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Functional communication training (FCT) provides an efficient means for reducing a variety of challenging behaviors while increasing social communication. After establishing a functional communication response (FCR), interventionists may need to design a program to thin the schedule of reinforcement to provide more durable effects and maintenance in the natural environment. Hanley et al. provided one means for thinning schedules that involved the systematic introduction of extinction (or denial trials) following a process of delaying reinforcement. This study evaluates a southeastern U.S. classroom application of this thinning process with three elementary age students who engaged in a variety of aggression, self-injury, disruption, and crying. The results indicate positive effects of the FCT program with systematic introduction of denial trials once the participants demonstrated proficiency with the FCR. In addition, data for two participants show prolonged effects post-intervention. Results are discussed in terms of extending Hanley et al. and how these procedures can be used within typical school-based settings.

Abstract Powerful associations that link drugs of abuse with cues in the drug-paired environment often serve as prepotent relapse triggers. Drug-associated contexts and cues activate ensembles of nucleus accumbens (NAc) neurons, including D1-class medium spiny neurons (MSNs) that typically promote, and D2-class MSNs that typically oppose, drug seeking. We found that in mice, cocaine conditioning upregulated transiently the activity-regulated transcription factor, Neuronal PAS Domain Protein 4 (NPAS4), in a small subset of NAc neurons. The NPAS4+ NAc ensemble was required for cocaine conditioned place preference. We also observed that NPAS4 functions within NAc D2-, but not D1-, MSNs to support cocaine-context associations and cue-induced cocaine, but not sucrose, seeking. Together, our data show that the NPAS4+ ensemble of NAc neurons is essential for cocaine-context associations in mice, and that NPAS4 itself functions in NAc D2-MSNs to support cocaine-context associations by suppressing drug-induced counteradaptations that oppose relapse-related behaviour.

The CDISC Standard for Exchange of Nonclinical Data (SEND) data standard has created new opportunities for collaborative development of open-source software solutions to facilitate cross-study analyses of toxicology study data. A public–private partnership between BioCelerate and the FDA/Center for Drug Evaluation and Research (CDER) was established in part to develop and publicize novel methods to facilitate cross-study analysis of SEND datasets. As part of this work in collaboration with the Pharmaceutical Users Software Exchange (PHUSE), an R package sendigR has been developed to enable users to construct a relational database from a collection of SEND datasets and then query that database to perform cross-study analyses. The sendigR package also includes an integrated Python package, xptcleaner, which can be used to harmonize the terminology used in SEND datasets by mapping to CDISC controlled terminologies. The sendigR R package is freely available on the comprehensive R Archive Network (CRAN) and at https://github.com/phuse-org/sendigR. An R Shiny web application was included in the R package to enable toxicologists with no coding experience to perform historical control analyses. Experienced R programmers will be able to integrate the package functions into their own custom scripts/packages and potentially contribute improvements to the functionality of sendigR.sendigR reference manual: https://phuse-org.github.io/sendigR/.sendigR R Shiny demo app: https://phuse-org.shinyapps.io/sendigR/.

Due to the rising annual incidence of lung cancer (LC), new treatment strategies are needed. While various options exist, many, if not all, remain suboptimal. Several studies have shown cryoablation to be a promising approach. Yet, a lack of basic information pertaining to LC response to freezing and requirement for percutaneous access has limited clinical use. In this study, we investigated the A549 lung carcinoma cell line response to freezing. The data show that a single 5 min freeze to −15 °C did not affect cell viability, whereas −20 °C and −25 °C result in a significant reduction in viability 1 day post freeze to

Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal disease that may be treated utilizing thermal therapies. Cryoablation is an effective, minimally invasive therapy that has been utilized for the treatment of various cancers, offering patients a quicker recovery and reduced side effects. Cryoablation has been utilized on a limited basis for the treatment of PDAC. With the recent reports on the success of cryoablation, there is a growing interest in the use of cryoablation as a standalone, minimally invasive procedure to treat PDAC. While offering a promising path, the application of cryoablation to PDAC is limited by current technologies. As such, there is a need for the development of new devices to support advanced treatment strategies for PDAC. To this end, this study investigated the performance of a new endoscopic ultrasound-compatible cryoablation catheter technology, FrostBite. We hypothesized that FrostBite would enable the rapid, effective, minimally invasive delivery of ultra-cold temperatures to target tissues, resulting in effective ablation via an endoscopic approach. Thermal properties and ablative efficacy were evaluated using a heat-loaded gel model, tissue-engineered models (TEMs), and an initial in vivo porcine study. Freeze protocols evaluated included single and repeat 3 and 5 min applications. Isotherm assessment revealed the generation of a 2.2 cm diameter frozen mass with the −20 °C isotherm reaching a diameter of 1.5 cm following a single 5 min freeze. TEM studies revealed the achievement of temperatures ≤ −20 °C at a diameter of 1.9 cm after a 5 min freeze. Fluorescent imaging conducted 24 h post-thaw demonstrated a uniformly shaped ellipsoidal ablative zone with a midline diameter of 2.5 cm, resulting in a total ablative volume of 6.9 cm3 after a single 5 min freeze. In vivo findings consistently demonstrated the generation of ablative areas measuring 2.03 cm × 3.2 cm. These studies demonstrate the potential of the FrostBite cryocatheter as an endoscopic ultrasound-based treatment option. The data suggest that FrostBite may provide for the rapid, effective, controllable freezing of cancerous pancreatic and liver tissues. This ablative power also offers the potential of improved safety margins via the minimally invasive nature of an endoscopic ultrasound-based approach or natural orifice transluminal endoscopic surgery (NOTES)-based approach. The results of this pre-clinical feasibility study show promise, affirming the need for further investigation into the potential of the FrostBite cryocatheter as an advanced, minimally invasive cryoablative technology.

Toxoplasma gondii is a widespread apicomplexan parasite that can cause severe disease in its human hosts. The ability of T. gondii and other apicomplexan parasites to invade into, egress from, and move between cells of the hosts they infect is critical to parasite virulence and disease progression. An unusual and highly conserved parasite myosin motor (TgMyoA) plays a central role in T. gondii motility. The goal of this work was to determine whether the parasite's motility and lytic cycle can be disrupted through pharmacological inhibition of TgMyoA, as an approach to altering disease progression in vivo. To this end, we first sought to identify inhibitors of TgMyoA by screening a collection of 50,000 structurally diverse small molecules for inhibitors of the recombinant motor's actin-activated ATPase activity. The top hit to emerge from the screen, KNX-002, inhibited TgMyoA with little to no effect on any of the vertebrate myosins tested. KNX-002 was also active against parasites, inhibiting parasite motility and growth in culture in a dose-dependent manner. We used chemical mutagenesis, selection in KNX-002, and targeted sequencing to identify a mutation in TgMyoA (T130A) that renders the recombinant motor less sensitive to compound. Compared to wild-type parasites, parasites expressing the T130A mutation showed reduced sensitivity to KNX-002 in motility and growth assays, confirming TgMyoA as a biologically relevant target of KNX-002. Finally, we present evidence that KNX-002 can slow disease progression in mice infected with wild-type parasites, but not parasites expressing the resistance-conferring TgMyoA T130A mutation. Taken together, these data demonstrate the specificity of KNX-002 for TgMyoA, both in vitro and in vivo, and validate TgMyoA as a druggable target in infections with T. gondii. Since TgMyoA is essential for virulence, conserved in apicomplexan parasites, and distinctly different from the myosins found in humans, pharmacological inhibition of MyoA offers a promising new approach to treating the devastating diseases caused by T. gondii and other apicomplexan parasites.

Chronic stress can produce reward system deficits (i.e., anhedonia) and other common symptoms associated with depressive disorders, as well as neural circuit hypofunction in the medial prefrontal cortex (mPFC). However, the molecular mechanisms by which chronic stress promotes depressive-like behavior and hypofrontality remain unclear. We show here that the neuronal activity-regulated transcription factor, NPAS4, in the mPFC is regulated by chronic social defeat stress (CSDS), and it is required in this brain region for CSDS-induced changes in sucrose preference and natural reward motivation in the mice. Interestingly, NPAS4 is not required for CSDS-induced social avoidance or anxiety-like behavior. We also find that mPFC NPAS4 is required for CSDS-induced reductions in pyramidal neuron dendritic spine density, excitatory synaptic transmission, and presynaptic function, revealing a relationship between perturbation in excitatory synaptic transmission and the expression of anhedonia-like behavior in the mice. Finally, analysis of the mice mPFC tissues revealed that NPAS4 regulates the expression of numerous genes linked to glutamatergic synapses and ribosomal function, the expression of upregulated genes in CSDS-susceptible animals, and differentially expressed genes in postmortem human brains of patients with common neuropsychiatric disorders, including depression. Together, our findings position NPAS4 as a key mediator of chronic stress-induced hypofrontal states and anhedonia-like behavior.

