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Membrane protein leucine–rich repeat containing 15 (LRRC15) is known to be expressed in several solid tumors including osteosarcoma. ABBV-085, an antibody–drug conjugate against LRRC15, conjugated to monomethyl auristatin E (MMAE), was studied in osteosarcoma patient-derived xenografts (PDXs) by the Pediatric Preclinical Testing Consortium (PPTC). LRRC15 expression data were obtained from PPTC RNA-sequencing data for the PDX models. The TARGET database was mined for LRRC15 expression in human osteosarcoma. Protein expression was confirmed via IHC in three PDX models. Seven osteosarcoma PDX models (OS1, OS9, OS33, OS34, OS42, OS55, and OS60) with varying LRRC15 gene expression were studied. ABBV-085 was administered at 3 mg/kg (OS33), 6 mg/kg (all seven PDXs), and 12 mg/kg (OS60) weekly for 4 consecutive weeks via intraperitoneal injection. Control cohorts included vehicle and an isotype MMAE-linked antibody. Tumor volumes and responses were reported using PPTC statistical analysis. OS1, OS33, OS42, OS55, and OS60 had high LRRC15 expression while OS9 and OS34 had low LRRC15 expression. ABBV-085 inhibited tumor growth in six of seven PDX models as compared with vehicle control and significantly improved event-free survival in five of seven models as compared with isotype controls. Two models showed maintained complete responses while all others showed progressive disease. Response correlated with LRRC15 expression. ABBV-085’s antitumor activity against osteosarcoma PDX suggests LRRC15 may be a rational target for pursuing clinical trials in patients with this disease.

Model Name, passage#, and time from implantation to testing

The need for assembled existing and new toxicity data has accelerated as the amount of chemicals introduced into commerce continues to grow and regulatory mandates require safety assessments for a greater number of chemicals. To address this evolving need, the ECOTOXicology Knowledgebase (ECOTOX) was developed starting in the 1980s and is currently the world's largest compilation of curated ecotoxicity data, providing support for assessments of chemical safety and ecological research through systematic and transparent literature review procedures. The recently released version of ECOTOX (Ver 5, www.epa.gov/ecotox) provides single-chemical ecotoxicity data for over 12,000 chemicals and ecological species with over one million test results from over 50,000 references. Presented is an overview of ECOTOX, detailing the literature review and data curation processes within the context of current systematic review practices and discussing how recent updates improve the accessibility and reusability of data to support the assessment, management, and research of environmental chemicals. Relevant and acceptable toxicity results are identified from studies in the scientific literature, with pertinent methodological details and results extracted following well-established controlled vocabularies and newly extracted toxicity data added quarterly to the public website. Release of ECOTOX, Ver 5, included an entirely redesigned user interface with enhanced data queries and retrieval options, visualizations to aid in data exploration, customizable outputs for export and use in external applications, and interoperability with chemical and toxicity databases and tools. This is a reliable source of curated ecological toxicity data for chemical assessments and research and continues to evolve with accessible and transparent state-of-the-art practices in literature data curation and increased interoperability to other relevant resources. Environ Toxicol Chem 2022;41:1520-1539. © 2022 SETAC. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

Conotruncal heart defects (CTDs) are heart malformations that affect the cardiac outflow tract and typically cause significant morbidity and mortality. Evidence from epidemiological studies suggests that maternal folate intake is associated with a reduced risk of heart defects, including CTD. However, it is unclear if folate-related gene variants and maternal folate intake have an interactive effect on the risk of CTDs. In this study, we performed targeted sequencing of folate-related genes on DNA from 436 case families with CTDs who are enrolled in the National Birth Defects Prevention Study and then tested for common and rare variants associated with CTD. We identified risk alleles in maternal MTHFS (ORmeta = 1.34; 95% CI 1.07 to 1.67), maternal NOS2 (ORmeta = 1.34; 95% CI 1.05 to 1.72), fetal MTHFS (ORmeta = 1.35; 95% CI 1.09 to 1.66), and fetal TCN2 (ORmeta = 1.38; 95% CI 1.12 to 1.70) that are associated with an increased risk of CTD among cases without folic acid supplementation. We detected putative de novo mutations in genes from the folate, homocysteine, and transsulfuration pathways and identified a significant association between rare variants in MGST1 and CTD risk. Results suggest that periconceptional folic acid supplementation is associated with decreased risk of CTD among individuals with susceptible genotypes.

