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Introduction: 30-40% patients with acute severe ulcerative colitis (ASUC) fail intravenous (IV) steroids requiring medical rescue therapy/colectomy. Low baseline albumin predicts steroid non-response, and exclusive enteral nutrition (EEN) has been shown to improve steroid response and albumin levels. Albumin infusion due to its anti-inflammatory and anti-oxidant properties might further improve steroid response in ASUC, which was evaluated in present study., Methods: In this open-label randomized controlled trial, patients with ASUC were randomized in 1:1 ratio to albumin + standard of care (SOC) + EEN vs. SOC + EEN (Jan2021 - Feb2023). Both arms received 5 days of EEN with 400 mg IV hydrocortisone/day. Patients in albumin arm were administered 5 days of 20% w/v intravenous albumin (100 ml). Primary outcome was 1) steroid failure (need for rescue medical therapy or colectomy) and 2) proportion of patients with adverse events., Results: Sixty-one patients (albumin-30, SOC-31)(mean age-31.6±0.4 years, male-57.4%), were included. Baseline characteristics were comparable. There was no difference in steroid failure between albumin and SOC arm(10/30(33.33 %) vs 13/31(41.94 %), p=0.49). No adverse events were reported with albumin infusions. Colectomy rate(10% vs 9.68%, P=1), response to salvage medical therapy (88.89% vs 76.92%, P=0.62) and median duration of hospitalization (10.5(7-16) vs 10(7-20), P=0.43) were also comparable. Long-term composite outcome of colectomy and re-admission rates was numerically higher in the albumin than SOC arm (37.04% vs 17.86%, p>0.05), although it did not reach statistical significance., Conclusion: There was no benefit of intravenous albumin infusion as an adjunct to IV steroids and EEN in patients with ASUC., (© The Author(s) 2024. Published by Oxford University Press on behalf of European Crohn’s and Colitis Organisation. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

This work reports that ligand engineering can modulate the triplet harvesting mechanism in iodide-bridged rhombic Cu 2 I 2 complexes. Complex-1, with a smaller Cu-Cu distance, exhibits phosphorescence from 3 (M+X)LCT and 3 CC states with 66% quantum yield, whereas an increased Cu-Cu distance in complex-2 results in a switch of the emission from phosphorescence to TADF, which occurs via 1/3 (M+X)LCT states with 83% quantum yield. The TADF property of complex-2 has been utilized for the fabrication of a pc-LED emitting efficient warm white light. Moreover, the high charge-transfer nature of these complexes leads to the emergence of third-harmonic generation (THG). Interestingly, complex-1 exhibits efficient third-harmonic generation with a χ (3) value of 1.15 × 10 -18 m 2 V -2 and LIDT value of 14.73 GW/cm 2 . This work aims to provide a structure-property relationship to achieve effective harvestation of triplet excitons in iodide-bridged rhombic Cu 2 I 2 complexes and their effective utilization in OLED device fabrication and nonlinear photon upconversion processes.

Background & Aims: Coconut water (CW) is anti-inflammatory, can manipulate the gut microbiome, and is a rich source of potassium. Gut microbiome modulation improves outcomes in ulcerative colitis (UC), and potassium possesses in vitro anti-inflammatory property. We evaluated the effect of CW as an adjunct therapy for patients with mild-moderate UC., Methods: This single-center, double-blind, placebo-controlled trial randomized patients with mild to moderate (Simple Clinical Colitis Activity Index [SCCAI]: 3-9) endoscopically active UC (Ulcerative Colitis Endoscopic Index of Severity [UCEIS] >1) in 1:1 ratio to CW + standard medical therapy (SMT) vs placebo + SMT. Four hundred mL of CW was administered for 8 weeks. Primary outcome measure was clinical remission (SCCAI ≤2), and secondary outcome measures were clinical response (SCCAI decline ≥3) and adverse events at 8 weeks. Microbiome was analyzed at baseline and 8 weeks., Results: Of 121 patients screened, 95 were included for modified intention to treat analysis (CW, n = 49; placebo, n = 46) (mean age, 37.2 ± 11.2 years; males, 54.1%; disease duration, 48 months [interquartile range (IQR), 24-90 months]; pancolitis, 26.1%; SCCAI, 5 [IQR, 4-6]; UCEIS, 4 [IQR, 3-5]). Clinical response (57.1% vs 28.3%; odds ratio [OR], 3.4; 95% confidence interval [CI], 1.4-7.9; P = .01), remission (53.1% vs 28.3%; OR, 2.9; 95% CI, 1.2-6.7; P = .02), and proportion of patients with fecal calprotectin (FCP) <150 μg/g (30.6% vs 6.5%; OR, 6.3; 95% CI, 1.7-23.6; P = .003) were significantly higher in CW. The relative abundance of bacterial taxa that had a significant or trend towards negative correlation with SCCAI, UCEIS, or FCP increased at 8 weeks in CW, and this effect was independent of disease activity and dietary fiber. Adverse events were comparable, and no patient developed hyperkalemia., Conclusions: CW was more effective than placebo for induction of clinical remission in patients with mild to moderate UC. The trial was prospectively registered on Clinical Trials Registry of India (ctri.nic.in, Number: CTRI/2019/03/01827)., (Copyright © 2024 AGA Institute. Published by Elsevier Inc. All rights reserved.)

