Statistical Considerations and Software for Designing Sequential, Multiple Assignment, Randomized Trials (SMART) with a Survival Final Endpoint.

Sequential, multiple assignment, randomized trial (SMART) designs are appropriate for comparing adaptive treatment interventions, in which intermediate outcomes (called tailoring variables) guide subsequent treatment decisions for individual patients. Within a SMART design, patients may be re-randomized to subsequent treatments following the outcomes of their intermediate assessments. In this paper, we provide an overview of statistical considerations necessary to design and implement a two-stage SMART design with a binary tailoring variable and a survival final endpoint. A chronic lymphocytic leukemia trial with a final endpoint of progression-free survival is used as an example for the simulations to assess how design parameters, including, choice of randomization ratios for each stage of randomization, and response rates of the tailoring variable affect the statistical power. We assess the choice of weights from restricted re-randomization on data analyses and appropriate hazard rate assumptions. Specifically, for a given first-stage therapy and prior to the tailoring variable assessment, we assume equal hazard rates for all patients randomized to a treatment arm. After the tailoring variable assessment, individual hazard rates are assumed for each intervention path. Simulation studies demonstrate that the response rate of the binary tailoring variable impacts power as it directly impacts the distribution of patients. We also confirm that when the first stage randomization is 1:1, it is not necessary to consider the first stage randomization ratio when applying the weights. We provide an R-shiny application for obtaining power for a given sample size for SMART designs.

Partial Least Squares, Experimental Design, and Near-Infrared Spectrophotometry for the Remote Quantification of Nitric Acid Concentration and Temperature.

Near-infrared spectrophotometry and partial least squares regression (PLSR) were evaluated to create a pleasantly simple yet effective approach for measuring HNO3 concentration with varying temperature levels. A training set, which covered HNO3 concentrations (0.1-8 M) and temperature (10-40 °C), was selected using a D-optimal design to minimize the number of samples required in the calibration set for PLSR analysis. The top D-optimal-selected PLSR models had root mean squared error of prediction values of 1.4% for HNO3 and 4.0% for temperature. The PLSR models built from spectra collected on static samples were validated against flow tests including HNO3 concentration and temperature gradients to test abnormal conditions (e.g., bubbles) and the model performance between sample points in the factor space. Based on cross-validation and prediction modeling statistics, the designed near-infrared absorption approach can provide remote, quantitative analysis of HNO3 concentration and temperature for production-oriented applications in facilities where laser safety challenges would inhibit the implementation of other optical techniques (e.g., Raman spectroscopy) and in which space, time, and/or resources are constrained. The experimental design approach effectively minimized the number of samples in the training set and maintained or improved PLSR model performance, which makes the described chemometric approach more amenable to nuclear field applications.

KDR genetic predictor of toxicities induced by sorafenib and regorafenib.

No biomarkers are available to predict toxicities induced by VEGFR TKIs. This study aimed to identify markers of toxicities induced by these drugs using a discovery-validation approach. The discovery set included 140 sorafenib-treated cancer patients (TARGET study) genotyped for SNPs in 56 genes. The most significant SNPs associated with grade ≥2 hypertension, diarrhea, dermatologic toxicities, and composite toxicity (any one of the toxicities) were tested for association with grade ≥2 toxicity in a validation set of 201 sorafenib-treated patients (Alliance/CALGB 80802). The validated SNP was tested for association with grade ≥2 toxicity in 107 (LCCC 1029) and 82 (Italian cohort) regorafenib-treated patients. SNP-toxicity associations were evaluated using logistic regression, and a meta-analysis between the studies was performed by inverse variance. Variant rs4864950 in KDR increased the risk of grade ≥2 composite toxicity in TARGET, Alliance/CALGB 80802, and the Italian cohort (meta-analysis p = 6.79 × 10-4, OR = 2.01, 95% CI 1.34-3.01). We identified a predictor of toxicities induced by VEGFR TKIs. CLINICALTRIALS.GOV IDENTIFIER: NCT00073307 (TARGET), NCT01015833 (Alliance/CALGB 80802), and NCT01298570 (LCCC 1029).

Initial results of a phase II trial of 18F-DOPA PET-guided re-irradiation for recurrent high-grade glioma.

