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Background: A sensitive and specific non-sputum-based test would be groundbreaking for the diagnosis of childhood tuberculosis. We assessed side by side the diagnostic accuracy of the urine-based lipoarabinomannan assays Fujifilm SILVAMP TB LAM (FujiLAM) and Alere Determine TB LAM Ag (AlereLAM) for detection of childhood tuberculosis., Methods: In this cross-sectional study, we tested urine samples from children younger than 15 years with presumed pulmonary tuberculosis. Children were consecutively recruited from four dedicated outpatient childhood tuberculosis clinics in The Gambia, Mali, Nigeria, and Tanzania. Biobanked urine samples were thawed and tested using FujiLAM and AlereLAM assays. We measured diagnostic performance against a microbiological reference standard (confirmed tuberculosis) and a composite reference standard (confirmed and unconfirmed tuberculosis). Sensitivity and specificity were estimated with bivariate random-effects meta-analyses., Findings: Between July 1, 2017, and Dec 1, 2018, we obtained and stored urine samples from 415 children. 63 (15%) children had confirmed tuberculosis, 113 (27%) had unconfirmed tuberculosis, and 239 (58%) were unlikely to have tuberculosis. 61 children were HIV-positive (prevalence 15%). Using the microbiological reference standard, the sensitivity of FujiLAM was 64·9% (95% CI 43·7-85·2; positive in 40 of 63 confirmed samples) and the sensitivity of AlereLAM was 30·7% (8·6-61·6; 19 of 63). The specificity of FujiLAM was 83·8% (95% CI 76·5-89·4; negative in 297 of 352 unconfirmed and unlikely samples) and the specificity of AlereLAM was 87·8% (79·0-93·7; 312 of 352). Against the composite reference standard, both assays had decreased sensitivity; the sensitivity of FujiLAM was 32·9% (95% CI 24·6-41·9; positive in 58 of 176 confirmed and unconfirmed samples) and the sensitivity of AlereLAM was 20·2% (12·3-29·4; 36 of 176). The specificity of FujiLAM was 83·3% (95% CI 71·8-91·7; negative in 202 of 239 unlikely samples) and the specificity of AlereLAM was 90·0% (81·6-95·6; 216 of 239)., Interpretation: By comparison with AlereLAM, FujiLAM showed higher sensitivity and similar specificity. FujiLAM could potentially add value to the rapid diagnosis of tuberculosis in children., Funding: German Federal Ministry of Education and Research, the Global Health Innovative Technology Fund, the UK Research and Innovation Global Challenges Research Fund, and the UK Medical Research Council., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Hepatocellular carcinoma (HCC) remains a leading cause of cancer death worldwide, and despite recent immunotherapeutic advances there remains a need for improved diagnostic, prognostic, and therapeutic tools. UL-16 binding protein 1 (ULBP1) is a ligand of the activatory receptor Natural Killer cell Group 2 receptor D (NKG2D) and is found as a cell-surface protein on some malignant cells including on human hepatocellular carcinomas. We aimed to explore the biological and clinical significance of NKG2D ligands in the circulation of patients with HCC. We measured ULBP1 in the serum of two retrospective cohorts of patients with HCC from the PROLIFICA cohort in The Gambia ( n = 43) and from a tertiary care setting in the UK ( n = 72) by sandwich ELISA. Exosome isolation by size exclusion was used to compare ULBP1 concentration in exosomes and as free protein. Survival analysis was performed and multiple linear regression and Poisson regression were used to assess the independent effect of ULBP1 concentration. ULBP1 was raised in both cohorts with HCC regardless of the underlying liver disease, and was not associated with markers of cirrhosis such as platelet count or serum albumin. ULBP1 was present predominantly as free protein rather than bound to exosomes. Serum ULBP1 > 2000 pg/ml was associated with a significantly reduced survival in both cohorts (hazard ratios in Gambian and UK cohorts 2.37 and 2.1, respectively). The effect remained significant after adjustment for BCLC staging ( p = 0.03). These data suggest that ULBP1 merits further investigation as a prognostic marker in HCC in diverse settings and should also be explored as a therapeutic target., (Copyright © 2020 Easom, Marks, Jobe, Gillmore, Meyer, Maini and Njie.)