The RNA-guided CRISPR-associated (Cas) enzyme Cas12a cleaves specific double-stranded (ds-) or single-stranded (ss-) DNA targets (in cis), unleashing non-specific ssDNA cleavage (in trans). Though this trans-activity is widely coopted for diagnostics, little is known about target determinants promoting optimal enzyme performance. Using quantitative kinetics, we show formation of activated nuclease proceeds via two steps whereby rapid binding of Cas12a ribonucleoprotein to target is followed by a slower allosteric transition. Activation does not require a canonical protospacer-adjacent motif (PAM), nor is utilization of such PAMs predictive of high trans-activity. We identify several target determinants that can profoundly impact activation times, including bases within the PAM (for ds- but not ssDNA targets) and sequences within and outside those complementary to the spacer, DNA topology, target length, presence of non-specific DNA, and ribose backbone itself, uncovering previously uncharacterized cleavage of and activation by RNA targets. The results provide insight into the mechanism of Cas12a activation, with direct implications on the role of Cas12a in bacterial immunity and for Cas-based diagnostics., (© The Author(s) 2024. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Planar 44π and 46π core-modified decaphyrins with ten thiophene units have been synthesized from short thiophene oligomers. They have been structurally characterized by single crystal X-ray diffraction with further support from spectroscopic analysis and quantum chemical calculations. Our analysis revealed diradicaloid characteristics for 46π species in contrast to the closed shell property of the 44π congener. Further, 44π and 46π undergo reversible two-electron chemical oxidation, as observed by spectro-electrochemical measurements., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Background and Aims: There are no prospective studies evaluating effect of non-alcoholic fatty liver disease (NAFLD) in patients with ulcerative colitis (UC). This prospective observational study assessed the prevalence of NAFLD, its predictors, and its effect on long-term outcomes in UC., Methods: Consecutive UC patients underwent transient elastography, body composition analysis, bone densitometry, anthropometry, and baseline demographic and subjective global assessment. NAFLD was diagnosed by controlled attenuation parameter of >260 dB/m. To evaluate predictors and outcomes, patients of UC with NAFLD (n = 29) were compared with age- and sex-matched patients of UC without NAFLD (n = 27)., Results: Among 107 patients of UC (mean age-29 ± 10.6 years; males = 56%, median disease duration-48 [interquartile range: 24-84] months, left sided/pancolitis = 84%), 27% (n = 29) had NAFLD. Patients with body mass index (BMI) > 23 kg/m 2 had higher proportion of NAFLD than with normal or low BMI (54.7% [23/42] vs 10% [5/50] vs 6.7% [1/15]). Patients with NAFLD had high BMI ( P < 0.001), waist circumference, and fat mass ( P < 0.001) but similar fat-free mass ( P  = 0.798) compared to patients without NAFLD. There was no difference in immunosuppressant and cumulative steroid exposure between two groups. Dietary parameters including daily energy, protein, fat, and carbohydrate intake were similar between the two groups. On multivariate analysis, high BMI was found to be predictive and low socioeconomic status as a protective factor of NAFLD. On long-term follow-up of three years, there was no difference in steroid, or biologic requirement, disease-related hospitalization, or composite of all three outcomes between two groups., Conclusion: The prevalence of NAFLD was found in nearly a quarter of patients of UC and was affected by metabolic parameters rather than disease activity., (© 2023 Indian National Association for Study of the Liver. Published by Elsevier B.V. All rights reserved.)

This research article explores the distinct TADF efficiency of three donor-acceptor based regio-isomers: DPAOCN ( ortho -isomer), DPAMCN ( meta -isomer), and DPAPCN ( para -isomer). DPAPCN exhibits maximum TADF efficiency in both solution and solid-state with an impressive reverse inter-system crossing (RISC) rate of ∼10 6 s -1 ; the underlying cause being the minimum singlet-triplet splitting energy or Δ E ST and maximum SOC (spin-orbit coupling) between the S 1 & T 1 states. Apart from TADF, differences in crystal packing of the regio-isomers result in intriguing bulk phase properties. DPAOCN, with its non-centrosymmetric P 2 1 2 1 2 1 space group and substantial crystal void volume, exhibits reversible tri-color mechanochromic luminescence behavior, while the meta and para isomers, due to their centrosymmetric packing and diminished crystal void volume, remain inert to mechanical pressure. Expanding the horizon of possibilities, the non-centrosymmetric nature of ortho -isomer further renders it an excellent SHG material, with a χ (2) value of 0.19 pm V -1 at 1220 nm and a laser-induced damage threshold (LIDT) value of 13.27 GW cm -2 . Overall, a comprehensive investigation into the regio-isomers has been carried out, encompassing their TADF, SHG, and mechanochromic luminescent properties., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Electronic and steric effects are known to greatly influence the structure, characteristics and reactivity of organic compounds. A typical π bond is weakened by oxidation (corresponding to the removal of electrons from bonding orbitals), by reduction (through addition of electrons to antibonding orbitals) and by unpairing of the bonding electrons, such as in the triplet state. Here we describe tetrafluorenofulvalene (TFF), a twisted, open-shell alkene for which these general rules do not hold. Through the synthesis, experimental characterization and computational analysis of its charged species spanning seven redox states, the central alkene bond in TFF is shown to become substantially stronger in the tri- and tetraanion, generated by chemical reduction. Furthermore, although its triplet state contains a weaker alkene bond than the singlet, in the quintet state its bond order increases substantially, yielding a flatter structure. This behaviour originates from the doubly bifurcated topology of the underlying spin system and can be rationalized by the balancing effects of benzenoid aromaticity and spin pairing., (© 2023. The Author(s).)

The electronic properties of a hexaphyrin was fine-tuned via core-modification leading to the formation of a Hückel aromatic 30π hexaphyrin which incorporates two pyrrole and four furan rings in the π-conjugated pathway. This Hückel aromatic hexaphyrin modified its conformation upon two-electron ring oxidation either with triflic acid or Meerwein salt [Et 3 O] + [SbCl 6 ] - to yield 28π Möbius aromatic dication species. Reversible aromatic transition was established by spectroscopic techniques and further supported by quantum chemical calculations., (© 2023 Wiley-VCH GmbH.)

The use of smartphone-based applications as a telenutrition tool could redefine the nutritional management of IBD. We developed and validated a digital health platform in the form of a smartphone application for the nutritional assessment of IBD patients. Our team of gastroenterologists and dieticians at the All-India Institute of Medical Sciences, New Delhi developed a smartphone application titled IBD NutriCare, which was made available in both Android and iOS interfaces in English and seven other Indian languages. The application includes >650 Indian recipes and provides subjective global assessment and IBD clinical activity scores in a patient-friendly manner. The utility of the smartphone app was validated in comparison with the traditional 24-h dietary recall method. A total of 49 IBD patients were enrolled in the study. The mean difference in energy intake between the two dietary assessment methods was −4.776 kJ (95% LOA, range −417.916−408.365 kJ). A total of 94% of patients found the smartphone application convenient and acceptable in comparison to the recall method for dietary assessment. Bland−Altman plots showed a good level of agreement for nutrients and food groups between the two methods. Telenutrition in the form of a smartphone application helps in real-time tracking of dietary details of IBD patients, thus making appropriate interventions and large-scale data acquisition feasible.

Herein, we describe the synthesis, structural diversity and diradicaloid characteristics of 38π core-modified aromatic expanded isophlorins with eight heterocyclic rings. The diradicaloid character of expanded isophlorinoid macrocycles was engineered by systematic structural modification. Depending on the nature of the link between the heteroatoms, they adopt planar and non-planar conformations. This large structural variation with a significant difference in the extent of aromaticity is correlated with the magnitude of their respective diradical character.

