bluuebunny/crossref_metadata_2025 huggingface dataset:
Hyperinsulinemia does not cause de novo capillary recruitment in rat skeletal muscle
Authors: Akerstrom Thorbjorn , Goldman Daniel , Nilsson Franciska , Milkovich Stephanie L. , Fraser Graham M. , Brand Christian Lehn , Hellsten Ylva , Ellis Christopher G.
Published: October 2019

Objective

The effect of insulin on blood flow distribution within muscle microvasculature has been suggested to be important for glucose metabolism. However, the “capillary recruitment” hypothesis is still controversial and relies on studies using indirect contrast‐enhanced ultrasound (CEU) methods.

Methods

We studied how hyperinsulinemia effects capillary blood flow in rat extensor digitorum longus (EDL) muscle during euglycemic hyperinsulinemic clamp using intravital video microscopy (IVVM). Additionally, we modeled blood flow and microbubble distribution within the vascular tree under conditions observed during euglycemic hyperinsulinemic clamp experiments.

Results

Euglycemic hyperinsulinemia caused an increase in erythrocyte (80 ± 25%, P < .01) and plasma (53 ± 12%, P < .01) flow in rat EDL microvasculature. We found no evidence of de novo capillary recruitment within, or among, capillary networks supplied by different terminal arterioles; however, erythrocyte flow became slightly more homogenous. Our computational model predicts that a decrease in asymmetry at arteriolar bifurcations causes redistribution of microbubble flow among capillaries already perfused with erythrocytes and plasma, resulting in 25% more microbubbles flowing through capillaries.

Conclusions

Our model suggests increase in CEU signal during hyperinsulinemia reflects a redistribution of arteriolar flow and not de novo capillary recruitment. IVVM experiments support this prediction showing increases in erythrocyte and plasma flow and not capillary recruitment.
Genome‐wide association mapping of QTLs implied in potato virus Y population sizes in pepper: evidence for widespread resistance QTL pyramiding
Authors: Tamisier Lucie , Szadkowski Marion , Nemouchi Ghislaine , Lefebvre Véronique , Szadkowski Emmanuel , Duboscq Renaud , Santoni Sylvain , Sarah Gautier , Sauvage Christopher , Palloix Alain , Moury Benoit
Published: October 2019

In this study, we looked for genetic factors in the pepper (Capsicum annuum) germplasm that control the number of potato virus Y (PVY) particles entering the plant (i.e. effective population size at inoculation) and the PVY accumulation at the systemic level (i.e. census population size). Using genotyping‐by‐sequencing (GBS) in a core collection of 256 pepper accessions, we obtained 10 307 single nucleotide polymorphisms (SNPs) covering the whole genome. Genome‐wide association studies (GWAS) detected seven SNPs significantly associated with the virus population size at inoculation and/or systemic level on chromosomes 4, 6, 9 and 12. Two SNPs on chromosome 4 associated with both PVY population sizes map closely to the major resistance gene pvr2 encoding the eukaryotic initiation factor 4E. No obvious candidates for resistance were identified in the confidence intervals for the other chromosomes. SNPs detected on chromosomes 6 and 12 colocalized with resistance quantitative trait loci (QTLs) previously identified with a biparental population. These results show the efficiency of GBS and GWAS in C. annuum, indicate highly consistent results between GWAS and classical QTL mapping, and suggest that resistance QTLs identified with a biparental population are representative of a much larger collection of pepper accessions. Moreover, the resistance alleles at these different loci were more frequently combined than expected by chance in the core collection, indicating widespread pyramiding of resistance QTLs and widespread combination of resistance QTLs and major effect genes. Such pyramiding may increase resistance efficiency and/or durability.
Th2 signals are not essential for the anti‐arthritic effects of Trichinella spiralis in mice
Authors: Osada Yoshio , Morita Kentaro , Tahara Sayaka , Ishihara Tsubasa , Wu Zhiliang , Nagano Isao , Maekawa Yoichi , Nakae Susumu , Sudo Katsuko , Kanazawa Tamotsu
Published: October 2019

Aims

Many parasitic helminths are known to alter host immune responses and consequently affect the progression of autoimmune and allergic diseases. The parasitic nematode Trichinella sp has been reported to suppress several experimental diseases in rodents, including experimental autoimmune encephalomyelitis, type 1 diabetes, colitis, airway inflammation and autoimmune arthritis. We tried to clarify requirement of Th2 cytokines in the anti‐arthritic effects of Trichinella spiralis (Ts) against collagen‐induced arthritis (CIA).

Methods and results

We infected Ts and then induced CIA in STAT6KO DBA/1 mice, comparing the disease progression with that in wild‐type (WT) DBA/1 mice, Ts significantly mitigated arthritis in WT mice, in addition to the impairment of anti–type II collagen (IIC) IgG production in a subclass‐independent manner. The genetic absence of STAT6 in the mice did not abrogate the anti‐arthritic effects of Ts. Alteration of splenic cytokines was not related to the anti‐arthritic effects of the parasite. Moreover, lack of IL‐10 did not abrogate the anti‐arthritic effects of Ts.

