HDM2 antagonist MI-219 (spiro-oxindole), but not Nutlin-3 (cis-imidazoline), regulates p53 through enhanced HDM2 autoubiquitination and degradation in human malignant B-cell lymphomas

Abstract Background Lymphomas frequently retain wild-type (wt) p53 function but overexpress HDM2, thereby compromising p53 activity. Therefore, lymphoma is a suitable model for studying the therapeutic value of disrupting the HDM2-p53 interaction by small-molecule inhibitors (SMIs). HDM2 have been developed and are under various stages of preclinical and clinical investigation. Previously, we examined the anti-lymphoma activity of MI-319, the laboratory grade of a new class of HDM2 SMI, the spiro-oxindole, in follicular lymphoma. Since then, MI-219, the clinical grade has become readily available. This study further examines the preclinical effects and mechanisms of MI-219 in a panel of human lymphoma cell lines as well as a cohort of patient-derived B-lymphcytes for its potential clinical use. Results Preclinical assessment of MI-219 was evaluated by means of an in vitro and ex vivo approach and compared to Nutlin-3, the gold standard. Characterization of p53 activity and stability were assessed by quantitative PCR, Western blot, and immunoprecipitation. Biological outcome was measured using Trypan blue exclusion assay, Annexin V/PI, PARP and caspase-3 cleavage. Surprisingly, the overall biological effects of Nutlin-3 were more delayed (48 h) while MI-219 triggered an earlier response (12-24 h), predominantly in the form of apoptotic cell death. Using a cell free autoubiquitination assay, neither agent interfered with HDM2 E3 ligase function. MI-219 was more effective in upregulating wt-p53 stabilization compared to Nutlin-3. MI-219, but not Nutlin-3, enhanced the autoubiquitination and degradation of HDM2. Conclusions Our data reveals unexpected differences between MI-219 and the well-studied Nutlin-3 in lymphoma cell lines and patient samples. We suggest a novel mechanism for MI-219 that alters the functional activity of HDM2 through enhanced autoubiquitination and degradation. Additionally, this mechanism appears to correspond to biological outcome. Our results provide evidence that different classes of HDM2 SMIs elicit molecular events that extend beyond HDM2-p53 dissociation which may be of biological and potentially therapeutic importance

Probing the Deuteron at Very Large Internal Momenta

We measure 2H(e,e′p)n cross sections at 4-momentum transfers of Q2 = 4.5 ± 0.5   (GeV/c)2 over a range of neutron recoil momenta pr, reaching up to ∼1.0  GeV/c. We obtain data at fixed neutron recoil angles θnq = 35°, 45°, and 75° with respect to the 3-momentum transfer →q. The new data agree well with previous data, which reached pr ∼ 500  MeV/c. At θnq = 35° and 45°, final state interactions, meson exchange currents, and isobar currents are suppressed and the plane wave impulse approximation provides the dominant cross section contribution. We compare the new data to recent theoretical calculations, where we observe a significant discrepancy for recoil momenta pr \u3e 700  MeV/c

Understanding plant invasions: An example of working with citizen scientists to collect environmental data

Citizen science programs are useful tools for collecting important environmental science data. To ensure data quality, however, it must be shown that data collected by volunteers can produce reliable results. We engaged 143 volunteers over four years to map and estimate abundance of invasive plants in New York and New Jersey parklands. We found that off trail abundance of only a few of our targeted invasive species were positively correlated with on trail abundance. Our results support that citizen science programs can be a useful and sometimes a much needed addition to environmental science protocols

Determination of the Argon Spectral Function From (e, e\u27p) Data

The E12-14-012 experiment, performed in Jefferson Lab Hall A, has measured the (e,e′p) cross section in parallel kinematics using a natural argon target. Here, we report the full results of the analysis of the data set corresponding to beam energy 2.222 GeV, and spanning the missing momentum and missing energy range 15 ≲ pm ≲ 300  MeV /c and 12 ≲ Em ≲ 80  MeV. The reduced cross section, determined as a function of pm and Em with ≈ 4% accuracy, has been fitted using the results of Monte Carlo simulations involving a model spectral function and including the effects of final state interactions. The overall agreement between data and simulations turns out to be quite satisfactory (χ2/d. o. f. =1.9). The resulting spectral function will provide valuable new information, needed for the interpretation of neutrino interactions in liquid argon detectors

Measurement of the Ar(e, e\u27 p) and Ti(e, e\u27 p) Cross Sections in Jefferson Lab Hall A