Background: Surge of SARS CoV-2 infections ascribed to omicron variant began in December 2021 in New Delhi. We determined the infection and reinfection density in a cohort of health care workers (HCWs) along with vaccine effectiveness (VE) against symptomatic infection within omicron transmission period (considered from December 01, 2021 to February 25, 2022., Methods: This is an observational study from the All India Institute of Medical Sciences, New Delhi. Data were collected telephonically. Person-time at risk was counted from November 30, 2021 till date of infection/ reinfection, or date of interview. Comparison of clinical features and severity was done with previous pandemic periods. VE was estimated using test-negative case-control design [matched pairs (for age and sex)]. Vaccination status was compared and adjusted odds ratios (OR) were computed by conditional logistic regression. VE was estimated as (1-adjusted OR)X100-., Findings: 11474 HCWs participated in this study. The mean age was 36⋅2 (±10⋅7) years. Complete vaccination with two doses were reported by 9522 (83%) HCWs [8394 (88%) Covaxin and 1072 Covishield (11%)]. The incidence density of all infections and reinfection during the omicron transmission period was 34⋅8 [95% Confidence Interval (CI): 33⋅5-36⋅2] and 45⋅6 [95% CI: 42⋅9-48⋅5] per 10000 person days respectively. The infection was milder as compared to previous periods. VE was 52⋅5% (95% CI: 3⋅9-76⋅5, p  = 0⋅036) for those who were tested within 14-60 days of receiving second dose and beyond this period (61-180 days), modest effect was observed., Interpretation: Almost one-fifth of HCWs were infected with SARS CoV-2 during omicron transmission period, with predominant mild spectrum of COVID-19 disease. Waning effects of vaccine protection were noted with increase in time intervals since vaccination., Funding: None., Competing Interests: We declare no competing interests., (© 2022 The Authors.)

With the onset of the SARS-CoV-2 pandemic, Kawasaki Disease (KD) has come to the fore with its many atypical manifestations. Atypical clinical neurological, ophthalmological, musculoskeletal, gastrointestinal and pulmonary manifestations in a febrile child with raised markers should prompt the clinician to think of Kawasaki disease. Peripheral gangrene is a rare atypical manifestation of KD reported in infancy. We present a three-and-a-half-year-old boy with extensive gangrene all four limbs and face along with purpura fulminans. He was successfully treated with two doses of intravenous immunoglobulin (IVIG) and infliximab, with no residual gangrene. This case highlights that very severe forms of Kawasaki disease require IVIG, pulse steroids as well as infliximab for adequate control and complete resolution of the disease.

Vector control strategies are among the most effective measures to combat mosquito-borne diseases, such as malaria. These strategies work by altering the mosquito age structure through increased mortality of the older female mosquitoes that transmit pathogens. However, methods to monitor changes to mosquito age structure are currently inadequate for programmatic implementation. Female mosquitoes generally mate a single time soon after emergence and draw down spermatozoa reserves with each oviposition cycle. Here, we demonstrate that measuring spermatozoa quantity in female Anopheles mosquitoes is an effective approach to assess mosquito age. Using multiplexed qPCR targeted at male spermatozoa, we show that Y-linked genes in female mosquitoes are exclusively found in the spermatheca, the organ that houses spermatozoa, and the quantity of these gene sequences significantly declines with age. The method can accurately identify mosquitoes more than 10 days old and thus old enough to potentially transmit pathogens harbored in the salivary glands during blood feeding. Furthermore, mosquito populations that differ by 10% in daily survivorship have a high likelihood of being distinguished using modest sample sizes, making this approach scalable for assessing the efficacy of vector intervention control programs., (© 2022. The Author(s).)

Current quantification methods of Escherichia coli ( E. coli ) contamination in water samples involve long incubation, laboratory equipment and facilities, or complex processes that require specialized training for accurate operation and interpretation. To address these limitations, we have developed a microfluidic device and portable instrument prototypes capable of performing a rapid and highly sensitive bacteriophage-based assay to detect E. coli cells with detection limit comparable to traditional methods in a fraction of the time. The microfluidic device combines membrane filtration and selective enrichment using T7-NanoLuc-CBM, a genetically engineered bacteriophage, to identify 4.1 E. coli CFU in 100 mL of drinking water within 5.5 hours. The microfluidic device was designed and tested to process up to 100 mL of real-world drinking water samples with turbidities below 10 NTU. Prototypes of custom instrumentation, compatible with our valveless microfluidic device and capable of performing all of the assay's units of operation with minimal user intervention, demonstrated similar assay performance to that obtained on the benchtop assay. This research is the first step towards a faster, portable, and semi-automated, phage-based microfluidic platform for improved in-field water quality monitoring in low-resource settings.

The next-generation flexible wearable electronics are among the most rapidly growing industries due to their extended use in everyday applications resulting in an increased demand for cheaper, safer, and flexible energy storage devices. This study aims to investigate and enhance the overall performance of a Zn-MnO 2 alkaline battery and make it suitable for safe and flexible wearable applications. To achieve high cyclability and performance of the cathode, issues of low active-material availability for redox reactions and inactive-phase formations are overcome by fabricating a binder-free hierarchical (increased surface area) additives (enabled reversible compound formation) based MnO 2 cathode. Furthermore, zinc/stainless steel composite anode (to reduce anode shape changes) and calcium hydroxide coated polymer electrolyte (to stop zincate ion transfer) are used to improve cyclability. By assembling the above mentioned layers, excellent rate capabilities, high-capacity utilization (487 mAh g -1 ), long cycling stabilities (1000 cycles with 70% retention), and high energy density (400 Wh kg -1 ) are achieved. Moreover, bending, hammering, puncturing, and lighting up an light emitting diode are conducted (under flat, bent, and cut) to demonstrate the cells' safety, flexibility, and robustness. The successful findings in this study can chart new pathways to the development of safe, flexible, and cost-effective next-generation energy storage sources for wearables., (© 2022 Wiley-VCH GmbH.)

Two dithienothiophene based 28π antiaromatic macrocycles with benzene and azulene units and their effects on local and global (anti)aromaticity have been described. Experimental and computational studies confirmed the presence of weak paratropic and strong diatropic ring current effects in neutral and dicationic states, respectively.

