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The concept of vivir bien (living well) has become ubiquitous in Bolivian state discourse and policy since the election of Evo Morales as Bolivia’s president in 2005. While Bolivia’s constitutional refounding as plurinational is supposed to facilitate indigenous peoples’ living according to their conception of living well, the state still appears to be attempting to implement its own conception through rural social programs promoted as enabling rural indigenous peoples to live well. The implementation of one such social program, a housing donation program in the municipality of Charazani (Department of La Paz), demonstrates differing notions of vivir bien between neighboring communities and suggests that a program designed to facilitate vivir bien may actually provide obstacles to the realization of an indigenous conception of living well.El concepto de vivir bien se ha vuelto omnipresente en el discurso y la política del estado boliviano desde la elección de Evo Morales como presidente en 2005. Si bien se supone que la refundación constitucional de Bolivia como estado plurinacional debe facilitar la vida de los pueblos indígenas de acuerdo a su concepción de vivir bien, el Estado aún parece estar tratando de implementar su propia concepción a través de programas sociales rurales promovidos como proyectos que permiten que los pueblos indígenas rurales vivan bien. La implementación de uno de estos programas sociales, un programa de donación de vivienda en el municipio de Charazani (Departamento de La Paz), muestra diferentes nociones de lo que implica vivir bien entre las comunidades vecinas y sugiere que un programa diseñado para facilitarlo puede, de hecho, generar obstáculos a la realización de una concepción indígena de vivir bien.

A red-letter day for RBC research The study of primary human red blood cell (huRBC) disorders such as sickle cell disease (SCD) and infectious diseases such as malaria has been hampered by a lack of in vivo models of human erythropoiesis. Song et al. transferred human fetal liver cells into MISTRG mice, which are immunodeficient and are genetically engineered with several human genes involved in hematopoiesis. This approach was unsuccessful because mature huRBCs are rapidly destroyed in the mouse liver. They then used CRISPR-Cas9 to mutate these mice into a fumarylacetoacetate hydrolase–deficient strain, allowing them to replace the mouse liver with engrafted human hepatocytes. These mice exhibited enhanced human erythropoiesis and circulating huRBC survival and could recapitulate SCD pathology when reconstituted with SCD-derived HSCs. Science , this issue p. 1019

This work was supported by The Leverhulme Trust (grant number ECF-2017-091), ESRC studentship (grant number 1211157), the UKRI Future Leaders Fellowship (grant number MR/V022261/1),and the UKRI Future Leaders Fellowship (grant number MR/S03501X/1). Coproduction brings together a diverse range of state and non-state actors to create and deliver public services. These processes, which occur across the Global South, have been widely studied. However, insufficient critical attention has been paid to their politics. We address this gap in the literature by analysing the politics of coproduction in Latin America during the ‘pink tide’ of the early twenty-first century. Drawing on original qualitative research, this article explores the coproduction of three distinct public services—water, housing, and waste—in three countries where left-leaning presidents and governments were elected into office—Ecuador, Bolivia, and Uruguay. We argue that coproduction is intrinsically political in these three cases; that is, the ‘political’ is internal to, and inherent in, coproduction. Our comparative analysis centres on two political dimensions—subject-making and collective autonomy—and shows that tensions around these two issues were central to coproduction in Ecuador, Bolivia, and Uruguay . The article demonstrates the importance of not treating politics as a mere contextual concern in coproduction analysis and shows that coproduction has the capacity to reshape political relationships and subjectivities. Taking politics into account is essential to understanding the dynamics and potential of coproduction in the Global South. Our comparative analysis also provides new insights into Latin American politics, especially concerning 'pink tide' governments and the provisioning of public services. Publisher PDF

By Alessandra Pellegrini Calderon. University of Arizona Press, Tucson, 2016. Deference Revisited: Andean Ritual in the Plurinational State. By Into Goudsmit. Carolina Academic Press, Durham, Nor...