Conclusion

Our results suggest that the anti‐arthritic effects of Ts do not require host Th2 signals.
Selective mineral transport barriers at Cuscuta‐host infection sites
Authors: Förste Frank , Mantouvalou Ioanna , Kanngießer Birgit , Stosnach Hagen , Lachner Lena Anna‐Maria , Fischer Karsten , Krause Kirsten
Published: October 2019

The uptake of inorganic nutrients by rootless parasitic plants, which depend on host connections for all nutrient supplies, is largely uncharted. Using X‐ray fluorescence spectroscopy (XRF), we analyzed the element composition of macro‐ and micronutrients at infection sites of the parasitic angiosperm Cuscuta reflexa growing on hosts of the genus Pelargonium. Imaging methods combining XRF with 2‐D or 3‐D (confocal) microscopy show that most of the measured elements are present at similar concentrations in the parasite compared to the host. However, calcium and strontium levels drop pronouncedly at the host/parasite interface, and manganese appears to accumulate in the host tissue surrounding the interface. Chlorine is present in the haustorium at similar levels as in the host tissue but is decreased in the stem of the parasite. Thus, our observations indicate a restricted uptake of calcium, strontium, manganese and chlorine by the parasite. Xylem‐mobile dyes, which can probe for xylem connectivity between host and parasite, provided evidence for an interspecies xylem flow, which in theory would be expected to carry all of the elements indiscriminately. We thus conclude that inorganic nutrient uptake by the parasite Cuscuta is regulated by specific selective barriers whose existence has evaded detection until now.
Trickle‐down debt: Infrastructure, development, and financialisation, Medellín 1960–2013
Authors: Furlong Kathryn
Published: October 2019

In many Latin American cities, infrastructure was largely financed through development lending over the second half of the 20th century. Exacerbated by debt crises and currency devaluations, public utilities became holders of significant levels of negative value. This encouraged public debt financialisation in order to mitigate the effects of shifting interest rates and devaluation. For David Harvey, negative value is the hallmark of contemporary capitalism whereby one must produce, not for profit, but to retire debt. This statement can be applied to indebted utilities, in the sense that the focus of utility governance – and its relationship towards those dependent on it for services – becomes reoriented towards debt management – or governing by debt. Full‐cost recovery emerges in this context as a mechanism to pay down the infrastructure debt held by utilities, which quickly led to increasing levels of user indebtedness. Service disconnection and pre‐paid metering emerge as processes to recover this user debt by enforcing a culture of payment through service exclusion. In these ways, the responsibility for infrastructure debt ‘trickles down’ in small – but individually significant – amounts to persons and households, enrolling them in the logic of debt (re)payment. This paper examines these issues through a case study of urban infrastructure financing, debt, and tariffs in Medellín, Colombia from 1960 to 2013.
The human central nervous system transmits common synaptic inputs to distinct motor neuron pools during non‐synergistic digit actions
Authors: Del Vecchio A. , Germer C. M. , Elias L. A. , Fu Q. , Fine J. , Santello M. , Farina D.
Published: October 2019

Key points

Neural connectivity between distinct motor neuronal modules in the spinal cord is classically studied through electrical stimulation or multi‐muscle EMG recordings.

We quantified the strength of correlation in the activity of two distinct populations of motor neurons innervating the thenar and first dorsal interosseous muscles during tasks that required the two hand muscles to exert matched or un‐matched forces in different directions.

We show that when the two hand muscles are concurrently activated, synaptic input to the two motor neuron pools is shared across all frequency bandwidths (representing cortical and spinal input) associated with force control.

The observed connectivity indicates that motor neuron pools receive common input even when digit actions do not belong to a common behavioural repertoire.

Neural connectivity between distinct motor neuronal modules in the spinal cord is classically studied through electrical stimulation or multi‐muscle EMG recordings. Here we quantify the strength of correlation in the activity of two distinct populations of motor neurons innervating the thenar and first dorsal interosseous muscles in humans during voluntary contractions. To remove confounds associated with previous studies, we used a task that required the two hand muscles to exert matched or un‐matched forces in different directions. Despite the force production task consisting of uncommon digit force coordination patterns, we found that synaptic input to motor neurons is shared across all frequency bands, reflecting cortical and spinal inputs associated with force control. The coherence between discharge timings of the two pools of motor neurons was significant at the delta (0–5 Hz), alpha (5–15 Hz) and beta (15–35 Hz) bands (P < 0.05). These results suggest that correlated input to motor neurons of two hand muscles can occur even during tasks not belonging to a common behavioural repertoire and despite lack of common innervation. Moreover, we show that the extraction of activity from motor neurons during voluntary force control removes cross‐talk associated with global EMG recordings, thus allowing direct in vivo interrogation of spinal motor neuron activity.
Then I Saw
Authors: Low U-Wen
Published: October 2019

In 1498, artist Albrecht Dürer published a series of woodcuts depicting scenes from the Book of Revelation, entitled Apocalypse. The impact and reach of Dürer’s work has been widely acknowledged by scholars, and its importance is difficult to overstate. Sixteen images were produced, each depicting scenes from the text of Revelation. This was a remarkable achievement, given the complexity and difficulty of the source material, and has influenced all subsequent depictions of the Apocalypse in the Western tradition. Apocalypse is therefore defined as a paratext, an artifact that affects the reading or interpretation of the original text, in this case the Book of Revelation. This essay will briefly discuss both the origins of Apocalypse and its paratextual influence, particularly with regards to Lucas Cranach’s illustrations for the Luther Bible. This essay will also briefly visually exegete selected Apocalypse illustrations and their engagement with the text of the Book of Revelation.
Alexander of Cotiaeum Teacher, Exegete, Diorthotes
Authors: Montana Fausto
Published: October 2019

