Quantifying the limits of CAR T-cell delivery in mice and men

CAR (Chimeric Antigen Receptor) T cells have demonstrated clinical success for the treatment of multiple lymphomas and leukaemias, but not for various solid tumours, despite promising data from murine models. Lower effective CAR T-cell delivery rates to human solid tumours compared to haematological malignancies in humans and solid tumours in mice might partially explain these divergent outcomes. We used anatomical and physiological data for human and rodent circulatory systems to calculate the typical perfusion of healthy and tumour tissues, and estimated the upper limits of immune cell delivery rates across different organs, tumour types and species. Estimated maximum delivery rates were up to 10 000-fold greater in mice than humans yet reported CAR T-cell doses are typically only 10–100-fold lower in mice, suggesting that the effective delivery rates of CAR T cells into tumours in clinical trials are far lower than in corresponding mouse models. Estimated delivery rates were found to be consistent with published positron emission tomography data. Results suggest that higher effective human doses may be needed to drive efficacy comparable to mouse solid tumour models, and that lower doses should be tested in mice. We posit that quantitation of species and organ-specific delivery and homing of engineered T cells will be key to unlocking their potential for solid tumours.     

The effect of fertilizer nitrogen input to grass-clover swards and calving date on the productivity of pasture-based dairy production

The objective of this systems-scale study was to investigate grazing season timeframes on pasture and milk production and on milk processability of dairy systems with compact spring-calving dairy cows grazing white clover (Trifolium repens L.) based grassland. Fifty-four primiparous and multiparous Holstein-Friesian dairy cows were used in a one-factor study with 3 systems (n = 18) and repeated over 2 yr (2008/09 and 2009/10). The 3 systems were: early spring calving with annual fertilizer N input of 100 kg·ha^?1 applied in spring (ES100N; 2.1 cows·ha^?1; grazing February to November), early spring calving without fertilizer N (ES0N; 1.6 cows·ha^?1; grazing February to November) and late spring calving without fertilizer N (LS0N; 1.53 cows·ha^?1; grazing April to January). Annual pasture production was affected by an interaction between grazing system and year: Mean annual pasture yields for 2008 and 2009 were ES100N; 10.35 and 9.88, ES0N; 8.88 and 8.63, LS0N; 9.18 and 10.31 t of dry matter (DM)·ha^?1 (SEM 0.39). LS0N had higher pasture DM yield in 2009 due to higher clover DM production and biological N fixation compared with the other systems. Clover stolon and root mass in the following February was correlated with stolon and root mass in the previous November with 64% of stolon mass present on LS0N in February (R² = 0.84). There were no detectable differences in per-lactation milk yield (6,335 kg·cow^?1), fat, protein and lactose yields (271, 226, 297 kg·cow^?1, respectively), cow liveweight (585 kg) or body condition score (3.02). Although winter grazing favored subsequent clover DM production, biological N fixation and pasture DM production, delaying calving date in spring and extending lactation into the following winter led to inefficient use of this pasture by the grazing herd and lowered the quality of late-lactation milk for processing purposes. Hence, a mean calving date in mid- to late-February is recommended for zero-fertilizer N input clover-based grassland.     

Deuterium kinetic isotopic study for hydrogenolysis of ethyl butyrate

The hydrogenation of ethyl butyrate, n-butyric acid, and n-butyraldehyde to their corresponding alcohol(s) has been studied over a γ-Al₂O₃-supported cobalt catalyst using a high-pressure fixed-bed reactor in the temperature range of 473–493 K. H₂–D₂–H₂ switching experiments show that ethyl butyrate and n-butyric acid follow an inverse kinetic isotope effect (KIE) (i.e. r^H/r^D = 0.50–0.54), whereas n-butyraldehyde did not display any KIE (i.e. r^H/r^D = 0.98). DRIFTS experiments were performed over the support and catalyst to monitor the surface species formed during the adsorption of ethyl butyrate and n-butyric acid at atmospheric pressure and the desired temperature. Butanoate and butanoyl species are the stable surface intermediates formed during hydrogenation of ethyl butyrate. Hydrogenation of butanoate to a partially hydrogenated intermediate is likely involved in the rate-determining step of ethyl butyrate and butyric acid hydrogenation.     

Time series shapelets: a novel technique that allows accurate, interpretable and fast classification

Classification of time series has been attracting great interest over the past decade. While dozens of techniques have been introduced, recent empirical evidence has strongly suggested that the simple nearest neighbor algorithm is very difficult to beat for most time series problems, especially for large-scale datasets. While this may be considered good news, given the simplicity of implementing the nearest neighbor algorithm, there are some negative consequences of this. First, the nearest neighbor algorithm requires storing and searching the entire dataset, resulting in a high time and space complexity that limits its applicability, especially on resource-limited sensors. Second, beyond mere classification accuracy, we often wish to gain some insight into the data and to make the classification result more explainable, which global characteristics of the nearest neighbor cannot provide. In this work we introduce a new time series primitive, time series shapelets, which addresses these limitations. Informally, shapelets are time series subsequences which are in some sense maximally representative of a class. We can use the distance to the shapelet, rather than the distance to the nearest neighbor to classify objects. As we shall show with extensive empirical evaluations in diverse domains, classification algorithms based on the time series shapelet primitives can be interpretable, more accurate, and significantly faster than state-of-the-art classifiers.     

