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We developed a novel adeno-associated virus 5 gene therapy (AAV5-GLA) expressing human alpha-galactosidase A (GLA) under the control of a novel, small and strong, liver-restricted promoter. We assessed the preclinical potential of AAV5-GLA for treating Fabry disease, an X-linked hereditary metabolic disorder resulting from mutations in the gene encoding GLA that lead to accumulation of the substrates globotriaosylceramide and globotriaosylsphingosine, causing heart, kidney, and central nervous system dysfunction. Effects of intravenously administered AAV5-GLA were evaluated in (1) GLA-knockout mice aged 7–8 weeks (early in disease) and 20 weeks (nociception phenotype manifestation) and (2) cynomolgus macaques during an 8-week period. In both species, AAV5-GLA was observed as safe, generated detectable vector DNA and mRNA levels in liver, and produced stable enzyme activity in liver and plasma. In mice, dose-dependent transgene enzyme activity, cross-correction (substrate reduction) in kidney and heart, and improved nociception lasted over 6 months. Moreover, after delayed administration when animals displayed the nociception phenotype, target organ enzyme activity was present, and accumulated substrates were reduced. Given the strong, durable expression of active GLA with this promoter and favorable profile of adeno-associated virus 5-based gene therapy in humans, AAV5-GLA warrants further investigation in clinical trials for Fabry disease.

There is a global upsurge in the use of cemented paste backfill (CPB) for various mining functions. However, the cost of the Portland cement binder is prohibitive, thus warranting strategies to reduce cement usage without overly diminishing the CPB quality. Since carbon dioxide is used for patented sand moulding processes, this study is premised on that physicochemical ability of CO2 to enhance the curing of consolidated inorganic materials. It evaluated the impact of carbon dioxide on the uniaxial compressive strength UCS and preparation cost of CPB standard samples (ASTM C109). The preparation cost was delimited to the purchase cost of the Portland cement. The backfill material was silica sand tailings with 4.5 wt.% Portland cement binder and a water-cement ratio of 7.6. Distilled water of pH 5.4 was used for the control samples while variable amounts of carbon dioxide were dissolved in distilled water to generate carbonated mixing water with pH values of 3.8; 4 and 4.2. The lower the carbonated water pH, the higher is the CO2 concentration. UCS tests were conducted on the samples after curing for 3, 7, 28, and 90 days. There was an observable increase in the UCSs and reduction in curing time with increasing carbon dioxide. Samples prepared with carbonated water of pH 3.8 had almost double the strength of those prepared with pure distilled water of pH 5.4, implying that more dissolved CO2 corresponds to higher CPB strength. This is supported by the trendline equations for the graphical simulation of strength on curing time. Thus, CPB with much less binder can be expected to attain the requisite UCS if carbon dioxide is incorporated. The average reduction in Portland cement consumption was 0.61 %, which translates to a cost saving of the same percentage points. If calculated over the operational life of a mine, this is a massive saving of millions of dollars.

The storage, transmission, and processing of data become significant problems when large digital data files or databases are involved, as in the case of decentralized online global databases such as blockchain. Here, we propose a novel method that allows for the scalability of digital assets, including blockchain databases in the download, validation, and confidentiality processes, by developing a lightweight blockchain technology called Entropic Blockchain. This is a computer-implemented mathematical method by which to generate an information-entropic numerical barcode representation of a digital asset. Using this technique, a 1–2 Mb block of digital data can be represented by a few bytes, significantly reducing the size of a blockchain. The entropic barcode file can be utilized on its own or as an optically machine-readable entropic barcode for secure data transmission, processing, labeling, identification, and one-way encryption, as well as for compression, validation, and digital tamper-proof checks. The mathematics of this process and all the steps involved in its implementation are discussed in detail in this article.

The simulation hypothesis is a philosophical theory, in which the entire universe and our objective reality are just simulated constructs. Despite the lack of evidence, this idea is gaining traction in scientific circles as well as in the entertainment industry. Recent scientific developments in the field of information physics, such as the publication of the mass-energy-information equivalence principle, appear to support this possibility. In particular, the 2022 discovery of the second law of information dynamics (infodynamics) facilitates new and interesting research tools at the intersection between physics and information. In this article, we re-examine the second law of infodynamics and its applicability to digital information, genetic information, atomic physics, mathematical symmetries, and cosmology, and we provide scientific evidence that appears to underpin the simulated universe hypothesis.

