A bioengineered artificial interstitium supports long-term islet xenograft survival in nonhuman primates without immunosuppression.

Cell-based therapies hold promise for many chronic conditions; however, the continued need for immunosuppression along with challenges in replacing cells to improve durability or retrieving cells for safety are major obstacles. We subcutaneously implanted a device engineered to exploit the innate transcapillary hydrostatic and colloid osmotic pressure generating ultrafiltrate to mimic interstitium. Long-term stable accumulation of ultrafiltrate was achieved in both rodents and nonhuman primates (NHPs) that was chemically similar to serum and achieved capillary blood oxygen concentration. The majority of adult pig islet grafts transplanted in non-immunosuppressed NHPs resulted in xenograft survival >100 days. Stable cytokine levels, normal neutrophil to lymphocyte ratio, and a lack of immune cell infiltration demonstrated successful immunoprotection and averted typical systemic changes related to xenograft transplant, especially inflammation. This approach eliminates the need for immunosuppression and permits percutaneous access for loading, reloading, biopsy, and recovery to de-risk the use of "unlimited" xenogeneic cell sources to realize widespread clinical translation of cell-based therapies.

Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.

Clinically available immunosuppression averts rejection but not systemic inflammation after porcine islet xenotransplant in cynomolgus macaques.

A safe, efficacious, and clinically applicable immunosuppressive regimen is necessary for islet xenotransplantation to become a viable treatment option for diabetes. We performed intraportal transplants of wild-type adult porcine islets in 25 streptozotocin-diabetic cynomolgus monkeys. Islet engraftment was good in 21, partial in 3, and poor in 1 recipient. Median xenograft survival was 25 days with rapamycin and CTLA4Ig immunosuppression. Adding basiliximab induction and maintenance tacrolimus to the base regimen significantly extended median graft survival to 147 days (p < .0001), with three animals maintaining insulin-free xenograft survival for 265, 282, and 288 days. We demonstrate that this regimen suppresses non-Gal anti-pig antibody responses, circulating effector memory T cell expansion, effector function, and infiltration of the graft. However, a chronic systemic inflammatory state manifested in the majority of recipients with long-term graft survival indicated by increased neutrophil to lymphocyte ratio, IL-6, MCP-1, CD40, and CRP expression. This suggests that this immunosuppression regimen fails to regulate innate immunity and resulting inflammation is significantly associated with increased incidence and severity of adverse events making this regimen unacceptable for translation. Additional studies are needed to optimize a maintenance regimen for regulating the innate inflammatory response.

A nonhuman primate model of vertical sleeve gastrectomy facilitates mechanistic and translational research in human obesity.

The obesity epidemic significantly contributes to overall morbidity and mortality. Bariatric surgery is the gold standard treatment for obesity and metabolic dysfunction, yet the mechanisms by which it exerts metabolic benefit remain unclear. Here, we demonstrate a model of vertical sleeve gastrectomy (VSG) in nonhuman primates (NHP) that mimics the complexity and outcomes in humans. We also show that VSG confers weight loss and durable metabolic benefit, where equivalent caloric intake in shams resulted in significant weight gain following surgery. Furthermore, we show that VSG is associated with early, weight-independent increases in bile acids, short-chain fatty acids, and reduced visceral adipose tissue (VAT) inflammation with a polarization of VAT-resident immunocytes toward highly regulatory myeloid cells and Tregs. These data demonstrate that this strongly translational NHP model can be used to interrogate factors driving successful intervention to unravel the interplay between physiologic systems and improve therapies for obesity and metabolic syndrome.

Molecular Characteristics, Ecology, and Zoonotic Potential of Escherichia coli Strains That Cause Hemorrhagic Pneumonia in Animals.

Hemorrhagic pneumonia (HP) is a rare but highly lethal disease, mainly of dogs and cats, caused by hemolytic Escherichia coli strains that contain cnf1 (encoding cytotoxic necrotizing factor 1). After encountering fatal HP in two dogs, we used contemporary molecular methods, including multilocus sequence typing and whole-genome sequencing, to compare the corresponding case isolates with published HP clinical isolates and newly obtained fecal E. coli isolates from 20 humans and animals in the index HP case household. We also compared the aggregated HP clinical isolates, which represented 13 discrete strains, by pulsotype with a large, private pulsotype library of diverse-source E. coli. The HP clinical isolates represented a narrow range of phylogenetic group B2 lineages (mainly sequence types 12 and 127), O types (mainly O4 and O6), and H types (mainly H5 and H31), but diverse fimH alleles (type-1 fimbriae adhesin). Their extensive, highly conserved virulence genotypes, which qualified as extraintestinal pathogenic E. coli (ExPEC), encoded diverse adhesins, toxins, iron uptake systems, and protectins. Household surveillance identified multiple HP-like fecal strains, plus abundant between-host strain sharing, including of the household's index HP strain. The pulsotype library search identified, for five HP clinical strains, same-pulsotype human and animal fecal and clinical (predominantly urine) isolates, from diverse locales and time periods. Thus, E. coli strains that cause HP derive from a narrow range of ExPEC lineages within phylogroup B2, contain multiple virulence genes other than cnf1, are shared extensively between hosts, and likely function in nature mainly as intestinal colonizers and uropathogens. IMPORTANCE This study clarifies the clonal background and extensive virulence genotypes of the E. coli strains that cause hemorrhagic pneumonia in domestic animals (mainly dogs and cats), shows that such strains circulate among animals and humans, identifies a substantial intestinal colonization component to their lifestyle, and extends their known clinical manifestations to include bacteremia and urinary tract infection. The findings place these strains better into context vis-à-vis current understandings of E. coli phylogeny, ecology, and pathogenesis; identify questions for future research; and may prove relevant for surveillance and prevention efforts.

