Development of an Attenuated Zika Virus by Editing the 5’ Untranslated Region

Zika Virus (ZIKV) is a mosquito-transmitted flavivirus that usually causes no symptoms to mild febrile in humans, and it has been regarded as an insignificant pathogen to public health. However, recent outbreaks of ZIKV infection have revealed that ZIKV can cause severe neurological effects in adults, such as Guillain-Barre Syndrome (GBS), and in infants whose mothers acquired the virus during pregnancy, causing Congenital Zika Syndrome (CZS). Currently, no approved vaccine is available and there is a critical need to develop an effective and safe vaccine. While most vaccine developmental strategies target the viral prM-E protein of ZIKV, we aimed to create an attenuated ZIKV by inserting several different nucleotide lengths into the 5’ untranslated region (UTR) of the viral genome. We generated a viable mutated ZIKV virus (Z7) was after inserting a 50-nt sequence into the 5’UTR. We then characterized Z7 by comparing growth kinetics in vivo replication by using qPCR and immunostaining assays in cell cultures. We also measured the infectivity and immunogenicity of Z7 in Ifnar1-/-mice. We found that this mutated virus developed a lower viremia compared to WT ZIKV control but induced a similar level of antibody response. Importantly, one dose of Z7 inoculation can protect the mice from a secondary high dose of ZIKV infection in Ifnar1-/-mice. Together, these results suggest that we have successfully developed an attenuated ZIKV strain, which induces protective immune responses against ZIKV infection in mice

Evaluating the Impact of Grafting on Local Tomato Production in Nebraska

Grafting has been successfully used in vegetable production for tomatoes, peppers, eggplants, cucumbers, and watermelon. Besides its usefulness for managing soil-borne diseases, grafting can improve nutrient uptake and yield. However, few studies have assessed the effects of grafting and soil fertility management on yield of open field-grown tomatoes in the Midwest. Therefore, the objective of this two-year research was to better document the effects of grafting heirloom and hybrid tomato cultivar onto hybrid tomato rootstocks on tomato yield and quality. The field experiments were located at the University of Nebraska Lincoln - East Campus in Lincoln, West Central Research and Extension Center in North Platte, and a farm trial at Perkarek’s Vegetable Farm near Dwight, Nebraska. Two determinant fresh market tomatoes, ‘Nebraska Wedding’ and ‘BHN-589’, were grafted onto two rootstocks, ‘Estamino’ and ‘Maxifort,’ with the non-grafted determinant market tomatoes as a control. During the second year of this study, fertilizer treatment was introduced with two N rates (0% and 100%). At the end of the growing season, ripe tomatoes were harvested on a weekly basis, and yield was determined by weighing all tomatoes from the five plants in each experimental unit. Overall, there was no consistent improvement in total yield for any of the grafting treatments, and the estimated total mean yield of BHN-589 was at least 50% more than Nebraska Wedding. Moreover, there was no interaction effect between grafting and fertilizing treatment within each location. Results from this study suggest the need for more assessment on the effect of tomato grafting under different environmental conditions

Current Understanding of West Nile Virus Clinical Manifestations, Immune Responses, Neuroinvasion, and Immunotherapeutic Implications

West Nile virus (WNV) is the most common mosquito-borne virus in North America. WNV-associated neuroinvasive disease affects all ages, although elderly and immunocompromised individuals are particularly at risk. WNV neuroinvasive disease has killed over 2300 Americans since WNV entered into the United States in the New York City outbreak of 1999. Despite 20 years of intensive laboratory and clinical research, there are still no approved vaccines or antivirals available for human use. However, rapid progress has been made in both understanding the pathogenesis of WNV and treatment in clinical practices. This review summarizes our current understanding of WNV infection in terms of human clinical manifestations, host immune responses, neuroinvasion, and therapeutic interventions