PURPOSE: In-field high-grade glioma (HGG) recurrence is a common challenge with limited treatment options, including re-irradiation. The radiotracer 3,4-dihydroxy-6-[18F]-fluoro-L-phenylalanine (18F-DOPA) crosses the blood brain barrier and demonstrates high uptake in tumor, but low uptake in normal tissue. This study investigated whether 18F-DOPA positron emission tomography (PET) and MRI guided re-irradiation for recurrent HGG may improve progression free survival (PFS). METHODS: Adults with recurrent or progressive HGG previously treated with radiation were eligible. The primary endpoint was a 20% improvement from the historical control PFS at 3 months (PFS3) of 20% with systemic therapy alone. Re-RT dose was 35 Gy in 10 fractions. The target volume was MRI T1 contrast-enhancement defined tumor plus 18F-DOPA PET defined tumor. RESULTS: Twenty patients completed treatment per protocol. Diagnosis was most commonly glioblastoma, IDH-wildtype (60%). MRI-defined volumes were expanded by a median 43% (0-436%) by utilizing 18F-DOPA PET. PFS3 was 85% (95% CI 63.2-95.8%), meeting the primary endpoint of PFS3 ≥ 40%. With 9.7 months median follow-up, 17 (85%) had progressed and 15 (75%) had died. Median OS from re-RT was 8.8 months. Failure following re-RT was within both the MRI and PET tumor volumes in 75%, MRI only in 13%, PET only in 0%, and neither in 13%. Four (20%) patients experienced grade 3 toxicity, including CNS necrosis (n = 2, both asymptomatic with bevacizumab initiation for radiographic findings), seizures (n = 1), fatigue (n = 1), and nausea (n = 1). No grade 4-5 toxicities were observed. CONCLUSION: 18F-DOPA PET-guided re-irradiation for progressive high-grade glioma appears safe and promising for further investigation.

Amplitude Sensing below the Zero-Point Fluctuations with a Two-Dimensional Trapped-Ion Mechanical Oscillator.

We present a technique to measure the amplitude of a center-of-mass (c.m.) motion of a two-dimensional ion crystal of ∼100 ions. By sensing motion at frequencies far from the c.m. resonance frequency, we experimentally determine the technique's measurement imprecision. We resolve amplitudes as small as 50 pm, 40 times smaller than the c.m. mode zero-point fluctuations. The technique employs a spin-dependent, optical-dipole force to couple the mechanical oscillation to the electron spins of the trapped ions, enabling a measurement of one quadrature of the c.m. motion through a readout of the spin state. We demonstrate sensitivity limits set by spin projection noise and spin decoherence due to off-resonant light scattering. When performed on resonance with the c.m. mode frequency, the technique demonstrated here can enable the detection of extremely weak forces (<1 yN) and electric fields (<1 nV/m), providing an opportunity to probe quantum sensing limits and search for physics beyond the standard model.

The effects of display size on performance.

We examined the systematic effects of display size on task performance as derived from a standard perceptual and cognitive test battery. Specifically, three experiments examined the influence of varying viewing conditions on response speed, response accuracy and subjective workload at four differing screen sizes under three different levels of time pressure. Results indicated a ubiquitous effect for time pressure on all facets of response while display size effects were contingent upon the nature of the viewing condition. Thus, performance decrement and workload elevation were evident only with the smallest display size under the two most restrictive levels of time pressure. This outcome generates a lower boundary threshold for display screen size for this order of task demand. Extrapolations to the design and implementation of all display sizes and forms of cognitive and psychomotor demand are considered.

Laser based accelerator for ultracold atoms.

We present our first results on our implementation of a laser based accelerator for ultracold atoms. Atoms cooled to a temperature of 420 nK are confined and accelerated by means of laser tweezer beams, and the atomic scattering is directly observed in laser absorption imaging. The optical collider has been characterized using 87Rb atoms in the |F=2, m(F)=2] state, but the scheme is not restricted to atoms in any particular magnetic substates and can readily be extended to other atomic species as well.

Immunohistochemical study of human costotransverse joints: A preliminary investigation.

The human costotransverse joint (CTJ) is the articulation between the posterior tubercle of the ribs with the first through tenth costal facet of the thoracic transverse processes. While the CTJ is well defined anatomically and considered a synovial joint, the human CTJ as a pain generating structure is controversial and not supported from a histological perspective. The objective of the present study was to investigate the histological pain producing properties of CTJ capsule tissue. Ten micron cross-sections at each level (1-10) were stained with H & E or immunostained with antisera against Substance P (SP), calcitonin-gene-related peptide (CGRP), and neuropeptide Y (NPY). Immunoreactivity was confirmed for SP, CGRP, and NPY within the CTJ tissue samples of two unembalmed male cadavers. The presence of previously mentioned neuropeptides suggests that human CTJ is capable of producing pain through somatic and autonomic nervous systems. Therefore, clinicians should consider the CTJ as a differential diagnostic possibility when examining and treating painful thoracic conditions.

