, and insufficient attention to the underlying causes of the greater civil conflict across both Iraq and Syria. This report assesses the current strategy and presents three options, derived from subject-matter-expert input. It recommends a new strategy to address root causes.

This report describes the likely evolution of Islamic State of Iraq and the Levant (ISIL) finances under three specific scenarios: a continuation of the current campaign, a negotiated settlement in Syria and political accommodation in Iraq, and military victory without negotiated settlement or political accommodation. Discussion of these scenarios offered ways to counter fundraising in ISIL territory and use financial information to damage ISIL.

This work describes the development of a novel microdevice for forensic DNA processing of reference swabs. This microdevice incorporates an enzyme-based assay for DNA preparation, which allows for faster processing times and reduced sample handling. Infrared-mediated PCR (IR-PCR) is used for STR amplification using a custom reaction mixture, allowing for amplification of STR loci in 45 min while circumventing the limitations of traditional block thermocyclers. Uniquely positioned valves coupled with a simple rotational platform are used to exert fluidic control, eliminating the need for bulky external equipment. All microdevices were fabricated using laser ablation and thermal bonding of PMMA layers. Using this microdevice, the enzyme-mediated DNA liberation module produced DNA yields similar to or higher than those produced using the traditional (tube-based) protocol. Initial microdevice IR-PCR experiments to test the amplification module and reaction (using Phusion Flash/SpeedSTAR) generated near-full profiles that suffered from interlocus peak imbalance and poor adenylation (significant -A). However, subsequent attempts using KAPA 2G and Pfu Ultra polymerases generated full STR profiles with improved interlocus balance and the expected adenylated product. A fully integrated run designed to test microfluidic control successfully generated CE-ready STR amplicons in less than 2 h (1 h of hands-on time). Using this approach, high-quality STR profiles were developed that were consistent with those produced using conventional DNA purification and STR amplification methods. This method is a smaller, more elegant solution than current microdevice methods and offers a cheaper, hands-free, closed-system alternative to traditional forensic methods.

Diffusion-weighted imaging (DWI) andIn this Health Insurance Portability and Accountability Act-compliant prospective study, with written informed consent and approval of the institutional review board and using simultaneously acquired FDG-PET and DWI, tissue diffusion (DSUV||DSimultaneously acquired and registered FDG-PET/DWI allowed quantifiable descriptions of breast cancer microenvironments that may provide a framework for monitoring and predicting response to treatment. Magn Reson Med 78:1147-1156, 2017. © 2016 International Society for Magnetic Resonance in Medicine.

The development of Unmanned Vehicles (UVs) with increasing autonomy has enabled a transition from teleoperation to Human Supervisory Control (HSC). In this demonstration, participants can test three innovative operator interfaces for HSC of UVs. The first system allows users to control a Micro Air Vehicle (MAV) via a hand-held device, such as an iPhone®, through high-level waypoint commands and fine-grained nudge controls. The second system enables a single operator to collaborate with an automated planner to control multiple heterogeneous UVs from a laptop-sized display for searching for, tracking and engaging moving ground targets. The third system is designed to aid in planning on naval aircraft carrier decks, serving as a decision support tool for a supervisor overseeing and scheduling the activity of people, vehicles and unmanned vehicles working in this complex and uncertain environment.

With recent regulatory efforts to reduce restrictions placed on the operation of Micro Air Vehicles (MAVs) in the United States, it is likely that in the next few years, these vehicles will become commonplace in the commercial marketplace as they are in military environments. In order to reduce the barrier to entry for operations of MAVs, customers of these systems will require ease of operation as well as minimal training time in order to reduce costs. To this end, a smartphone application was developed to control a quadrotor remotely in the exploration of an unknown environment, and tested for users with only three minutes of training. Initial motion capture room tests produced encouraging results for localization and target identification tasks, however, such environments are inherently artificial and the extensibility of such results is limited. A follow-on outdoor field study was conducted in order to compare the indoor and outdoor results and to assess operator performance in a realistic environment. Performance on the outdoor localization tasks was comparable to the indoor study, however, participants generally performed slightly worse on the target identification task in the outdoor experiment, attributed to camera image quality and GPS localization issues. Other issues such as wind and flight safety considerations are discussed.

Most major medical organizations recommend routine screening for breast, cervical, and colorectal cancers. Screening can lead to early detection of these cancers, resulting in reduced mortality. Yet, not all people who should be screened are screened regularly or, in some cases, ever. This report presents results of systematic reviews of effectiveness, applicability, economic efficiency, barriers to implementation, and other harms or benefits of provider reminder/recall interventions to increase screening for breast, cervical, and colorectal cancers. These interventions involve using systems to inform healthcare providers when individual clients are due (reminder) or overdue (recall) for specific cancer screening tests. Evidence in this review of studies published from 1986 through 2004 indicates that reminder/recall systems can effectively increase screening with mammography, Pap, fecal occult blood tests, and flexible sigmoidoscopy. Additional research is needed to determine if provider reminder/recall systems are effective in increasing colorectal cancer screening by colonoscopy. Specific areas for further research are also suggested.

In this article I wish to elucidate a form of receptivity in our relation to televisual images that cannot be reduced to the postmodern formula of a closed circuit of desire through which the self(same) infinitely reproduces itself. Through analysis of the peculiar experience of viewing Gore Verbinski’s 2003 film The Ring (a remake of Hideo Nakata’s Ring), I argue for the possibility of an experience of alterity in and through the televisual. I pair this event with the religious experience of viewing the icon that Jean-Luc Marion opposes to the televisual in his recent works collected in The Crossing of the Visible. The purpose of the comparison is not to conflate the two experiences but rather to suggest that the possibility for reclaiming a space for difference in a postmodern world lies within the televisual itself, the very medium that Marion credits with the bankruptcy of the aesthetic. In The Ring, the process through which the viewer expects to see her own desire reproduced is violently interrupted by a vengeful image that shatters the mirror of reflection between viewer and viewed and threatens to cross over that threshold — to cross the visible, in Marion’s terms.