Abstract Minnesota is the state with the strongest winter warming in the contiguous United States. We performed regional climate projections at 10 km horizontal resolution using the Weather Research Forecasting model forced with eight CMIP5 GCMs. The selected GCMs have previously been found to be in relatively good agreement with observations over Minnesota compared to other members of the CMIP5 model ensemble. Our projections suggest ongoing warming in all seasons, especially in winter, as well as shallower snow depth and fewer days with snow cover. We expect significant increases in spring and early summer heavy precipitation events. Our comparisons between different time slices and two different emission scenarios indicate a climate for the state of Minnesota near the end of the 21st century that is significantly different from what has been observed by the end of the 20th century. Winters and summers are expected to be up to 6 and 4°C warmer, respectively, over northern and central Minnesota, and spring precipitation may increase by more than 1 mm d−1 over northern Minnesota. Especially over the central part of the state, winter snow depth is projected to decrease by more than 12 cm, and the number of days per year with snow depth of more than 2.54 cm (one inch) is expected to decrease by up to 55.

Solar radiation management (SRM) and carbon dioxide removal (CDR) are geoengineering methods that have been proposed to mitigate global warming in the event of insufficient greenhouse gas emission reductions. Here, we have studied temperature and precipitation responses to CDR and SRM with the Representative Concentration Pathway 4.5 (RCP4.5) scenario using the MPI-ESM and CESM Earth system models (ESMs). The SRM scenarios were designed to meet one of the two different long-term climate targets: to keep either global mean (1) surface temperature or (2) precipitation at the 2010–2020 level via stratospheric sulfur injections. Stratospheric sulfur fields were simulated beforehand with an aerosol–climate model, with the same aerosol radiative properties used in both ESMs. In the CDR scenario, atmospheric CO2 concentrations were reduced to keep the global mean temperature at approximately the 2010–2020 level. Results show that applying SRM to offset 21st century climate warming in the RCP4.5 scenario leads to a 1.42 % (MPI-ESM) or 0.73 % (CESM) reduction in global mean precipitation, whereas CDR increases global precipitation by 0.5 % in both ESMs for 2080–2100 relative to 2010–2020. In all cases, the simulated global mean precipitation change can be represented as the sum of a slow temperature-dependent component and a fast temperature-independent component, which are quantified by a regression method. Based on this component analysis, the fast temperature-independent component of the changed atmospheric CO2 concentration explains the global mean precipitation change in both SRM and CDR scenarios. Based on the SRM simulations, a total of 163–199 Tg S (CESM) or 292–318 Tg S (MPI-ESM) of injected sulfur from 2020 to 2100 was required to offset global mean warming based on the RCP4.5 scenario. To prevent a global mean precipitation increase, only 95–114 Tg S was needed, and this was also enough to prevent global mean climate warming from exceeding 2∘ above preindustrial temperatures. The distinct effects of SRM in the two ESM simulations mainly reflected differing shortwave absorption responses to water vapour. Results also showed relatively large differences in the individual (fast versus slow) precipitation components between ESMs.

This article explores the possibilities and limitations of online teaching, based on our experience transforming a study abroad program to Argentina into an online class, amidst the COVID-19 pandemic and uprisings for racial justice. In a moment when radical educators and activists are moving online, the article considers the spatial politics involved in teaching about protest and resistance online, and in establishing transnational solidarities between U.S. and Latin American scholars, artists, activists, and students. We introduce the theory and practice of a trasnational feminist pedagogy that incorporates embodied knowledge, fosters transnational collaborations, and promotes liberatory learning practices. Drawing on autoethnography, participant observation, visual and media analysis, student interaction with course materials, and interviews with transnational feminist scholars, we investigate how educators and students adapt their teaching and learning practices to an online environment. Transnational feminist pedagogy is a flexible method that allows for transformative teaching, is attentive to power dynamics in and out of the classroom, and maintains commitments to antiracist, feminist and socialist pedagogy.

