Illite polytype quantification allows the differentiation of diagenetic and detrital illite components. In Paleozoic shales from the Illinois Basin, we observe 3 polytypes: 1Md, 1M and 2M1. 1Md and 1M are of diagenetic origin and 2M1 is of detrital origin. In this paper, we compare experimental X-ray diffraction (XRD) traces with traces calculated using WILDFIRE© and quantify mixtures of all 3 polytypes, adjusting the effects of preferred orientation and overlapping peaks. The broad intensity (“illite hump”) around the illite 003, which is very common in illite from shales, is caused by the presence of 1Md illite and mixing of illite polytypes and is not an artifact of sample preparation or other impurities in the sample. Illite polytype quantification provides a tool to extrapolate the K/Ar age and chemistry of the detrital and diagenetic end-members by analysis of different size fractions containing different proportions of diagenetic and detrital illite polytypes.

Small scale fabrication spaces have shown their potential to support our local supply chains during the collapse of many global supply chain networks at the onset of the COVID-19 pandemic. In anticipation of these spaces becoming more significant in local supply chains, it is increasingly important to reduce their environmental impacts. This work investigates the life cycle of small scale fabrication spaces by interviewing 18 participants from these spaces in the United States. Key insights from the interviews include the following: a) material selection, robust inventory management, and user support for material disposal are factors influencing optimal flow of materials and equipment through a fabrication space; b) lack of information from manufacturers and suppliers is a critical obstacle to achieve optimal use of materials and equipment, and informed decision making related to environmental sustainability and ethical labor practices; c) there are opportunities to take advantage of where financial and sustainability goals align; d) individual motivators for fabrication influences sustainable behaviors; and e) effective education about material and equipment use helps fabrication space users with more sustainable decision making.

The remarkable archaeological record of Neolithic Orkney has ensured that these islands play a prominent role in narratives of European late prehistory, yet knowledge of the subsequent Bronze Age is comparatively poor. The Bronze Age settlement and cemetery at the Links of Noltland, on the island of Westray, offers new evidence, including aDNA, that points to a substantial population replacement between the Late Neolithic and Bronze Age. Focusing on funerary practice, the authors argue for interconnecting identities centred on household and community, patrilocality and inheritance. The findings prompt a reconsideration of the Orcadian Bronze Age, with wider implications for population movement and the uptake of cultural innovations more widely across prehistoric north-western Europe.

Increasingly digital products and services make cybersecurity a crucial issue for designers. However, human-centered designers struggle to consider it in their work, partially a consequence of the high psychological distance between designers and cybersecurity. In this work, we build on the Design for Cybersecurity (DfC) Cards, an intervention to help designers consider cybersecurity, and examine a project-based design course to understand how and why specific DfC cards were used. Three findings result. First, designers found the intervention useful across all design phases and activities. Second, the cards helped design teams refocus their attention on the problem domain and project outcome. Third, we identify a need for support in framing and converging during user research, opportunity identification, and prototyping. We argue that the psychological distance between designers and the problem space of cybersecurity partially explains these findings, and ultimately exacerbates existing challenges in the design process. These findings suggest that design interventions must consider the psychological distance between designer and problem space, and have application in design practice across many complex problem domains.