Importance: A surge of COVID-19 occurred from March to June 2021, in New Delhi, India, linked to the B.1.617.2 (Delta) variant of SARS-CoV-2. COVID-19 vaccines were rolled out for health care workers (HCWs) starting in January 2021., Objective: To assess the incidence density of reinfection among a cohort of HCWs and estimate the effectiveness of the inactivated whole virion vaccine BBV152 against reinfection., Design, Setting, and Participants: This was a retrospective cohort study among HCWs working at a tertiary care center in New Delhi, India., Exposures: Vaccination with 0, 1, or 2 doses of BBV152., Main Outcomes and Measures: The HCWs were categorized as fully vaccinated (with 2 doses and ≥15 days after the second dose), partially vaccinated (with 1 dose or 2 doses with <15 days after the second dose), or unvaccinated. The incidence density of COVID-19 reinfection per 100 person-years was computed, and events from March 3, 2020, to June 18, 2021, were included for analysis. Unadjusted and adjusted hazard ratios (HRs) were estimated using a Cox proportional hazards model. Estimated vaccine effectiveness (1 - adjusted HR) was reported., Results: Among 15 244 HCWs who participated in the study, 4978 (32.7%) were diagnosed with COVID-19. The mean (SD) age was 36.6 (10.3) years, and 55.0% were male. The reinfection incidence density was 7.26 (95% CI: 6.09-8.66) per 100 person-years (124 HCWs [2.5%], total person follow-up period of 1696 person-years as time at risk). Fully vaccinated HCWs had lower risk of reinfection (HR, 0.14 [95% CI, 0.08-0.23]), symptomatic reinfection (HR, 0.13 [95% CI, 0.07-0.24]), and asymptomatic reinfection (HR, 0.16 [95% CI, 0.05-0.53]) compared with unvaccinated HCWs. Accordingly, among the 3 vaccine categories, reinfection was observed in 60 of 472 (12.7%) of unvaccinated (incidence density, 18.05 per 100 person-years; 95% CI, 14.02-23.25), 39 of 356 (11.0%) of partially vaccinated (incidence density 15.62 per 100 person-years; 95% CI, 11.42-21.38), and 17 of 1089 (1.6%) fully vaccinated (incidence density 2.18 per 100 person-years; 95% CI, 1.35-3.51) HCWs. The estimated effectiveness of BBV152 against reinfection was 86% (95% CI, 77%-92%); symptomatic reinfection, 87% (95% CI, 76%-93%); and asymptomatic reinfection, 84% (95% CI, 47%-95%) among fully vaccinated HCWs. Partial vaccination was not associated with reduced risk of reinfection., Conclusions and Relevance: These findings suggest that BBV152 was associated with protection against both symptomatic and asymptomatic reinfection in HCWs after a complete vaccination schedule, when the predominant circulating variant was B.1.617.2.

Since 2009, the mechanical engineering (ME) scholarship-science technology engineering and mathematics (S-STEM) Program at the University of Maryland Baltimore County (UMBC) has provided financial support and program activities to ME undergraduate students aiming at improving their retention and graduation rates. The objective of this study is to identify program activities that were most effective to help students for improvements. Current ME S-STEM scholars were asked to complete a survey that measures their scientific efficacy, engineering identity, expectations, integration, and sense of belonging, as well as how program activities impact their attitudes and perceptions. Analyses of 36 collected surveys showed that scholars reported high levels of engineering identity, expectations, and sense of belonging. However, further improvements were needed to help students in achieving scientific efficacy and academic integration into the program. Results demonstrated that pro-active mentoring was the most effective method contributing to positive attitudes and perceptions. The implemented S-STEM research-related activities and internship were viewed favorably by the scholars in helping them establish their scientific efficacy and engineering identity, and understand their expectations and goals. Community building activities were considered helpful for them to integrate into campus life and improve their sense of belonging to the campus and program. Scholars identified mentoring, research related activities, internships, and social interaction with faculty and their peers as important factors for their retention and graduation. Although the sample size was small in the study, we believe that the cost-effective activities identified could be adopted by other institutions to further improve students' retention and graduation rates in engineering programs., (Copyright © 2021 by ASME.)

Multifunctional organic luminogens exhibiting simultaneous aggregation induced emission (AIE), room-temperature phosphorescence (RTP), and mechanochromism have recently attracted considerable attention owing to their potential applications in optoelectronics and bioimaging. However, a comprehensive correlation among these three distinguished properties is yet to be unveiled, which will help to decipher defined methodologies to design future generation multifunctional organic materials. Herein, we have demonstrated a route to obtain a multifunctional organic luminogen, starting from an ACQphore (TPANDI) by simple structural engineering. We have shown that a slight reduction in length of the planar acceptor moieties can effectively inhibit the undesirable π-π stacking interaction between molecules in the condensed state and thereby cause an ACQ to AIE type transformation from TPANDI to TPANMI and TPAPMI. Both TPANMI and TPAPMI exhibit RTP properties (even in ambient condition) because of the presence of a reasonably low singlet-triplet energy gap (Δ E ST ). In our study, these two luminogens were found to be mechano-inactive. Interestingly, an insertion of cyano-ethylene group and benzene linker in between the triphenylamine and phthalimide moieties introduced another luminogen TPACNPMI, which can simultaneously exhibit AIE, RTP, and mechanochromic properties.

Bacterial CRISPR systems provide acquired immunity against invading nucleic acids by activating RNA-programmable RNases and DNases. Cas13a and Cas12a enzymes bound to CRISPR RNA (crRNA) recognize specific nucleic acid targets, initiating cleavage of the targets as well as non-target ( trans ) nucleic acids. Here, we examine the kinetics of single-turnover target and multi-turnover trans -nuclease activities of both enzymes. High-turnover, non-specific Cas13a trans -RNase activity is coupled to rapid binding of target RNA. By contrast, low-turnover Cas12a trans -nuclease activity is coupled to relatively slow cleavage of target DNA, selective for DNA over RNA, indifferent to base identity, and preferential for single-stranded substrates. Combining multiple crRNA increases detection sensitivity of targets, an approach we use to quantify pathogen DNA in samples from patients suspected of Buruli ulcer disease. Results reveal that these enzymes are kinetically adapted to play distinct roles in bacterial adaptive immunity and show how kinetic analysis can be applied to CRISPR-based diagnostics., Competing Interests: Some methods described herein have patents pending on which E.A.N., P.Y.J.L., G.J.N., J.A.D, A.-L.M.L.N, and D.M are inventors. The Regents of the University of California have patents issued and pending for CRISPR technologies on which G.J.K. and J.A.D. are inventors. J.A.D. is a cofounder of Caribou Biosciences, Editas Medicine, Scribe Therapeutics, and Mammoth Biosciences; is a scientific advisory board member of Caribou Biosciences, Intellia Therapeutics, eFFECTOR Therapeutics, Scribe Therapeutics, Mammoth Biosciences, Synthego, and Inari; is a director at Johnson & Johnson; and has research projects sponsored by 10.13039/100005614Biogen and 10.13039/100004319Pfizer., (© 2021 The Authors.)