Copanlisib is a pan-class I phosphoinositide 3-kinase (PI3K) inhibitor, with activity against all four PI3K class I isoforms (PI3Kα, PI3Kβ, PI3Kγ, and PI3Kδ). Whole-genome and RNA sequencing data have revealed several PI3K aberrations in osteosarcoma tumor samples. The in vivo anticancer effects of copanlisib were assessed in a panel of six osteosarcoma models. Copanlisib induced prolonged event-free survival in five of six osteosarcoma models; however, all models demonstrated progressive disease suggesting minimal activity. While copanlisib did not result in tumor regression, more data are needed to fully explore the role of the PI3K pathway in the pathogenesis of osteosarcoma.

Coupled harmonic oscillators are ubiquitous in physics and play a prominent role in quantum science. They are a cornerstone of quantum mechanics and quantum field theory, where second quantization relies on harmonic oscillator operators to create and annihilate particles. Descriptions of quantum tunneling, beamsplitters, coupled potential wells, ``hopping terms'', decoherence, and many other phenomena rely on coupled harmonic oscillators. However, the ability to couple separate quantum harmonic oscillators directly, with coupling rates that substantially exceed decoherence rates, has remained elusive. Here, we realize high-fidelity coherent exchange of single motional quanta between harmonic oscillators -- in this case, spectrally separated harmonic modes of motion of a trapped ion crystal -- where the timing, strength, and phase of the coupling are controlled through an oscillating electric potential with suitable spatial variation. The coupling rate can be made much larger than the decoherence rates, enabling demonstrations of high-fidelity quantum state transfer, entanglement of motional modes, and Hong-Ou-Mandel-type interference. We also project a harmonic oscillator into its ground state by measurement and preserve that state during repetitions of the projective measurement, an important prerequisite for non-destructive syndrome measurement in continuous-variable quantum error correction. Controllable coupling between harmonic oscillators has potential applications in quantum information processing with continuous variables, quantum simulation, and precision measurements. It can also enable cooling and quantum logic spectroscopy involving motional modes of trapped ions that are not directly accessible.

Background The PhageDx™Cronobacter Assay is based on the infection of Cronobacter spp. by specific bacteriophages and expression of a luciferase reporter gene. Results are generated in as little as 18.5 h for powdered infant formula (PIF). Objective An AOAC Performance Tested MethodsSM (PTM) study was conducted to validate the PhageDx Cronobacter Assay for the detection of Cronobacter in 10, 100, and 300 g milk- and soy-based PIF test portions. Method The performance of the PhageDx method was compared to the ISO 22964:2006/2017 Microbiology of the Food Chain—Horizontal Method for the Detection of Cronobacter spp. and the U.S. Food and Drug Administration (FDA) Bacteriological Analytical Manual (BAM) Chapter 29 Cronobacter: 2012. Inclusivity/exclusivity, product consistency and stability, and robustness testing also were conducted. Results There was no significant difference between the 10, 100, or 300 g test portions for the milk and soy PIF matrixes between the PhageDx Cronobacter Assay, the ISO 22964:2006/2017, and the FDA BAM Chapter 29 Cronobacter: 2012 methods. The reporter bacteriophages were specific for Cronobacter and infected 75 strains in inclusivity testing. They did not infect 35 non-Cronobacter bacteria in exclusivity testing. Robustness testing showed that the method performed well with specific deviations from the standard protocol. Consistency and stability testing demonstrated that the recombinant phage gave consistent results across three production lots and was stable when stored under appropriate conditions for at least 3 months. Conclusions Work in the submitting and independent laboratories demonstrated that the PhageDx Cronobacter Assay meets the qualifications for PTM status. Highlights The PhageDx Cronobacter Assay is a rapid, simple, and specific test that has shown equivalence to both the FDA BAM and ISO reference methods for detecting Cronobacter spp. in PIF.