Association of Adiponectin and Vitamin D With Tumor Infiltrating Lymphocytes and Survival in Stage III Colon Cancer.

Background: Adipocyte-derived adiponectin may play a role in the host inflammatory response to cancer. We examined the association of plasma adiponectin with the density of tumor-infiltrating lymphocytes (TILs) in colon cancers and with vitamin D, clinicopathological features, and patient survival. Methods: Plasma adiponectin and 25-hydroxyvitamin D [25(OH)D] were analyzed by radioimmunoassay in 600 patients with stage III colon cancer who received FOLFOX-based adjuvant chemotherapy (NCCTG N0147 [Alliance]). TIL densities were determined in histopathological sections. Associations with disease-free survival (DFS), time to recurrence, and overall survival were evaluated by multivariable Cox regression adjusting for potential confounders (ie, body mass index, race, TILs, and N stage). All statistical tests were 2-sided. Results: We found a statistically significant reduction in adiponectin, but not 25(OH)D, levels in tumors with high vs low TIL densities (median = 6845 vs 8984 ng/mL; P = .04). A statistically significant reduction in adiponectin was also observed in obese (body mass index >30 kg/m2) vs nonobese patients (median = 6608 vs 12 351 ng/mL; P < .001), in men vs women (median = 8185 vs 11 567 ng/mL; P < .001), in Blacks vs Whites or Asians (median = 6412 vs 8847 vs 7858 ng/mL; P < .03), and in those with fewer lymph node metastases (N1 vs N2: median = 7768 vs 9253 ng/mL; P = .01). Insufficiency of 25(OH)D (<30 ng/mL) was detected in 291 (48.5%) patients. In multivariable analyses, neither adiponectin nor 25(OH)D were associated with a statistically significant difference in DFS, overall survival , or time to recurrence in models adjusted for potential confounders. We found a statistically significant association of TILs with prognosis, yet no such interaction was observed for the association of adiponectin with TILs for DFS. Conclusions: Lower circulating adiponectin levels were associated with a statistically significant increase in TIL densities in colon cancers, indicating an enhanced antitumor immune response. In contrast to TILs, neither adiponectin nor 25(OH)D was independently prognostic.

A laser-microfabricated electrohydrodynamic thruster for centimeter-scale aerial robots.

To date, insect scale robots capable of controlled flight have used flapping-wings for generating lift, but this requires a complex and failure-prone mechanism. A simpler alternative is electrohydrodynamic (EHD) thrust, which requires no moving mechanical parts. In EHD, corona discharge generates a flow of ions in an electric field between two electrodes; the high-velocity ions transfer their kinetic energy to neutral air molecules through collisions, accelerating the gas and creating thrust. We introduce a fabrication process for EHD thruster based on 355 nm laser micromachining, which potentially allows for greater materials selection, such as fiber-based composites, than is possible with semiconductor-based lithographic processing. Our four-thruster device measures 1.8 × 2.5 cm and is composed of steel emitters and a lightweight carbon fiber mesh. We measured the electrical current and thrust of each thruster of our four-thruster design, showing agreement with the Townsend relation. The peak thrust of our device, at 5.2 kV, was measured to be 3.03 times its 37 mg (363.0 µN) mass using a precision balance. In free flight, we demonstrated liftoff at 4.6 kV.

A bio-hybrid odor-guided autonomous palm-sized air vehicle.

Biohybrid systems integrate living materials with synthetic devices, exploiting their respective advantages to solve challenging engineering problems. One challenge of critical importance to society is detecting and localizing airborne volatile chemicals. Many flying animals depend their ability to detect and locate the source of aerial chemical plumes for finding mates and food sources. A robot with comparable capability could reduce human hazard and drastically improve performance on tasks such as locating disaster survivors, hazardous gas leaks, incipient fires, or explosives. Three advances are needed before they can rival their biological counterparts: (1) a chemical sensor with a much faster response time that nevertheless satisfies the size, weight, and power constraints of flight, (2) a design, sensor suite, and control system that allows it to move toward the source of a plume fully autonomously while navigating obstacles, and (3) the ability to detect the plume with high specificity and sensitivity among the assortment of chemicals that invariably exist in the air. Here we address the first two, introducing a human-safe palm-sized air vehicle equipped with the odor-sensing antenna of an insect, the first odor-sensing biohybrid robot system to fly. Using this sensor along with a suite of additional navigational sensors, as well as passive wind fins, our robot orients upwind and navigates autonomously toward the source of airborne plumes. Our robot is the first flying biohybrid system to successfully perform odor localization in a confined space, and it is able to do so while detecting and avoiding obstacles in its flight path. We show that insect antennae respond more quickly than metal oxide gas sensors, enabling odor localization at an improved speed over previous flying robots. By using the insect antennae, we anticipate a feasible path toward improved chemical specificity and sensitivity by leveraging recent advances in gene editing.