Oral swab analysis (OSA) has been shown to detect Mycobacterium tuberculosis (MTB) DNA in patients with pulmonary tuberculosis (TB). In previous analyses, qPCR testing of swab samples collected from tongue dorsa was up to 93% sensitive relative to sputum GeneXpert, when 2 swabs per patient were tested. The present study modified sample collection methods to increase sample biomass and characterized the viability of bacilli present in tongue swabs. A qPCR targeting conserved bacterial ribosomal rRNA gene (rDNA) sequences was used to quantify bacterial biomass in samples. There was no detectable reduction in total bacterial rDNA signal over the course of 10 rapidly repeated tongue samplings, indicating that swabs collect only a small portion of the biomass available for testing. Copan FLOQSwabs collected ~2-fold more biomass than Puritan PurFlock swabs, the best brand used previously (p = 0.006). FLOQSwabs were therefore evaluated in patients with possible TB in Uganda. A FLOQSwab was collected from each patient upon enrollment (Day 1) and, in a subset of sputum GeneXpert Ultra-positive patients, a second swab was collected on the following day (Day 2). Swabs were tested for MTB DNA by manual IS6110-targeted qPCR. Relative to sputum GeneXpert Ultra, single-swab sensitivity was 88% (44/50) on Day 1 and 94.4% (17/18) on Day 2. Specificity was 79.2% (42/53). Among an expanded sample of Ugandan patients, 62% (87/141) had colony-forming bacilli in their tongue dorsum swab samples. These findings will help guide further development of this promising TB screening method., Competing Interests: At the time of study design and evaluation SB, CMB, DB, and AS received salary support from the Global Good Fund. At the time of study design and evaluation GH, KPN, AML, CO, DM, and KJM were employed by Intellectual Ventures Laboratory. There are no patents, products in development or marketed products to declare. This does not alter our adherence to PLOS ONE policies on sharing data and materials. A number of the authors are currently affiliated with GH Labs and Roche Molecular Systems, but were not affiliated with them during the study, and these companies had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Steric hindrance induced by thiophene molecules in predesigned precursors favors the exclusive formation of a three dimensional (3D) π-conjugated cage and quasi-cage like molecules instead of a porphyrinoid macrocycle. Herein we report the synthesis of a tetrapod 3D fully π-conjugated molecular cage using a simple acid catalysed reaction. The X-Ray crystallography analysis confirmed the tetrapod cage structure and intermediates, which resemble three-fourths or half of the cage structures.

Purpose: To measure sensitivity and specificity of vision screeners in identifying children with visual impairment and positive signs and symptoms and assess association of effectiveness with individual characteristics of screeners and type of schools screened., Methods: A total of 1096 children from age 5 to 15 years of age were screened. A total of 396 children were screened from a municipal school, 200 children from a government-aided school, and 500 children from a private school were screened. Four persons with basic 12th standard science qualification willing to be a part of school eye health program were selected who carried out screening in school children after receiving appropriate training., Results: The two vision screeners who had a background of conducting community eye health programs and worked in eye hospital had 100% sensitivity and specificity for presenting visual acuity, squint detection, and blurring. The screening by these screeners was done in private and semi-private schools, respectively. The other two screeners with no such background conducting screening in government schools had 60% and 75% sensitivity in detecting presenting visual acuity, respectively., Conclusion: People with a background of organizing community eye health programs and those working in eye hospitals are the best candidates for being trained as new cadre of vision screeners with best results being obtained in private and government-aided schools., Competing Interests: None

East Coast fever (ECF) is an often fatal, economically important cattle disease that predominantly affects eastern, central, and southern Africa. ECF is controlled through vaccination by means of simultaneous injection of oxytetracycline and cryogenically preserved stabilate containing live, disease-causing parasites. Storage and transportation of the stabilate requires liquid nitrogen, a commodity that is commonly unreliable in low-resource settings. Here we show that storage of conventionally prepared stabilate at -80 °C for up to 30 days does not significantly affect its ability to infect cultured peripheral blood mononucleated cells or live cattle, suggesting an alternative cold chain that maintains these temperatures could be used to effectively manage ECF., (Copyright © 2020. Published by Elsevier B.V.)

The objectives of this study were to evaluate the current status of exposure to bio-engineering research in community college (CC) students and University of Maryland Baltimore County (UMBC) students, and to estimate relationships between research activities sponsored by the Mechanical Engineering (ME) S-STEM Scholarship Program and improvement in student enrollment/diversification, retention rates, and graduation rates. The analysis drew on data from ME undergraduate academic records at UMBC from 2008 to 2019. A survey was designed to assess the research exposure of CC and UMBC students and their evaluation of the research components included in recruitment and curriculum activities. Results show that exposure to research measured by attending a research seminar was low for the participants, around 37% for CC students and 21% for ME students at UMBC. The survey results indicate the positive impact of the scholarship programs at UMBC on the research exposure and research experience. The impact is more evident in students who originally transferred from a CC. The large increase in recruited female and CC students over the past 10 years indicated that the research-related activities of the ME S-STEM program played an instrumental role in those increases. Because of the research-related activities, the ME S-STEM program achieved retention and graduation rates higher than those in the ME undergraduate program (89% versus 60% for the 6 year graduation rate), as well a higher percentage of students enrolled in graduate school (30% versus 10%). We conclude that there is still a need to implement research-related activities in the ME undergraduate program, starting with student recruitment and continuing through the academic program. Results suggest that there is a positive impact of ME S-STEM research activities on student diversification, retention rates, and percentage of our graduates who are pursuing graduate degree., (Copyright © 2020 by ASME.)

Purpose: To evaluate skills transfer in short-term phacoemulsification surgery training program by International Council of Ophthalmology -Ophthalmology Surgical Competency Assessment Rubrics (ICO-OSCAR) and assessment of efficacy of ICO-OSCAR for objective evaluation of skills transfer., Methods: Prospective analysis of total 1300 phacoemulsification surgeries performed by 52 phacoemulsification trainees divided into three groups based on the previous cataract surgical experience. The pretraining and posttraining average mean scores, independent completion rates (ICR), and complication rates among the groups were calculated and statistically analyzed., Results: Mean age of trainees (N = 52) was found to be 30.98 ± 2.44 years. Mean pretraining scores in the three groups were 42.8 ± 0.2, 46.6 ± 0.2, and 50.1 ± 0.1, respectively, and posttraining mean scores of the groups were 88.4 ± 0.2, 92.3 ± 0.2, and 94.3 ± 0.2, respectively, out of 100. Pretraining independent completion rates (%) were 22.1 ± 2.5, 52.1 ± 2.5, and 62 ± 3.7, respectively, and posttraining independent completion rates (%) were 74.8 ± 3, 79.7 ± 3, and 90.6 ± 3.5, respectively. Learning curves of all groups were noted to be steep. Rate of complications was within acceptable range, overall complication rate being 12.30% (160 out of 1300 cases). OSCAR scores correlate significantly (P = 0.0004) with ICR., Conclusion: Steep learning curve for phacoemulsification can be made easy by the use of standardized rubrics, proper techniques, expert guidance, and adequate surgical volume. The previous SICS experience provides smoother learning curve. ICO-OSCAR is indicator of proficiency and provides constructive feedback and objective evaluation immediately after surgery as well as at the end of training. Short-term (1 Month) training program is an effective, efficient, and safe approach to progressively master phacoemulsification skills for trainees with SICS experience., Competing Interests: None

Competing Interests: There are no conflicts of interest.

