Of all methods exercised in modern molecular biology, modification of cellular properties through the introduction or removal of nucleic acids is one of the most fundamental. As such, several methods ...have arisen to promote this process; these include the condensation of nucleic acids with calcium, polyethylenimine or modified lipids, electroporation, viral production, biolistics, and microinjection. An ideal transfection method would be (1) low cost, (2) exhibit high levels of biological safety, (3) offer improved efficacy over existing methods, (4) lack requirements for ongoing consumables, (5) work efficiently at any scale, (6) work efficiently on cells that are difficult to transfect by other methods, and (7) be capable of utilizing the widest array of existing genetic resources to facilitate its utility in research, biotechnical and clinical settings. To address such issues, we describe here Pressure-jump-poration (PJP), a method using rapid depressurization to transfect even difficult to modify primary cell types such as embryonic stem cells. The results demonstrate that PJP can be used to introduce an array of genetic modifiers in a safe, sterile manner. Finally, PJP-induced transfection in primary versus transformed cells reveals a surprising dichotomy between these classes which may provide further insight into the process of cellular transformation.
Full text
Available for:
IZUM, KILJ, NUK, PILJ, PNG, SAZU, UL, UM, UPUK
Entangling detectors in anti-de Sitter space Henderson, Laura J.; Hennigar, Robie A.; Mann, Robert B. ...
The journal of high energy physics,
05/2019, Volume:
2019, Issue:
5
Journal Article
Peer reviewed
Open access
A
bstract
We examine in (2+1)-dimensional anti-de Sitter (AdS) space the phenomena of entanglement harvesting — the process in which a pair of detectors (two-level atoms) extract entanglement from a ...quantum field through local interactions with the field. We begin by reviewing the Unruh-DeWitt detector and its interaction with a real scalar field in the vacuum state, as well as the entanglement harvesting protocol in general. We then examine how the entanglement harvested by a pair of such detectors depends on their spacetime trajectory, separation, spacetime curvature, and boundary conditions satisfied by the field. The harvested entanglement is interpreted as an indicator of field entanglement between the localized regions where the detectors interact with the field, and thus this investigation allows us to probe indirectly the entanglement structure of the AdS vacuum. We find an island of separability for specific values of the detectors’ energy gap and separation at intermediate values of the AdS length for which entanglement harvesting is not possible; an analogous phenomena is observed in AdS
4
, to which we compare and contrast our results. In the process we examine how the transition probability of a single detector, as a proxy for local fluctuations of the field, depends on spacetime curvature, its location in AdS space, and boundary conditions satisfied by the field.
We implement the entanglement harvesting protocol, in which two Unruh-DeWitt detectors become entangled through local interactions with a quantum field, for the first time in the vicinity of a black ...hole. Our study, focusing on the BTZ black hole, reveals that black holes inhibit entanglement harvesting. For two detectors with fixed proper separation along the radial direction, we demonstrate that the entanglement harvested rapidly falls to zero as the detectors move closer to the horizon. This effect is a combination of black hole radiation and gravitational redshift, both generic properties of horizons, suggesting it is a general result for black holes.
The thesis of this article is that multilevel interventions based on ecological models and targeting individuals, social environments, physical environments, and policies must be implemented to ...achieve population change in physical activity. A model is proposed that identifies potential environmental and policy influences on four domains of active living: recreation, transport, occupation, and household. Multilevel research and interventions require multiple disciplines to combine concepts and methods to create new transdisciplinary approaches. The contributions being made by a broad range of disciplines are summarized. Research to date supports a conclusion that there are multiple levels of influence on physical activity, and the active living domains are associated with different environmental variables. Continued research is needed to provide detailed findings that can inform improved designs of communities, transportation systems, and recreation facilities. Collaborations with policy researchers may improve the likelihood of translating research findings into changes in environments, policies, and practices.
Objective
To evaluate the safety and efficacy of belimumab as adjunctive therapy to maintain remission in antineutrophil cytoplasmic antibody (ANCA)–associated vasculitis (AAV).
Methods
In this ...multicenter, double‐blind, placebo‐controlled study, patients with AAV (ages ≥18 years) were randomized 1:1 to receive azathioprine (2 mg/kg/day), low‐dose oral glucocorticoids (≤10 mg/day), and either intravenous belimumab (10 mg/kg) or placebo, following remission induction with rituximab or cyclophosphamide along with glucocorticoids. The primary end point was time to first protocol‐specified event (PSE), with first PSE defined as a Birmingham Vasculitis Activity Score (BVAS) of ≥6, presence of ≥1 major BVAS item, or receipt of prohibited medications for any reason, resulting in treatment failure (adjusted for ANCA type proteinase 3 (PR3) or myeloperoxidase (MPO), disease stage at induction, and induction regimen). Vasculitis relapse was defined as the PSE of either a BVAS activity score of ≥6 or receipt of prohibited medications for vasculitis. Changes in treatment practice led to truncation of the study population from ~300 patients to ~100 patients.