Alexander of Cotiaeum, the cultivated sophistes and one among the teachers of Aelius Aristides and Marcus Aurelius, distinguished himself in linguistic and literary studies, teaching, and cultural communication. Though without achieving brilliant results, he also engaged in some of the questions previously discussed by the most learned scholars. This cultural figure displays some typicality with respect to the average educated personalities (grammatikoi) of the Antonine renaissance. However, current studies are revealing a possible specificity of Alexander’s role: his influence, by way of educational approach, on the making of literary trends and models (canons) of the concurrent high culture, between New Sophistic and Atticism. This paper focuses on the very philological side (diorthosis, or textual criticism) of the composite and complex intellectual profile of Alexander.
The Rhetorical Commentary in Late Antiquity
Authors: Pepe Cristina
Published: October 2019

This paper examines the tradition of rhetorical exegesis on technical works – namely on Hermogenes’ treatises – flourished from the third century AD onwards. A focus on the evidence preserved proves the exegetes’ preference for the commentary format and the significant similarities in the structure and arrangement of the material with other exegetical literature of the same period. Moreover, by discussing further the content and scope of these commentaries, their relationship with teaching practice will be argued.
Micro-CT measurements of within-ring variability in longitudinal hydraulic pathways in Norway spruce
Authors: Adey-Johnson Robin , Mclean J. Paul , Van den Bulcke Jan , Van Acker Joris , McDonald Peter J.
Published: October 2019

This study aimed to define the variability in the microstructure of Norway spruce within an annual ring by examining differences between earlywood and latewood. In particular, we were interested in obtaining new information on bordered pit occurrence and locations relative to tracheid ends, plus the lumina dimensions and longitudinal overlap of tracheids that collectively define the longitudinal hydraulic pathways. A stacked series of X-ray micro-CT scans of an annual ring of Norway spruce were made and stitched together longitudinally to form a three-dimensional volume. The imaging resolution was carefully chosen to capture both longitudinal and transverse anatomical details. Measurements of tracheid length, overlap, radial lumen diameter, and bordered pit location were made semi-automatically using image analysis. The distribution of radial lumen diameter was used to define earlywood and latewood. Then bordered pit linear density and spatial distribution, tracheid length and overlap were analysed, presented and contrasted for earlywood and latewood. Further differences between earlywood and latewood were found only in bordered pit linear density. Clear trends in radial lumen diameter and pit linear density were observed with radial position within the growth ring. These results provide new information on the variability of the Norway spruce microstructure within an annual ring.
Direct exposure to solar radiation causes radial growth eccentricity at the beginning of the growing season in Robinia pseudoacacia
Authors: Miodek Adam , Gizińska Aldona , Klisz Marcin , Wojda Tomasz , Ukalski Krzysztof , Kojs Paweł
Published: October 2019

Our study investigated the effect of stem temperature increase on xylem formation inRobinia pseudoacaciatree-trunks, caused by direct exposure to solar radiation. It is important to determine factors which may improve the concentricity of deposited wood tissue and intensify xylogenesis because a strong irregularity of wood tissue deposited in the radial direction in mature trees ofR. pseudoacaciareduces the commercial value of the wood. Samples of vascular cambium along with adjacent tissues were collected from the southern (illuminated) and northern (shaded) side of tree-trunks growing in the inner and peripheral (thus exposed to direct sunlight) zones of the research plot. Sampling was performed several times during the growing season. The collected material was examined by epifluorescence microscopy and the thickness of deposited tissue comprising cambial xylem derivatives was measured. Deposition of a markedly greater amount of xylem on the southern side of tree-trunks in the peripheral zone of the plot was observed before full leaf development. Instrumental climatic data confirmed that in the early stage of the growing season, temperature on the southern side of the peripheral zone tree-trunk was higher than on the northern side. No clear response in terms of directional deposition of xylem was noticed in the inner zone trees and in peripheral zone trees after full leaf development. This study highlights the importance of temperature increase, caused by solar radiation, forR. pseudoacaciaxylogenesis, which may be considered as a factor that affects the course of the radial growth before full leaf development.
After Darwin
Authors: O’Connell Jeff , Ruse Michael
Published: October 2019

In the second half of the nineteenth century, many people lost their faith in the Christian God. Nevertheless, they were eager to show that this move towards a secular world picture did not mean the end of morality and that it could continue as much before. In a Darwinian age this was not possible and the Christian cherishing of the virtue of meekness was replaced by a moral respect for vigor and effort directed both towards self-realization and to the well-being of society. We compare the British moves to those promoted by the German philosopher Friedrich Nietzsche. There are significant similarities but also differences that reflect the British industrialized notion of progress versus the German idealistic notion of progress.
Stress and Deformation Analysis of Piezoresistive Square Diaphragm Nano Pressure Sensor
Authors: Devi Rekha , Gill Sandeep Singh
Published: October 2019

