Thermal Transport in Nanoelectronic Devices Cooled by On-Chip Magnetic Refrigeration.

On-chip demagnetization refrigeration has recently emerged as a powerful tool for reaching microkelvin electron temperatures in nanoscale structures. The relative importance of cooling on-chip and off-chip components and the thermal subsystem dynamics are yet to be analyzed. We study a Coulomb blockade thermometer with on-chip copper refrigerant both experimentally and numerically, showing that dynamics in this device are captured by a first-principles model. Our work shows how to simulate thermal dynamics in devices down to microkelvin temperatures, and outlines a recipe for a low-investment platform for quantum technologies and fundamental nanoscience in this novel temperature range.

Neutron imaging of an operational dilution refrigerator.

The invention of the 3He/4He dilution refrigerator opened a new chapter in experimental ultra-low temperature physics. Dilution refrigerators became essential for providing ultra-low temperature environments for nuclear demagnetisation experiments, superconducting-qubit quantum processors and highly sensitive bolometers used in fundamental physics experiments. Development of dilution refrigeration technology requires thorough understanding of the quantum mechanical processes that take place in liquid helium at ultra-low temperatures. For decades the quantum fluids research community provided valuable information to engineers and designers involved in the development of advanced dilution refrigerators. However, the lack of methods that allow the measurement of physical parameters of liquid helium during the operation of a dilution refrigerator was hindering development of the technology. Here we show direct imaging of an operational dilution refrigerator using neutron radiography. This allows direct observation of the dilution process in 3He/4He mixtures and opens an opportunity for direct measurement of the 3He concentration. We observe the refrigerator behaviour in different regimes, such as continuous circulation and single shot, and show that our method allows investigation of various failure modes. Our results demonstrate that neutron imaging applied to the study of dilution refrigeration processes can provide essential information for developers of ultra-low temperature systems. We expect that neutron imaging will become instrumental in the research and development of advanced dilution refrigerators.

Trichomoniasis and adverse birth outcomes: a systematic review and meta-analysis.

BACKGROUND: Trichomoniasis commonly affects women of childbearing age and has been linked to several adverse birth outcomes. OBJECTIVE: To elucidate the association between trichomoniasis in pregnant women and adverse birth outcomes, including preterm delivery, prelabour rupture of membranes and low birthweight. SEARCH STRATEGY: MEDLINE, EMBASE and ClinicalTrials.gov were systematically searched in December 2020 without time or language restrictions. SELECTION CRITERIA: Original research studies were included if they assessed at least one of the specified adverse birth outcomes in pregnant women with laboratory-diagnosed trichomoniasis. DATA COLLECTION AND ANALYSIS: Estimates from included articles were either extracted or calculated and then pooled to produce a combined estimate of the association of trichomoniasis with each adverse birth outcome using the random effects model. Heterogeneity was assessed using the I2 statistic and Cochran's Q test. MAIN RESULTS: Literature search produced 1658 publications after removal of duplicates (n = 770), with five additional publications identified by hand search. After screening titles and abstracts for relevance, full text of 84 studies was reviewed and 19 met inclusion criteria for meta-analysis. Significant associations were found between trichomoniasis and preterm delivery (OR 1.27; 95% CI 1.08-1.50), prelabour rupture of membranes (OR 1.87; 95% CI 1.53-2.29) and low birthweight (OR 2.12; 95% CI 1.15-3.91). CONCLUSIONS: Trichomoniasis in pregnant women is associated with preterm delivery, prelabour rupture of membranes and low birthweight. Rigorous studies are needed to determine the impact of universal trichomoniasis screening and treatment during pregnancy on reducing perinatal morbidity. TWEETABLE ABSTRACT: This systematic review and meta-analysis found that in the setting of pregnancy, trichomoniasis is significantly associated with multiple adverse birth outcomes, including preterm delivery, low birthweight, and prelabour rupture of membranes.

Progress in Cooling Nanoelectronic Devices to Ultra-Low Temperatures.