Three-dimensional (3D) printing is capable of transforming intricate digital models into tangible objects, allowing geoscientists to replicate the geometry of 3D pore networks of sedimentary rocks. We provide a refined method for building scalable pore-network models ("proxies") using stereolithography 3D printing that can be used in repeated flow experiments (e.g., core flooding, permeametry, porosimetry). Typically, this workflow involves two steps, model design and 3D printing. In this study, we explore how the addition of post-processing and validation can reduce uncertainty in the 3D-printed proxy accuracy (difference of proxy geometry from the digital model). Post-processing is a multi-step cleaning of porous proxies involving pressurized ethanol flushing and oven drying. Proxies are validated by: (1) helium porosimetry and (2) digital measurements of porosity from thin-section images of 3D-printed proxies. 3D printer resolution was determined by measuring the smallest open channel in 3D-printed "gap test" wafers. This resolution (400 µm) was insufficient to build porosity of Fontainebleau sandstone (∼13%) from computed tomography data at the sample's natural scale, so proxies were printed at 15-, 23-, and 30-fold magnifications to validate the workflow. Helium porosities of the 3D-printed proxies differed from digital calculations by up to 7% points. Results improved after pressurized flushing with ethanol (e.g., porosity difference reduced to ∼1% point), though uncertainties remain regarding the nature of sub-micron "artifact" pores imparted by the 3D printing process. This study shows the benefits of including post-processing and validation in any workflow to produce porous rock proxies., (© 2017, National Ground Water Association.)

Background: Replication of intervention research is reported infrequently, limiting what we know about external validity and generalisability. The Smoke Free Homes Program, a minimal intervention, increased home smoking bans by United Way 2-1-1 callers in randomised controlled trials in Atlanta, Georgia and North Carolina., Objective: Test the programme's generalisability-external validity in a different context., Methods: A randomised controlled trial (n=508) of English-speaking callers from smoking-discordant households (≥1 smoker and ≥1 non-smoker). 2-1-1 Texas/United Way HELPLINE call specialists serving the Texas Gulf Coast recruited callers and delivered three mailings and one coaching call, supported by an online tracking system. Data collectors, blind to study assignment, conducted telephone interviews 3 and 6 months postbaseline., Results: At 3 months, more intervention households reported a smoke-free home (46.6% vs 25.4%, p<0.0001; growth model intent-to-treat OR=1.48, 95% CI 1.241 to 1.772, p<0.0001). At 6 months, self-reported full bans were 62.9% for intervention participants and 38.4% for controls (OR=2.19). Texas trial participants were predominantly women (83%), single-smoker households (76%) and African-American (65%); half had incomes ≤US$10 000/year (50%). Texas recruitment was <50% of the other sites. Fewer callers reported having a smoker in the household. Almost twice the callers with a household smoker declined interest in the programme/study., Conclusions: Our findings in a region with lower smoking rates and more diverse callers, including English-speaking Latinos, support programme generalisability and convey evidence of external validity. Our recruitment experience indicates that site-specific adjustments might improve recruitment efficiency and reach., Trial Registration Number: NCT02097914, Results., (Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/.)