Background The PhageDx™ Listeria Assay is a simple, specific, and sensitive assay based on the infection of Listeria spp. by selected bacteriophages and the resultant expression of a luciferase reporter gene. Results are generated in as little as 24.5 h for stainless steel and ceramic environmental surfaces. Objective An AOAC Performance Tested MethodsSM (PTM) study was conducted to validate the PhageDx Listeria Assay for the detection of Listeria on stainless steel and ceramic surfaces. Method The performance of the PhageDx method was compared to that of the U.S. Food and Drug Administration (FDA) Bacterial Analytical Manual (BAM) Ch. 10. Inclusivity/exclusivity, product consistency and stability, and robustness testing also were conducted. Results Inclusivity testing demonstrated that the reporter bacteriophages were specific for Listeria ssp. and detected 58/61 Listeria strains tested, including all 34 L. monocytogenes strains. The reporter bacteriophage also was shown to not detect 46/47 non-Listeria bacteria in exclusivity testing. Robustness testing showed that the method performed well with specific deviations from the standard protocol. Consistency and stability testing demonstrated that the recombinant phage gave consistent results across three production lots and was stable when stored under appropriate conditions for at least 6 months. Matrix studies on stainless steel and ceramic surfaces showed that there was no significant difference between the PhageDx Listeria Assay and the FDA/BAM Chapter 10 reference method. Conclusions and Highlights The validation study demonstrates that the PhageDx Listeria Assay is an effective method for the detection of Listeria spp. on stainless steel and ceramic environmental surfaces and meets the qualifications for AOAC PTM status.

Salmonella spp. is one of the most common foodborne pathogens worldwide, therefore, its control is highly relevant for the food industry. Phages of the Felixounavirus genus have the characteristic that one phage can infect a large number of different Salmonella serovars and, thus, are proposed as an alternative to antimicrobials in food production. Here, we describe two new members of the Felixounavirus genus named vB_Si_35FD and vB_Si_DR94, which can infect Salmonella Infantis. These new members were isolated and sequenced, and a subsequent comparative genomic analysis was conducted including 23 publicly available genomes of Felixounaviruses that infect Salmonella. The genomes of vB_Si_35FD and vB_Si_DR94 are 85,818 and 85,730 bp large and contain 129 and 125 coding sequences, respectively. The genomes did not show genes associated with virulence or antimicrobial resistance, which could be useful for candidates to use as biocontrol agents. Comparative genomics revealed that closely related Felixounavirus are found in distinct geographical locations and that this genus has a conserved genomic structure despite its worldwide distribution. Our study revealed a highly conserved structure of the phage genomes, and the two newly described phages could represent promising biocontrol candidates against Salmonella spp. from a genomic viewpoint.

Background ThePhageDx™ E. coli O157:H7 Assay originally required a 5 h enrichment period and a 10 mL test sample for 25 g ground beef. The proposed method modifications seek to include beef trim (375 g) matrix and a new method procedure for 25 g ground beef comprised of 6–7 h enrichment and 1 mL test sample. Objective To validate the method modifications for PhageDx E. coli O157:H7 Assay to include beef trim (375 g) matrix and modify enrichment time and test sample size for 25 g ground beef. Method For both matrixes, pre-warmed tryptic soya broth (TSB; 42 ± 1°C) was added in a 3:1 (media:sample size) ratio, blended, and enriched for 9–10 h (beef trim, 375 g) or 6 h (ground beef, 25 g) at 42 ± 1°C. One milliliter samples were transferred to a microfuge tube, centrifuged, and the supernatant removed. The pellet was resuspended in 0.2 mL of TSB and phage reagent was added. Samples were incubated for 2 h at 37 ± 1°C. After infection, the sample was centrifuged, and 150 µl of the supernatant was transferred to a 96-well plate. Then, lysis buffer and luciferase substrate were added and the sample read on a luminometer to determine the presence or absence of E. coli O157:H7. Results No significant differences were found between the PhageDx E. coli O157:H7 Assay method modifications and the U.S. Department of Agriculture Food Safety and Inspection Service (FSIS) Microbiology Laboratory Guidebook (MLG) reference method. Conclusions The independent study demonstrated that the PhageDx E. coli O157:H7 Assay method modifications meet the qualifications for Performance Tested Method SM status. Highlights The PhageDx E. coli O157:H7 Assay is a rapid detection method capable of detecting E. coli O157:H7 in 25 g ground beef and 375 g beef trim samples in as little as 8.5 h and 11.5 h total turn around time, respectively.