Here we review recent progress in cooling micro-/nanoelectronic devices significantly below 10 mK. A number of groups worldwide are working to produce sub-millikelvin on-chip electron temperatures, motivated by the possibility of observing new physical effects and improving the performance of quantum technologies, sensors and metrological standards. The challenge is a longstanding one, with the lowest reported on-chip electron temperature having remained around 4 mK for more than 15 years. This is despite the fact that microkelvin temperatures have been accessible in bulk materials since the mid-twentieth century. In this review, we describe progress made in the last 5 years using new cooling techniques. Developments have been driven by improvements in the understanding of nanoscale physics, material properties and heat flow in electronic devices at ultralow temperatures and have involved collaboration between universities and institutes, physicists and engineers. We hope that this review will serve as a summary of the current state of the art and provide a roadmap for future developments. We focus on techniques that have shown, in experiment, the potential to reach sub-millikelvin electron temperatures. In particular, we focus on on-chip demagnetisation refrigeration. Multiple groups have used this technique to reach temperatures around 1 mK, with a current lowest temperature below 0.5 mK.

Epidemiology of functional stroke mimic patients: a systematic review and meta-analysis.

Stroke mimics form a significant proportion of cases in acute stroke services and some present with functional neurological disorder. Little is known about the prevalence or clinical characteristics of functional stroke mimics (FSMs). A systematic literature search and meta-analysis were carried out on published studies reporting suspected stroke and stroke mimic rates; 114 papers met the inclusion criteria of which 70 provided an FSM rate. Random-effects models estimated prevalence rates across settings and moderators of FSM rate. Pooled proportions indicated that 25% [95% confidence intervals (CI), 22-27%] of suspected stroke cases were stroke mimics. Within the 67 studies providing positive FSM rates, FSMs represented 15% (95% CI, 13-18%) of stroke mimics and 2% (95% CI, 2-3%) of suspected strokes. FSMs were younger and more likely to be female, and presented more with weakness/numbness but less with reduced consciousness or language problems. Stratified analyses suggested higher stroke mimic rates in primary care versus acute settings (38% vs. 12%) but higher FSM rates in stroke units compared with primary care (24% vs. 12%). Functional rates were higher in studies that were descriptive, retrospective and in patients receiving thrombolysis. Several studies reported the proportion of functional stroke patients presenting to stroke services. FSMs have discernible demographic and clinical characteristics, but there is a conspicuous lack of evidence on their presentation or guidance for treatment. The social and psychological mechanisms underlying FSM presentations need more accurate quantification to help inform stroke pathways and improve care for these patients.

EJP18 peptide derived from the juxtamembrane domain of epidermal growth factor receptor represents a novel membrane-active cell-penetrating peptide.

Membrane-active peptides have been extensively studied to probe protein-membrane interactions, to act as antimicrobial agents and cell-penetrating peptides (CPPs) for the delivery of therapeutic agents to cells. Hundreds of membrane-active sequences acting as CPPs have now been described including bioportides that serve as single entity modifiers of cell physiology at the intracellular level. Translation of promising CPPs in pre-clinical studies have, however, been disappointing as only few identified delivery systems have progressed to clinical trials. To search for novel membrane-active peptides a sequence from the EGFR juxtamembrane region was identified (named EJP18), synthesised, and examined in its L- and D-form for its ability to mediate the delivery of a small fluorophore and whole proteins to cancer cell lines. Initial studies identified the peptide as being highly membrane-active causing extensive and rapid plasma membrane reorganisation, blebbing, and toxicity. At lower, non-toxic concentrations the peptides outperformed the well-characterised CPP octaarginine in cellular delivery capacity for a fluorophore or proteins that were associated with the peptide covalently or via ionic interactions. EJP18 thus represents a novel membrane-active peptide that may be used as a naturally derived model for biophysical protein-membrane interactions or for delivery of cargo into cells for therapeutic or diagnostic applications.

LEGO® Block Structures as a Sub-Kelvin Thermal Insulator.

We report measurements of the thermal conductance of a structure made from commercial Acrylonitrile Butadiene Styrene (ABS) modules, known as LEGO® blocks, in the temperature range from 70 mK to 1.8 K. A power law for the sample's thermal conductivity κ = (8.7 ± 0.3) × 10-5 T 1.75±0.02 WK-1 m-1 was determined. We conclude that this ABS/void compound material provides better thermal isolation than well-known bulk insulator materials in the explored temperature range, whilst maintaining solid support. LEGO blocks represent a cheap and superlative alternative to materials such as Macor or Vespel. In our setup, <400 nW of power can heat an experimental area of 5 cm2 to over 1 K, without any significant change to the base temperature of the dilution refrigerator. This work suggests that custom-built modular materials with even better thermal performance could be readily and cheaply produced by 3D printing.