In vivo models of human erythropoiesis with generation of circulating mature human red blood cells (huRBC) have remained elusive, limiting studies of primary human red cell disorders. In our prior study, we have generated the first combined cytokine-liver humanized immunodeficient mouse model (huHepMISTRG-Fah) with fully mature, circulating huRBC when engrafted with human CD34+ hematopoietic stem and progenitor cells (HSPCs)1. Here we present for the first time a humanized mouse model of human sickle cell disease (SCD) which replicates the hallmark pathophysiologic finding of vaso-occlusion in mice engrafted with primary patient-derived SCD HSPCs. SCD is an inherited blood disorder caused by a single point mutation in the beta-globin gene. Murine models of SCD exclusively express human globins in mouse red blood cells in the background of murine globin knockouts2 which exclusively contain murine erythropoiesis and red cells and thus fail to capture the heterogeneity encountered in patients. To determine whether enhanced erythropoiesis and most importantly circulating huRBC in engrafted huHepMISTRG-Fah mice would be sufficient to replicate the pathophysiology of SCD, we engrafted it with adult SCD BM CD34+ cells as well as age-matched control BM CD34+ cells. Overall huCD45+ and erythroid engraftment in BM (Fig. a, b) and PB (Fig. c, d) were similar between control or SCD. Using multispectral imaging flow cytometry, we observed sickling huRBCs (7-11 sickling huRBCs/ 100 huRBCs) in the PB of SCD (Fig. e) but not in control CD34+ (Fig. f) engrafted mice. To determine whether circulating huRBC would result in vaso-occlusion and associated findings in SCD engrafted huHepMISTRG-Fah mice, we evaluated histological sections of lung, liver, spleen, and kidney from control and SCD CD34+ engrafted mice. SCD CD34+ engrafted mice lungs showed an increase in alveolar macrophages (arrowheads) associated with alveolar hemorrhage and thrombosis (arrows) but not observed control engrafted mice (Fig. g). Spleens of SCD engrafted mice showed erythroid precursor expansion, sickled erythrocytes in the sinusoids (arrowheads), and vascular occlusion and thrombosis (arrows) (Fig. h). Liver architecture was disrupted in SCD engrafted mice with RBCs in sinusoids and microvascular thromboses (Fig. i). Congestion of capillary loops and peritubular capillaries and glomeruli engorged with sickled RBCs was evident in kidneys (Fig. j) of SCD but not control CD34+ engrafted mice. SCD is characterized by ineffective erythropoiesis due to structural abnormalities in erythroid precursors3. As a functional structural unit, erythroblastic islands (EBIs) represent a specialized niche for erythropoiesis, where a central macrophage is surrounded by developing erythroblasts of varying differentiation states4. In our study, both SCD (Fig. k) and control (Fig. l) CD34+ engrafted mice exhibited EBIs with huCD169+ huCD14+ central macrophages surrounded by varying stages of huCD235a+ erythroid progenitors, including enucleated huRBCs (arrows). This implies that huHepMISTRG-Fah mice have the capability to generate human EBIs in vivo and thus represent a valuable tool to not only study the effects of mature RBC but also to elucidate mechanisms of ineffective erythropoiesis in SCD and other red cell disorders. In conclusion, we successfully engrafted adult SCD patient BM derived CD34+ cells in huHepMISTRG-Fah mice and detected circulating, sickling huRBCs in the mouse PB. We observed pathological changes in the lung, spleen, liver and kidney, which are comparable to what is seen in the established SCD mouse models and in patients. In addition, huHepMISTRG-Fah mice offer the opportunity to study the role of the central macrophage in human erythropoiesis in health and disease in an immunologically advantageous context. This novel mouse model could therefore serve to open novel avenues for therapeutic advances in SCD. Reference 1. Song Y, Shan L, Gybli R, et. al. In Vivo reconstruction of Human Erythropoiesis with Circulating Mature Human RBCs in Humanized Liver Mistrg Mice. Blood. 2019;134:338. 2. Ryan TM, Ciavatta DJ, Townes TM. Knockout-transgenic mouse model of sickle cell disease. Science. 1997;278(5339):873-876. 3. Blouin MJ, De Paepe ME, Trudel M. Altered hematopoiesis in murine sickle cell disease. Blood. 1999;94(4):1451-1459. 4. Manwani D, Bieker JJ. The erythroblastic island. Curr Top Dev Biol. 2008;82:23-53. Disclosures Xu: Seattle Genetics: Membership on an entity's Board of Directors or advisory committees. Flavell:Zai labs: Consultancy; GSK: Consultancy.