A method combining on-line immunoextraction microcolumns with high-performance affinity chromatography (HPAC) was developed and tested for use in examining drug-protein interactions with normal or modified proteins. Normal human serum albumin (HSA) and glycated HSA were used as model proteins for this work. High-performance immunoextraction microcolumns with sizes of 1.0-2.0 cm × 2.1mm i.d. and containing anti-HSA polyclonal antibodies were developed and tested for their ability to bind normal HSA or glycated HSA. These microcolumns were able to extract up to 82-93% for either type of protein at 0.05-0.10 mL/min and had a binding capacity of 0.34-0.42 nmol HSA for a 1.0 cm × 2.1mm i.d. microcolumn. The immunoextraction microcolumns and their adsorbed proteins were tested for use in various approaches for drug binding studies. Frontal analysis was used with the adsorbed HSA/glycated HSA to measure the overall affinities of these proteins for the drugs warfarin and gliclazide, giving comparable values to those obtained previously using similar protein preparations that had been covalently immobilized within HPAC columns. Zonal elution competition studies with gliclazide were next performed to examine the specific interactions of this drug at Sudlow sites I and II of the adsorbed proteins. These results were also comparable to those noted in prior work with covalently immobilized samples of normal HSA or glycated HSA. These experiments indicated that drug-protein binding studies can be carried out by using on-line immunoextraction microcolumns with HPAC. The same method could be used in the future with clinical samples and other drugs or proteins of interest in pharmaceutical studies or biomedical research., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Background: Research is needed to understand parental factors influencing human papillomavirus (HPV) vaccination, particularly in groups with a higher burden of cervical cancer., Purpose: To determine correlates of HPV vaccination among a sample of low-income parents of age-eligible daughters (aged 9-17 years) who called the 2-1-1 Helpline. Secondary analyses describe potential differences in HPV vaccination correlates by Hispanic and black parent groups, in particular., Methods: This 2009 cross-sectional feasibility survey of cancer prevention needs was conducted in Houston at the 2-1-1 Texas/United Way Helpline. In 2012, to examine the association between parental psychosocial, cognitive, and decisional factors and HPV vaccination uptake (one or two doses), bivariate and multivariable logistic regression analyses were conducted for minority parents and for Hispanic and black parent groups, separately., Results: Lower rates of HPV vaccination uptake were reported among minority daughters of 2-1-1 callers (29% overall) compared with national and Texas rates. In final adjusted analysis, factors positively associated with HPV vaccination uptake included being offered the vaccination by a doctor or nurse, belief that the vaccine would prevent cervical cancer, and Hispanic ethnicity. Secondary analyses detected differences in factors associated with vaccination in Hispanic and black groups., Conclusions: Findings indicate low levels of vaccination among 2-1-1 callers. Increased understanding of determinants of HPV vaccination in low-income minority groups can guide interventions to increase coverage. Because 2-1-1 informational and referral services networks reach populations considered medically underserved, 2-1-1 can serve as a community hub for informing development of and implementing approaches aimed at hard-to-reach groups., (Copyright © 2012 American Journal of Preventive Medicine. Published by Elsevier Inc. All rights reserved.)

Innovative interventions are needed to connect underserved populations to cancer control services. With data from Missouri, North Carolina, Texas, and Washington this study a) estimated the cancer control needs of callers to 2-1-1, an information and referral system used by underserved populations, b) compared rates of need with state and national data, and c) examined receptiveness to needed referrals. From October 2009 to March 2010 callers' (N=1,408) cancer control needs were assessed in six areas: breast, cervical, and colorectal cancer screening, HPV vaccination, smoking, and smoke-free homes using Behavioral Risk Factor Surveillance System (BRFSS) survey items. Standardized estimates were compared with state and national rates. Nearly 70% of the sample had at least one cancer control need. Needs were greater for 2-1-1 callers than for state and national rates, and callers were receptive to referrals. 2-1-1 could be a key partner in efforts to reduce cancer disparities.