Developmental role of adenosine kinase for the expression of sex-dependent neuropsychiatric behavior.

Deficits in social memory, cognition, and aberrant responses to stimulants are common among persons affected by schizophrenia and other conditions with a presumed developmental etiology. We previously found that expression changes in the adenosine metabolizing enzyme adenosine kinase (ADK) in the adult brain are associated with deficits in various cognitive domains. To distinguish between developmental and adult functions of ADK, we used two transgenic mouse lines with widespread disruption of ADK expression in the adult brain, but differences in the onset of ADK deletion. Specifically, we compared Nestin-Cre+/-:ADK-floxfl/fl (ADKΔBrain) mice with global loss of ADK in the whole brain, beginning in mid-gestation and persisting for life, with Gfa2-Cre+/-:ADK-floxfl/fl (ADKΔAstro) mice that have normal ADK expression throughout development, but lose astrocyte-specific ADK-expression in young adulthood. Because ADK-expression in adulthood is generally confined to astrocytes, adult ADKΔAstro mice show a similar expression profile of ADK in key areas of the brain related to neuropsychiatric behavior, compared to adult ADKΔBrain mice. We sought to determine a neurodevelopmental role of ADK on the expression of psychiatric behaviors in adult male and female mice. Adult ADKΔBrain mice showed significant deficits in social memory in males, significant contextual learning impairments in both sexes, and a hyper-responsiveness to amphetamine in males. In contrast, ADKΔAstro mice showed normal social memory and contextual learning but hypo-responsiveness to amphetamine in males. Our results demonstrate a key developmental role of ADK in mediating behaviors in adulthood related to neuropsychiatric disease and support the greater prevalence of these disorders among males.

Contrasting roles for actin in the cellular uptake of cell penetrating peptide conjugates.

The increased need for macromolecular therapeutics, such as peptides, proteins and nucleotides, to reach intracellular targets necessitates more effective delivery vectors and a higher level of understanding of their mechanism of action. Cell penetrating peptides (CPPs) can transport a range of macromolecules into cells, either through direct plasma membrane translocation or endocytosis. All known endocytic pathways involve cell-cortex remodelling, a process shown to be regulated by reorganisation of the actin cytoskeleton. Here using flow cytometry, confocal microscopy and a variety of actin inhibitors we identify how actin disorganisation in different cell types differentially influences the cellular entry of three probes: the CPP octaarginine - Alexa488 conjugate (R8-Alexa488), octaarginine conjugated Enhanced Green Fluorescent Protein (EGFP-R8), and the fluid phase probe dextran. Disrupting actin organisation in A431 skin epithelial cells dramatically increases the uptake of EGFP-R8 and dextran, and contrasts strongly to inhibitory effects observed with transferrin and R8 attached to the fluorophore Alexa488. This demonstrates that uptake of the same CPP can occur via different endocytic processes depending on the conjugated fluorescent entity. Overall this study highlights how cargo influences cell uptake of this peptide and that the actin cytoskeleton may act as a gateway or barrier to endocytosis of drug delivery vectors.

Fluorescence labelling of extracellular vesicles using a novel thiol-based strategy for quantitative analysis of cellular delivery and intracellular traffic.

Extracellular vesicles, including exosomes, are naturally derived nanovesicles generated in and released by numerous cell types. As extracellular entities they have the capacity to interact with neighbouring cells and distant tissues and affect physiological processes as well as being implicated in numerous diseases including tumorigenesis and neurodegeneration. They are also under intense investigation as delivery vectors for biotherapeutics. The ways in which EVs interact with recipient cells to influence cell physiology and deliver a macromolecular payload are at the early stages of exploration. A significant challenge within these studies is the ability to label EVs directly or indirectly with fluorescent probes to allow visualization without compromising functionality. Here, we present a thiol-based fluorescence labelling method allowing comprehensive analysis of the cellular uptake of prostate cancer derived EVs in live cells using confocal microscopy. Labelling of the EVs in this way did not influence their size and had no effect on their ability to induce differentiation of lung fibroblasts to myofibroblasts. For endocytosis analyses, depletion of key endocytic proteins and the use of chemical inhibitors (Dynasore, EIPA, Rottlerin and IPA-3) indicated that fluid-phase endocytosis and/or macropinocytosis was involved in EV internalisation. Over a period of six hours EVs were observed to increasingly co-localise with lysosomes, indicating a possible termination point following internalisation. Overall this method provides new opportunities for analysing the cellular dynamics of EVs as biological entities affecting cell and whole body physiology as well as investigating their potential as drug delivery vectors.