BACKGROUND: Evidence is accumulating that coronavirus disease 2019 increases the risk of hospitalization and mechanical ventilation in pregnant patients and for preterm delivery. However, the impact on maternal mortality and whether morbidity is differentially affected by disease severity at delivery and trimester of infection are unknown. OBJECTIVE: This study aimed to describe disease severity and outcomes of severe acute respiratory syndrome coronavirus 2 infections in pregnancy across the Washington State, including pregnancy complications and outcomes, hospitalization, and case fatality. STUDY DESIGN: Pregnant patients with a polymerase chain reaction-confirmed severe acute respiratory syndrome coronavirus 2 infection between March 1, 2020, and June 30, 2020, were identified in a multicenter retrospective cohort study from 35 sites in Washington State. Sites captured 61% of annual state deliveries. Case-fatality rates in pregnancy were compared with coronavirus disease 2019 fatality rates in similarly aged adults in Washington State using rate ratios and rate differences. Maternal and neonatal outcomes were compared by trimester of infection and disease severity at the time of delivery. RESULTS: The principal study findings were as follows: (1) among 240 pregnant patients in Washington State with severe acute respiratory syndrome coronavirus 2 infections, 1 in 11 developed severe or critical disease, 1 in 10 were hospitalized for coronavirus disease 2019, and 1 in 80 died; (2) the coronavirus disease 2019-associated hospitalization rate was 3.5-fold higher than in similarly aged adults in Washington State (10.0% vs 2.8%; rate ratio, 3.5; 95% confidence interval, 2.3-5.3); (3) pregnant patients hospitalized for a respiratory concern were more likely to have a comorbidity or underlying conditions including asthma, hypertension, type 2 diabetes mellitus, autoimmune disease, and class III obesity; (4) 3 maternal deaths (1.3%) were attributed to coronavirus disease 2019 for a maternal mortality rate of 1250 of 100,000 pregnancies (95% confidence interval, 257-3653); (5) the coronavirus disease 2019 case fatality in pregnancy was a significant 13.6-fold (95% confidence interval, 2.7-43.6) higher in pregnant patients than in similarly aged individuals in Washington State with an absolute difference in mortality rate of 1.2% (95% confidence interval, -0.3 to 2.6); and (6) preterm birth was significantly higher among women with severe or critical coronavirus disease 2019 at delivery than for women who had recovered from coronavirus disease 2019 (45.4% severe or critical coronavirus disease 2019 vs 5.2% mild coronavirus disease 2019; P

Characterization and suppression of noise are essential for the control of harmonic oscillators in the quantum regime. We measure the noise spectrum of a quantum harmonic oscillator from low frequency to near the oscillator resonance by sensing its response to amplitude modulated periodic drives with a qubit. Using the motion of a trapped ion, we experimentally demonstrate two different implementations with combined sensitivity to noise from 500 Hz to 600 kHz. We apply our method to measure the intrinsic noise spectrum of an ion trap potential in a previously unaccessed frequency range., 6 + 10 pages, 4 + 7 figures

A mixed-species geometric phase gate has been proposed for implementing quantum logic spectroscopy on trapped ions, which combines probe and information transfer from the spectroscopy to the logic ion in a single pulse. We experimentally realize this method, show how it can be applied as a technique for identifying transitions in currently intractable atoms or molecules, demonstrate its reduced temperature sensitivity, and observe quantum-enhanced frequency sensitivity when it is applied to multi-ion chains. Potential applications include improved readout of trapped-ion clocks and simplified error syndrome measurements for quantum error correction., Physical Review X, 10 (2), ISSN:2160-3308