Experimental Evidence of an Age-Specific Shift in Chemical Detection of Predators in a Lizard

The risk posed by predation is one of the most fundamental aspects of an animal's environment. Avoidance of predators implies an ability to obtain reliable information about the risk of predation, and for many species, chemosensory cues are likely to be an important source of such information. Chemosensory cues reliably reveal the presence of predators or their presence in the recent past. We used retreat site selection experiments to test whether the Australian scincid lizard Eulamprus heatwoleiM uses chemical cues for predator detection and avoidance. Both adult and juvenile lizards were given the choice of retreat sites treated with scents from invertebrate predators, as well as sympatric and allopatric snake predators. Some of the snake predators were known to eat E. heatwolei, while others did not pose a predation threat. All invertebrate predators posed a risk to juveniles, but not adults because of their size. We found that juvenile E. heatwolei avoided predator odors more strongly than adults. Juveniles avoided both invertebrate predators and snakes, and the strongest response was toward the funnelweb spider, the only ambush predator used in this experiment. This result may demonstrate the importance of predator ecology in the evolution of predator detection mechanisms, with chemical cues being more useful in detecting sedentary predators than active predators. Adult lizards showed no avoidance behavior toward predator odors. This result suggests an age specific shift in predator avoidance behavior as lizards get older and become too large for many predators. However, adults showed no response to the odor from the red-bellied black snake, a known predator of adult E. heatwolei. This finding further demonstrates the importance of predator ecology when examining communication between predators and prey. Chemical cues, which are persistent long after predators have vacated the area, may not be useful in detecting the red-bellied black snake, a wide-ranging active forager.     

Supporting exact indexing of arbitrarily rotated shapes and periodic time series under Euclidean and warping distance measures

Shape matching and indexing is important topic in its own right, and is a fundamental subroutine in most shape data mining algorithms. Given the ubiquity of shape, shape matching is an important problem with applications in domains as diverse as biometrics, industry, medicine, zoology and anthropology. The distance/similarity measure for used for shape matching must be invariant to many distortions, including scale, offset, noise, articulation, partial occlusion, etc. Most of these distortions are relatively easy to handle, either in the representation of the data or in the similarity measure used. However, rotation invariance is noted in the literature as being an especially difficult challenge. Current approaches typically try to achieve rotation invariance in the representation of the data, at the expense of discrimination ability, or in the distance measure, at the expense of efficiency. In this work, we show that we can take the slow but accurate approaches and dramatically speed them up. On real world problems our technique can take current approaches and make them four orders of magnitude faster without false dismissals. Moreover, our technique can be used with any of the dozens of existing shape representations and with all the most popular distance measures including Euclidean distance, dynamic time warping and Longest Common Subsequence. We further show that our indexing technique can be used to index star light curves, an important type of astronomical data, without modification. Reproducible Research Statement: All datasets and images used in this work are freely available at .     

Derivation of HLA-A11/Kb transgenic mice: functional CTL repertoire and recognition of human A11-restricted CTL epitopes

Transgenic mice expressing chimeric human (alpha1 and alpha2 HLA-A11 domains) and murine (alpha3, transmembrane, and cytoplasmic H-2Kb domains) class I molecules were derived. These mice were used as a model system to study the immunogenicity of human CTL epitopes and also to examine the aspects of Ag processing differences of mice vs man. Immunization of these mice with seven known HLA-A11-restricted CTL epitopes emulsified in IFA resulted in vigorous specific CTL responses. A larger panel of 45 A11-binding peptides was used to examine the relationship between immunogenicity in the HLA-A11/Kb transgenic mice and HLA-A11 binding capacity. Twenty-one of 28 (75%) peptides with high binding affinities (50% inhibitory concentration (IC50), 2-50 nM) and 7 of 13 (54%) intermediate binding peptides (IC50, 50-500 nM range) were immunogenic. In parallel, 19 of these peptides were used for in vitro primary immunizations of PBMC derived from HLA-A11 healthy human donors. It was found that 8 of 8 peptides that were able to elicit CTL in primary human in vitro cultures were also immunogenic in HLA-A11/Kb mice. Finally, HLA-A11/Kb transgenic mice were found to generate an A11/Kb restricted CTL response following immunization with influenza virus A/PR/8/34, suggesting that, at least to some extent, A11 epitopes are generated by transgenic mice as a result of natural in vivo processing and presentation.     

Alterations in the expression of P2X1 receptors in failing and nondiseased human atria

This is the first report of the analysis of the ATP-specific P2X1 receptor subunit in human hearts. We have examined homogenate samples of human left atria for the presence of P2X1 receptors using Western blots. Anti-P2X1 immunoreactivity was detected in populations of nondiseased atria as well as in atria from explanted hearts from patients with terminally failing heart conditions such as dilated cardiomyopathy. At least three groups of P2X1 immunoreactive proteins were detected in the Western blots with approximate molecular mass values of 50, 70, and 160 kDa. We report changes in expression of their 50 and 70 kDa components. These changes may be related to the type of deficit in these hearts since the changes have been observed in hearts with decreased ejection fractions characteristic of dilated cardiomyopathy.