Comparing a Novel Neuroanimation Experience to Conventional Therapy for High-Dose Intensive Upper-Limb Training in Subacute Stroke: The SMARTS2 Randomized Trial.

BACKGROUND: Evidence from animal studies suggests that greater reductions in poststroke motor impairment can be attained with significantly higher doses and intensities of therapy focused on movement quality. These studies also indicate a dose-timing interaction, with more pronounced effects if high-intensity therapy is delivered in the acute/subacute, rather than chronic, poststroke period. OBJECTIVE: To compare 2 approaches of delivering high-intensity, high-dose upper-limb therapy in patients with subacute stroke: a novel exploratory neuroanimation therapy (NAT) and modified conventional occupational therapy (COT). METHODS: A total of 24 patients were randomized to NAT or COT and underwent 30 sessions of 60 minutes time-on-task in addition to standard care. The primary outcome was the Fugl-Meyer Upper Extremity motor score (FM-UE). Secondary outcomes included Action Research Arm Test (ARAT), grip strength, Stroke Impact Scale hand domain, and upper-limb kinematics. Outcomes were assessed at baseline, and days 3, 90, and 180 posttraining. Both groups were compared to a matched historical cohort (HC), which received only 30 minutes of upper-limb therapy per day. RESULTS: There were no significant between-group differences in FM-UE change or any of the secondary outcomes at any timepoint. Both high-dose groups showed greater recovery on the ARAT (7.3 ± 2.9 points; P = .011) but not the FM-UE (1.4 ± 2.6 points; P = .564) when compared with the HC. CONCLUSIONS: Neuroanimation may offer a new, enjoyable, efficient, and scalable way to deliver high-dose and intensive upper-limb therapy.

Comparative analysis of the boundary layer filtering of odor signals in the amblypygid (whip spider) species Paraphrynus laevifrons and Phrynus marginemaculatus.

Amblypygids use a pair of modified walking legs (antenniform) as chemosensory and mechanosensory appendages. At the tip of these legs are covered in chemosensory sensilla, which the animals use to sample odor stimuli in their environment by moving the antenniform leg through the air. We designed a set of experiments to measure the filtering effect that aerodynamic boundary layers have on the temporal and spatial structure of chemical stimuli. In addition, two different species of amblypygids (Paraphrynus laevifrons and Phrynus marginemaculatus) that live in two distinct habitats were used for a comparative analysis. Pulses of a tracer molecule were quantified at different distances and flow velocities using an electrochemical detection system. Temporal attributes of the chemical pulses were extracted and were statistically compared across velocities, distances from the appendage, and the two species. Overall, the boundary layer significantly decreased the concentration and increased the duration of pulses for both species. This filtering effect was more pronounced for P. marginemaculatus than P. laevifrons, as the chemical signal was lower in concentration and longer in duration at any distance from the antenniform leg. It is speculated that the difference in boundary layer filtering, as a function of appendage morphology, is tuned to the different types of odor plumes in these animals' native habitats.

Examination of Homing Behaviors in Two Species of Crayfish Following Translational Displacements.

Crayfish have been model systems for examining complex behaviors and the underlying neural mechanisms that guide these behaviors. While spatial learning has been examined in a subset of crayfish species, homing behaviors remained largely unexamined. Here we examined homing behavior following translational displacements in a primary burrowing (Creaserinus fodiens) and tertiary burrowing species (Faxonius rusticus). Individuals of both species were placed in an arena with artificial burrows embedded within the arena floor. The arena floor was fitted with a panel, which served as a treadmill belt to allow for translational displacement. Individuals were displaced after they had left the burrows. The movement pathways of displaced crayfish were compared with those in two control groups, one which underwent no displacement and the second in which the treadmill belt was displaced but returned to its original position almost immediately. Homing success for displaced individuals of both species was considerably reduced in comparison to the control groups. Moreover, displaced primary burrowers had significantly lower homing success in comparison to displaced tertiary burrowers. Primary burrowers exhibited greater homing error and significantly impaired homing behaviors compared with tertiary burrowers. Furthermore, heading angles in displaced groups (of both species) were significantly higher than the control group of both species. Species-specific differences in homing success and homing error indicate that primary burrowers were more negatively impacted by translational displacements. These homing differences indicate that these two species of crayfish have differing homing strategies.