Artificial insemination (AI) is widely used in livestock industries to breed for desirable characteristics and increase yields. The standard practice of storing and transporting bovine semen uses liquid nitrogen (LN), a scarce commodity in many regions of the world. This study explored the feasibility of using dry ice, a more readily available alternative. We developed equipment that dispenses dry ice from widely available liquid carbon dioxide (LCO 2 ) tanks into an easily transportable device. In vivo fertility results with a dry ice cold chain showed no statistical difference to the conventional LN method. In vitro bovine semen analyses also showed that storage under these conditions minimally affects characteristics associated with fertility. A dry ice cold chain system could leverage the global availability of LCO 2 to expand the reach of AI and other cold storage applications of biological materials in low-resource settings., (© The Author(s) 2020. Published by Oxford University Press on behalf of the American Society of Animal Science.)

Background: Literature remains constrained to case reports with respect to epilepsy-associated takotsubo cardiomyopathy (TC) or stress-induced cardiomyopathy and its impact on in-hospital outcomes remains largely obscure., Methods: The National Inpatient Sample databases (2010-2014) were queried to identify and compare baseline characteristics and outcomes in adult hospitalizations for epilepsy with and without secondary TC using ICD-9-CM codes and propensity-matching. Primary outcomes were the frequency of TC, ensuing all-cause mortality, and complications. Secondary outcome was healthcare resource utilization., Results: Of 981,571 epilepsy-related hospitalizations, 854 (0.1%, 1 in 1000) admissions (unspecified, 49.1%; grand mal/status epilepticus, 28.1% and generalized convulsive 11.7%) revealed associated in-hospital TC. Of the propensity-matched cohorts of epilepsy (TC = 793; mean 61.1 ± 15.0 yrs. & 82.4% females vs. non-TC = 795; mean 60.7 ± 14.2 yrs. & 84.2% females), the TC group consisted more often white (83.7% vs. 78.0%, p < 0.02) patients with higher cardiovascular risk factors. The all-cause inpatient mortality (3.7% vs <11; p = 0.002), arrhythmia (22.7% vs. 18.7%, p = 0.05), cardiac arrest (3.9% vs <11; p = 0.001), cardiogenic shock (3.2% vs <11, p < 0.001), stroke (3.5% vs 1.9%, p = 0.04), venous thromboembolism (4.4% vs. 1.9%, p = 0.004), and respiratory failure (29.4% vs. 14.8%, p < 0.001) were significantly higher in the TC cohort. The mean LOS (6.3 ± 5.6 vs. 5.1 ± 7.1 days), hospital charges ($77,908 vs. $45,881), transfers to other facilities (3.8% vs. 3.2%), and need of home healthcare (19.4% vs. 9.9%) were higher in the TC group (p < 0.001)., Conclusion: In this nationwide population-based study, 1 in every 1000 epilepsy-related hospitalizations was associated with secondary TC which resulted in poor inpatient outcomes and higher healthcare resource utilization., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Peritoneal dialysis solution (PDS) dilates peritoneal microvessels predominantly by the activation of the endothelial nitric oxide (NO) pathway. We made an incidental observation of decreased PDS-induced, NO-dependent peritoneal microvascular vasoreactivity in elderly rats naïve to PDS exposure. We hypothesized that this subordinate NO-mediated peritoneal microvascular vasoreactivity is caused by increased oxidative stress in the aged endothelium, which compromises NO bioavailability in the elderly, and that peritoneal microvascular vasoreactivity can be improved by the supplementation of antioxidant glycine to PDS. Methods: We studied PDS-mediated vasoreactivity of four intestinal visceral arterioles of different orders by in vivo intravital microscopy in weaned, adult, and elderly rats to (i) confirm subordinate vasoreactivity to PDS in elderly rats; (ii) restore vasoreactivity by glycine supplementation; and (iii) establish age as an independent risk factor for endothelial cell dysfunction. Results: In a crossover series, peritoneal microvascular vasoreactivity to PDS exposure was remarkably decreased in elderly rats. This subordinate vasoreactivity was completely restored by the supplementation of glycine to PDS. In a separate series, we assessed in situ endothelial cell function in weaned and adult rats using the cumulative acetylcholine concentration-response curves. Unlike the adults, the weaned rats demonstrated remarkable sensitivity and reactivity to cumulative acetylcholine concentrations, suggesting the dependency of endothelial cell function on age. Conclusion: Aging is an independent risk factor for peritoneal microvascular endothelial cell dysfunction. Endothelial function in the elderly can be recovered by reinforcing the bioavailability of endothelial-derived NO through glycine. Dietary glycine supplementation is a potential therapeutic strategy to decrease the burden of oxidative stress on the aged endothelium., (© 2019 Zakaria, Joseph, Hamidi, Zeeshan, Algamal, Sartaj, Althani, Fadl, Madan, licensee HBKU Press.)

Pathogen cell-free DNA (pcfDNA) in blood and urine is an attractive biomarker; however, the impact of preanalytical factors is not well understood. Blood and urine samples from healthy donors spiked with cfDNA from Mycobacterium tuberculosis , Salmonella enterica , Aspergillus fumigatus , and Epstein-Barr virus (EBV) and samples from tuberculosis patients were used to evaluate the impact of blood collection tube, urine preservative, processing delay, processing method, freezing and thawing, and sample volume on pcfDNA. The PCR cycle threshold ( C T ) was used to measure amplifiable cfDNA. In spiked samples, the median C T values for M. tuberculosis , S. enterica , and EBV cfDNA were significantly lower in blood collected in K 2 EDTA tubes than those in Streck and PAXgene blood collection tubes, and they were was significantly lower in urine preserved with EDTA (EDTA-urine) than in urine preserved with Streck reagent (Streck-urine). Blood and urine samples from TB patients preserved with K 2 EDTA and Tris-EDTA, respectively, showed significantly lower median M. tuberculosis C T values than with the Streck blood collection tube and Streck urine preservative. Processing delay increased the median pathogen C T values for Streck and PAXgene but not K 2 EDTA blood samples and for urine preserved with Streck reagent but not EDTA. Double-spin compared with single-spin plasma separation increased the median pathogen C T regardless of blood collection tube. No differences were observed between whole urine and supernatant and between fresh and thawed plasma and urine after 24 weeks at -80°C. Larger plasma and urine volumes in contrived and patient samples showed a significantly lower median M. tuberculosis C T These findings suggest that large-volume single-spin K 2 EDTA-plasma and EDTA-whole urine with up to a 24-h processing delay may optimize pcfDNA detection., (Copyright © 2019 Murugesan et al.)