Highly-mass-loaded hot galactic winds are unstable to cool filament formation

When cool clouds are ram-pressure accelerated by a hot supersonic galactic wind, some of the clouds may be shredded by hydrodynamical instabilities and incorporated into the hot flow. Recent one-dimensional steady-state calculations show how cool cloud entrainment directly affects the bulk thermodynamics, kinematics, and observational characteristics of the hot gas. In particular, mass-loading decelerates the hot flow and changes its entropy. Here, we investigate the stability of planar and spherical mass-loaded hot supersonic flows using both perturbation analysis and three-dimensional time-dependent radiative hydrodynamical simulations. We show that mass-loading is stable over a broad range of parameters and that the 1D time-steady analytic solutions exactly reproduce the 3D time-dependent calculations, provided that the flow does not decelerate sufficiently to become subsonic. For higher values of the mass-loading, the flow develops a sonic point and becomes thermally unstable, rapidly cooling and forming elongated dense cometary filaments. We explore the mass-loading parameters required to reach a sonic point and the radiative formation of these filaments. For certain approximations, we can derive simple analytic criteria. In general a mass-loading rate similar to the initial mass outflow rate is required. In this sense, the destruction of small cool clouds by a hot flow may ultimately spontaneously generate fast cool filaments, as observed in starburst superwinds. Lastly, we find that the kinematics of filaments is sensitive to the slope of the mass-loading function. Filaments move faster than the surrounding wind if mass-loading is over long distances whereas filaments move slower than their surroundings if mass-loading is abrupt.Comment: 12 pages, 15 figures, submitted to MNRAS (21 July 2023

Congenital Zika Virus Infection in Immunocompetent Mice Causes Postnatal Growth Impediment and Neurobehavioral Deficits

A small percentage of babies born to Zika virus (ZIKV)-infected mothers\u27 manifest severe defects at birth, including microcephaly. Among those who appeared healthy at birth, there are increasing reports of postnatal growth or developmental defects. However, the impact of congenital ZIKV infection in postnatal development is poorly understood. Here, we report that a mild congenital ZIKV-infection in pups born to immunocompetent pregnant mice did not display apparent defects at birth, but manifested postnatal growth impediments and neurobehavioral deficits, which include reduced locomotor and cognitive deficits that persisted into adulthood. We found that the brains of these pups were smaller, had a thinner cortical layer 1, displayed increased astrogliosis, decreased expression of microcephaly- and neuron development- related genes, and increased pathology as compared to mock-infected controls. In summary, our results showed that even a mild congenital ZIKV infection in immunocompetent mice could lead to postnatal deficits, providing definitive experimental evidence for a necessity to closely monitor postnatal growth and development of presumably healthy human infants, whose mothers were exposed to ZIKV infection during pregnancy

Osteopontin Facilitates West Nile Nirus Neuroinvasion via Neutrophil Trojan Horse Transport

West Nile virus (WNV) can cause severe human neurological diseases including encephalitis and meningitis. The mechanisms by which WNV enters the central nervous system (CNS) and host-factors that are involved in WNV neuroinvasion are not completely understood. The proinflammatory chemokine osteopontin (OPN) is induced in multiple neuroinflammatory diseases and is responsible for leukocyte recruitment to sites of its expression. In this study, we found that WNV infection induced OPN expression in both human and mouse cells. Interestingly, WNV-infected OPN deficient (Opn −/−) mice exhibited a higher survival rate (70%) than wild type (WT) control mice (30%), suggesting OPN plays a deleterious role in WNV infection. Despite comparable levels of viral load in circulating blood cells and peripheral organs in the two groups, WNV-infected polymorphonuclear neutrophil (PMN) infiltration and viral burden in brain of Opn −/− mice were significantly lower than in WT mice. Importantly, intracerebral administration of recombinant OPN into the brains of Opn −/− mice resulted in increased WNV-infected PMN infiltration and viral burden in the brain, which was coupled to increased mortality. The overall results suggest that OPN facilitates WNV neuroinvasion by recruiting WNV-infected PMNs into the brain

Osteopontin Facilitates West Nile Virus Neuroinvasion via Neutrophil “Trojan Horse” Transport

West Nile virus (WNV) can cause severe human neurological diseases including encephalitis and meningitis. The mechanisms by which WNV enters the central nervous system (CNS) and host factors that are involved in WNV neuroinvasion are not completely understood. The proinflammatory chemokine osteopontin (OPN) is induced in multiple neuroinflammatory diseases and is responsible for leukocyte recruitment to sites of its expression. In this study, we found that WNV infection induced OPN expression in both human and mouse cells. Interestingly, WNV-infected OPN deficient (Opn−/−) mice exhibited a higher survival rate (70%) than wild type (WT) control mice (30%), suggesting OPN plays a deleterious role in WNV infection. Despite comparable levels of viral load in circulating blood cells and peripheral organs in the two groups, WNV-infected polymorphonuclear neutrophil (PMN) infiltration and viral burden in brain of Opn−/− mice were significantly lower than in WT mice. Importantly, intracerebral administration of recombinant OPN into the brains of Opn−/− mice resulted in increased WNV-infected PMN infiltration and viral burden in the brain, which was coupled to increased mortality. The overall results suggest that OPN facilitates WNV neuroinvasion by recruiting WNV-infected PMNs into the brain