Examen del Instinto de Regresar al Hogar en Dos Especies de Cangrejos de Río Después de Desplazamientos Translacionales (Examination of Homing Behaviors in Two Species of Crayfish Following Translational Displacements) Los cangrejos de río han sido sistemas modelo para examinar comportamientos complejos y los mecanismos neuronales subyacentes que guían estos comportamientos. Si bien el aprendizaje espacial se ha examinado en un subconjunto de especies de cangrejos de río, los comportamientos de regresar al hogar permanecieron en gran parte sin examinar. Aquí se examinó el comportamiento de regresar al hogar luego de los desplazamientos traslacionales en una especie de madriguera primaria (Creaserinus fodiens) y de madriguera terciaria (Faxonius rusticus). Los individuos de ambas especies se colocaron en una arena con madrigueras artificiales incrustadas dentro del piso de la arena. El piso de arena estaba equipado con un panel, que servía como cinta de correr para permitir el desplazamiento traslacional. Los individuos fueron desplazados después de haber abandonado las madrigueras. Las vías de movimiento de los cangrejos de río desplazados se compararon con las de dos grupos de control, uno que no experimentó ningún desplazamiento y el segundo en el que se desplazó el cinturón de la cinta de corer pero fue devuelto a su posición original casi de inmediato. El éxito de regresar a casa para los individuos desplazados de ambas especies se redujo considerablemente en comparación con los grupos de control. Además, los excavadores primarios desplazados tuvieron significativamente menos éxito de regresar al hogar en comparación con los excavadores terciarios desplazados. Los excavadores primarios exhibieron un mayor error de regresar al hogar y el desempeño de ese comportamiento también fue significativamente deteriorado en comparación con los excavadores terciarios. Además, los ángulos de encabezamiento en los grupos desplazados (de ambas especies) fueron significativamente más altos que el grupo de control de ambas especies. Las diferencias específicas de la especie en el éxito de regresar al hogar y el error de la orientación indican que las excavaciones primarias se vieron afectadas más negativamente por los desplazamientos traslacionales. Estas diferencias de orientación indican que estas dos especies de cangrejos de río tienen diferentes estrategias de localización. translated to Spanish by Y.E. Jimenez (yordano_jimenez@brown.edu).

Examinando Comportamentos de Entocamento em Duas Espécies de Lagostins Após Deslocamentos Translationais (Examination of Homing Behaviors in Two Species of Crayfish Following Translational Displacements) Os lagostins têm sido usados como sistemas-modelo para examinar comportamentos complexos e os mecanismos neurais subjacentes que ditam esses comportamentos. Embora a aprendizagem espacial tenha sido examinada em um subconjunto de espécies de lagostins, os comportamentos de entocamento permaneceram em grande parte sem estudo. Aqui, foram examinados os comportamentos de encotamento após deslocamentos translacionais em espécies fossoriais primárias (Creaserinus fodiens) e terciárias (Faxonius rusticus). Indivíduos de ambas as espécies foram colocados em uma arena com tocas artificiais embutidas no chão da arena. O piso foi equipado com um painel que serviu como uma esteira para permitir o deslocamento translacional. Indivíduos foram deslocados depois de terem deixado as tocas. As rotas de movimentação dos lagostins deslocados foram comparadas com as de dois grupos-controle: um que não sofreu deslocamento e um segundo cuja esteira foi deslocada, mas retornou à sua posição original quase imediatamente. O sucesso no alojamento de indivíduos deslocados de ambas as espécies foi consideravelmente reduzido em comparação com os grupos-controle. Além disso, os indivíduos fossoriais primários deslocados tiveram um sucesso significativamente menor em comparação com os fossoriais terciários deslocados. Fossoriais primários exibiram maior erro de entocamento e tiveram os comportamentos de entocamento significativamente prejudicados em comparação aos fossoriais terciários. Ademais, os ângulos de direção nos grupos deslocados (de ambas as espécies) foram significativamente maiores que os do grupo-controle de ambas as espécies. Diferenças específicas para cada espécie no sucesso e no erro de entocamento indicam que as espécies fossoriais primárias foram mais impactadas negativamente pelos deslocamentos translacionais. Essas diferenças indicam que as duas espécies de lagostins têm diferentes estratégias de orientação. translated to Portuguese by G. Sobral (gabisobral@gmail.com).