Control of targeting ligand display by pH-responsive polymers on gold nanoparticles mediates selective entry into cancer cells.

Selective targeting of cells for intracellular delivery of therapeutics represents a major challenge for pharmaceutical intervention in disease. Here we show pH-triggered receptor-mediated endocytosis of nanoparticles via surface ligand exposure. Gold nanoparticles were decorated with two polymers: a 2 kDa PEG with a terminal folate targeting ligand, and a di-block copolymer including a pH-responsive and a hydrophilic block. At the normal serum pH of 7.4, the pH-responsive block (apparent pKa of 7.1) displayed a hydrophilic extended conformation, shielding the PEG-folate ligands, which inhibited cellular uptake of the nanoparticles. Under pH conditions resembling those of the extracellular matrix around solid tumours (pH 6.5), protonation of the pH-responsive polymer triggered a coil-to-globule polymer chain contraction, exposing folate residues on the PEG chains. In line with this, endocytosis of folate-decorated polymer-coated gold nanoparticles in cancer cells overexpressing folate receptor was significantly increased at pH 6.5, compared with pH 7.4. Thus, the tumour acidic environment and high folate receptor expression were effectively exploited to activate cell binding and endocytosis of these nanoparticles. These data provide proof-of-concept for strategies enabling extracellular pH stimuli to selectively enhance cellular uptake of drug delivery vectors and their associated therapeutic cargo.

On-chip magnetic cooling of a nanoelectronic device.

We demonstrate significant cooling of electrons in a nanostructure below 10 mK by demagnetisation of thin-film copper on a silicon chip. Our approach overcomes the typical bottleneck of weak electron-phonon scattering by coupling the electrons directly to a bath of refrigerated nuclei, rather than cooling via phonons in the host lattice. Consequently, weak electron-phonon scattering becomes an advant- age. It allows the electrons to be cooled for an experimentally useful period of time to temperatures colder than the dilution refrigerator platform, the incoming electrical connections, and the host lattice. There are efforts worldwide to reach sub-millikelvin electron temperatures in nanostructures to study coherent electronic phenomena and improve the operation of nanoelectronic devices. On-chip magnetic cooling is a promising approach to meet this challenge. The method can be used to reach low, local electron temperatures in other nanostructures, obviating the need to adapt traditional, large demagnetisation stages. We demonstrate the technique by applying it to a nanoelectronic primary thermometer that measures its internal electron temperature. Using an optimised demagnetisation process, we demonstrate cooling of the on-chip electrons from 9 mK to below 5 mK for over 1000 seconds.

Can estimates of genetic effective population size contribute to fisheries stock assessments?

Sustainable exploitation of fisheries populations is challenging to achieve when the size of the population prior to exploitation and the actual numbers removed over time and across fishing zones are not clearly known. Quantitative fisheries' modeling is able to address this problem, but accurate and reliable model outcomes depend on high quality input data. Much of this information is obtained through the operation of the fishery under consideration, but while this seems appropriate, biases may occur. For example, poorly quantified changes in fishing methods that increase catch rates can erroneously suggest that the overall population size is increasing. Hence, the incorporation of estimates of abundance derived from independent data sources is preferable. We review and evaluate a fisheries-independent method of indexing population size; inferring adult abundance from estimates of the genetic effective size of a population (Ne ). Recent studies of elasmobranch species have shown correspondence between Ne and ecologically determined estimates of the population size (N). Simulation studies have flagged the possibility that the range of Ne /N ratios across species may be more restricted than previously thought, and also show that declines in Ne track declines in the abundance of model fisheries species. These key developments bring this new technology closer to implementation in fisheries science, particularly for data-poor fisheries or species of conservation interest.