The murine host has remained a readily available and ethically acceptable model for the study of human diseases and therapeutic testing. Immunodeficient mouse models support engraftment of human hematopoietic stem cells (HSC) but with limitation in efficiency and mature lineage representation. Combined knock-in of several non-crossreactive human cytokines (M-CSF, IL3/GM-CSF, and Thrombopoietin) into the corresponding murine loci in the SRG strain (in short termed "MISTRG") has enhanced engraftment and maintenance of human HSCs with multi-lineage differentiation (Rongvaux et al. Annu Rev Immunol 2013, Deng et al. Nature 2015, Saito et al. Blood 2016, Theocharides et al. Haematologica 2016). Despite robust HSC engraftment and myelo- and erythropoiesis in bone marrow (BM), all humanized immunodeficient mouse models lack of mature human red blood cells (RBC), platelets, and myeloid cells in peripheral blood (PB) (Rahmig et al. Stem Cell Reports 2016, Yurino et al. Stem Cell Reports 2016, Song et al. Nat Commun 2019). Yet, full maturation and representation of all myeloid lineages in PB is essential to study diseases of the HSC, such as MDS, and of the RBC, such as sickle cell anemia or malaria. With universal absence of a murine adaptive immune system the culprit is likely the murine host's innate immune system. Previous studies have shown that treatment of engrafted mice with liposomal clodronate that abrogates murine (and human) macrophages, with or without cobra venom factor that eliminates complement, can increase mature human circulating RBC, but only transiently and with significant toxicity. We first sought to determine the site of huRBC sequestration and destruction. Intravital imaging after injection of CFSE labelled huRBC identified the murine liver as the major site of RBC destruction. While muRBC rapidly circulate through the liver circulation, huRBC have greatly increased transit times and are sequestered in liver vessels. We hypothesized that humanization of the murine host's liver could potentially alleviate huRBC sequestration and significantly increase circulating huRBC. In previous studies deletion of fumarylacetoacetate hydrolase (Fah) in the Rag-/-Il2rg-/- background has allowed humanization of the liver and served to study diseases such as malaria (Vaughan et al. J Clin Invest 2012). The liver is the site of synthesis of numerous proteins, some of which directly impact hematopoiesis and blood cells, such as complement. We deleted the Fah gene via CRISPR/Cas9 in MISTRG mice and crossed MISTRG-Fah-/- mice to homozygosity (MISTRGFah). MISTRGFah are viable, fertile, and healthy when maintained on drinking water supplemented with 2-(2-nitro-4-trifluoromethylbenzoyl)-1, 3-cyclohexanedione (NTBC), that blocks tyrosine metabolism upstream of Fah and prevents buildup of hepatotoxic metabolites. At 8 weeks of age we implanted MISTRGFah mice with commercially available, adult human hepatocytes (HuHep) via direct injection into the splenic vein, followed by gradual withdrawal of NTBC water. Regulated buildup of intracellular fumarylacetoacetate results in death of murine Fah-/- hepatocytes and regeneration with HuHep with up to 90% repopulation by HuHep (Azuma et al. Nature biotechnology 2007). When plasma human albumin levels reached 2mg/dL, indicative of significant (~80%) HuHep repopulation, we sublethally (80cGy) irradiated HuHepMISTRGFah mice and engrafted each mouse with 105 fetal liver (FL) derived CD34+ cells. 10 weeks post transplantation, mice were analyzed for engraftment and specifically erythroid maturation in PB. HuHepMISTRGFah mice had significantly higher levels of BM and interestingly spleen erythropoiesis and circulating huRBC in PB (Fig.1 a). CD235a+ huRBC in HuHepMISTRGFah mice are enucleated (Hoechst neg) and mature as evident by loss of CD49d (ITGA4) and gain of Band3 staining (Hu et al. Blood 2013) (Fig.1 b). Interestingly, human erythroid cells in MISTRG but not HuHepMISTRGFah mice are coated with murine complement C3 (muC3) (Fig.1 c) suggesting that liver humanization results in loss of muC3 expression. In conclusion, we have generated the first humanized mouse model with fully mature, circulating huRBC when engrafted with human CD34+ stem and progenitor cells. Ongoing studies are testing the applicability of this model to MDS and sickle cell disease. Disclosures Flavell: SMOC: Equity Ownership; Zai labs: Consultancy; GSK: Consultancy; Artizan Biosciences: Equity Ownership; Troy: Equity Ownership; Rheos Biomedicines: Equity Ownership.