Pressure sensors are among the most widely used MEMS product as sensors in commercial and industrial applications. The diaphragm structure, thickness, and dimensions along with the placement, size, and shape of the piezo resistors are important in the process of designing a pressure
sensor. This paper presents the design and simulation of the square diaphragm to analysis the deformation and stress of various materials as well as for the different thickness of the diaphragms at low pressure in the range of 0.1 to 1.5 bar. The design of diaphragm is as a 10 μm
thick square diaphragm with 800 μm edge length are considered, which were analyzed by using ANSYS 17 software. The results show that deformation and stress reduction as in the increase in the thickness, which is directly related to the performance of the diaphragm. CNT based diaphragm
also showed the improved performance of the pressure sensor as compared to the silicon and polysilicon material.
Electromagnetic Sensor to Detect Objects by Classification and Neural Networks
Authors: Gharamohammadi Ali , Darestani Mohammad Reza Yousefi , Taghavirashidizadeh Ali , Abbasi Ahmad , Shokouhmand Arash
Published: October 2019

The backscatter signal analysis, as the landmine material could vary, has to be as much advanced as possible. One major problem with the conventional methods is that they are not able to detect new plastic landmines. In the recent research, the classification techniques and neural networks
(NNs) were exploited for detection. In NNs-based method, a network is trained based on the feature extracted from the data, which leads to landmine detection. Other conventional classification methods, attempts to classify the objects sharing common characteristics. In this letter, an algorithm
is introduced based on classification, data reduction and neural networks. Indeed, this algorithm employs neural network and classification method, simultaneously. The simple methods using either neural network or classification separately usually suffer from high rate of risk. In this letter,
a novel classifier is proposed such that the data is classified based on similarity. It will be shown that the similarity between signals in a class is more than 90%, which proves the method's efficiency. Moreover, the scattering parameter, having magnitude and phase parts, is used to create
an algorithm with parallel process.
Factors Affecting Consumer Adoption of Mobile Apps in NCR: A Qualitative Study
Authors: Arora Neerja , Malik Garima , Chawla Deepak
Published: October 2019

The recent years have seen an exponential rise in the mobile application space. Organizations strive to be present in this space for interacting with the customer. In this light, it is important to study the current usage of mobile applications and factors which affect their adoption across age groups. This research is a qualitative study, which investigates the pattern of usage, the user friendliness, the liked and disliked features of mobile apps and the beliefs associated with them. Mobile applications being a fairly recent technological advancement, it has been important to study their adoption in the light of the diffusion of innovation and investigate the effect of social influence on decisions to use mobile apps. The findings of this study are based on a total of 29 responses (19 through semi-structured interviews and 10 through focus group discussion) in National Capital Region of India (NCR). The responses were analyzed using content analysis. This study reveals differences in patterns of usage and beliefs among Gen X and Gen Y users. It was found that Gen Y users used mobile apps much more extensively than Gen X users. Though both categories of users believed that usage of mobile apps made life easier, Gen X users had negative beliefs about mobile apps. The findings also reveal the presence of the trait of innovativeness and uniqueness among Gen Y users, which leads to adoption of mobile applications.
Gender’s ontoformativity, or refusing to be spat out of reality: reclaiming queer women’s solidarity through experimental writing
Authors: Rudy Susan
Published: October 2019

In this article, I argue that queer women – especially cis and trans lesbians – have more in common than contemporary fissures either allow for or acknowledge. Lesbians who recognised their queer sexuality in the 1970s have in common with trans women the shared condition of being, in the words of the 1970s radical feminist Marilyn Frye, ‘spat summarily out of reality’. We also share the experience of refusing to accept this condition. I make this argument by manoeuvring away from questions of gender identity and focusing instead on gender’s ontoformativity: the astonishing, welcome and transformative fact that new social realities are brought into being by new social practices. I turn to experimental writing to explore this matter. Through this medium, the cis lesbian poet Nicole Brossard and the trans lesbian poet Trace Peterson wrote themselves into worlds, languages and social orders that refused to acknowledge their existence. Brossard was writing in 1970s Montreal, Peterson in early twenty-first-century New York, but what they have in common, indeed what radical lesbian theory from the 1970s shares with contemporary theorising by trans women, is the insight that identifying with men is expected. It is in identifying with women that we are most at risk.
Of lizards and ideological entrepreneurs: Alex Jones and Infowars in the relationship between populist nationalism and the post-global media ecology
Authors: Van den Bulck H , Hyzen A
Published: October 2019

This contribution analyses the nexus between contemporary US populist nationalism and the post-global media ecology through the case of US radio show host and ‘most paranoid man in America’ Alex Jones and his Infowars. It evaluates the role of Alt Right alternative/activist media and global digital platforms in the success of Jones as ideological entrepreneur. To this end, it looks at Jones’ and Infowars' message (mostly Falls Flag conspiracy theories and pseudo-science-meets-popular-culture fantasy), persona as celebrity populist spectacle, business model, political alliances with Alt Right and Trump, audience as diverse mix of believers and ironic spectators and, most of all, media. In particular, we analyse the mix of legacy and social media and their respective role in his rise and alleged downfall. We evaluate Jones’ efforts as effective ideological entrepreneur, pushing his counter-hegemonic ideology from the fringes to the mainstream.
Burden of premature atrial beats in middle-aged endurance athletes with and without lone atrial fibrillation versus sedentary controls
Authors: Cipriani Alberto , Vio Riccardo , Mastella Giulio , Ciarmatori Nicolò , Del Monte Alvise , Trovato Domenico , Iliceto Sabino , Schiavon Maurizio , Bertaglia Emanuele , Corrado Domenico , Zorzi Alessandro
Published: October 2019