Correction: A laser-microfabricated electrohydrodynamic thruster for centimeter-scale aerial robots.

[This corrects the article DOI: 10.1371/journal.pone.0231362.].

Sindbis virus mutants selected for rapid growth in cell culture display attenuated virulence in animals.

Mutants of Sindbis virus were selected for rapid growth in baby hamster kidney (BHK) cell cultures and screened for attenuation of virulence in suckling mice. Comparisons among independently isolated virulent and attenuated strains, as well as a classical reversion analysis, showed that accelerated penetration of BHK cells was correlated with attenuation in vivo. Both phenotypic changes resulted from a reorganization of virion structure as detected by monoclonal antibodies. These results suggest that mutants selected for rapid growth in cell culture may be useful as attenuated vaccines and for studies of the molecular basis of virus pathogenesis.

Inattentiveness and Language Abilities in Preschoolers: A Latent Profile Analysis.

Growing evidence suggests that early symptoms of inattentiveness may affect the language development and academic success of young children. In the present study, we examined the extent to which profiles of inattentiveness and language could be discerned within a heterogeneous group of preschoolers attending early childhood special education programs (n = 461). Based on parent-reported observations of children's symptoms of inattentiveness and direct assessments of children's language skills (grammar, vocabulary, and narrative ability), three distinct profiles were identified. The three groups, representing levels of severity (at risk, almost average, above average), differed not only by their end of year performance, but also with respect to which their abilities changed over the course of the academic year. Children in the poorest performing profile had poorer mean scores in the spring of their preschool year on all measures, but exhibited patterns of gain that exceeded or equaled their peers in higher-performing groups, in the domains of vocabulary and grammar. Examination of subsequent kindergarten reading skills suggested that profile differences remained consistent. Findings underscore the associations between early symptoms of inattentiveness and language difficulties, and further indicate that these relations extend to the acquisition of early reading skills. Future research is needed to corroborate these findings with more robust measures of attention, and to understand the long-term associations between inattentiveness, language and literacy, and potential effects on these associations from early intervention.

Endothelial progenitor cells are recruited into resolving venous thrombi.

BACKGROUND: The purpose of this study was to determine whether endothelial cells of bone marrow origin are involved in thrombus recanalization. METHODS AND RESULTS: Irradiated mice were reconstituted with bone marrow from transgenic donors expressing green fluorescent protein (GFP) linked to the Tie2 promoter. Thrombi were formed in 2 groups of 6 mice. GFP-expressing cells were located and quantified in sections of the thrombi taken after 7 and 14 days. The cell markers Mac-3, F4/80, CD68 (macrophage), and vascular endothelial growth factor receptor 2 (VEGFR2; endothelial cells) were used to determine colocalization with GFP expression in tissue sections and peritoneal macrophages. The markers CD34 and VEGFR2 were used to quantify changes in circulating endothelial cells by flow cytometry of blood from 3 cohorts of wild-type animals that had either a thrombus induced (n=18), a sham operation (n=18), or no operation (n=10). The number of GFP-expressing cells was found to increase by approximately 3-fold in thrombi formed in transplanted animals between 7 and 14 days after induction (P=0.0022). No GFP-expressing cells were found lining the new vascular channels that formed at either time interval, but many of the GFP-expressing cells also expressed Mac-3, CD68, and VEGFR2. Approximately twice as many circulating CD34+/VEGFR2+ cells were found by day 3 in animals with thrombus compared with sham controls (CD45-, P=0.046 and CD45(+), P=0.016). CONCLUSIONS: Bone marrow-derived, Tie2-expressing cells were recruited into the thrombus during resolution but did not line the new vessels. Many of these cells expressed a macrophage phenotype and may represent a population of plastic stem cells that orchestrate thrombus recanalization.

Short-term morbidity in transdiaphragmatic cardiophrenic lymph node resection for advanced stage gynecologic cancer.