ABSTRACT Enterovirus D68 (EV-D68) is an RNA virus that can cause outbreaks of acute flaccid paralysis (AFP), a polio-like disease. Before 2010, EV-D68 was a rare pathogen associated with mild respiratory symptoms, but the recent EV-D68 related increase in severe respiratory illness and outbreaks of AFP is not yet understood. An explanation for the rise in severe disease is that it may be due to changes in the viral genome resulting in neurotropism. In this regard, in addition to sialic acid, binding to heparan sulfate proteoglycans (HSPGs) has been identified as a feature for viral entry of some EV-D68 strains in cell lines. Studies in human primary organotypic cultures that recapitulate human physiology will address the relevance of these HSPG-binding mutations for EV-D68 infection in vivo. Therefore, in this work, we studied the replication and neurotropism of previously determined sialic acid-dependent and HSPG-dependent strains using primary human airway epithelial (HAE) cultures and induced human pluripotent stem cell (iPSC)-derived brain organoids. All three strains (B2/2042, B2/947, and A1/1348) used in this study infected HAE cultures and human brain organoids (shown for the first time). Receptor-blocking experiments in both cultures confirm that B2/2042 infection is solely dependent on sialic acid, while B2/947 and A1/1348 (HSPG to a lesser extent) binds to sialic acid and HSPG for cell entry. Our data suggest that HSPG-binding can be used by EV-D68 for entry in human physiological models but offers no advantage for EV-D68 infection of brain cells. IMPORTANCE Recent outbreaks of enterovirus D68, a nonpolio enterovirus, is associated with a serious neurological condition in young children, acute flaccid myelitis (AFM). As there is no antiviral treatment or vaccine available for EV-D68 it is important to better understand how EV-D68 causes AFM and why only recent outbreaks are associated with AFM. We investigated if a change in receptor usage of EV-D68 increases the virulence of EV-D68 in the airway or the central nervous system and thus could explain the increase in AFM cases. We studied this using physiologically relevant human airway epithelium and cerebral organoid cultures that are physiologically relevant human models. Our data suggest that heparan sulfate proteoglycans can be used by EV-D68 as an additional entry receptor in human physiological models but offers no advantage for EV-D68 infection of brain cells, and our data show the potential of these 46 innovative models for virology.

One of the most powerful laws in physics is the second law of thermodynamics, which states that the entropy of any system remains constant or increases over time. In fact, the second law is applicable to the evolution of the entire universe and Clausius stated, “The entropy of the universe tends to a maximum.” Here, we examine the time evolution of information systems, defined as physical systems containing information states within Shannon’s information theory framework. Our observations allow the introduction of the second law of information dynamics (infodynamics). Using two different information systems, digital data storage and a biological RNA genome, we demonstrate that the second law of infodynamics requires the information entropy to remain constant or to decrease over time. This is exactly the opposite to the evolution of the physical entropy, as dictated by the second law of thermodynamics. The surprising result obtained here has massive implications for future developments in genomic research, evolutionary biology, computing, big data, physics, and cosmology.

The development of gene therapies for central nervous system disorders is challenging because it is difficult to translate preclinical data from current in vitro and in vivo models to the clinic. Therefore, we developed induced pluripotent stem cell (iPSC)-derived cerebral organoids as a model for recombinant adeno-associated virus (rAAV) capsid selection and for testing efficacy of AAV-based gene therapy in a human context. Cerebral organoids are physiological 3D structures that better recapitulate the human brain compared with 2D cell lines. To validate the model, we compared the transduction efficiency and distribution of two commonly used AAV serotypes (rAAV5 and rAAV9). In cerebral organoids, transduction with rAAV5 led to higher levels of vector DNA, transgenic mRNA, and protein expression as compared with rAAV9. The superior transduction of rAAV5 was replicated in iPSC-derived neuronal cells. Furthermore, rAAV5-mediated delivery of a human sequence-specific engineered microRNA to cerebral organoids led to a lower expression of its target ataxin-3. Our studies provide a new tool for selecting and deselecting AAV serotypes, and for demonstrating therapeutic efficacy of transgenes in a human context. Implementing cerebral organoids during gene therapy development could reduce the usage of animal models and improve translation to the clinic.