Background

The burden of premature atrial beats (PABs) at 24-h electrocardiographic (ECG) monitoring correlates with the risk of atrial fibrillation. It is unknown whether prolonged and intense exercise increases the burden of PABs, thus contributing to the higher prevalence of atrial fibrillation observed in middle-aged athletes.

Methods

We compared the burden of PABs at 24-h ECG monitoring off therapy in 134 healthy middle-aged (30–60-year-old) competitive athletes who had practised 9 (7-11) h of endurance sports for 8 (4-15) consecutive years, 134 age- and gender-matched healthy sedentary individuals, and 66 middle-aged patients (20 athletes and 46 non-athletes) with ‘lone’ paroxysmal atrial fibrillation.

Results

More than 50 PABs/24 h or ≥1 run of ≥3 PABs were recorded in 23/134 (17%) healthy athletes and in 29/134 (22%) sedentary controls ( p = 0.61). Healthy athletes with frequent or repetitive PABs were older (median 50 years vs. 43 years, p < 0.01) and had practised sport for a longer time (median 10 years vs. 6 years, p = 0.03). At multivariable analysis only age (odds ratio 1.11, 95% confidence interval 1.04–1.20, p < 0.01) remained an independent predictor of a higher burden of PABs. Also among patients with ‘lone’ paroxysmal atrial fibrillation, there was no difference in the prevalence of >50 PABs/24 h or ≥1 run of ≥3 PABs between athletes (40%) and controls (48%, p = 0.74) .

Conclusions

Middle-aged endurance athletes, with or without paroxysmal atrial fibrillation, did not show a higher burden of PABs at 24-h ECG monitoring than sedentary controls. Age, but not intensity and duration of sports activity, predicted a higher burden of PABs among healthy athletes.
Strategy and Strategic Alignment
Authors: Mukherjee Indrajit
Published: October 2019

The article is written to elaborate the meaning of the term ‘strategy’ and the domain it covers. It presents the common views about what strategy is and explains the meaning of each of them. It also argues that one of the views, strategy as an overarching ‘alignment’, is a common thread running through most of the views. Two examples are given to explain this view better. The article is expected to help the reader appreciate the domain of strategy and link it with the other functional areas.
Involvement and functional impairment of the CD34+CD38−Thy-1+ hematopoietic stem cell pool in myelodysplastic syndromes with trisomy 8
Authors: Nilsson Lars , Åstrand-Grundström Ingbritt , Anderson Kristina , Arvidsson Ingrid , Hokland Peter , Bryder David , Kjeldsen Lars , Johansson Bertil , Hellström-Lindberg Eva , Hast Robert , Jacobsen Sten Eirik W.
Published: October 2019

Clonality studies of mature cells suggest that the primary transformation event in myelodysplastic syndrome (MDS) most frequently occurs in a myeloid-restricted progenitor, a hypothesis supported by recent studies of purified CD34+Thy1+hematopoietic stem cells (HSCs) in cases with trisomy 8 (+8). In contrast, we recently demonstrated that a lymphomyeloid HSC is the target for transformation in MDS cases with del(5q), potentially reflecting heterogeneity within MDS. However, since +8 is known to frequently be a late event in the MDS transformation process, it remained a possibility that CD34+CD38−Thy1+ HSC disomic for chromosome 8 might be part of the MDS clone. In the present studies, although a variable fraction of CD34+CD38−Thy1+ cells were disomic for chromosome 8, they did not possess normal HSC activity in long-term cultures and nonobese diabetic–severe combined immunodeficiency (NOD-SCID) mice. Mixing experiments with normal CD34+CD38− cells suggested that this HSC deficiency was intrinsic and not mediated by indirect mechanisms. Furthermore, investigation of 4 MDS cases with combined del(5q) and +8 demonstrated that the +8 aberration was always secondary to del(5q). Whereas del(5q) invariably occurs in CD34+CD38−Thy-1+ HSCs, the secondary +8 event might frequently arise in progeny of MDS HSCs. Thus, CD34+CD38−Thy1+ HSCs are invariably part of the MDS clone also in +8 patients, and little HSC activity can be recovered from the CD34+ CD38−Thy1+ HSC. Finally, in advanced cases of MDS, the MDS reconstituting activity is exclusively derived from the minor CD34+CD38−HSC population, demonstrating that MDS stem cells have a similar phenotype as normal HSCs, potentially complicating the development of autologous transplantation for MDS.
Baseline PET as prognostic marker for Hodgkin?
Authors: Zijlstra Josée M. , Boellaard Ronald
Published: October 2019