Impact of an electronic health record alert in primary care on increasing hepatitis c screening and curative treatment for baby boomers.

Despite effective treatment for chronic hepatitis C, deficiencies in diagnosis and access to care preclude disease elimination. Screening of baby boomers remains low. The aims of this study were to assess the impact of an electronic health record-based prompt on hepatitis C virus (HCV) screening rates in baby boomers in primary care and access to specialty care and treatment among those newly diagnosed. We implemented an electronic health record-based "best practice advisory" (BPA) that prompted primary care providers to perform HCV screening for patients seen in primary care clinic (1) born between 1945 and 1965, (2) who lacked a prior diagnosis of HCV infection, and (3) who lacked prior documented anti-HCV testing. The BPA had associated educational materials, order set, and streamlined access to specialty care for newly diagnosed patients. Pre-BPA and post-BPA screening rates were compared, and care of newly diagnosed patients was analyzed. In the 3 years prior to BPA implementation, 52,660 baby boomers were seen in primary care clinics and 28% were screened. HCV screening increased from 7.6% for patients with a primary care provider visit in the 6 months prior to BPA to 72% over the 1 year post-BPA. Of 53 newly diagnosed patients, all were referred for specialty care, 11 had advanced fibrosis or cirrhosis, 20 started treatment, and 9 achieved sustained virologic response thus far. CONCLUSION: Implementation of an electronic health record-based prompt increased HCV screening rates among baby boomers in primary care by 5-fold due to efficiency in determining needs for HCV screening and workflow design. Streamlined access to specialty care enabled patients with previously undiagnosed advanced disease to be cured. This intervention can be easily integrated into electronic health record systems to increase HCV diagnosis and linkage to care. (Hepatology 2017;66:1805-1813).

Self-efficacy Mediates the Relationship between Balance/Walking Performance, Activity, and Participation after Stroke.

BACKGROUND: Many outcome measures (OM) that assess individuals' ability or beliefs in their ability to perform tasks exist to evaluate activity and participation after stroke; however, the relationship between various OM and activity/participation is unclear. OBJECTIVE: The purpose of this study was to explore the relationships between different OM and activity and participation in people after stroke. METHODS: Fifty-nine subjects post stroke participated in an assessment including self-selected walking speed, 6 minute walk test, Timed "Up and Go" test, Berg Balance Scale, Functional Gait Assessment, Walk 12, and Activity-specific Balance Confidence Scale. Step Watch Activity Monitoring (SAM) was used as a measure of activity and Stroke Impact Scale-Participation (SIS-P) as a measure of participation. Exploratory Factor Analysis was performed including all measures except SAM and SIS-P. Two factors were extracted and termed performance based (PB) and self-efficacy (SE). A path analysis assessed the role of SE as a mediator in the relationships of PB and SAM/SIS-P. RESULTS: In the path analysis, PB significantly predicts SE (p < 0.001, b = 0.44), but not SAM or SIS-P (p > 0.05, b = 0.25, and b = 0.11, respectively). SE significantly predicts both SAM and SIS-P (p < 0.001, b = 0.46, and b = 0.59, respectively). The indirect effects of PB on SAM and SIS-P were significant (p < 0.001; b = 0.20, and b = 0.26, respectively). CONCLUSION: These results suggest that SE mediates the relationship between PB and activity and participation after stroke, reinforcing that improving activity and participation is more complicated than only targeting performance. Clinicians should administer SE and PB measures to determine the most accurate view of patients after stroke and seek to improve SE through interventions.

Investigation of maternal and fetal exposure to an IgG2 monoclonal antibody following biweekly intravaginal administration to cynomolgus monkeys throughout pregnancy.

To assess the potential for male-mediated drug transfer to their female partner and/or developing conceptus, vaginal uptake of a monoclonal antibody (mAb) biotherapeutic was assessed in cynomolgus monkeys. A human IgG2 mAb (IgG2X; bound human and cynomolgus monkey neonatal Fc-receptor, FcRn, with similar high affinity) was administered intravaginally (IvG; 100mg/dose) to 5 pregnant cynomolgus monkeys biweekly from gestation day (gd) 21 to gd133. In all maternal samples collected before gd119, IgG2X plasma concentrations were below the limit of quantification (BLQ; <25ng/mL). After dosing on gd119 and 133, maternal IgG2X plasma concentrations remained BLQ in 3/5 monkeys and were very low in 2/5 (up to 116ng/mL; ∼0.01% of the IvG dose). IgG2X was BLQ in all fetal plasma samples. These data indicate that male-mediated mAb drug transfer via seminal fluid does not present a health risk to the female partner and is not bioavailable to the developing conceptus.