The mass–energy–information equivalence principle proposed in 2019 and the information content of the observable matter in the universe estimated in 2021 represent two important conjectures, called the information conjectures. Combining information theory and physical principles of thermodynamics, these theoretical proposals made specific predictions about the mass of information as well as the most probable information content per elementary particle. Here, we propose an experimental protocol that allows for empirical verification of the information conjectures by confirming the predicted information content of elementary particles. The experiment involves a matter–antimatter annihilation process. When an electron–positron annihilates, in addition to the two 511 keV gamma photons resulting from the conversion of their rest masses into energy, we predict that two additional low energy photons should be detected, resulting from their information content erasure. At room temperature, a positron–electron annihilation should produce two ∼50 µm wavelength infrared photons due to the information erasure. This experiment could, therefore, confirm both information conjectures and the existence of information as the fifth state of matter in the universe.

There is growing interest in the role that morphological knowledge plays in literacy acquisition, but there is no research directly comparing the efficacy of different forms of morphological instruction. Here we compare two methods of teaching English morphology in the context of a memory experiment when words were organized by affix during study (e.g., a list of words was presented that all share an affix, such as , , , , etc.) or by base during study (e.g., a list of words was presented that all share a base, such as , , , ). We show that memory for morphologically complex words is better in both conditions compared to a control condition that does not highlight the morphological composition of words, and most importantly, show that studying words in a base-centric format improves memory further still. We argue that the morphological matrix that organizes words around a common base may provide an important new tool for literacy instruction.

The information capacity of the universe has been a topic of great debate since the 1970s and continues to stimulate multiple branches of physics research. Here, we used Shannon’s information theory to estimate the amount of encoded information in all the visible matter in the universe. We achieved this by deriving a detailed formula estimating the total number of particles in the observable universe, known as the Eddington number, and by estimating the amount of information stored by each particle about itself. We determined that each particle in the observable universe contains 1.509 bits of information and there are ∼6 × 1080 bits of information stored in all the matter particles of the observable universe.

Huntington disease (HD) is a fatal neurodegenerative genetic disorder, thought to reflect a toxic gain of function in huntingtin (Htt) protein. Adeno-associated viral vector serotype 5 (AAV5)- microRNA targeting huntingtin (miHTT) is a HD gene-therapy candidate that efficiently lowers HTT using RNAi. This study analyzed the efficacy and potential for off-target effects with AAV5-miHTT in neuronal and astrocyte cell cultures differentiated from induced pluripotent stem cells (iPSCs) from two individuals with HD (HD71 and HD180). One-time AAV5-miHTT treatment significantly reduced human HTT mRNA by 57% and Htt protein by 68% in neurons. Small RNA sequencing showed that mature miHTT was processed correctly without off-target passenger strand. No cellular microRNAs were dysregulated, indicating that endogenous RNAi machinery was unaffected by miHTT overexpression. qPCR validation of in silico-predicted off-target transcripts, next-generation sequencing, and pathway analysis confirmed absence of dysregulated genes due to sequence homology or seed-sequence activity of miHTT. Minor effects on gene expression were observed in both AAV5-miHTT and AAV5-GFP-treated samples, suggesting that they were due to viral transduction rather than miHTT. This study confirms the efficacy of AAV5-miHTT in HD patient iPSC-derived neuronal cultures and lack of off-target effects in gene expression and regulation in neuronal cells and astrocytes. Keywords: AAV5, AAV5-miHTT, gene therapy, Huntington disease, Huntingtin, microRNA, RNA-seq, iPSC, off-target, patient-derived neuronal cultures