Cryoablation (CA) is unique as the singular energy deprivation therapy that impacts all cellular processes. CA is independent of cell cycle stage and degree of cellular stemness. Importantly, CA is typically applied as a non-repetitive (single session) treatment that does not support adaptative mutagenesis as do many repetitive therapies. CA is characterized by the launch of multiple forms of cell death including (a) ice-related physical damage, (b) initiation of cellular stress responses (kill switch activation) and launch of necrosis and apoptosis, (c) vascular stasis, and (d) likely activation of ablative immune responses. CA is not without limitation related to the thermal gradient formed between cryoprobe surface (∼−185°C) and the distal surface of the freeze zone (∼0°C) requiring freeze margin extension beyond the tumor boundary (up to ∼1 cm). This limitation is mitigated in part by commonly applied dual freeze thaw cycles and the use of freeze sensitizing adjuvants. This review will (1) identify the cascade of damaging effects of the freeze–thaw process, its physical and molecular-based relationships, (2) a likely immunological involvement (abscopic effect), and (3) explore the use of freeze-sensitizing adjuvants necessary to limit freezing beyond the tumor margin.

Undescended parathyroid adenoma is a rare cause of primary hyperparathyroidism that happens < 1% of cases. If not suspected, it can lead to a negative bilateral parathyroid exploration and extensive iatrogenic trauma. We propose that with proper imaging the correct diagnosis can be established to simplify surgical management. We describe two cases of patients who underwent a targeted neck exploration due to an undescended parathyroid adenoma diagnosed with an appropriate preoperative imaging protocol. With an appropriate imaging protocol for primary hyperparathyroidism and parathyroid hormone aspirates, an undescended parathyroid adenoma can be primarily diagnosed to guide a focused parathyroidectomy.

Introduction: Breast cancer is the most prominent form of cancer and the second leading cause of death in women behind lung cancer. The primary modes of treatment today include surgical excision (lumpectomy, mastectomy), radiation, chemoablation, anti-HER2/neu therapy, and/or hormone therapy. The severe side effects associated with these therapies suggest a minimally invasive therapy with fewer quality of life issues would be advantageous for treatment of this pervasive disease. Cryoablation has been used in the treatment of other cancers, including prostate, skin, and cervical, for decades and has been shown to be a successful minimally invasive therapeutic option. To this end, the use of cryotherapy for the treatment of breast cancer has increased over the last several years. Although successful, one of the challenges in cryoablation is management of cancer destruction in the periphery of the ice ball as the tissue within this outer margin may not experience ablative temperatures. In breast cancer, this is of concern due to the lobular nature of the tumors. As such, in this study, we investigated the level of cell death at various temperatures associated with the margin of a cryogenic lesion as well as the impact of repetitive freezing and thawing methods on overall efficacy. Methods: Human breast cancer cells, MCF-7, were exposed to temperatures of −5°C, −10°C, −15°C, −20°C, or −25°C for 5-minute freeze intervals in a single or repeat freeze-thaw cycle. Samples were thawed with either passive or active warming for 5 or 10 minutes. Samples were assessed at 1, 2, and 3 days post-freeze to assess cell survival and recovery. In addition, the modes of cell death associated with freezing were assessed over the initial 24-hour post-thaw recovery period. Results: Exposure of MCF-7 cells to −5°C and −10°C resulted in minimal cell death regardless of the freeze/thaw conditions. Freezing to a temperature of −25°C resulted in complete cell death 1 day post-thaw with no cell recovery in all freeze/thaw scenarios evaluated. Exposure to a single freeze event resulted in a gradual increase in cell death at −15°C and −20°C. Application of a repeat freeze-thaw cycle (dual 5-minute freeze) resulted in an increase in cell death with complete destruction at −20°C and near complete death at −15°C (day 1 survival: single −15°C freeze/thaw = 20%; repeated −15°C freeze/thaw = 4%). Analysis of thaw interval time (5 vs 10 minute) demonstrated that the shorter 5-minute thaw interval between freezes resulted in increased cell destruction. Furthermore, investigation of thaw rate (active vs passive thawing) demonstrated that active thawing resulted in increased cell survival thereby less effective ablation compared with passive thawing (eg, −15°C 5/10/5 procedure survival, passive thaw: 4% vs active thaw: 29%). Conclusions: In summary, these in vitro findings suggest that freezing to temperatures of 25°C results in a high degree of breast cancer cell destruction. Furthermore, the data demonstrate that the application of a repeat freeze procedure with a passive 5-minute or 10-minute thaw interval between freeze cycles increases the minimal lethal temperature to the −15°C to −20°C range. The data also demonstrate that the use of an active thawing procedure between freezes reduces ablation efficacy at temperatures associated with the iceball periphery. These findings may be important to improving future clinical applications of cryoablation for the treatment of breast cancer.