Background During the early months of the coronavirus disease of 2019 (COVID-19) pandemic, risks to pregnant women of a severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection were uncertain. Pregnant patients can serve as a model for the success of the clinical and public health response during public health emergencies as they are typically in frequent contact with the medical system. Population-based estimates of SARS-CoV-2 infections in pregnancy are unknown due to incomplete ascertainment of pregnancy status or inclusion of only single centers or hospitalized cases. Whether pregnant women were protected by the public health response or through their interactions with obstetrical providers in the early pandemic is poorly understood. Objective(s) To estimate the SARS-CoV-2 infection rate in pregnancy and examine disparities by race/ethnicity and English-language proficiency in Washington State. Study design Pregnant patients with a polymerase chain reaction (PCR)-confirmed severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection diagnosed between March 1-June 30, 2020 were identified within 35 hospitals/clinic systems capturing 61% of annual deliveries in Washington State. Infection rates in pregnancy were estimated overall and by Washington State Accountable Community of Health (ACH) region and cross-sectionally compared to SARS-CoV-2 infection rates in similarly aged adults in Washington State. Race/ethnicity and language used for medical care among the pregnant patients were compared to recent data from Washington State. Results A total of 240 pregnant patients with SARS-CoV-2 infections were identified during the study period with 70.7% from minority racial and ethnic groups. The principal findings in our study are: 1) The SARS-CoV-2 infection rate in pregnancy was 13.9/1,000 deliveries (95% confidence interval [CI], 8.3-23.2) compared to 7.3/1,000 (95%CI 7.2-7.4) in 20-39 year old adults in Washington State (Rate Ratio [RR] 1.7, 95%CI 1.3-2.3), 2) the SARS-CoV-2 infection rate reduced to 11.3/1000 (95%CI 6.3-20.3) when excluding 45 cases of SARS-CoV-2 detected through asymptomatic screening (RR 1.3, 95%CI 0.96-1.9), 3) the proportion of SARS-CoV-2 cases in pregnancy among most non-white racial/ethnic groups was 2-4 fold higher than the race and ethnicity distribution of women in Washington State who delivered live births in 2018, and 5) the proportion of SARS-CoV-2 infected pregnant patients receiving medical care in a non-English language was higher than estimates of limited English proficiency in Washington State (30.4% versus 7.6%). Conclusions The SARS-CoV-2 infection rate in pregnant people was 70% higher than similarly aged adults in Washington State, which could not be completely explained by universal screening at delivery. Pregnant patients from nearly all racial/ethnic minority groups and patients receiving medical care in a non-English language were overrepresented. Pregnant women were not protected from COVID-19 in the early months of the pandemic with the greatest burden of infections occurring in nearly all racial/ethnic minority groups. This data coupled with a broader recognition that pregnancy is a risk factor for severe illness and maternal mortality strongly suggests that pregnant people should be broadly prioritized for COVID-19 vaccine allocation in the U.S. similar to some states.

Rationale & Objective Although maintaining high fluid intake is an effective low-risk intervention for the secondary prevention of urinary stone disease, many patients with stones do not increase their fluid intake. Study Design We describe the rationale and design of the Prevention of Urinary Stones With Hydration (PUSH) Study, a randomized trial of a multicomponent behavioral intervention program to increase and maintain high fluid intake. Participants are randomly assigned (1:1 ratio) to the intervention or control arm. The target sample size is 1,642 participants. Setting & Participants Adults and adolescents 12 years and older with a symptomatic stone history and low urine volume are eligible. Exclusion criteria include infectious or monogenic causes of urinary stone disease and comorbid conditions precluding increased fluid intake. Interventions All participants receive usual care and a smart water bottle with smartphone application. Participants in the intervention arm receive a fluid intake prescription and an adaptive program of behavioral interventions, including financial incentives, structured problem solving, and other automated adherence interventions. Control arm participants receive guideline-based fluid instructions. Outcomes The primary end point is recurrence of a symptomatic stone during 24 months of follow-up. Secondary end points include changes in radiographic stone burden, 24-hour urine output, and urinary symptoms. Limitations Periodic 24-hour urine volumes may not fully reflect daily behavior. Conclusions With its highly novel features, the PUSH Study will address an important health care problem. Funding National Institute of Diabetes and Digestive and Kidney Diseases. Trial Registration Registered at ClinicalTrials.gov with study number NCT03244189 .