Spinocerebellar ataxia type 3 (SCA3), or Machado-Joseph disease (MJD), is a progressive neurodegenerative disorder caused by a CAG expansion in the ATXN3 gene. The expanded CAG repeat is translated into a prolonged polyglutamine repeat in the ataxin-3 protein and accumulates within inclusions, acquiring toxic properties, which results in degeneration of the cerebellum and brain stem. In the current study, a non-allele-specific ATXN3 silencing approach was investigated using artificial microRNAs engineered to target various regions of the ATXN3 gene (miATXN3). The miATXN3 candidates were screened in vitro based on their silencing efficacy on a luciferase (Luc) reporter co-expressing ATXN3. The three best miATXN3 candidates were further tested for target engagement and potential off-target activity in induced pluripotent stem cells (iPSCs) differentiated into frontal brain-like neurons and in a SCA3 knockin mouse model. Besides a strong reduction of ATXN3 mRNA and protein, small RNA sequencing revealed efficient guide strand processing without passenger strands being produced. We used different methods to predict alteration of off-target genes upon AAV5-miATXN3 treatment and found no evidence for unwanted effects. Furthermore, we demonstrated in a large animal model, the minipig, that intrathecal delivery of AAV5 can transduce the main areas affected in SCA3 patients. These results proved a strong basis to move forward to investigate distribution, efficacy, and safety of AAV5-miATXN3 in large animals.

Huntington disease (HD) is a fatal neurodegenerative disorder caused by an autosomal dominant CAG repeat expansion in the huntingtin (HTT) gene. The translated expanded polyglutamine repeat in the HTT protein is known to cause toxic gain of function. We showed previously that strong HTT lowering prevented neuronal dysfunction in HD rodents and minipigs after single intracranial injection of adeno-associated viral vector serotype 5 expressing a microRNA targeting human HTT (AAV5-miHTT). To evaluate long-term efficacy, AAV5-miHTT was injected into the striatum of knockin Q175 HD mice, and the mice were sacrificed 12 months post-injection. AAV5-miHTT caused a dose-dependent and sustained HTT protein reduction with subsequent suppression of mutant HTT aggregate formation in the striatum and cortex. Functional proof of concept was shown in transgenic R6/2 HD mice. Eight weeks after AAV5-miHTT treatment, a significant improvement in motor coordination on the rotarod was observed. Survival analysis showed that a single AAV5-miHTT treatment resulted in a significant 4-week increase in median survival compared with vehicle-treated R6/2 HD mice. The combination of long-term HTT lowering, reduction in aggregation, prevention of neuronal dysfunction, alleviation of HD-like symptoms, and beneficial survival observed in HD rodents treated with AAV5-miHTT supports the continued development of HTT-lowering gene therapies for HD. Keywords: Huntington disease, gene silencing, HD mouse model, gene therapy, therapeutic microRNAs, adeno-associated viral vector serotype 5

A hexanucleotide GGGGCC expansion in intron 1 of chromosome 9 open reading frame 72 (C9orf72) gene is the most frequent cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). The corresponding repeat-containing sense and antisense transcripts cause a gain of toxicity through the accumulation of RNA foci in the nucleus and deposition of dipeptide-repeat (DPR) proteins in the cytoplasm of the affected cells. We have previously reported on the potential of engineered artificial anti-C9orf72-targeting miRNAs (miC) targeting C9orf72 to reduce the gain of toxicity caused by the repeat-containing transcripts. In the current study, we tested the silencing efficacy of adeno-associated virus (AAV)5-miC in human-derived induced pluripotent stem cell (iPSC) neurons and in an ALS mouse model. We demonstrated that AAV5-miC transduces different types of neuronal cells and can reduce the accumulation of repeat-containing C9orf72 transcripts. Additionally, we demonstrated silencing of C9orf72 in both the nucleus and cytoplasm, which has an added value for the treatment of ALS and/or FTD patients. A proof of concept in an ALS mouse model demonstrated the significant reduction in repeat-containing C9orf72 transcripts and RNA foci after treatment. Taken together, these findings support the feasibility of a gene therapy for ALS and FTD based on the reduction in toxicity caused by the repeat-containing C9orf72 transcripts. Keywords: C9orf72, ALS, FTD, miRNA, gene therapy, AAV