In this issue of Blood, Akhtari et al present a retrospective analysis of baseline positron emission tomography-computed tomography (PET-CT) in Hodgkin lymphoma (HL) in relation to the prognostic significance of the metabolic tumor volume (MTV) in risk classification of early-stage HL.1 These measurements yield estimates of the total tumor burden, which has previously been demonstrated to be one of the most significant prognostic factor in HL.2 A more precise discrimination between low-risk vs high-risk HL using baseline PET-CT characteristics could be clinically useful and might inform treatment decisions.
Aurora-B Kinase Translocates to the Midzone in Endomitotic Megakaryocytes.
Authors: Geddis Amy E. , Kaushansky Kenneth
Published: October 2019

Megakaryocyte (MK) differentiation is marked by the development of progressive polyploidy, facilitating platelet production by the creation of a large cytoplasmic volume. MKs become polyploid through repeated cycles of endomitosis (EnM), in which mitosis is initiated but subsequently aborted in late anaphase with failure to complete karyokinesis and cytokinesis. However, the mechanisms underlying EnM remain poorly understood. Recent hypotheses explored in the literature have focused on the possible absence or mislocalization of the chromosomal passenger protein Aurora-B kinase, as it has a pivotal role in many aspects of cytokinesis. Along with the other passenger proteins, Aurora-B kinase transits from the centromeres of metaphase chromosomes to the bundled microtubules of the spindle midzone and overlying cortex between separating chromosomes in anaphase. The midzone and its associated proteins, are thought to be critical for determining the position of the cleavage furrow. One of these proteins, the kinesin MKLP-2, is required for the translocation of Aurora-B kinase to the midzone, where it co-localizes with the GTPase MgcRacGAP and stimulates its activity towards RhoA, potentially regulating actin dynamics at the cleavage furrow. We have previously demonstrated that several chromosomal passenger proteins including Aurora-B kinase are normally expressed and localized to centromeres in EnM MKs. In this work, we use deconvolution microscopy in primary murine and human MKs to extend those findings and demonstrate that EnM MKs form midzone structures that are characteristic of late anaphase; in addition, Aurora-B kinase is clearly present on the spindle midzone, as are MKLP-2 and MgcRacGAP. Although we found images suggestive of initial cleavage furrow formation with cortical localization of Aurora-B kinase in late phase cells, we were unable to demonstrate enhanced localization of actin or anillin to the furrow in EnM cells, despite their normal localization in diploid control cells. Therefore, many of the components of the central spindle are intact during MK EnM, but the formation of the cleavage furrow appears to be incomplete. These data add to our understanding of the possible mechanisms underlying EnM and offer an alternative hypothesis to that of failed expression or localization of the chromosomal passenger proteins. Ongoing studies will focus on the assembly and function of the cleavage furrow in this enigmatic process.
Oligoclonal Expansion of Revertant T Cells Leads to Omenn Syndrome in a Patient with RAG1-Deficient Severe Combined Immunodeficiency.
Authors: Wada Taizo , Okamoto Hiroyuki , Toma Tomoko , Kasahara Yoshihito , Koizumi Shoichi , Kato Masahiko , Yachie Akihiro
Published: October 2019

Omenn syndrome (OS) is an autosomal recessive primary immunodeficiency characterized by early-onset generalized erythroderma, lymphadenopathy, hepatosplenomegaly, hypogammaglobulinemia, and elevated serum IgE levels. OS patients carry mutations of the recombination activating genes (RAG1 and RAG2) that are essential for V(D)J recombination. OS mutations maintain a residual recombination activity that allows limited T-cell receptor (TCR) gene rearrangements in the thymus, whereas null mutations cause a complete block of T- and B-cell development and lead to severe combined immune deficiency with absence of mature T and B lymphocytes (T-B- SCID). Revertant mosaicism due to back mutations or second-site mutations has been described in an increasing number of primary immunodeficiencies. The occurrence of reversion is considered rare and revertant cells have been shown to carry a single revertant sequence. Here we describe somatic mosaicism due to multiple events of second-site mutations in a patient affected with Omenn syndrome. We found that the patient is homozygous for a single base C deletion after nucleotide 2113 of the RAG1 gene in DNA obtained from his granulocytes. This novel mutation leads to a frameshift at codon 668 and a premature termination, and likely abrogates RAG function. The patient, however, showed typical features of OS including expansion of activated and oligoclonal T cells in peripheral blood. Molecular analysis using various sorted samples revealed that several different second-site mutations, all of which restored the RAG1 reading frame and resulted in missense mutations located in the RAG2 interacting domain, were simultaneously demonstrated in DNA from CD3+ T cells. On the contrary, such sequences were not detectable in DNA obtained from granulocytes, CD14+ monocytes and CD56+ NK cells. We also showed that TCRVβ8+ and TCRVβ1+ T cells, which were predominantly expanded within CD4+ and CD8+ T cells respectively, had different second-site mutations. The presence of multiple second-site mutations in a single patient is an unprecedented finding that supports the possibility that genetic reversions may be more common than previously thought. In addition, these observations indicate that T-B- SCID with reverse mutations that impair but do not abolish the V(D)J recombination process is a novel mechanism for the pathogenesis of OS.
Self-Reactivity in a Lymphopenic Mouse: A Model of Acute Graft-versus-Host Disease.
Authors: Knoechel Birgit , Lohr Jens G. , Kahn Estelle , Abbas Abul K.
Published: October 2019

