Low Energy Electron Irradiation Is a Potent Alternative to Gamma Irradiation for the Inactivation of (CAR-)NK-92 Cells in ATMP Manufacturing

Background: With increasing clinical use of NK-92 cells and their CAR-modified derivatives in cancer immunotherapy, there is a growing demand for efficient production processes of these “off-the-shelf” therapeutics. In order to ensure safety and prevent the occurrence of secondary tumors, (CAR-)NK-92 cell proliferation has to be inactivated before transfusion. This is commonly achieved by gamma irradiation. Recently, we showed proof of concept that low energy electron irradiation (LEEI) is a new method for NK-92 inactivation. LEEI has several advantages over gamma irradiation, including a faster reaction time, a more reproducible dose rate and much less requirements on radiation shielding. Here, LEEI was further evaluated as a promising alternative to gamma irradiation yielding cells with highly maintained cytotoxic effector function. Methods: Effectiveness and efficiency of LEEI and gamma irradiation were analyzed using NK-92 and CD123-directed CAR-NK-92 cells. LEE-irradiated cells were extensively characterized and compared to gamma-irradiated cells via flow cytometry, cytotoxicity assays, and comet assays, amongst others. Results: Our results show that both irradiation methods caused a progressive decrease in cell viability and are, therefore, suitable for inhibition of cell proliferation. Notably, the NKmediated specific lysis of tumor cells was maintained at stable levels for three days postirradiation, with a trend towards higher activities after LEEI treatment as compared to gamma irradiation. Both gamma irradiation as well as LEEI led to substantial DNA damage and an accumulation of irradiated cells in the G2/M cell cycle phases. In addition, transcriptomic analysis of irradiated cells revealed approximately 12-fold more differentially expressed genes two hours after gamma irradiation, compared to LEEI. Analysis of surface molecules revealed an irradiation-induced decrease in surface expression of CD56, but no changes in the levels of the activating receptors NKp46, NKG2D, or NKp30. Conclusions: The presented data show that LEEI inactivates (CAR-)NK-92 cells as efficiently as gamma irradiation, but with less impact on the overall gene expression. Due to logistic advantages, LEEI might provide a superior alternative for the manufacture of (CAR-)NK-92 cells for clinical application

Schmidt-hammer exposure ages from periglacial patterned ground (sorted circles) in Jotunheimen, Norway, and their interpretative problems

© 2016 Swedish Society for Anthropology and Geography Periglacial patterned ground (sorted circles and polygons) along an altitudinal profile at Juvflya in central Jotunheimen, southern Norway, is investigated using Schmidt-hammer exposure-age dating (SHD). The patterned ground surfaces exhibit R-value distributions with platycurtic modes, broad plateaus, narrow tails, and a negative skew. Sample sites located between 1500 and 1925 m a.s.l. indicate a distinct altitudinal gradient of increasing mean R-values towards higher altitudes interpreted as a chronological function. An established regional SHD calibration curve for Jotunheimen yielded mean boulder exposure ages in the range 6910 ± 510 to 8240 ± 495 years ago. These SHD ages are indicative of the timing of patterned ground formation, representing minimum ages for active boulder upfreezing and maximum ages for the stabilization of boulders in the encircling gutters. Despite uncertainties associated with the calibration curve and the age distribution of the boulders, the early-Holocene age of the patterned ground surfaces, the apparent cessation of major activity during the Holocene Thermal Maximum (HTM) and continuing lack of late-Holocene activity clarify existing understanding of the process dynamics and palaeoclimatic significance of large-scale sorted patterned ground as an indicator of a permafrost environment. The interpretation of SHD ages from patterned ground surfaces remains challenging, however, owing to their diachronous nature, the potential for a complex history of formation, and the influence of local, non-climatic factors

Landscape evolution in Val Mulix, eastern Swiss Alps – soil chemical and mineralogical analyses as age proxies