We have measured the x-ray power and energy radiated by a tungsten-wire-array z pinch as a function of the peak pinch current and the width of the anode-cathode gap at the base of the pinch. The measurements were performed at 13- and 19-MA currents and 1-, 2-, 3-, and 4-mm gaps. The wire material, number of wires, wire-array diameter, wire-array length, wire-array-electrode design, normalized-pinch-current time history, implosion time, and diagnostic package were held constant for the experiments. To keep the implosion time constant, the mass of the array was increased as I2 (i.e., the diameter of each wire was increased as I), where I is the peak pinch current. At 19 MA, the mass of the 300-wire 20-mm-diam 10-mm-length array was 5.9 mg. For the configuration studied, we find that to eliminate the effects of gap closure on the radiated energy, the width of the gap must be increased approximately as I. For shots unaffected by gap closure, we find that the peak radiated x-ray power P(r) proportional to I1.24+/-0.18, the total radiated x-ray energy E(r) proportional to I1.73+/-0.18, the x-ray-power rise time tau(r) proportional to I0.39+/-0.34, and the x-ray-power pulse width tau(w) proportional to demonstrate that the internal energy and radiative opacity of the pinch are not responsible for the observed subquadratic power scaling. Heuristic wire-ablation arguments suggest that quadratic power scaling will be achieved if the implosion time tau(i) is scaled as I(-1/3). The measured 1sigma shot-to-shot fluctuations in P(r), E(r), tau(r), tau(w), and tau(i) are approximately 12%, 9%, 26%, 9%, and 2%, respectively, assuming that the fluctuations are independent of I. These variations are for one-half of the pinch. If the half observed radiates in a manner that is statistically independent of the other half, the variations are a factor of 2(1/2) less for the entire pinch. We calculate the effect that shot-to-shot fluctuations of a single pinch would have on the shot-success probability of the double-pinch inertial-confinement-fusion driver proposed by Hammer et al. [Phys. Plasmas 6, 2129 (1999)]. We find that on a given shot, the probability that two independent pinches would radiate the same peak power to within a factor of 1+/-alpha (where 0< or =alpha<<1) is equal to erf(alpha/2sigma), where sigma is the 1sigma fractional variation of the peak power radiated by a single pinch. Assuming alpha must be < or =7% to achieve adequate odd-Legendre-mode radiation symmetry for thermonuclear-fusion experiments, sigma must be <3% for the shot-success probability to be > or =90%. The observed (12/2(1/2))%=8.5% fluctuation in P(r) would provide adequate symmetry on 44% of the shots. We propose that three-dimensional radiative-magnetohydrodynamic simulations be performed to quantify the sensitivity of the x-ray emission to various initial conditions, and to determine whether an imploding z pinch is a spatiotemporal chaotic system.

Absorption spectroscopy measurements of the time-dependent heating of thin foils exposed to intense z-pinch radiation sources are presented. These measurements and their analysis provide valuable benchmarks for, and insights into, the radiative heating of matter by x-ray sources. Z-pinch radiation sources with peak powers of up to 160 TW radiatively heated thin plastic-tamped aluminum foils to temperatures approximately 60 eV. The foils were located in open slots at the boundary of z-pinch hohlraums surrounding the pinch. Time-resolved Kalpha satellite absorption spectroscopy was used to measure the evolution of the Al ionization distribution, using a geometry in which the pinch served as the backlighter. The time-dependent pinch radius and x-ray power were monitored using framing camera, x-ray diode array, and bolometer measurements. A three-dimensional view factor code, within which one-dimensional (1D) radiation-hydrodynamics calculations were performed for each surface element in the view factor grid, was used to compute the incident and reemitted radiation flux distribution throughout the hohlraum and across the foil surface. Simulated absorption spectra were then generated by postprocessing radiation-hydrodynamics results for the foil heating using a 1D collisional-radiative code. Our simulated results were found to be in good general agreement with experimental x-ray spectra, indicating that the spectral measurements are consistent with independent measurements of the pinch power. We also discuss the sensitivity of our results to the spectrum of the radiation field incident on the foil, and the role of nonlocal thermodynamic equilibrium atomic kinetics in affecting the spectra.

The JC virus (JCV) is a ubiquitous human polyomavirus that frequently resides in the kidneys of healthy individuals and is excreted in the urine of a large percentage of the population. Geographic-specific JCV variants, isolated from urine and from brain of progressive multifocal leukoencephalopathy (PML) patients, have been grouped into seven distinct genotypes based on whole genome analysis and by individual polymorphic nucleotides (typing sites) in the VP1 coding region. Mutations in the archetypal regulatory region, sometimes consisting of deletions and/or duplications, are also useful taxonomic characters for further characterizing and subdividing genotypes. Investigation of JCV variation in Papua New Guinea (PNG) revealed three distinct variants called PNG- 1, PNG-2, and PNG-3. These variants exhibited consistent coding region and regulatory region mutations. Evolutionary analysis of 32 complete JCV genomes including six new viral genomes from the western Pacific suggests that the new PNG JCV variants are closely associated with the broad group of Type 2 strains of JCV found throughout Asia, forming a monophyletic group with the Northeast Asian strains (Type 2A). Within the Type 2 clade, however, the PNG JCV variants cluster as two distinct groups and are therefore described here as new JCV genotypes designated Type 2E and Type 8.