Core-modified hexaphyrin and octaphyrin display four- and six-electron reversible redox reactions respectively, to alternate between aromatic and anti-aromatic states of a given macrocycle. We have identified and isolated a hexaphyrin in three discrete states with 26π, 28π and 30π electrons that are inter-convertible with each other. Its higher congener, octaphyrin, can exist as four discrete species with 34π, 36π, 38π and 40π electrons. A difference of two-electrons between each stable redox state is reflected by the significant variation in their electronic and structural properties as characterized in both solution and solid states. The observed redox inter-conversions were achieved by a combination of both proton coupled electron-transfer (PCET) and electron-transfer (ET) processes respectively.

Thermoelectric generators (TEGs) fabricated using additive manufacturing methods are attractive because they offer the advantages of scalability, lower cost, and potentially higher power density than conventional TEGs. Additive manufacturing of TEGs requires active thermoelectric particles to be dispersed in a polymer binder to synthesize printable slurries, and printed films to be subsequently subjected to a long and high temperature curing to enhance their thermoelectic properties. A large amount of polymer binder present in composite films results in a sizable loss in the electrical conductivity. In addition, a long and high-temperature film curing results is a slow and energy intensive fabrication process. In this work, we demonstrate the feasibility of using a small amount (≤10 -3 wt ratio) of novel nanofiber cellulose (NFC) as a binder to provide sufficient adhesion strength to hold the TE particles together in the composite films. We also demonstrate a pressure induced densification process to enhance the thermoelectic properties of printed composite films. This novel approach has the potential to fundamentally transform the manufacting method for printing TEGs by eliminating the need of long-duration and high-temperature curing. A higher applied pressure leads to a compact packing and densification of films resulting in an improvement in the electrical conductivity. The highest power factor achieved for best performing p-type thermoelectric-NFC composite film subjected to pressure induced densification is 611 μW/m-K 2 .

We investigated alternatives to whole blood for blood feeding of mosquitoes with a focus on improved stability and compatibility with mass rearing programs. In contrast to whole blood, an artificial blood diet of ATP-supplemented plasma was effective in maintaining mosquito populations and was compatible with storage for extended periods refrigerated, frozen, and as a lyophilized powder. The plasma ATP diet supported rearing of both Anopheles and Aedes mosquitoes. It was also effective in rearing Wolbachia-infected Aedes mosquitoes, suggesting compatibility with vector control efforts.

Air-stable and soluble tetrabutylammonium fluoride (TBAF) is demonstrated as an efficient n-type dopant for the conjugated polymer ClBDPPV. Electron transfer from F - anions to the π-electron-deficient ClBDPPV through anion-π electronic interactions is strongly corroborated by the combined results of electron spin resonance, UV-vis-NIR, and ultraviolet photoelectron spectroscopy. Doping of ClBDPPV with 25 mol% TBAF boosts electrical conductivity to up to 0.62 S cm -1 , among the highest conductivities that have been reported for solution-processed n-type conjugated polymers, with a thermoelectric power factor of 0.63 µW m -1 K -2 in air. Importantly, the Seebeck coefficient agrees with recently published correlations to conductivity. Moreover, the F - -doped ClBDPPV shows significant air stability, maintaining the conductivity of over 0.1 S cm -1 in a thick film after exposure to air for one week, to the best of our knowledge the first report of an air-stable solution-processable n-doped conductive polymer with this level of conductivity. The result shows that using solution-processable small-anion salts such as TBAF as an n-dopant of organic conjugated polymers possessing lower LUMO (lowest unoccupied molecular orbital), less than -4.2 eV) can open new opportunities toward high-performance air-stable solution-processable n-type thermoelectric (TE) conjugated polymers., (© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Ever since the discovery of the porphyrin ring in "pigments of life", such as chlorophyll and hemoglobin, it has become a prime synthetic target for optoelectronic properties and in the design of metal complexes. During one such early expedition on the synthesis of porphyrin, Woodward proposed that condensing pyrrole with an aldehyde under acidic conditions yields the "precursor" porphyrinogen macrocycle. Its four-electron oxidation leads to the "transitory" 20π isophlorin, which undergoes subsequent two-electron oxidation to form the 18π "porphyrin". Due to its fleeting lifetime, it has been a synthetic challenge to stabilize the tetrapyrrolic isophlorin. This macrocycle symbolizes the antiaromatic character of a porphyrin-like macrocycle. In addition, the pyrrole NH also plays a key role in the proton-coupled, two-electron oxidation of isophlorin to the aromatic porphyrin. However, a major aspect of its unstable nature was attributed to its antiaromatic character, which is understood to destabilize the macrocycle upon conjugation. Antiaromaticity in general has not gained significant attention mainly due to the lack of stable 4nπ systems. In this regard, a stable isophlorin and its derivatives provide a glimmering hope to peek into the world of antiaromatic systems. This review will focus on the attempted synthesis of antiaromatic isophlorin ever since its conception. Based on recent synthetic advances, the chemistry of isophlorins can be expected to blossom into expanded derivatives of this antiaromatic macrocycle. Along with the synthetic details, the structural, electronic, and redox properties of isophlorin and its expanded derivatives will be elaborated.