Uniaxial ferroelectric quantum criticality in multiferroic hexaferrites BaFe12O19 and SrFe12O19.

BaFe12O19 is a popular M-type hexaferrite with a Néel temperature of 720 K and is of enormous commercial value ($3 billion/year). It is an incipient ferroelectric with an expected ferroelectric phase transition extrapolated to lie at 6 K but suppressed due to quantum fluctuations. The theory of quantum criticality for such uniaxial ferroelectrics predicts that the temperature dependence of the electric susceptibility χ diverges as 1/T(3), in contrast to the 1/T(2) dependence found in pseudo-cubic materials such as SrTiO3 or KTaO3. In this paper we present evidence of the susceptibility varying as 1/T(3), i.e. with a critical exponent γ = 3. In general γ = (d + z - 2)/z, where the dynamical exponent for a ferroelectric z = 1 and the dimension is increased by 1 from deff = 3 + z to deff = 4 + z due to the effect of long-range dipole interactions in uniaxial as opposed to multiaxial ferroelectrics. The electric susceptibility of the incipient ferroelectric SrFe12O19, which is slightly further from the quantum phase transition is also found to vary as 1/T(3).

Improved confidence intervals for the linkage disequilibrium method for estimating effective population size.

The linkage disequilibrium method is currently the most widely used single sample estimator of genetic effective population size. The commonly used software packages come with two options, referred to as the parametric and jackknife methods, for computing the associated confidence intervals. However, little is known on the coverage performance of these methods, and the published data suggest there may be some room for improvement. Here, we propose two new methods for generating confidence intervals and compare them with the two in current use through a simulation study. The new confidence interval methods tend to be conservative but outperform the existing methods for generating confidence intervals under certain circumstances, such as those that may be encountered when making estimates using large numbers of single-nucleotide polymorphisms.

Differentiation of tumour-promoting stromal myofibroblasts by cancer exosomes.

Activation of myofibroblast rich stroma is a rate-limiting step essential for cancer progression. The responsible factors are not fully understood, but TGFß1 is probably critical. A proportion of TGFß1 is associated with extracellular nano-vesicles termed exosomes, secreted by carcinoma cells, and the relative importance of soluble and vesicular TGFß in stromal activation is presented. Prostate cancer exosomes triggered TGFß1-dependent fibroblast differentiation, to a distinctive myofibroblast phenotype resembling stromal cells isolated from cancerous prostate tissue; supporting angiogenesis in vitro and accelerating tumour growth in vivo. Myofibroblasts generated using soluble TGFß1 were not pro-angiogenic or tumour-promoting. Cleaving heparan sulphate side chains from the exosome surface had no impact on TGFß levels yet attenuated SMAD-dependent signalling and myofibroblastic differentiation. Eliminating exosomes from the cancer cell secretome, targeting Rab27a, abolished differentiation and lead to failure in stroma-assisted tumour growth in vivo. Exosomal TGFß1 is therefore required for the formation of tumour-promoting stroma.

Distal phenylalanine modification for enhancing cellular delivery of fluorophores, proteins and quantum dots by cell penetrating peptides.

For cell penetrating peptides (CPPs) to fulfil their promise as effective delivery vectors we need a better understanding of their mechanisms of cell binding and uptake. This is especially the case when they are linked to different types of cargo. Here we describe new studies based on our previous findings suggesting that, for peptide-CPP chimeras, distal hydrophobic residues upstream of the CPP sequence can have profound effects on the way they interact with cells. We studied peptides bearing an N-terminal Glycine or Phenylalanine linked via a neutral and flexible bridging group, SGSGSGSG, to three well-studied CPPs: octaarginine, penetratin and TP10. Using a combination of flow cytometry, live-cell imaging and image analysis we examined the effects of this single amino acid change on binding and uptake of Alexa488-fluorophore, bovine serum albumin and quantum dot cargoes. The influence of the glycine-phenylalanine switch for fluorophore delivery was most dramatic in TP10, increasing cellular uptake by 4.4 and 9.9 fold in non-adherent and adherent cells, respectively. Only penetratin showed effective uptake of bovine serum albumin with the phenylalanine variant showing an increase of 1.6 fold over the glycine variant. The uptake of quantum dots was most efficiently demonstrated by octaarginine, with the glycine variant increasing uptake 4.8 fold and the phenylalanine variant increasing uptake 9.5 fold over quantum dots alone. Overall the data demonstrate that hydrophobicity distal to the CPP could be utilised to enhance their capacity to bind to the cell membrane and deliver a range of macromolecules to the insides of cells.

siRNA and pharmacological inhibition of endocytic pathways to characterize the differential role of macropinocytosis and the actin cytoskeleton on cellular uptake of dextran and cationic cell penetrating peptides octaarginine (R8) and HIV-Tat.