Transfer of naive antigen-specific T cells into self-antigen expressing transgenic mice leads to the development of hyporesponsiveness and deletion of the self-reactive T cells. This form of functional tolerance is characterized by decreased recall responses both in vivo and in vitro. Although the development of tolerance has been studied extensively, few studies have analyzed the factors that determine the breakdown of tolerance and how host cell populations influence the process. Using transgenic mice that express a secreted form of ovalbumin (sOVA Tg) in the serum either on a wildtype (WT) or a T-cell deficient (TCRa−/−), or B- and T-cell deficient (Rag−/−) background, we investigate tolerance development of OVA-specific DO11.10 CD4+ T cells that have been transferred into these mice. We report kinetics, phenotypes and cytokine production of the antigen-specific T cells after transfer. We further describe the histological, immunohistochemical and clinical picture and the effects of cytokine blockade by administration of antibodies.

Naïve OVA-specific T cells that encounter OVA as a self-antigen in lymphocyte-sufficient recipients undergo some expansion, followed by a contraction phase, and become functionally hyporesponsive to the antigen. In contrast, adoptive transfer of DO11 cells into Rag-deficient sOVA Tg animals results in rapid development of wasting and death, phenotypically and histologically resembling cytokine release syndrome or acute GvHD. Histologic examination reveals inflammatory infiltrates predominantly in the skin and gut. Under these circumstances the transferred DO11 cells undergo massive expansion and produce abundant amounts of IL-2 and IFN-g. In contrast, transfer into TCRa-deficient sOVA Tg animals, in which B cells are present, leads to similar T cell expansion but substantially reduced IFN-g production and no death.

Our data suggest that T cell tolerance in vivo is critically dependent on an intact, lymphocyte-sufficient host. In the absence of endogenous lymphocytes, T cells specific for a systemic self-antigen are activated and cause lethal immune reactions.
Cancer Immunotherapy: Cytotoxic Activity of γδ T-Cells Expanded Ex Vivo by the Third Generation Bisphosphonate Zoledronate.
Authors: Sato Kiyoshi , Kimura Shinya , Yuasa Takeshi , Wada Hiromi , Maekawa Taira
Published: October 2019

Bisphosphonates (BPs), widely used to treat bone diseases, have recently been attracted much interest for their antitumor activity and have been reported to exert direct antitumor effects on several cancer cell lines via the inactivation of Ras proteins. BPs inhibit farnesyl pyrophosphate (FPP) synthetase in the mevalonate pathway and deplete cellular pools of PPs such as farnesyl PP and geranylgeranyl PP which are indispensable for the activation of Ras proteins. In addition to their direct antitumor activity, BPs expand γδ T-cells which are potent effector cells and also induce the accumulation of isopentenyl PP as a tumor antigen in target cancer cells. The purpose of this study was to clarify the cytotoxic activity of γδ T-cells expanded ex vivo by the potent third generation BP zoledronate (ZOL). Peripheral blood mononuclear cells of five healthy donors were incubated with different concentrations of ZOL and interleukin-2. After 14 days incubation, 1 μM ZOL increased the absolute number of γδ T-cells 500–800 fold. Expanded γδ T-cells were of the Vγ9Vδ2 subset and cytokine levels of IL-2, -4, -5, -10, TNF-α and IFN-γ were not elevated at resting i.e. before contact to target cancer cells. In vitro cytotoxic activities of γδ T-cells against the luciferase-labeled small cell lung cancer (SCLC) cell line SBC-5 were examined by a newly developed cytotoxic assay using an in vivo imaging system (Xenogen, Alameda, CA) and video microscopy, Leica AS MDW (Leica Microsystems Inc., Bannockburn, IL). γδ T-cells killed SBC-5 cells pre-treated with 5 μM ZOL for 12 h after 1.5–3.0 h contact with the target cells whereas untreated SBC-5 were rarely killed. SBC-5 cells pretreated with 5 μM ZOL showed a marked increase in their sensitivity to lysis by γδ T-cells, percentages of specific lysis were 42% and 55% at effector/target (E/T) ratios of 5:1 and 10:1, respectively, while those of untreated SBC-5 cells were 8% and 13% at E/T ratios of 5:1 and 10:1, respectively. In vivo efficacy of γδ T-cells was investigated in mice xenografted subcutaneously with SBC-5 cells. Pretreatment with 80 μg/kg ZOL enhanced significantly antitumor activity of γδ T-cells also in vivo. These findings showed that ZOL stimulated the proliferation of γδ T-cells significantly and that the cytotoxic activity of γδ T-cells required pre-treatment of target cells with ZOL, indicating the potential use of autologous ex vivo expanded γδ T-cells for cancer immunotherapy.
Birth Order, Atopy, and Risk of Non-Hodgkin Lymphoma.
Authors: Grulich Andrew E. , Vajdic Claire , Kaldor John M. , Hughes Anne-Maree , Kricker Anne , Fritschi Lin , Turner Jennifer J. , Milliken Sam , Benke Geza , Armstrong Bruce K.
Published: October 2019