The most common pathogenic mutation in amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) is an intronic GGGGCC (G4C2) repeat in the chromosome 9 open reading frame 72 (C9orf72) gene. Cellular toxicity due to RNA foci and dipeptide repeat (DPR) proteins produced by the sense and antisense repeat-containing transcripts is thought to underlie the pathogenesis of both diseases.RNA sequencing (RNA-seq) data of C9orf72-ALS patients and controls were analyzed to better understand the sequence conservation of C9orf72 in patients. MicroRNAs were developed in conserved regions to silence C9orf72 (miC), and the feasibility of different silencing approaches was demonstrated in reporter overexpression systems. In addition, we demonstrated the feasibility of a bidirectional targeting approach by expressing two concatenated miC hairpins. The efficacy of miC was confirmed by the reduction of endogenously expressed C9orf72 mRNA, in both nucleus and cytoplasm, and an ∼50% reduction of nuclear RNA foci in (G4C2)44-expressing cells. Ultimately, two miC candidates were incorporated in adeno-associated virus vector serotype 5 (AAV5), and silencing of C9orf72 was demonstrated in HEK293T cells and induced pluripotent stem cell (iPSC)-derived neurons. These data support the feasibility of microRNA (miRNA)-based and AAV-delivered gene therapy that could alleviate the gain of toxicity seen in ALS and FTD patients. Keywords: C9orf72, ALS, FTD, gene therapy, microRNA

The current scientific consensus is that genetic mutations are random processes. According to the Darwinian theory of evolution, only natural selection determines which mutations are beneficial in the course of evolution, and there is no deterministic correlation between any parameter and the probability that these mutations will occur. Here, we investigate RNA genetic sequences of the SARS-CoV-2 virus using Shannon’s information theory, and we report a previously unobserved relationship between the information entropy of genomes and their mutation dynamics. Based on the analysis presented here, we are able to formulate a governing law of genetic mutations, stating that genomes undergo genetic mutations over time driven by a tendency to reduce their overall information entropy, challenging the existing Darwinian paradigm.

Coal-fired power plants utilising fluidised bed technologies emit copious amounts of fly-ash, which is harmful to people owing to its particulate nature. A planned 300 MW power plant will have an electrostatic precipitator (ESP) for fly-ash emissions control, in line with power generation industry best practices. This ESP should meet a fly-ash emission limit value ≤ 50 mg/Nm3. This paper details the design process and resultant technical specifications of a cold-side, single-stage, and plate-wire dry ESP designed for the power plant. The ESP will consist of twin-chambers with quadruple-fields (2 4 315 m3) and octonary bus-sections independently energised by individual high-frequency three-phase switched integrated rectifiers (70 kV, 800 mA) to maximise ionisation. Dynamically balanced, single-impact, tumbling hammer rappers (857.5 rpm) will dislodge fly-ash from the collector plates into mass-flow wedge-shaped hoppers. A specific collection area (≥ 56.9 m2/m3/s) and an ESP index (≥ 709.2 (kV/cm)2m2/m3/s) should guarantee a collection efficiency ( ≥ 99.5 %) and the specified emission limit value at 100 % boiler capacity and normal operating conditions (gas velocity ≤ 2.4 m/s; gas temperature ≤ 137 ℃; fly-ash loading ≤ 10 000 kg/Nm3; resistivity ≤ 1010 Ω‧cm). The design was successfully verified in principle using the validation square method, in conjunction with the leading comparable historical case studies approach. It is recommended to simulate ESP designs using suitable programs like Comsol Multiphysics and construct a pilot plant before attempting scaled-up construction and commissioning. Highlights • A cold-side, single-stage plate-wire dry ESP system designed. • Theoretical collection efficiency in excess of 99.5%. • High-frequency, three-phase switched integrated rectifiers to be used for ionisation.

Currently, we produce ∼1021 digital bits of information annually on Earth. Assuming a 20% annual growth rate, we estimate that after ∼350 years from now, the number of bits produced will exceed the number of all atoms on Earth, ∼1050. After ∼300 years, the power required to sustain this digital production will exceed 18.5 × 1015 W, i.e., the total planetary power consumption today, and after ∼500 years from now, the digital content will account for more than half Earth’s mass, according to the mass-energy–information equivalence principle. Besides the existing global challenges such as climate, environment, population, food, health, energy, and security, our estimates point to another singular event for our planet, called information catastrophe.