As the incidence of pancreatic ductal adenocarcinoma (PDAC) continues to grow, so does the need for new strategies for treatment. One such area being evaluated is cryoablation. While promising, studies remain limited and questions surrounding basic dosing (minimal lethal temperature) coupled with technological issues associated with accessing PDAC tumors and tumor proximity to vasculature and bile ducts, among others, have limited the use of cryoablation. Additionally, as chemotherapy remains the first-line of attack for PDAC, there is limited information on the impact of combining freezing with chemotherapy. As such, this study investigated the in vitro response of a PDAC cell line to freezing, chemotherapy, and the combination of chemotherapy pre-treatment and freezing. PANC-1 cells and PANC-1 tumor models were exposed to cryoablation (freezing insult) and compared to non-frozen controls. Additionally, PANC-1 cells were exposed to varying sub-clinical doses of gemcitabine or oxaliplatin alone and in combination with freezing. The results show that freezing to −10 °C did not affect viability, whereas −15 °C and −20 °C resulted in a reduction in 1 day post-freeze viability to 85% and 20%, respectively, though both recovered to controls by day 7. A complete cell loss was found following a single freeze below −25 °C. The combination of 100 nM gemcitabine (1.1 mg/m2) pre-treatment and a single freeze at −15 °C resulted in near-complete cell death (2) pre-treatment and a single −15 °C freeze resulted in a similar trend of increased PANC-1 cell death. In summary, these in vitro results suggest that freezing alone to temperatures in the range of −25 °C results in a high degree of PDAC destruction. Further, the data support a potential combinatorial chemo/cryo-therapeutic strategy for the treatment of PDAC. These results suggest that a reduction in chemotherapeutic dose may be possible when offered in combination with freezing for the treatment of PDAC.

The development and use of complex cell-based products in clinical and discovery science continues to grow at an unprecedented pace. To this end, cryopreservation plays a critical role, serving as an enabling process, providing on-demand access to biological material, facilitating large scale production, storage, and distribution of living materials. Despite serving a critical role and substantial improvements over the last several decades, cryopreservation often remains a bottleneck impacting numerous areas including cell therapy, tissue engineering, and tissue banking. Studies have illustrated the impact and benefit of controlling cryopreservation-induced delayed-onset cell death (CIDOCD) through various “front end” strategies, such as specialized media, new cryoprotective agents, and molecular control during cryopreservation. While proving highly successful, a substantial level of cell death and loss of cell function remains associated with cryopreservation. Recently, we focused on developing technologies (RevitalICE™) designed to reduce the impact of CIDOCD through buffering the cell stress response during the post-thaw recovery phase in an effort to improve the recovery of previously cryopreserved samples. In this study, we investigated the impact of modulating apoptotic caspase activation, oxidative stress, unfolded protein response, and free radical damage in the initial 24 h post-thaw on overall cell survival. Human hematopoietic progenitor cells in vitro cryopreserved in both traditional extracellular-type and intracellular-type cryopreservation freeze media were utilized as a model cell system to assess impact on survival. Our findings demonstrated that through the modulation of several of these pathways, improvements in cell recovery were obtained, regardless of the freeze media and dimethyl sulfoxide concentration utilized. Specifically, through the use of oxidative stress inhibitors, an average increase of 20% in overall viability was observed. Furthermore, the results demonstrated that by using the post-thaw recovery reagent on samples cryopreserved in intracellular-type media (Unisol™), improvements in overall cell survival approaching 80% of non-frozen controls were attained. While improvements in overall survival were obtained, an assessment on the impact of specific cell subpopulations and functionality remains to be completed. While work remains, these results represent an important step forward in the development of improved cryopreservation processes for use in discovery science, and commercial and clinical settings.