People with congenital or acquired immune deficiency have rates of non-Hodgkin lymphoma (NHL) that are raised 50 fold or more above population rates, but it is unknown whether risk of NHL is associated with other forms of immune dysregulation. We performed a population based case-control study of risk factors for NHL in adults aged 20–74 years in New South Wales and the Australian Capital Territory, Australia. We investigated the association of NHL risk with atopy, which is associated with a Th2 dominant immune response. In addition, we investigated the association of NHL with birth order and childhood crowding, which are known to predict atopy. Cases with NHL were selected from a cancer register (n=704), and controls (n=694) were randomly selected from state electoral rolls and frequency-matched to cases by age, sex and area. Cases with clinically apparent immune deficiency were excluded. Birth order, childhood crowding and history of atopic conditions (hayfever, asthma, eczema, and specific allergies) were assessed by questionnaire and by interview. Logistic regression models of NHL risk included the three matching variables as covariates. Being an only child or first born child of a larger family was strongly inversely associated with risk of NHL. Compared to a fourth or later born child, the odds ratios (OR) for development of NHL were 0.52 (95% CI 0.32–0.84) for an only child, 0.55 (95% CI 0.40–0.75) for a first-born child, and 0.70 (95% CI 0.51–0.96) and 0.81 (95 % CI 0.57–1.14) respectively for second and third born children (p trend < 0.0001). Indicators of crowding in later childhood, such as ever sharing a bed or bedroom, and number of years of sharing, were not associated with NHL risk. Diagnosis of atopic conditions was also inversely associated with NHL risk. Self-reported histories of hayfever, asthma, eczema and food allergies were each associated with reduced NHL risk; this reduction was significant for hayfever (OR 0.65, 95% CI 0.52–0.82) and food allergies (OR 0.29, 95% CI 0.20–0.42), but not significant for asthma (OR 0.88, 95% CI 0.67–1.17) and eczema (OR 0.79, 95% CI 0.57–1.10). In summary, being an only or other first born child and reporting a history of atopic conditions were associated with reduced NHL risk. Taken together, these data suggest a hypothesis that late exposure to infections in early life and atopic conditions, both of which are associated with a predominant Th2 response, are associated with a reduced risk of NHL.
UV-Induced B-Cell Lymphomas in p53 Heterozygous Mice.
Authors: Miyahara Yasuko , Kazimi Nasser , Multani Asha S. , Pathak Sen , Chang Sandy , Angkasekwinai Yee , Qin Xiao-Feng , Ullrich Stephen E.
Published: October 2019

The association between ultraviolet (UV) irradiation and skin cancer induction is well known. Recently, UV exposure was associated with the induction of internal lymphoid malignancies in UV-irradiated p53 heterozygous mice. Here we characterize the non-skin cancer that develops in the lymphoid organs of UV-irradiated mice and found that UV exposure promotes the induction of a B-cell lymphoma. p53 heterozygous mice were exposed to UVB irradiation (10KJ/m2; three times a week) supplied by a bank of FS40 sunlamps. After thirty weeks, we collected spleen cells from 5 irradiated mice, prepared single suspensions, pooled the cells and injected 2X106 cells/flank (subcutaneous injection) into Rag2−/−mice. The subcutaneous tumors that developed in the Rag2−/−mice were successfully transplanted into other Rag2−/−mice and adapted to grow in vitro. Flow cytometric analysis indicated that enlarged cells were positive for CD19, B220, IgM, CD24, CD40, B7-1, B7-2, but negative for CD43, BP-1, CD127, CD3, CD4, CD8, CD49b, NK1.1, Thy1.2, Gr-1. This phenotype is compatible with a mature B cell but also express CD5, CD11b, CD117. Immunohistchemical analysis confirmed that the tumor cells were of B-cell origin and southern blot analysis demonstrated rearrangement of Immunoglobulin heavy chain. Chromosomal analysis revealed a translocations, t (14;19). Subcutaneous injection of the tumor cells into Rag2−/− mice resulted in an aggressive tumor that infiltrated into the lymph nodes, the spleen and the bone marrow. After establishing the cell line from this B-cell lymphoma, we transfected GFP by using lentiviral vector to mark and injected into Rag2−/− mice. First, the cells bearing GFP infiltrated into the lymph nodes, and then into the spleen and the bone marrow. We also found that this cell line produced IL-10, which can promote lymphoma growth and can cause systemic immunosuppression (Masood et al., 1995Blood85:3423–30; Rivas et al., 1994J Leukoc Biol.59:769–75). The B-cell lymphoma, which grew progressively in immune compromised Rag2−/−mice, was rejected when injected into wild type mice and syngeneic UV-irradiated C57Bl/6 mice. This data indicates that UV induced B-cell lymphoma in p53 heterozygous mice does not express UV common antigen found on UV-induced skin cancer because UV irradiated mice are unable to reject UV induced tumor cells bearing this antigen as previously reported (Kripke et al., 1974J.Natl Cancer Inst.53:1333–6). Since sunlight exposure has been associated with the development of non-Hodgkin’s lymphomas in humans, tumor development in UV-irradiated p53 heterozygous mice may provide a useful animal model for lymphoma development in humans.