Acute Skeletal Muscle Wasting and Dysfunction Predict Physical Disability at Hospital Discharge in Patients with Critical Illness

BACKGROUND: Patients surviving critical illness develop muscle weakness and impairments in physical function; however, the relationship between early skeletal muscle alterations and physical function at hospital discharge remains unclear. The primary purpose of this study was to determine whether changes in muscle size, strength and power assessed in the intensive care unit (ICU) predict physical function at hospital discharge. METHODS: Study design is a single-center, prospective, observational study in patients admitted to the medicine or cardiothoracic ICU with diagnosis of sepsis or acute respiratory failure. Rectus femoris (RF) and tibialis anterior (TA) muscle ultrasound images were obtained day one of ICU admission, repeated serially and assessed for muscle cross-sectional area (CSA), layer thickness (mT) and echointensity (EI). Muscle strength, as measured by Medical Research Council-sum score, and muscle power (lower-extremity leg press) were assessed prior to ICU discharge. Physical function was assessed with performance on 5-times sit-to-stand (5STS) at hospital discharge. RESULTS: Forty-one patients with median age of 61 years (IQR 55-68), 56% male and sequential organ failure assessment score of 8.1 ± 4.8 were enrolled. RF muscle CSA decreased significantly a median percent change of 18.5% from day 1 to 7 (F = 26.6, p = 0.0253). RF EI increased at a mean percent change of 10.5 ± 21% in the first 7 days (F = 3.28, p = 0.081). At hospital discharge 25.7% of patients (9/35) met criteria for ICU-acquired weakness. Change in RF EI in first 7 days of ICU admission and muscle power measured prior to ICU were strong predictors of ICU-AW at hospital discharge (AUC = 0.912). Muscle power at ICU discharge, age and ICU length of stay were predictive of performance on 5STS at hospital discharge. CONCLUSION: ICU-assessed muscle alterations, specifically RF EI and muscle power, are predictors of diagnosis of ICU-AW and physical function assessed by 5x-STS at hospital discharge in patients surviving critical illness

Long-range coupling and scalable architecture for superconducting flux qubits

Constructing a fault-tolerant quantum computer is a daunting task. Given any design, it is possible to determine the maximum error rate of each type of component that can be tolerated while still permitting arbitrarily large-scale quantum computation. It is an underappreciated fact that including an appropriately designed mechanism enabling long-range qubit coupling or transport substantially increases the maximum tolerable error rates of all components. With this thought in mind, we take the superconducting flux qubit coupling mechanism described in PRB 70, 140501 (2004) and extend it to allow approximately 500 MHz coupling of square flux qubits, 50 um a side, at a distance of up to several mm. This mechanism is then used as the basis of two scalable architectures for flux qubits taking into account crosstalk and fault-tolerant considerations such as permitting a universal set of logical gates, parallelism, measurement and initialization, and data mobility.Comment: 8 pages, 11 figure

A Randomized Controlled Trial of a Mindfulness-Based Weight Loss Intervention on Cardiovascular Reactivity to Social-Evaluative Threat Among Adults with Obesity.

Objective:Mindfulness-based interventions have been found to reduce psychological and physiological stress reactivity. In obesity, however, stress reactivity is complex, with studies showing both exaggerated and blunted physiological responses to stressors. A nuanced view of stress reactivity is the "challenge and threat" framework, which defines adaptive and maladaptive patterns of psychophysiological stress reactivity. We hypothesized that mindfulness training would facilitate increased challenge-related appraisals, emotions, and cardiovascular reactivity, including sympathetic nervous system activation paired with increased cardiac output (CO) and reduced total peripheral resistance (TPR) compared to a control group, which would exhibit an increased threat pattern of psychophysiological reactivity to repeated stressors.Methods:Adults (N=194) with obesity were randomized to a 5.5-month mindfulness-based weight loss intervention or an active control condition with identical diet-exercise guidelines. Participants were assessed at baseline and 4.5 months later using the Trier Social Stress Task. Electrocardiogram, impedance cardiography, and blood pressure were acquired at rest and during the speech and verbal arithmetic tasks to assess pre-ejection period (PEP), CO, and TPR reactivity.Results:Mindfulness participants showed significantly greater maintenance of challenge-related emotions and cardiovascular reactivity patterns (higher CO and lower TPR) from pre to post-intervention compared to control participants, but groups did not differ in PEP. Findings were independent of changes in body mass index.Conclusions:Mindfulness training may increase the ability to maintain a positive outlook and mount adaptive cardiovascular responses to repeated stressors among persons with obesity though findings need to be replicated in other populations and using other forms of mindfulness interventions