The objective was to assess the efficacy of sonography with and without contrast medium enhancement in guiding and monitoring percutaneous ethanol ablation of tumors in an animal model. VX-2 carcinoma was implanted into the thighs of New Zealand white rabbits and examined by grey-scale ultrasound, color, power, and pulse Doppler, before and after injection of 95% ethanol into the tumors. Injections of ethanol were guided by ultrasound to sites of tumor vascularity, until all tumor vascularity had been obliterated. Microbubble contrast medium or saline was injected IV prior to each of the ultrasonic interrogations. Arteriography was performed before and after ablation. Selected tumor samples were submitted for histologic examination. Contrast enhanced tumor vascularity over saline controls in all cases. In some, incompletely ablated foci of tumor could only be identified with contrast medium enhancement. Arteriography showed complete ablation of all but 1 tumor. We conclude that ultrasound enhanced by contrast better shows the presence or absence of tumor vascularity. Ultrasound enhanced by contrast might offer an accurate means of guiding and monitoring percutaneous ethanol injection for tumor ablation.

Rationale and objectives The authors conduct technical feasibility and performance studies on a new 0.009-inch nitinol microguidewire for potential neuroendovascular applications. Methods In vivo microcatheterizations of brachiocephalic arteries were performed in four pigs using the 0.009-inch nitinol microguidewire with a commonly used microcatheter and compared with the performance of commercially available microguidewires. Vessels catheterized using the 0.009-inch wire were evaluated by histopathological analysis. A standardized, bench-top method of evaluating basic mechanical properties of microguidewires also was used to compare the 0.009-inch nitinol microguidewire with two commonly used microguidewires. Results The 0.009-inch microguidewire had similar steerability, tractability, torque control, and distal tip flexibility to the commonly used microguidewires in the in vivo simulations. Bench top testing showed the 0.009-inch microguidewire to have comparable distal tip flexibility and objective mechanical properties to the commonly used microguidewires. Conclusions The comparable subjective and objective mechanical properties of the 0.009-inch nitinol microguidewire to that of commonly used microguidewires further establishes the possibility of clinical implementation.

The measurement of volumetric blood flow in small vessels in vitro and in vivo poses a significant technological challenge. In this study, two pulsatile flow models were developed, one with a 3.2-mm lumen diameter and one with a 12.7-mm lumen diameter, to assess the accuracy of volumetric flow estimation of two pulsed-Doppler devices, a Crystal Biotech VF1 20-MHz system with either a cuff-mounted or a needle-mounted probe and an Advanced Technology Laboratories Ultramark 9 High Definition Imaging system with a 5-MHz linear array transducer. The VF1 volumetric flow error was measured in the 3.2-mm phantom over a variety of pulsatile and continuous waveforms. The accuracy of the VF1 was also tested in porcine femoral and renal arteries. VF1 volumetric flow error ranged from 4.8% to 54.3% in the in vivo studies. The ATL demonstrated similar volumetric flow errors in the porcine femoral artery (approximately 3.2 mm diameter), but these errors were reduced toor = 17.4% in the 12.7-mm-diameter in vitro flow model.

mRNA transfection is a useful approach for temporal cell reprogramming with minimal risk of transgene-mediated mutagenesis. We applied this to redirect lymphocyte cytotoxicity toward malignant cells. Using the chimeric immune receptor (CIR) constructs anti-CD19 CIR and 8H9 CIR, we achieved uniform expression of CIRs on virtually the entire population of lymphocytes. We reprogrammed CD3+ CD8+, CD3+ CD4+, and natural killer (NK ) cells toward autologous and allogeneic targets such as B cells, Daudi lymphoma, primary melanoma, breast ductal carcinoma, breast adenocarcinoma, and rhabdomyosarcoma. The reprogramming procedure is fast. Although most of the experiments were performed on lymphocytes obtained after 7-day activation, only 1-day activation of T cells with anti-CD3, anti-CD28 antibodies, and interleukin-2 is sufficient to develop both lymphocyte cytotoxicity and competence for mRNA transfer. The entire procedure, which includes lymphocyte activation and reprogramming, can be completed in 2 days. The efficiency of mRNA-modified human T cells was tested in a murine xenograft model. Human CD3+CD8+ lymphocytes expressing anti-CD19 CIR mRNA inhibited Daudi lymphoma growth in NOD/SCID mice. These results demonstrate that a mixed population of cytotoxic lymphocytes, including T cells together with NK cells, can be quickly and simultaneously reprogrammed by mRNA against autologous malignancies. With relatively minor modifications the described method of lymphocyte reprogramming can be scaled up for cancer therapy.