This paper examines the extent of misalignment of the real effective exchange rate (REER) of South African rand. With South Africa being an open emerging market economy closely linked with global markets, the country’s economy is susceptible to external shocks and changes in global trade patterns. Based on the behavioural equilibrium exchange rate framework of Clark and MacDonald () the study uses a cointegration technique, which caters for endogeneity to estimate the equilibrium value of the REER of the rand. Next, the study uses a Markov regime-switching (MSM) method to determine whether the exchange rate’s departure from the equilibrium level is meaningful enough to be considered as either over- or undervalued. The results show that long-run equilibrium relationship between the rand’s REER and economic variables inclusive of terms of trade, external openness, capital flows and government expenditure. The MSM correctly captures exchange rate misalignment as distinct episodes of exchange rate overvaluation and undervaluation. Most of the exchange rate undervaluation episodes identified in the study have been in response to either idiosyncratic shocks emanating from internal economic/political challenges or systemic global factors transmitted through either the nominal exchange rate or capital flows and not policy-induced behaviour.

Pathogenesis of viral infections of the central nervous system (CNS) is poorly understood, and this is partly due to the limitations of currently used preclinical models. Brain organoid models can overcome some of these limitations, as they are generated from human derived stem cells, differentiated in three dimensions (3D), and can mimic human neurodevelopmental characteristics. Therefore, brain organoids have been increasingly used as brain models in research on various viruses, such as Zika virus, severe acute respiratory syndrome coronavirus 2, human cytomegalovirus, and herpes simplex virus. Brain organoids allow for the study of viral tropism, the effect of infection on organoid function, size, and cytoarchitecture, as well as innate immune response; therefore, they provide valuable insight into the pathogenesis of neurotropic viral infections and testing of antivirals in a physiological model. In this review, we summarize the results of studies on viral CNS infection in brain organoids, and we demonstrate the broad application and benefits of using a human 3D model in virology research. At the same time, we describe the limitations of the studies in brain organoids, such as the heterogeneity in organoid generation protocols and age at infection, which result in differences in results between studies, as well as the lack of microglia and a blood brain barrier.

Abstract Background Spinocerebellar ataxia type 3 (SCA3) is a progressive neurodegenerative disorder caused by expansion of the polyglutamine repeat in the ataxin-3 protein. Expression of mutant ataxin-3 is known to result in transcriptional dysregulation, which can contribute to the cellular toxicity and neurodegeneration. Since the exact causative mechanisms underlying this process have not been fully elucidated, gene expression analyses in brains of transgenic SCA3 mouse models may provide useful insights. Methods Here we characterised the MJD84.2 SCA3 mouse model expressing the mutant human ataxin-3 gene using a multi-omics approach on brain and blood. Gene expression changes in brainstem, cerebellum, striatum and cortex were used to study pathological changes in brain, while blood gene expression and metabolites/lipids levels were examined as potential biomarkers for disease. Results Despite normal motor performance at 17.5 months of age, transcriptional changes in brain tissue of the SCA3 mice were observed. Most transcriptional changes occurred in brainstem and striatum, whilst cerebellum and cortex were only modestly affected. The most significantly altered genes in SCA3 mouse brain were Tmc3, Zfp488, Car2, and Chdh. Based on the transcriptional changes, α-adrenergic and CREB pathways were most consistently altered for combined analysis of the four brain regions. When examining individual brain regions, axon guidance and synaptic transmission pathways were most strongly altered in striatum, whilst brainstem presented with strongest alterations in the pi-3 k cascade and cholesterol biosynthesis pathways. Similar to other neurodegenerative diseases, reduced levels of tryptophan and increased levels of ceramides, di- and triglycerides were observed in SCA3 mouse blood. Conclusions The observed transcriptional changes in SCA3 mouse brain reveal parallels with previous reported neuropathology in patients, but also shows brain region specific effects as well as involvement of adrenergic signalling and CREB pathway changes in SCA3. Importantly, the transcriptional changes occur prior to onset of motor- and coordination deficits.

The multicaloric effect is defined as the adiabatic reversible temperature change in multiferroic materials induced by the application of an external electric or magnetic field, and it was first theoretically proposed in 2012. The multicaloric effects in multiferroics, as well as other similar caloric effects in single ferroics, such as magnetocaloric, elastocaloric, barocaloric, and electrocaloric, have been the focus of much research due to their potential commercialization in solid-state refrigeration. In this short communication article, we examine the thermodynamics of the multicaloric effect for solid-state heating applications. A possible thermodynamic multicaloric heating cycle is proposed and then implemented to estimate the solid-state heating effect for a known electrocaloric system. This work offers a path to implementing caloric and multicaloric effects to efficient heating systems, and we offer a theoretical estimate of the upper limit of the temperature change achievable in a multicaloric cooling or heating effect.