Recently, medical examiners reported two cases of a 21-year-old male and 24-year-old male with high amounts of morphine in their blood at autopsy. It was suspected that the decedents ingested lethal amounts of morphine from home-brewed poppy seed tea. No studies to date have investigated opium alkaloid content extracted from poppy seeds by home-brewing methods. Various poppy seed products were purchased from online sources and extracted with four home-brewing methods representative of recipes found on drug user forums. Morphine, codeine, and thebaine were quantified in the tea extracts by liquid chromatography-tandem mass spectrometry using a validated analytical method. Morphine, codeine, and thebaine concentrations from seeds were

Iron-containing ferritin has been used for light harvesting and as a photocatalyst. In this study, we test the hypothesis that changing the iron mineral core composition can alter the light harvesting and photocatalytic properties of ferritin, by co-depositing iron in the presence of halides or oxo-anions. This caused the anions to be incorporated into the iron mineral. We report that some of these new iron minerals possess different band gaps than the original ferrihydrite within ferritin. We found an increase in band gap of up to 0.288 eV or a decrease by as much as 0.104 eV, depending on the type of anion and amount of anions incorporated into the ferrihydrite mineral.

We report correlation measurements on two ${^{9}\mathrm{Be}}^{+}$ ions that violate a chained Bell inequality obeyed by any local-realistic theory. The correlations can be modeled as derived from a mixture of a local-realistic probabilistic distribution and a distribution that violates the inequality. A statistical framework is formulated to quantify the local-realistic fraction allowable in the observed distribution without the fair-sampling or independent-and-identical-distributions assumptions. We exclude models of our experiment whose local-realistic fraction is above 0.327 at the 95% confidence level. This bound is significantly lower than 0.586, the minimum fraction derived from a perfect Clauser-Horne-Shimony-Holt inequality experiment. Furthermore, our data provide a device-independent certification of the deterministically created Bell states.

The Nottingham Galley, and its captain John Deane, are remembered today for having survived a grim winter wreck and cannibalism on Boon Island, Maine, in 1710. But that episode is only part of a larger, compelling story involving mutiny, politics, class conflict, reputation, and the power of the written and printed word.

Low copy number proteins within macromolecular complexes, such as viruses, can be critical to biological function while comprising a minimal mass fraction of the complex. The Bacillus subtilis double-stranded DNA bacteriophage ϕ29 gene 13 product (gp13), previously undetected in the virion, was identified and localized to the distal tip of the tail knob. Western blots and immuno-electron microscopy detected a few copies of gp13 in ϕ29, DNA-free particles, purified tails, and defective particles produced in suppressor-sensitive ( sus ) mutant sus 13(330) infections. Particles assembled in the absence of intact gp13 ( sus 13(342) and sus 13(330)) had the gross morphology of ϕ29 but were not infectious. gp13 has predicted structural homology and sequence similarity to the M23 metalloprotease LytM. Poised at the tip of the ϕ29 tail knob, gp13 may serve as a plug to help restrain the highly pressurized packaged genome. Also, in this position, gp13 may be the first virion protein to contact the cell wall in infection, acting as a pilot protein to depolymerize the cell wall. gp13 may facilitate juxtaposition of the tail knob onto the cytoplasmic membrane and the triggering of genome injection.

This paper investigates the comproportionation reaction of MnII with [Formula: see text] as a route for manganese oxide nanoparticle synthesis in the protein ferritin. We report that [Formula: see text] serves as the electron acceptor and reacts with MnII in the presence of apoferritin to form manganese oxide cores inside the protein shell. Manganese loading into ferritin was studied under acidic, neutral, and basic conditions and the ratios of MnII and permanganate were varied at each pH. The manganese-containing ferritin samples were characterized by transmission electron microscopy, UV/Vis absorption, and by measuring the band gap energies for each sample. Manganese cores were deposited inside ferritin under both the acidic and basic conditions. All resulting manganese ferritin samples were found to be indirect band gap materials with band gap energies ranging from 1.01 to 1.34 eV. An increased UV/Vis absorption around 370 nm was observed for samples formed under acidic conditions, suggestive of MnO2 formation inside ferritin.

We use a non-perturbative, space-time resolved, simulation of quantum electrodynamics to explore the properties of electrons dressed with photons. We observe differences in the dynamics of dressed and bare electron wave packets, including differences in dispersion, speed, and mass.