A comparison of sea level pressure (SLP) trends in a subset of seven Coupled Model Intercomparison Project (CMIP) phase 5 general circulation models (GCM), namely decadal simulations with CCSM4, CanCM4, MPI-ESM-LR, FGOALS-g2, MIROC4h, MIROC5, and MRI-CGCM3, to their CMIP3 counterparts reveals an unrealistically strong forecast skill in CMIP3 models for trend predictions for 2001–2011 when using the 1979–2000 period to train the forecast. Boreal-winter SLP trends over five high-, mid-, and low-latitude zones were calculated over the 1979–2000 initialization period for each ensemble member and then ranked based on their performance relative to HadSLP2 observations. The same method is used to rank the ensemble members during the 2001–2011 period. In CMIP3, 17 out of 38 ensemble members retain their rank in the 2001–2011 hindcast period and 3 retain the neighboring rank. However, numbers are much lower in more recent CMIP5 decadal predictions over the similar 2001–2010 period when using the 1981–2000 period as initialization with the same number of ensembles. Different periods were used for CMIP3 and CMIP5 because although the 1979–2000 initialization is widely used for CMIP3, CMIP5 decadal predictions are only available for 30-year periods. The conclusion to consider the forecast skill in CMIP3 predictions during 2001–2011 as unrealistic is corroborated by comparisons to earlier periods from the 1960s to the 1980s in both CMIP3 and CMIP5 simulations. Thus, although the 2001–2011 CMIP3 predictions show statistically significant forecast skill, this skill should be treated as a spurious result that is unlikely to be reproduced by newer more accurate GCMs.

This study is aimed at exploring the optimal ELISA standard curve fitting process for reducing measurement uncertainty. Using an ELISA kit for measuring cyanobacterial toxin (microcystin), we show that uncertainty associated with the estimated microcystin concentrations can be reduced by defining the standard curve as a four-parameter logistic function on the natural log concentration scale, instead of the current approach of defining the curve on the concentration scale. The model comparison method is outlined in this paper, allowing it to be transferable to test different statistical models for other ELISA test kits. Keywords: Cyanobacteria, Drinking water, Harmful algal bloom, Nonlinear regression

Objectives: Cryoablation is an effective alternative treatment for cardiac arrhythmias offering shortened recovery and reduced side effects. As the use of cryoablation increases, the need for new devices and procedures has emerged. This has been driven by technological limitations including lengthy periods to generate a single lesion (3–5 min), uncertain transmurality, and differential efficacy. Furthermore, due to limited ablation capacity under high heat loads, cryo has had limited success in the treatment of ventricular arrhythmias. To this end, in this study we evaluated a new cryoablation catheter, ICEolate , for the targeted ablation of cardiac tissue. Methods: Performance assessment included calorimetry, freeze zone isothermal distribution characterization and catheter ablation capacity in a submerged, circulating, heat-loaded ex vivo tissue model. A pilot in vivo study was also conducted to assess ablative capacity of the cryocatheter in a fully beating heart. Results: Ex vivo studies demonstrated ice formation at the tip of a cryocatheter within 5 s and a tip temperature of ~−150°C within 10 s. The device repeatedly generated freeze zones of 2 cm × 3 cm in less than 2 min. Tissue model studies revealed the generation of a full thickness (5–10 mm) cryogenic lesion within 1 min with an opposite (transmural) surface temperature of

The prevalence of gestational diabetes mellitus (GDM) is increasing because of the worldwide obesity/diabetes epidemic. The complications of untreated GDM affect both the mother and baby and include complications during pregnancy as well as increased risk of subsequent type-2 diabetes in mothers and offspring. Standard tests for hyperglycemia in diabetes, such as fasting glucose and hemoglobin (HbA1c), are currently not recommended for GDM screening. Instead, an oral glucose tolerance test is specified, which is invasive, time-consuming, and not easily accessible to many at-risk populations. In this study, we describe a multi-analyte maternal serum profile test that incorporates novel glycoprotein biomarkers and previously described GDM-associated markers. In screening for GDM by multi-analyte panel, the detection rate was 87% at a false-positive rate of 1%.