A 73-year-old man with history of pulmonary sarcoidosis was found to have runs of non-sustained bidirectional ventricular tachycardia (BVT) with two different QRS morphologies on a Holter monitor. Cardiac magnetic resonance delayed gadolinium imaging revealed a region of patchy mid-myocardial enhancement within the left ventricular basal inferolateral myocardium. An 18-fluorodeoxyglucose positron emission tomography (FDG-PET) showed increased uptake in the same area, consistent with active sarcoid, with no septal involvement. Follow-up FDG-PET one year later showed disease progression with new septal involvement. Cardiac sarcoidosis, characterized by myocardial inflammation and interstitial fibrosis that can lead to conduction system disturbance and macro re-entrant arrhythmias, should be considered in differential diagnosis of BVT. BVT may indicate septal involvement with sarcoidosis before the lesions are large enough to be detected radiologically.

Landauer’s principle formulated in 1961 states that logical irreversibility implies physical irreversibility and demonstrated that information is physical. Here we formulate a new principle of mass-energy-information equivalence proposing that a bit of information is not just physical, as already demonstrated, but it has a finite and quantifiable mass while it stores information. In this framework, it is shown that the mass of a bit of information at room temperature (300K) is 3.19 × 10-38 Kg. To test the hypothesis we propose here an experiment, predicting that the mass of a data storage device would increase by a small amount when is full of digital information relative to its mass in erased state. For 1Tb device the estimated mass change is 2.5 × 10-25 Kg.

Background Long‐term corticosteroid therapy is the standard of care for treatment of cardiac sarcoidosis (CS). The efficacy of long‐term corticosteroid‐sparing immunosuppression in CS is unknown. The goal of this study was to assess the efficacy of methotrexate with or without adalimumab for long‐term disease suppression in CS, and to assess recurrence and adverse event rates after immunosuppression discontinuation. Methods and Results Retrospective chart review identified treatment‐naive CS patients at a single academic medical center who received corticosteroid‐sparing maintenance therapy. Demographics, cardiac uptake of 18‐fluorodeoxyglucose, and adverse cardiac events were compared before and during treatment and between those with persistent or interrupted immunosuppression. Twenty‐eight CS patients were followed for a mean 4.1 (SD 1.5) years. Twenty‐five patients received 4 to 8 weeks of high‐dose prednisone (>30 mg/day), followed by taper and maintenance therapy with methotrexate±low‐dose prednisone (low‐dose prednisone,

Huntington disease (HD) is a fatal, neurodegenerative genetic disorder with aggregation of mutant Huntingtin protein (mutHTT) in the brain as a key pathological mechanism. There are currently no disease modifying therapies for HD; however, HTT-lowering therapies hold promise. Recombinant adeno-associated virus serotype 5 expressing a microRNA that targets HTT mRNA (AAV5-miHTT) is in development for the treatment of HD with promising results in rodent and minipig HD models. To support a clinical trial, toxicity studies were performed in non-human primates (NHP, Macaca fascicularis) and Sprague-Dawley rats to evaluate the safety of AAV5-miHTT, the neurosurgical administration procedure, vector delivery and expression of the miHTT transgene during a 6-month observation period. For accurate delivery of AAV5-miHTT to the striatum, real-time magnetic resonance imaging (MRI) with convection-enhanced delivery (CED) was used in NHP. Catheters were successfully implanted in 24 NHP, without neurological symptoms, and resulted in tracer signal in the target areas. Widespread vector DNA and miHTT transgene distribution in the brain was found, particularly in areas associated with HD pathology. Intrastriatal administration of AAV5-miHTT was well tolerated with no clinically relevant changes in either species. These studies demonstrate the excellent safety profile of AAV5-miHTT, the reproducibility and tolerability of intrastriatal administration, and the delivery of AAV5-miHTT to the brain, which support the transition of AAV5-miHTT into clinical studies.