This project was designed to detect the development of tumor neovascularity and determine if intravenous microbubble contrast improves visualization of otherwise undetectable tumors in an animal model. VX-2 carcinoma was implanted into one thigh of 10 New Zealand white rabbits. Tumors were assessed without and with contrast at 1- to 4-day intervals from day 3-19 postimplantation, using gray scale, color flow, pulse Doppler and power Doppler imaging. Tumor vascularity was compared with the contralateral thigh muscle, so each animal was its own control. Contrast injection improved visualization of tumor neovascularity. Early tumors had homogeneous vasculature but, with time, the centers became less vascular, while the periphery increased. Following contrast injection, color gain was decreased by 40% without compromising color intensity. Neovascularity was detected by contrast injection before the tumor could be palpated or visualized by gray scale. Based on these data, we conclude that enhancement of neovascularity by intravenous contrast permits earlier detection and improved visualization of soft tissue tumors in rabbits.

The purpose of this study was to assess the utility of an intravenous contrast agent (FS069) for visualization of normal visceral perfusion compared with perfusion after segmental infarction in a canine model. Six mongrel dogs were used as subjects. Splenic, renal, hepatic, and small bowel perfusion was assessed without and with intravenous microbubble contrast material using gray scale, color Doppler, pulsed Doppler, and color power Doppler sonography. Each organ was then reassessed after ligation of a segmental vessel. Imaging was again performed without and with contrast material using all four ultrasonographic modalities. In all organs color and spectral Doppler signals were significantly enhanced from normally perfused tissue after intravenous contrast agent injection. Ischemic areas were more conspicuous after contrast medium injection except in the liver. Hepatic perfusion was maintained by portal flow in the liver despite arterial ligation. Ligation of collateral arcades was required to produce bowel ischemia. Intravenous injection of FS069 improves evaluation of visceral perfusion and identification of focal visceral ischemia in dogs. These results suggest that this agent may increase sensitivity for detection of blood flow in small and deep vessels.

OBJECTIVE: To determine, using an animal model of blood loss, (1) if stroke distance, derived non-invasively from the time integral of the maximum velocity of red cells in the aorta, changed to a greater extent than heart rate and mean arterial pressure (MAP), which are recognised to be unreliable indicators of blood loss; (2) if changes in stroke distance reflected changes in stroke volume derived from thermodilution cardiac output measurements. METHODS: Eight anaesthetised swine had baseline measurements of heart rate, MAP, stroke volume, and stroke distance and were then exsanguinated at a rate of 1 ml/kg/min. Percentage changes from baseline of heart rate, MAP, stroke volume, and stroke distance were compared after 10, 20, and 30 ml/kg blood loss. The animal's blood was then reinfused at the rate of 2 ml/kg/min for 15 min, followed by normal saline 1 ml/kg/min. Percentage changes from baseline measurement of stroke volume and stroke distance over the whole experiment were evaluated by regression analysis. RESULTS: Heart rate, MAP, and stroke distance changed +7.9%, -22.5%, and -18.1% respectively (from baseline values) after 10 ml/kg blood loss; +23.2%, -44.0%, and -47.4% after 20 ml/kg blood loss; and +55.7%, -62.0%, and -69.8% after 30 ml/kg blood loss. Regression analysis of percentage changes in stroke volume and stroke distance from their baseline values at experimental time zero is stroke volume = 1.014 x stroke distance -2.156, r = 0.92, n = 54, P < 0.0001. CONCLUSIONS: (1) At maximal blood loss, stroke distance changes to a greater extent than heart rate and MAP. (2) Changes in stroke distance reflected changes in stroke volume but with less variability at lower values. Stroke distance may be a more useful measure of blood loss than heart rate and MAP.

Using a continuous haemorrhage model, 8 anaesthetised swine were bled 1 ml/kg per min for 30 min. The resistance index (RI) of the main renal artery, interlobar and arcuate vessels all significantly increased. Cortical Doppler signals were lost in 4 animals at a mean arterial pressure of 26 mmHg. After reinfusion of blood and normal saline only the RI of the interlobar vessels was significantly different from baseline readings. Ultrasound demonstrated non-invasively changes in regional blood flow within the kidney in response to hypovolaemic shock.