The structural genes of the prohead of phage phi 29 of Bacillus subtilis and a small phi 29 RNA (sRNA) were cloned and expressed in Escherichia coli individually or in combination to study the role of the sRNA in prohead assembly and the mechanism of prohead morphogenesis. The genes coding for the proteins of the scaffold (gp7), the capsid (gp8), the portal vertex (gp10), and the dispensable head fiber (gp8.5) were expressed in E. coli and the gene products were assembled, with and without the presence of the sRNA, into uniform and prolate particles that resembled the typical native phi 29 prohead. No differences in particle size and shape were found between the particles of 7-8-8.5-10 (scaffold-capsid-fiber-portal vertex) and 7-8-8.5-10-RNA (scaffold-capsid-fiber-portal vertex-RNA), suggesting that the phi 29 sRNA was not required for phi 29 prohead assembly. The 7-8-8.5-10 particles produced in E. coli in the absence of phi 29 sRNA were fully competent to package phi 29 DNA in the defined in vitro DNA packaging system by the addition of purified sRNA. Moreover, these DNA-filled heads were assembled into infectious virions in extracts. Without the addition of the sRNA, the 7-8-8.5-10 particles were incompetent while the 7-8-8.5-10-RNA particles were competent in DNA packaging. Bacterial sRNA present in E. coli cannot substitute for the phi 29 sRNA. The assembly of prohead particles in E. coli indicated that host factors unique to B. subtilis were not required. The evidence that the phi 29 sRNA was not required for phi 29 prohead assembly and was not a fixed structural component of the phi 29 prohead favors the conclusion that the phi 29 sRNA is a specific enzyme or morphogenetic factor in DNA packaging.

Bacteriophage φ29 of Bacillus subtilis packages its double-stranded DNA into a preformed prohead during morphogenesis. The prohead is composed of the scaffold protein gp7, the capsid protein gp8, the portal protein gpt 0, and the dispensable head fiber protein gp8.5. Our objective was to elucidate the φ29 prohead assembly pathway and to define the factors that determine prohead shape and size. The structural genes of the φ29 prohead were cloned and expressed in Escherichia coli individually or in combination to study form determination. The scaffold protein was purified from E. coli as a soluble monomer. In vivo and in vitro studies showed that the scaffolding protein interacted with both the portal vertex and capsid proteins. When the scaffold protein interacted only with the capsid protein in vivo , particles were formed with variable size and shape. However, in the presence of the portal vertex protein, particles with uniform size and shape were produced in vivo . SDS-PAGE analysis showed that the latter particles contained the proteins of the scaffold, capsid, head fiber, and portal vertex. These results suggest that the scaffolding protein serves as the linkage between the portal vertex and the capsid proteins, and that the portal vertex plays a crucial role in regulating the size and shape of the prohead.

Alcohol, a potent teratogen, has been suggested as an etiologic agent in attention deficit disorder with hyperactivity (ADHD), which is often diagnosed in children with fetal alcohol syndrome (FAS) and in children of alcoholics. We studied attentional and behavioral factors associated with diagnosis of this disorder in children selected from a predominantly low-income, black population who were tested as part of a longitudinal follow-up of children with prenatal alcohol exposure. Sixty-eight children with a mean age of 5 years 10 months, born to three groups of mothers, were assessed. These groups consisted of: a) women who reported not drinking during pregnancy (n = 21), b) women who reported drinking throughout pregnancy (n = 25), and c) women who reported drinking an equivalent amount but who stopped drinking after educational intervention during the second trimester (n = 22). Dimensions assessed included factors related to attention on a computerized task, impulsivity, and the presence of psychiatrically significant internalizing and externalizing behaviors. In addition, free play and mother-child interactions were video-taped, and evidence of overactive and noncompliant behaviors were noted. Hyperactivity and impulsive behavior were not evident. Results indicated that children exposed throughout pregnancy showed deficits in the ability to sustain attention and were more often described by teachers, although not by their mothers, as showing attentional and behavioral problems. Problems in both internalizing and externalizing behaviors also were noted by teachers. However, when current drinking was controlled, only externalizing behaviors remained different by group.(ABSTRACT TRUNCATED AT 250 WORDS)