The E12-14-012 experiment, performed in Jefferson Lab Hall A, has collected exclusive electron-scattering data (e, e\u27p) in parallel kinematics using natural argon and natural titanium targets. Here we report the first results of the analysis of the data set corresponding to beam energy 2222 GeV, electron scattering angle 21.5 degrees, and proton emission angle -50°. The differential cross sections, measured with ≈ 4% uncertainty, have been studied as a function of missing energy and missing momentum, and compared to the results of Monte Carlo simulations, obtained from a model based on the distorted-wave impulse approximation

Surface-based constraints on target selection and distractor rejection: Evidence from preview search

In preview search when an observer ignores an early appearing set of distractors, there can subsequently be impeded detection of new targets that share the colour of this preview. This “negative carry-over effect” has been attributed to an active inhibitory process targeted against the old items and inadvertently their features. Here we extend negative carry-over effects to the case of stereoscopically defined surfaces of coplanar elements without common features. In Experiment 1 observers previewed distractors in one surface (1000 ms), before being presented with the target and new distractors divided over the old and a new surface either above or below the old one. Participants were slower and less efficient to detect targets in the old surface. In Experiment 2 in both the first and second display the items were divided over two planes in the proportion 66/33% such that no new planes appeared following the preview, and there was no majority of items in any one plane in the final combined display. The results showed that participants were slower to detect the target when it occurred in the old majority surface. Experiment 3 held constant the 2D properties of the stimuli while varying the presence of binocular depth cues. The carry-over effect only occurred in the presence of binocular depth cues, ruling out any account of the results in terms of 2-D cues. The results suggest well formed surfaces in addition to simple features may be targets for inhibition in search

Novel plasma treatment for preparation of laser sintered nanocomposite parts

Polymer Laser Sintering (LS) is a well-known Additive Manufacturing process, capable of producing highly complex geometries with little or no cost penalty. However, the restricted range of materials currently available for this process has limited its applications. Whilst it is common to modify the properties of standard LS polymers with the inclusion of fillers e.g. nanoclays, achieving effective dispersions can be difficult. The work presented here investigates the use of plasma treatment as a method of enhancing dispersion with an expectation of improving consistency and surface quality of laser sintered nanocomposite parts. To enable the preparation of polyamide 12 nanocomposite powder for applications in LS, plasma surface modification using Low Pressure Air Plasma Treatment was carried out on two nanoclays: Cloisite 30B (C30B) and Nanomer I.34TCN (I.34TCN). Plasma treatment strongly reduced the aggregation of the nanoclay (C30B and I.34TCN) particles, and powders displayed higher decomposition temperatures than those without plasma treatment. LS parts from neat polyamide 12, untreated I.34TCN and plasma treated I.34TCN composites were successfully produced with different complex shapes. The presence of well dispersed plasma treated nanoclays was observed and found to be essential for an improved surface quality of LS fabricated which was achieved only for plasma treated I.34TCN. Likewise, some mechanical properties could be improved above that of PA12 by incorporation of treated I.34TCN. For example, the elastic modulus of plasma treated composites was higher than that of polyamide 12 and the untreated composite. In the case of the ultimate strain, the plasma treated composite performed better than untreated and results had a reduced variation between samples. This illustrates the feasibility of the use of plasma treatments on nanoclays to improve the properties of LS parts, even though further studies will be required to exploit the full potential

Cost-effectiveness of Routine Provider-Initiated Testing and Counseling for Children With Undiagnosed HIV in South Africa

Background: We compared the cost-effectiveness of pediatric provider-initiated HIV testing and counseling (PITC) vs no PITC in a range of clinical care settings in South Africa. Methods: We used the Cost-Effectiveness of Preventing AIDS Complications Pediatric model to simulate a cohort of children, aged 2-10 years, presenting for care in 4 settings (outpatient, malnutrition, inpatient, tuberculosis clinic) with varying prevalence of undiagnosed HIV (1.0%, 15.0%, 17.5%, 50.0%, respectively). We compared "PITC" (routine testing offered to all patients; 97% acceptance and 71% linkage to care after HIV diagnosis) with no PITC. Model outcomes included life expectancy, lifetime costs, and incremental cost-effectiveness ratios (ICERs) from the health care system perspective and the proportion of children with HIV (CWH) diagnosed, on antiretroviral therapy (ART), and virally suppressed. We assumed a threshold of $3200/year of life saved (YLS) to determine cost-effectiveness. Sensitivity analyses varied the age distribution of children seeking care and costs for PITC, HIV care, and ART. Results: PITC improved the proportion of CWH diagnosed (45.2$710 to $1240/YLS. PITC remained cost-effective unless undiagnosed HIV prevalence was <0.2%. Conclusions: Routine testing improves HIV clinical outcomes and is cost-effective in South Africa if the prevalence of undiagnosed HIV among children exceeds 0.2%. These findings support current recommendations for PITC in outpatient, inpatient, tuberculosis, and malnutrition clinical settings