Towards the end of the last glacial cycle, repeated re-advances of valley glaciers in the European Alps combined with periglacial processes led to the formation of a variety of climate-related landforms. Independent age measurements of moraines and rock glacier lobes using both in-situ produced and meteoric 10Be allows for the use of soil formation as an age proxy. In this complementary study we present chemical and mineralogical data from five Podzols from Val Mulix in Eastern Switzerland. Two of them developed on granitic Lateglacial moraines (14.9 ka and 10.7 ka, respectively) and three were sampled on lobes of a morphostratigraphically connected relict rock glacier, covering an age range of approx. 10.7 ka to 8.6 ka. Besides the evaluation of the suitability of selected pedosignatures for a relative age separation, we hypothesised that these pedosignatures should give further information about the evolution of the specific sites. Although the soils had a high skeleton content and the oldest soil started its development in a slightly colder climatic phase, typical weathering trends could still be detected. Whereas weathering indices such as the (K + Ca)/Ti ratio or the B-index reflect time trends reasonably well, the mineralogical composition of the fine earth and clay fraction yielded a slightly more inconsistent picture; to a lesser extent, some inconsistencies were also exhibited when using the weathering mass balance approach. This is especially true for the relict rock glacier and it supports the suggested complex development history of these soils as well as the presence of pre-weathered material. Techniques that include several surface soil horizons and the soil skeleton such as the (K + Ca)/Ti ratio, the B-index and the mass balance approach gave more robust results (in terms of the expected chronology) than the ones that only referred to single horizons (clay mineralogy). Errors or variations due to potential reallocation processes within the soil horizons but without a prominent change of the overall soil characteristics are minimised using such an approach. Weathering indexes and the clay mineral assemblage provided a differentiation of soils even within a relatively narrow time range and gave insight into processes that have occurred at the specific sites. The combined relative-numerical dating approach used here not only enables an extended interpretation and mutual control, but ultimately leads to a better understanding of landscape reconstruction and evolution

10Be inventories in Alpine soils and their potential for dating land surfaces

To exploit natural sedimentary archives and geomorphic landforms it is necessary to date them first. Landscape evolution of Alpine areas is often strongly related to the activities of glaciers in the Pleistocene and Holocene. At sites where no organic matter for radiocarbon dating exists and where suitable boulders for surface exposure dating (using in situ produced cosmogenic nuclides) are absent, dating of soils could give information about the timing of landscape evolution. This paper explores the applicability of soil dating using the inventory of meteoric 10Be in Alpine soils. For this purpose, a set of 6 soil profiles in the Swiss and Italian Alps was investigated. The surface at these sites had already been dated (using the radiocarbon technique or the surface exposure determination using in situ produced 10Be). Consequently, a direct comparison of the ages of the soils using meteoric 10Be and other dating techniques was made possible. The estimation of 10Be deposition rates is subject to severe limitations and strongly influences the obtained results. We tested three scenarios using a) the meteoric 10Be deposition rates as a function of the annual precipitation rate, b) a constant 10Be input for the Central Alps, and c) as b) but assuming a pre-exposure of the parent material. The obtained ages that are based on the 10Be inventory in soils and on scenario a) for the 10Be input agreed reasonably well with the age using surface exposure or radiocarbon dating. The ages obtained from soils using scenario b) produced ages that were mostly too old whereas the approach using scenario c) seemed to yield better results than scenario b). Erosion calculations can, in theory, be performed using the 10Be inventory and 10Be deposition rates. An erosion estimation was possible using scenario a) and c), but not using b). The calculated erosion rates using these scenarios seemed to be plausible with values in the range of 0-57�mm/ky. The dating of soils using 10Be has several potential error sources. Analytical errors as well as errors from other parameters such as bulk soil density and soil skeleton content have to be taken into account. The error range was from 8 up to 21. Furthermore, uncertainties in estimating 10Be deposition rates substantially influence the calculated ages. Relative age estimates and, under optimal conditions, absolute dating can be carried out. Age determination of Alpine soils using 10Be gives another possibility to date surfaces when other methods fail or are not possible at all. It is, however, not straightforward, quite laborious and may consequently have some distinct limitations

Exploring the potential of luminescence methods for dating Alpine rock glaciers

Rock glaciers contain valuable information about the spatial and temporal distribution of permafrost. The wide distribution of these landforms in high mountains promotes them as useful archives for the deciphering of the environmental conditions during their formation and evolution. However, age constraints are needed to unravel the palaeoclimatic context of rock glaciers, but numerical dating is difficult. Here, we present a case study assessing the potential of luminescence techniques (OSL, IRSL) to date the inner sand-rich layer of active rock glaciers. We focus on the signal properties and the resetting of the signal prior to deposition by investigating single grains. While most quartz shows low signal intensities and problematic luminescence characteristics, K-feldspar exhibits much brighter and well-performing signals. Most signals from plagioclases do not show suitable properties. Luminescence signals far below saturation indicate distinct but differential bleaching. The finite mixture model was used to determine the prominent populations in the equivalent dose distributions. The luminescence ages represent travel times of grains since incorporation into the rock glacier and hence, minimum ages of rock glacier formation. Luminescence ages between 3 ka and 8 ka for three rock glaciers from the Upper Engadine and Albula region (Swiss Alps) agree well with independent age estimates from relative and semi-quantitative approaches. Therefore, luminescence seems to have the potential of revealing age constraints about processes related to the formation of rock glaciers, but further investigations are required for solving some of the problems remaining and reducing the dating uncertainties