Detection of borreliacidal antibodies is an accurate serodiagnostic test for confirmation of Lyme disease in humans. In this study, 13 pathogen-free beagles, 12 to 26 weeks old, were infected with Borrelia burgdorferi by tick challenge. Dogs were monitored for clinical signs and symptoms of Lyme disease along with borreliacidal antibody production against B. burgdorferi sensu stricto isolates 297 and 50772. Ten (77%) dogs developed lameness in one or more legs within 210 days after attachment of Ixodes scapularis ticks. Eight (80%) of the lame animals had concurrent fever of > or =38 degrees C. Spirochetes were also recovered from the skin and joints of 12 (92%) dogs, but rarely from other organs. Borreliacidal antibodies against B. burgdorferi isolate 297 were detected in only four (31%) dogs, and the levels of killing antibodies remained low for the duration of the infection. In contrast, borreliacidal antibodies against B. burgdorferi isolate 50772 were detected in 13 (100%) dogs within 21 days of infection. Furthermore, the borreliacidal antibody levels correlated with the severity of B. burgdorferi infection. Detection of borreliacidal antibodies, especially against B. burgdorferi isolate 50772, is also a reliable serodiagnostic test for detection of Lyme disease in dogs.

A recombinant Lyme borreliosis vaccine consisting of outer surface protein A (OspA) is commercially available for vaccination of humans against infection with Borrelia burgdorferi. Vaccination with OspA induces an antibody response that makes serologic interpretation of infection with B. burgdorferi difficult, especially by screening tests based on whole-cell preparations of B. burgdorferi. We show that an enzyme-linked immunosorbent assay with B. burgdorferi sensu stricto 50772, which lacks the plasmid encoding OspA and OspB, or a full-length recombinant OspC protein can identify patients infected with B. burgdorferi. We found that 69 and 65% of serum samples from patients with case-defined early Lyme borreliosis had anti-B. burgdorferi sensu stricto 50772 and anti-OspC reactivities, respectively. In addition, little or no reactivity was detected with sera obtained from individuals vaccinated with OspA. Unfortunately, 51 and 33% of sera from healthy patients and sera from patients with other illnesses were also reactive against B. burgdorferi sensu stricto 50772 and OspC, respectively. Although these assays can discriminate B. burgdorferi infection from vaccination with OspA, their lack of specificity highlights the necessity for confirming equivocal or positive reactivities with more specific serodiagnostic tests.

The borreliacidal-antibody test has been used for the serological detection and confirmation of Lyme borreliosis. However, the presence of antimicrobial agents in serum can confound the accurate detection of borreliacidal antibodies. In this study, we developed a Bacillus subtilis agar diffusion bioassay to detect small concentrations of antimicrobial agents in serum. We also used XAD-16, a nonionic polymeric resin, to adsorb and remove high concentrations of amoxicillin, cefotaxime, ceftriaxone, cefuroxime, doxycycline, and erythromycin without significantly affecting even small concentrations of immunoglobulin M (IgM) or IgG borreliacidal antibodies. High concentrations of penicillin could also be removed by adding 1 U of penicillinase without significantly influencing the levels of borreliacidal antibodies. These simple procedures greatly enhance the clinical utility of the borreliacidal-antibody test.