Ovarian cancer is commonly diagnosed at an advanced stage, with disease involving the upper abdomen. The finding of enlarged cardiophrenic lymph nodes (CPLNs) on pre-operative imaging often indicates the presence of malignant spread to the mediastinum. Surgical resection of CPLN through a transdiaphragmatic approach can help to achieve cytoreduction to no gross residual. A retrospective chart review was conducted on all patients who underwent transdiaphragmatic cardiophrenic lymph node resection from 8/1/11 through 2/1/15. All relevant pre-, intra-, and post-operative characteristics and findings were recorded. A brief description of the surgical technique is included for reference. Eleven patients were identified who had undergone transdiaphragmatic resection of cardiophrenic lymph nodes. Malignancy was identified in 18/21 (86%) of total lymph nodes submitted. The median number of post-operative days was 7. The overall post-operative morbidity associated with CPLN resection was low, with the most common finding being a small pleural effusion present on chest x-ray between POD# 3-5 (55%). Transdiaphragmatic CPLN resection is a feasible procedure with relatively minor short-term post-operative morbidities that can be used to achieve cytoreduction to no gross residual disease.

Effects of calcitonin and parathyroid hormone on the metabolism of chondrocytes in culture.

Rabbit articular chondrocytes in suspension culture synthesize Type II collagen [3alpha1(II)] in the absence of extracellular Ca2+ and Type I collagen [2alpha1(I) - alpha2] in the complete medium. As a result of pre-treatment in monolayer culture with calcitonin or parathyroid hormone in the complete medium, an influx of Ca2+ into the cells occurs. These cells produce mainly Type I collagen when transferred to suspension cultures in the medium devoid of CaCl2. If added directly to the suspension culture medium containing no CaCl2, calcitonin stimulates an active efflux of Ca2+ from the cells into the medium and leads the cells to synthesize Type I collagen. Under similar conditions, parathyroid hormone does not change the collagen-phenotype.

The melanocytotoxic action of 4-hydroxyanisole.

Cultured normal mammalian melanocytes exposed to a variety of antioxidants in the presence of millimolar concentrations of 4-hydroxyanisole exhibit dose-dependent modifications of cytotoxicity. While some antioxidants reduced the extent of damage produced by 4-hydroxyanisole, others appeared to increase it. Similar effects were found in a model system using lysis of human erythrocytes as an index of cell damage. Estimations on rat liver microsomes in the presence of tyrosinase and 4-hydroxyanisole showed increased peroxidation only at low substrate concentrations.

Further evidence that serotonin is a neurotransmitter involved in the control of prolactin secretion.

Administration of a new specific serotonin uptake inhibitor, fluoxetine, depressed the firing rate of raphe neurons. A highly significant increase in serum prolactin levels was observed after ip injection of 30 mg/kg of 5-hydroxytryptophan (5-HTP) in male or female rats pretreated with 10 mg/kg (ip) of fluoxetine. Neither 5-HTP nor fluoxetine given separately had any effect on serum prolactin levels. In animals pretreated with methysergide the combination of fluoxetine and 5-HTP did not increase significantly serum levels of prolactin. In addition, the serotonin agonist quipazine elevated significantly serum prolactin levels in male and female rats. The results of this study strengthen the idea that 5-HTP is acting via serotonin-containing neurons that influence anterior pituitary prolactin release, and that serotonin receptor activation leads to prolactin release.

Studies on the role of the preoptic area in the control of reproductive function in the rat.

The present study was designed to determine the effects of bilateral lesions restricted to the medial preoptic area on reproductive function in rats. Adult female rats were placed in a stereotaxic instrument, and bilateral lesions were made in the medial preoptic area of the diencephalon using an anodic current and platinum or stainless steel electrodes. Both types of lesions produced identical effects. Most animals with lesions restricted to the medial preoptic area showed repeated periods of pseudopregnancy for the duration of their lives. Ova were detected in the oviducts on the day of estrus between pseudopregnancies, and deciduomata could be produced by uterine traumatization during pseudopregnancy. Daily administration of lergotrile mesylate, a dopamine agonist, for about two weeks, to the pseudopregnant rats with lesions resulted in the appearance of normal 4 or 5-day estrous cycles. These results suggest that the medial preoptic area may not be necessary for ovulation in the rat, and that the normal function of the preoptic area in the control of reproductive cyclicity may be mediated via dopaminergic neurons. In agreement with earlier studies, we found that lesions that extended into the anterior hypothalamus resulted in constant estrus.