A combination of relative-numerical dating methods indicates two high Alpine rock glacier activity phases after the glacier advance of the Younger Dryas

To exploit the potential of rock glaciers as indicators of past climate condition it is first necessary to date them. The combined application of both relative and absolute dating techniques is a promising approach. In this study, we present Schmidt-hammer rebound value measurements and weathering rind thicknesses on four active and one relict rock glacier in the Albula area of the eastern Swiss Alps. Associated landforms such as the moraines in front of rock glaciers and glacially polished bedrock also were used to set up the temporal framework. This was done using soil chemical analyses, radiocarbon dating of the stable fraction of soil organic matter and surface exposure dating of boulders. Schmidt-hammer and weathering rind measurements showed, in most cases, well-pronounced trends with increasing surface ages. These values are in line with measurements from other nearby rock glaciers with comparable lithologies. Use of this information together with the numeric ages makes it possible to derive two main activity phases: one started soon after the ice retreat following the Younger Dryas, the main activity occurred most likely in the early Holocene and lasted approximately until the Holocene climate optimum. The second activity phase continues today and had an unclear start between 10 to 6 cal ky BP

Comparison of exposure ages and spectral properties of rock surfaces in steep, high alpine rock walls of Aiguille du Midi, France

Among various factors, permafrost and frost-thaw cycles play an important role for the stability of steep rock slopes in high alpine regions. Climate change in general and local temperature and precipitation trends in particular are likely to influence permafrost and, consequently, also the stability of rock walls. As stress relief following deglaciation can be excluded at Aguille du Midi (France), rockfall activity is mainly related to changes in permafrost and frost-thaw cycles. To put modern observations of possible climate-induced rockfalls into perspective, information on past rockfall activity is required. In this study, we investigated a combination of surface exposure dating and spectrometry to derive a correlation between rock surface ages and their spectral properties in homogenous lithology. The surface ages found varied from less than 2,000 years to around 40,000 years, and showed a clear correlation with reflectance behavior in the range 380–580 nm. These results may be a first step towards the possible generation of spatial data fields of age distribution in steep rock walls. This may provide deeper insights into spatial and temporal rock-wall development of permafrost in high alpine permafrost environments

To what extent do water isotope records from low accumulation Alpine ice cores reproduce instrumental temperature series?

Among Alpine ice core drilling sites, the Colle Gnifetti glacier saddle situated in the Monte Rosa summit range is the only one whose net snow accumulation rate is low enough to offer climate records back to some 1000 yr. It is demonstrated that the strong snow erosion at this site particularly hampers the interpretation of stable water isotope records δ18O, δD in terms of atmospheric temperature changes. We evaluate the δ18O records from four Colle Gnifetti cores for their common variability to extract a composite isotope record that may be compared with the instrumental temperature evidence. Time series analyses over the last 120 yr reveal that the common δ18O signal is mainly reflected in the low frequency variability, starting at the decadal scale. Comparing the correspondingly smoothed composite record to the high-elevation temperature time series (specifically adjusted to the seasonality of the net snow accumulation) reveals the following findings: On the decadal scale, the isotope variability correlates with the temperature record at around R=0.65 but is interrupted by three, ca. 10-yr long mismatch periods. The multidecadal isotope signal closely reflects the strong overall 20th century temperature increase, thereby showing an up to three-fold higher isotope temperature sensitivity than commonly assumed. Over the entire instrumental period back to 1760, five more such mismatch periods are embedded in the generally coherent pattern of the δ18O and instrumental temperature records (including the strong overestimate of the temperature around 1850 by the isotope temperature proxy). For the early instrumental period (1890–1760) characterized by a comparably weak long-term temperature trend, the isotope signal generally suggests warmer conditions of about 0.4°C compared to instrumental data