Cell penetrating peptides (CPPs) have been extensively studied as vectors for cellular delivery of therapeutic macromolecules. It is widely accepted that they can enter cells directly across the plasma membrane but also gain access through endocytic pathways that are yet to be fully defined. Here we developed siRNA methods in epithelial cell lines, HeLa and A431, to inhibit endocytic pathways regulated by clathrin heavy chain, flotillin-1, caveolin-1, dynamin-2 and Pak-1. In each case, functional uptake assays were developed to characterize the requirement for these proteins, and the pathways they regulate, in the internalisation of defined endocytic probes and also the CPPs octaarginine and HIV-Tat. Peptide uptake was only inhibited in A431 cells depleted of the macropinocytosis regulator Pak-1, but experimental variables including choice of cell line, pharmacological inhibitor, macropinocytic probe and serum starvation significantly influence our ability to assess and assign this pathway as an important route for CPP uptake. Actin disruption with Cytochalasin D inhibited peptide entry in both cell lines but the effects of this agent on dextran uptake was cell line dependent, reducing uptake in HeLa cells and increasing uptake in A431 cells. This was further supported in experiments inducing actin stabilisation by Jasplakinolide, emphasising that the actin cytoskeleton can both promote and hinder endocytosis. Overall the data identify important aspects regarding the comparative mechanisms of CPP uptake and macropinocytosis, and accentuate the significant methodological challenges of studying this pathway as an endocytic portal and an entry route for drug delivery vectors.

An analysis of subdomain orientation, conformational change and disorder in relation to crystal packing of aspartic proteinases.

The analysis reported here describes detailed structural studies of endothiapepsin (the aspartic proteinase from Endothia parasitica), with and without bound inhibitors, and human pepsin 3b. Comparison of multiple crystal structures of members of the aspartic proteinase family has revealed small but significant differences in domain orientation in different crystal forms. In this paper, it is shown that these differences in domain orientation do not necessarily correlate with the presence or absence of bound inhibitors, but appear to stem at least partly from crystal contacts mediated by sulfate ions. However, since the same inherent flexibility of the structure is observed for other enzymes in this family such as human pepsin, the native structure of which is also reported here, the observed domain movements may well have implications for the mechanism of catalysis.

Prostate cancer in BRCA2 germline mutation carriers is associated with poorer prognosis.

BACKGROUND: The germline BRCA2 mutation is associated with increased prostate cancer (PrCa) risk. We have assessed survival in young PrCa cases with a germline mutation in BRCA2 and investigated loss of heterozygosity at BRCA2 in their tumours. METHODS: Two cohorts were compared: one was a group with young-onset PrCa, tested for germline BRCA2 mutations (6 of 263 cases had a germline BRAC2 mutation), and the second was a validation set consisting of a clinical set from Manchester of known BRCA2 mutuation carriers (15 cases) with PrCa. Survival data were compared with a control series of patients in a single clinic as determined by Kaplan-Meier estimates. Loss of heterozygosity was tested for in the DNA of tumour tissue of the young-onset group by typing four microsatellite markers that flanked the BRCA2 gene, followed by sequencing. RESULTS: Median survival of all PrCa cases with a germline BRCA2 mutation was shorter at 4.8 years than was survival in controls at 8.5 years (P=0.002). Loss of heterozygosity was found in the majority of tumours of BRCA2 mutation carriers. Multivariate analysis confirmed that the poorer survival of PrCa in BRCA2 mutation carriers is associated with the germline BRCA2 mutation per se. CONCLUSION: BRCA2 germline mutation is an independent prognostic factor for survival in PrCa. Such patients should not be managed with active surveillance as they have more aggressive disease.