Results
The intent‐to‐treat population totaled 105 patients with AAV, of whom 52 (40 with PR3‐ANCAs, 12 with MPO‐ANCAs) received placebo and 53 (41 with PR3‐ANCAs, 12 with MPO‐ANCAs) received belimumab; 27 of the patients were in rituximab‐induced disease remission, while 78 were in cyclophosphamide‐induced disease remission at baseline. Compared with placebo, treatment with belimumab did not reduce the risk of a PSE (adjusted hazard ratio HR 1.07, 95% confidence interval 95% CI 0.44–2.59; P = 0.884) or vasculitis relapse (adjusted HR 0.88, 95% CI 0.29–2.65; P = 0.821). The overall rate of PSEs was low (11 21.2% of 52 patients receiving placebo, 10 18.9% of 53 patients receiving belimumab). Vasculitis relapse in the placebo group (n = 8) occurred independent of the induction regimen, disease stage, or ANCA type. All vasculitis relapses in the belimumab group (n = 6) occurred in patients who had PR3‐ANCA–associated vasculitis with cyclophosphamide‐induced disease remission. Adverse events occurred in 49 (92.5%) of 53 patients receiving belimumab and 43 (82.7%) of 52 patients receiving placebo, with no new safety concerns.
Conclusion
Belimumab plus azathioprine and glucocorticoids for the maintenance of remission in AAV did not reduce the risk of relapse.
Full text
Available for:
BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
Abstract
Background
Immunosuppressant drugs reduce proteinuria and anti-phospholipase A2 receptor autoantibodies (PLA2R-Ab) in primary membranous nephropathy (PMN) with varying success and associated ...toxicities. This study aimed to evaluate the effect of belimumab on proteinuria and PLA2R-Ab in participants with PMN.
Methods
In this prospective, open-label, experimental medicine study, 14 participants with PMN and persistent nephrotic-range proteinuria received up to 2 years belimumab monotherapy (10 mg/kg, every 4 weeks). Changes in proteinuria (urinary protein:creatinine ratio), PLA2R-Ab, albumin, cholesterol, B-cell subsets and pharmacokinetics were analysed during treatment and up to 6 months after treatment.
Results
Eleven participants completed to the primary endpoint (Week 28) and nine participants completed the study. In the intention-to-treat population population, baseline proteinuria of 724 mg/mmol 95% confidence interval (CI) 579–906 decreased to 498 mg/mmol (95% CI 383–649) and 130 mg/mmol (95% CI 54–312) at Weeks 28 and 104, respectively, with changes statistically significant from Week 36 (n = 11, P = 0.047). PLA2R-Ab decreased from 174 RU/mL (95% CI 79–384) at baseline to 46 RU/mL (95% CI 16–132) and 4 RU/mL (95% CI 2–6) at Weeks 28 and 104, respectively, becoming statistically significant by Week 12 (n = 13, P = 0.02). Nine participants achieved partial (n = 8) or complete (n = 1) remission. Participants with abnormal albumin and/or cholesterol at baseline gained normal/near normal levels by the last follow-up. Adverse events were consistent with those expected in this population.
Conclusions
Belimumab treatment in participants with PMN can reduce PLA2R-Ab and subsequently proteinuria, important preludes to remission induction.
Neutrophils are the first immune cells to migrate into infected tissue sites. Therefore an important step in the initiation of an immune response is the synthesis of the neutrophil-recruiting ...chemokines. In this in vivo study in mice, we show that resident tissue macrophages are the source of the major neutrophil chemoattractants, KC and MIP-2. Synthesis of these chemokines is rapidly regulated at the transcriptional level by signaling through TLR2, TLR3, and TLR4 that have diverse specificities for pathogens. The major and alternative TLR signaling pathways are characterized by the adaptor proteins MyD88 or TRIF, respectively. KC and MIP-2 are both produced by signaling through MyD88. However MIP-2, but not KC, is also synthesized through the TRIF adaptor protein, identifying it as a new product of this alternative pathway. Use of both pathways by TLR4 ensures maximal levels of KC and MIP-2 that lead to robust neutrophil recruitment. However the MIP-2 generated exclusively by the TRIF pathway is still sufficient to cause an influx of neutrophils. In summary we show that TLR signaling by tissue macrophages directly controls the synthesis of neutrophil-attracting chemokines that are essential for the earliest recruitment step in the innate immune response to microbial challenge.
Anti-Hawking phenomena Henderson, Laura J.; Hennigar, Robie A.; Mann, Robert B. ...
Physics letters. B,
10/2020, Volume:
809
Journal Article
Peer reviewed
Open access
The Unruh effect is a surprising prediction of quantum field theory that asserts accelerating observers perceive a thermal spectrum of particles with a temperature proportional to their acceleration. ...However, it has recently been shown that particle detectors can click less often or even cool down as their acceleration increases, in contrast to the heating one would expect. This leads to the so called anti-Unruh phenomena. Here we consider detectors outside a BTZ black hole and demonstrate the existence of black hole analogues of these effects, which we dub anti-Hawking phenomena.
Full text
Available for:
GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Abstract
The quantum vacuum has long been known to be characterized by field correlations between spacetime points. We show that such correlations—vacuum entanglement—in the environment of ...near-extremal black holes is significantly amplified (up to ten-fold) relative to their slowly rotating counterparts. We demonstrate this effect for rotating Banados–Teitelboim–Zanelli black holes by measuring the entanglement through the concurrence extracted from the vacuum via two-level quantum systems (Unruh–DeWitt detectors). The effect is manifest at intermediate distances from the horizon, and is most pronounced for near-extremal small mass black holes. The effect is also robust, holding for all boundary conditions of the field and at large spacelike detector separations. Smaller amplification occurs near the horizon, where we find that the entanglement shadow—a region near the black hole from which entanglement cannot be extracted—is diminished in size as the black hole’s angular momentum increases.
PDF