Purpose: The purpose of this study was to evaluate functional and anatomical outcomes of posterior iris-claw intraocular lens (IOL) implant for correction of aphakia in eyes with inadequate capsular support., Materials and Methods: Prospective case series of 108 aphakic eyes with inadequate capsular support which underwent posterior iris-claw IOL with a 6-month follow-up period was conducted. The cases belonged to two clinical settings: elective secondary implantation and those with intraoperative posterior dislocation of cataractous lens or IOL. Main outcome measures were visual acuity, anterior chamber reaction, stability of IOL, endothelial cell count, intraocular pressure (IOP), and cystoid macular edema (CME)., Results: The mean best-corrected visual acuity was LogMAR 0.25. None had chronic anterior chamber inflammation. The mean difference in central endothelial counts before surgery and 1 month after surgery was 104.21 cell/mm2 (4.92%). There was no statistically significant difference in central endothelial cell count at 1 and 6 months (P = 0.91) and also in the central macular thickness at preoperative and after 6 months suggestive of CME (P = 0.078). Three eyes had raised IOP which were managed with neodymium-doped yttrium aluminum garnet laser peripheral iridotomy. There were no IOL dislocations or other adverse events in our series., Conclusion: Posterior chamber iris-claw lenses are a good option in eyes with inadequate posterior capsular support. Chronic inflammation, poor lens stability, or significant central endothelial cell loss was not observed during the 6-month follow-up period.

Artificial insemination of dairy cattle is a common practice in the developing world that can improve farmer incomes and food security. Maintaining the fertilizing potential of frozen semen as it is manipulated, transported and stored is crucial to the success of this process. Here we describe simple technological improvements to protect semen from inadvertent thermal fluctuations that occur when users mishandle semen using standard equipment. We show that when frozen semen is mishandled, characteristics of semen biology associated with fertility are negatively affected. We describe several design modifications and results from thermal performance tests of several improved prototypes. Finally, we compare semen that has been mishandled in standard and improved equipment. The data suggest that our canister improvements can better maintain characteristics of semen biology that correlate with fertility when it is mishandled.

Autotaxin (ATX) is an enzyme discovered in the conditioned medium of cultured melanoma cells and identified as a protein that strongly stimulates motility. This unique ectonucleotide pyrophosphatase and phosphodiesterase facilitates the removal of a choline headgroup from lysophosphatidylcholine (LPC) to yield lysophosphatidic acid (LPA), which is a potent lipid stimulator of tumorigenesis. Thus, ATX has received renewed attention because it has a prominent role in malignant progression with significant translational potential. Specifically, we sought to develop active site-targeted irreversible inhibitors as anti-cancer agents. Herein we describe the synthesis and biological activity of an LPC-mimetic electrophilic affinity label that targets the active site of ATX, which has a critical threonine residue that acts as a nucleophile in the lysophospholipase D reaction to liberate choline. We synthesized a set of quaternary ammonium derivative-containing vinyl sulfone analogs of LPC that function as irreversible inhibitors of ATX and inactivate the enzyme. The analogs were tested in cell viability assays using multiple cancer cell lines. The IC50 values ranged from 6.74 to 0.39 μM, consistent with a Ki of 3.50 μM for inhibition of ATX by the C16H33 vinyl sulfone analog CVS-16 (10b). A phenyl vinyl sulfone control compound, PVS-16, lacking the choline-like quaternary ammonium mimicking head group moiety, had little effect on cell viability and did not inhibit ATX. Most importantly, CVS-16 (10b) significantly inhibited melanoma progression in an in vivo tumor model by preventing angiogenesis. Taken together, this suggests that CVS-16 (10b) is a potent and irreversible ATX inhibitor with significant biological activity both in vitro and in vivo., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

This work presents a novel method to synthesize p-type composite thermoelectric materials to print scalable thermoelectric generator (TEG) devices in a cost-effective way. A maximum ZT of 0.2 was achieved for mechanically alloyed (MA) p-type Bi0.5Sb1.5Te3 (8 wt % extra Te additive)-epoxy composite films cured at 250 °C. A 50% increase in Seebeck coefficient as a result of adding 8 wt % extra Te in stoichiometric Bi0.5Sb1.5Te3 contributed to the increase in ZT. To demonstrate cost-effective and scalable manufacturing, we fabricated a sixty element thermoelectric generator prototype with 5.0 mm × 600 μm × 120 μm printed dimensions on a custom designed polyimide substrate with thick metal contacts. The prototype TEG device produced a power output of 20.5 μW at 0.15 mA and 130 mV for a temperature difference of 20 K resulting in a device areal power density of 152 μW/cm(2). This power is sufficient for low power applications such as wireless sensor network (WSN) devices.

Autotaxin (ATX), an autocrine motility factor that is highly upregulated in metastatic cancer, is a lysophospholipase D enzyme that produces the lipid second messenger lysophosphatidic acid (LPA) from lysophosphatidylcholine (LPC). Dysregulation of the lysolipid signaling pathway is central to the pathophysiology of numerous cancers, idiopathic pulmonary fibrosis, rheumatoid arthritis, and other inflammatory diseases. Consequently, the ATX/LPA pathway has emerged as an important source of biomarkers and therapeutic targets. Herein we describe development and validation of a fluorogenic analog of LPC (AR-2) that enables visualization of ATX activity in vivo. AR-2 exhibits minimal fluorescence until it is activated by ATX, which substantially increases fluorescence in the near-infrared (NIR) region, the optimal spectral window for in vivo imaging. In mice with orthotopic ATX-expressing breast cancer tumors, ATX activated AR-2 fluorescence. Administration of AR-2 to tumor-bearing mice showed high fluorescence in the tumor and low fluorescence in most healthy tissues with tumor fluorescence correlated with ATX levels. Pretreatment of mice with an ATX inhibitor selectively decreased fluorescence in the tumor. Together these data suggest that fluorescence directly correlates with ATX activity and its tissue expression. The data show that AR-2 is a non-invasive and selective tool that enables visualization and quantitation of ATX-expressing tumors and monitoring ATX activity in vivo.

Rheumatoid arthritis (RA) is a destructive arthropathy with systemic manifestations, characterized by chronic synovial inflammation. Under the influence of the pro-inflammatory milieu synovial fibroblasts (SFs), the main effector cells in disease pathogenesis become activated and hyperplastic while releasing a number of signals that include pro-inflammatory factors and tissue remodeling enzymes. Activated RA SFs in mouse or human arthritic joints express significant quantities of autotaxin (ATX), a lysophospholipase D responsible for the majority of lysophosphatidic acid (LPA) production in the serum and inflamed sites. Conditional genetic ablation of ATX from SFs resulted in attenuation of disease symptoms in animal models, an effect attributed to diminished LPA signaling in the synovium, shown to activate SF effector functions. Here we show that administration of 1-bromo-3(S)-hydroxy-4-(palmitoyloxy)butyl-phosphonate (BrP-LPA), a metabolically stabilized analog of LPA and a dual function inhibitor of ATX and pan-antagonist of LPA receptors, attenuates collagen induced arthritis (CIA) development, thus validating the ATX/LPA axis as a novel therapeutic target in RA.