Patients who receive high-dose chemotherapy, such as induction regimens followed by high dose melphalan (HD-MEL) and autologous stem cell transplant (ASCT) to treat multiple myeloma (MM), often develop oral mucositis (OM) that impairs adequate intake of fluids and nutrition, serves as a breeding ground for bacteria, leads to increased complexity and cost of care, and may interrupt or delay cancer treatment. Previous work by members of our team demonstrated that renal function and melphalan dose contribute to risk predisposition for oral mucositis. In this study we explored whether genetic variants along with baseline clinical characteristics contribute to risk for oral mucositis by conducting a genome-wide associations study (GWAS). Following strict data quality screening and genotyping using Illumina’s HumanOmni1-Quad v1.0 BeadChip, 892,589 SNPs were tested for association to OM. The analyses evaluated oral mucositis risk in three steps: 1) baseline clinical characteristics; 2) genetic factors (SNPs); and then 3) clinical characteristics combined with genetic factors. Among 972 Caucasian patients who participated in Total Therapy clinical trials (TT2, TT3, TT4) with HD-MEL and ASCT for newly diagnosed myeloma, 353 developed grades 2-4 OM and 619 developed grades 0-1 OM. The baseline sample characteristics included mean age 57.65 (SD 9.16); gender (male 63.68); mean estimated GFR 80.44 (SD 28.40); mean serum albumin 3.85 (SD 0.50); and mean melphalan dose 4.44 (SD 0.79) The combination of estimated glomerular filtration rate (GFR) with melphalan dose (mg/kg), serum albumin, and female gender predicted 67.2% of the risk for oral mucositis (grades 2-4). Analysis of the genetic data identified 11 SNPs significantly associated with Grade 2-4 oral mucositis. The SNPs are located in or near MMP13, JPH3, DHRS7C, CEP 192, CPEB1/CEBP, FBN2, ALDH1A1, DMRTA1/FLJ35282 and have functions related to gene regulation and transcription. We surmise that inflammatory pathways underlie the significance of these SNPs; for example, expression of MMP13 is upregulated in oral epithelial cells during inflammation and FBN2 contributes to wound healing, possibly related to TGFβ signaling and availability. Cyclophospamide (CTX) was part of the total therapy regimens and is known to be detoxified by dehydrogenases. SNP variations associated with aldehyde dehydrogenase 1A1 (ALDH1A1) may alter susceptibility to chemotherapy induced oral mucositis in the presence of HD-MEL, CTX, and other regimens. We postulate that SNPs associated with oral mucositis may influence gene expression through non-coding RNA, thus providing a pathway and novel drug target to prevent this deleterious treatment effect. We will test this hypothesis in future work. Disclosures: No relevant conflicts of interest to declare.

The arena of social program implementation has been divided over the issue of whether new programs should be disseminated and implemented with fidelity (i.e., with close correspondence to the original validated model) or implemented according to the idiosyncratic needs, values, and resources of the local adopting organization. To empirically examine this issue, seven innovative programs were examined. Each of the four educational and three criminal justice programs had been carefully evaluated and widely disseminated. Fidelity and effectiveness criteria were developed for each program. A multimethod data collection strategy included lengthy phone interviews, on-site interviews, and archival analysis of program adopters. The results indicated that well-operationalized programs are usually adopted and implemented with a degree of fidelity acceptable to program developers. Furthermore, this fidelity is positively correlated with the effectiveness of the adopted program. Implications for educational policy and construct measurement are discussed.

A small RNA of Bacillus subtilis bacteriophage phi 29 is shown to have a novel and essential role in viral DNA packaging in vitro. This requirement for RNA in the encapsidation of viral DNA provides a new dimension of complexity to the attendant protein-DNA interactions. The RNA is a constituent of the viral precursor shell of the DNA-packaging machine but is not a component of the mature virion. Studies of the sequential interactions involving this RNA molecule are likely to provide new insight into the structural and possible catalytic roles of small RNA molecules. The phi 29 assembly in extracts and phi 29 DNA packaging in the defined in vitro system were strongly inhibited by treatment with the ribonucleases A or T1. However, phage assembly occurred normally in the presence of ribonuclease A that had been treated with a ribonuclease inhibitor. An RNA of approximately 120 nucleotides co-purified with the phi 29 precursor protein shell (prohead), and this particle was the target of ribonuclease action. Removal of RNA from the prohead by ribonuclease rendered it inactive for DNA packaging. By RNA-DNA hybridization analysis, the RNA was shown to originate from a viral DNA segment very near the left end of the genome, the end packaged first during in vitro assembly.