OBJECTIVES/SPECIFIC AIMS: The goal for this project is to determine the feasibility of using a novel multi-photon fiber-coupled microscope to aid surgeons in localizing STN during surgeries. In order to accomplish this goal, we needed to identify the source of a strong autofluorescent signal in the STN and determine whether we could use image classification methods to automatically distinguish STN from surrounding brain regions. METHODS/STUDY POPULATION: We acquired 3 cadaveric brains from the University of Colorado Anschutz Medical Campus, Department of Pathology. Two of these brains were non-PD controls whereas 1 was diagnosed with PD. We dissected a 10 square centimeter region of midbrain surrounding STN, then prepared this tissue for slicing on a vibratome or cryostat. Samples were immuno-labeled for various cellular markers for identification, or left unlabeled in order to observe the autofluorescence for image classification. RESULTS/ANTICIPATED RESULTS: The border of STN is clearly visible based on the density of a strong autofluorescent signal. The autofluorescent signal is visible using 2-photon (850–1040 nm excitation) and conventional confocal microscopy (488–647 nm excitation). We were also able to visualize blood vessels with second harmonic generation. The autofluorescent signal is quenched by high concentrations of Sudan-black B (0.5%–5%), and is primarily localized in microtubule-associated protein-2 (MAP2)+ cells, indicating that it is likely lipofuscin accumulation in neurons. Smaller lipofuscin particles also accumulate in microglia, identified based on ionized calcium binding adopter 1 (Iba1)+ labeling. We anticipate that colocalization analysis will confirm these qualitative observations. Using 2-photon images of the endogenous autofluorescent signal in these samples, we trained a logistic regression-based image classifier using features derived from gray-level co-occurrence matrices. Preliminary testing indicates that our classifier performed well, with a mean accuracy of 0.89 (standard deviation of 0.11) and a Cohen’s Kappa value of 0.76 (standard deviation of 0.24). We are currently using coherent anti-Stokes Raman scattering and third harmonic imaging to identify different features of myelin that can be used to distinguish between these regions and expect similar results. DISCUSSION/SIGNIFICANCE OF IMPACT: Traditional methods for localizing STN during DBS surgery include the use of stereotactic coordinates and multi-electrode recording (MER) during implantation. MERs are incredibly useful in DBS surgeries, but require penetration of brain structures in order to infer location. Using multi-photon microscopy techniques to aid identification of STN during DBS surgeries offers a number of advantages over traditional methods. For example, blood vessels can be clearly identified with second harmonic generation, something that is not possible with MER. Multi-photon microscopy also allows visualization deep into tissue without actually penetrating it. This ability to look within a depth of field is useful for detection of STN borders based on autofluorescent cell density. When combined with traditional stereotactic information, our preliminary image classification methods are a fast, reliable way to provide surgeons with extra information concerning their location in the midbrain. We anticipate that future advancements and refinements to our image classifier will only increase accuracy and the potential applications and value. In summary, these preliminary data support the feasibility of multi-photon microscopy to aid in the identification of target brain regions during DBS surgeries. The techniques described here complement and enhance current stereotactic and electrophysiological methods for DBS surgeries.

The final effort of the CLIMAP project was a study of the last interglaciation, a time of minimum ice volume some 122,000 yr ago coincident with the Substage 5e oxygen isotopic minimum. Based on detailed oxygen isotope analyses and biotic census counts in 52 cores across the world ocean, last interglacial sea-surface temperatures (SST) were compared with those today. There are small SST departures in the mid-latitude North Atlantic (warmer) and the Gulf of Mexico (cooler). The eastern boundary currents of the South Atlantic and Pacific oceans are marked by large SST anomalies in individual cores, but their interpretations are precluded by no-analog problems and by discordancies among estimates from different biotic groups. In general, the last interglacial ocean was not significantly different from the modern ocean. The relative sequencing of ice decay versus oceanic warming on the Stage 6/5 oxygen isotopic transition and of ice growth versus oceanic cooling on the Stage 5e/5d transition was also studied. In most of the Southern Hemisphere, the oceanic response marked by the biotic census counts preceded (led) the global ice-volume response marked by the oxygen-isotope signal by several thousand years. The reverse pattern is evident in the North Atlantic Ocean and the Gulf of Mexico, where the oceanic response lagged that of global ice volume by several thousand years. As a result, the very warm temperatures associated with the last interglaciation were regionally diachronous by several thousand years. These regional lead-lag relationships agree with those observed on other transitions and in long-term phase relationships; they cannot be explained simply as artifacts of bioturbational translations of the original signals.

Amphibian populations have been declining worldwide, with multiple potential causes. At La Selva field station in north-eastern Costa Rica, previous work has shown that populations of many amphibians have decreased significantly since the 1970s, especially in primary forest. Starting in 1998, we investigated one of the most common frog species at La Selva, the poison-dart frog Oophaga pumilio (= Dendrobates pumilio). In a survey of 50 plots of 100 m2 in 1998, adult frogs were 4.6 times more abundant in secondary forest than in primary forest. Tadpoles were found only in secondary-forest plots. Almost all (89%) of the tadpoles were found in leaf axils of Dieffenbachia spp., which were much more abundant in secondary-forest than in primary-forest plots. The greater abundance of Dieffenbachia spp. in secondary forest was confirmed in a broad survey of ~11 km of trails within La Selva in 2002. When the same trails were resampled in 2012, Dieffenbachia spp. had been extirpated from 72% of the 50-m segments where plants were present in 2002; abundance was greatly reduced in the few trail segments where any Dieffenbachia spp. remained in 2012. The loss of Dieffenbachia spp., especially in secondary forest, removed the species most often used by O. pumilio for tadpole rearing. Based on counts of calling frogs in 2010, there was no difference in O. pumilio abundance in primary versus secondary forest, in striking contrast to multiple earlier surveys that found much greater frog abundance in secondary forest. We propose that the reason for the rapid decline in Dieffenbachia spp. is herbivory by the collared peccary (Pecari tajacu), which has increased in abundance at La Selva in recent years. A likely consequence is continued reduction in O. pumilio populations.