Early Lyme borreliosis sera with significant titers of anti-outer surface protein C (OspC) borreliacidal antibodies were identified. Human anti-OspC borreliacidal antibodies could be either IgM or IgG. Significant concentrations of borreliacidal activity were detected after vaccination of mice with OspC. Detection of anti-OspC borreliacidal activity was dependent on surface expression of OspC by Borrelia burgdorferi isolate 50772. The ability of OspC to induce borreliacidal antibodies in vivo and after vaccination offers another possible explanation for the ability of vaccination with OspC to protect against infection with B. burgdorferi. Furthermore, detection of anti-OspC borreliacidal antibodies, especially IgM antibodies, in early Lyme borreliosis sera provides additional evidence that borreliacidal antibody detection may be useful for the serodiagnosis of early Lyme borreliosis.

The serodiagnosis of early Lyme disease has been plagued with problems of sensitivity and specificity. We found that the flow-cytometric borreliacidal-antibody test had a sensitivity of 72% for the detection of patients with early Lyme disease. By contrast, the sensitivity of the enzyme immunofluorescence assay was 28%. The enhanced sensitivity of the borreliacidal-antibody test was due to the use of Borrelia burgdorferi 50772, which lacks OspA and OspB. When B. burgdorferi 297, which expresses both OspA and OspB, was used, the sensitivity of the borreliacidal-antibody test was 15%. Our results also showed that the borreliacidal-antibody test was specific. No borreliacidal activity was detected in normal sera or in sera from patients with mononucleosis, rheumatoid factor, or syphilis. These results demonstrate that the flow-cytometric borreliacidal-antibody test may be the laboratory "gold standard" for the serodiagnosis of Lyme disease.

Background: Borreliacidal antibodies can be detected in serum samples from patients with early or late Lyme disease symptoms. When these serum samples are incubated with Borrelia burgdorferi and complement, spirochetes are rapidly killed. Detection of these antibodies can be used as a serodiagnostic test., Methods: Individual serum samples containing IgM or IgG borreliacidal antibodies were Used to develop a method for detection using flow cytometry. An additional 10 case-defined Lyme disease serum samples and 10 normal serum samples were used to confirm appropriate flow cytometric parameters. To determine specificity, 157 normal serum samples and 104 potential cross-reactive serum samples were tested for borreliacidal activity and antibodies to B burgdorferi using indirect fluorescent antibody or enzyme immunoassay., Results: Flow cytometry can be used to detect borreliacidal activity within 16 to 24 hours after incubation of B burgdorferi organisms. Lyme disease serum, and complement. Significant borreliacidal activity was detected in all Lyme disease serum samples. The percentages of positive normal serum samples were comparable (6% to 10%) using all three assays. In addition, the indirect fluorescent antibody and enzyme immunoassay identified 41 (39%) and 47 (45%) potential cross-reactive serum samples as positive, respectively. In contrast, significant borreliacidal activity was not detected in any potential cross-reactive serum samples., Conclusion: Detection of borreliacidal antibody, unlike indirect fluorescent antibody and enzyme immunoassay, is an accurate, highly specific serodiagnostic test for detection of Lyme disease.

Although identified less than 20 years ago, Lyme disease has proved to be the most common tick-borne disease in the United States: some 10,000 cases were reported in 1992. In some cases the disease may be transitory and of little consequence but in others it may become chronic and severely disabling. Accurate diagnosis is, therefore, of great importance but, as this article shows, laboratory testing techniques still need improvement.

A total of 150 extracted maxillary and mandibular molars separated into six groups of 25 teeth each were prepared to the size of a 30 endodontic file. Numbers 2, 3, and 4 Peeso drills were used to produce a post cavity preparation in the root canals of these teeth. After use of each size of Peeso drill, the thickness of the canal walls was measured at two levels: the middle level, 4 mm from the canal orifice; and the apical level, 7 mm from the canal orifice. Mean thickness of all walls was determined for each tooth and drill size. The incidence of root perforations and strippings was recorded also. We found that distal roots of mandibular molars and palatal roots of maxillary molars are best suited for post cavity preparation. Most perforations and strippings occurred on the distal walls of mesial roots of mandibular molars or on the proximal walls of the buccal roots of maxillary molars.