As schools across the US and around the world closed their physical buildings to stop the spread of COVID-19, students, teachers, families, and communities grappled with the shift to online learning. While emerging research examines how educators navigated this shift, little is known about how students experienced schooling as the pandemic unfolded and how their experiences may contribute to emerging calls for reimagining education. In this essay, Joanne E. Marciano, Lee Melvin M. Peralta, and Ji Soo Lee look at how youth navigated online learning in the sixteen months after their schools closed. Using weekly interviews with youth conducted between March 2020 and June 2021, they center the voices of black, Lebanese American, and white students who experienced limited access to economic resources and call attention to the (post)pandemic schooling experiences they needed and desired.

Animal models and cell lines are invaluable for virology research and host–pathogen interaction studies. However, it is increasingly evident that these models are not sufficient to fully understand human viral diseases. With the advent of three-dimensional organotypic cultures, it is now possible to study viral infections in the human context. This perspective explores the potential of these organotypic cultures, also known as organoids, for virology research, antiviral testing, and shaping the virology landscape.

Systemic lupus erythematosus (SLE) is an autoimmune disease with a strong genetic component and etiology characterized by chronic inflammation and autoantibody production. The purpose of this study was to ascertain copy number variation (CNV) in SLE using a case-control design in an admixed Brazilian population. The whole-genome detection of CNV was performed using Cytoscan HD array in SLE patients and healthy controls. The best CNV candidates were then evaluated by quantitative real-time PCR in a larger cohort or validated using droplet digital PCR. Logistic regression models adjusted for sex and ancestry covariates was applied to evaluate the association between CNV with SLE susceptibility. The data showed a synergistic effect between the FCGR3B and ADAM3A loci with the presence of deletions in both loci significantly increasing the risk to SLE (5.9-fold) compared to the deletion in the single FCGR3B locus (3.6-fold). In addition, duplications in these genes were indeed more frequent in healthy subjects, suggesting that high FCGR3B/ADAM3A gene copy numbers are protective factors against to disease development. Overall, 21 rare CNVs were identified in SLE patients using a four-step pipeline created for identification of rare variants. Furthermore, heterozygous deletions overlapping the CFHR4, CFHR5 and HLA-DPB2 genes were described for the first time in SLE patients. Here we present the first genome-wide CNV study of SLE patients in a tri-hybrid population. The results show that novel susceptibility loci to SLE can be found once the distribution of structural variants is analyzed throughout the whole genome.

Spatial metaphors are used to represent and reason about time. Such metaphors are typically arranged along the sagittal axis in most languages. For example, in English, “The future lies ahead of us” and “We look back on our past.” This is less straightforward for Chinese. Specifically, both the past and future can either be behind or ahead. The present study aims to explore these cross-linguistic differences by priming auditory targets (e.g., tomorrow) with either a congruent (i.e., pointing forwards) or incongruent (i.e., pointing backwards) gesture. Two groups of college-age young adult participants (English and Chinese speakers) made temporal classifications of words after watching a gestural prime. If speakers represent time along the sagittal axis, they should respond faster if the auditory target is preceded with a gesture indicating a congruent vs. incongruent spatial location. Results showed that English speakers responded faster to congruent gesture-word pairs than to incongruent pairs, mirroring spatio-temporal metaphors commonly recruited to talk about time in their native language. However, such an effect of congruency was not found for Chinese speakers. These findings suggest that while the spatio-temporal metaphors commonly recruited to talk about time help to structure the mental timelines of English speakers, the varying instances in how time is represented along the sagittal axis in Chinese may lead to a more variable mental timeline as well. In addition, our findings demonstrate that gestures may not only be a means of accessing concrete concepts in the mind, as shown in previous studies, but may be used to access abstract ones as well.

Long-QT syndrome mutations can cause syncope and sudden death by prolonging the cardiac action potential (AP). Ion channels affected by mutations are various, and the influences of cellular calcium cycling on LQTS cardiac events are unknown. To better understand LQTS arrhythmias, we performed current-clamp and intracellular calcium ([Ca2+]i) measurements on cardiomyocytes differentiated from patient-derived induced pluripotent stem cells (iPS-CM). In myocytes carrying an LQT2 mutation (HERG-A422T), APs and [Ca2+]i transients were prolonged in parallel. APs were abbreviated by nifedipine exposure and further lengthened upon releasing intracellularly stored Ca2+. Validating this model, control iPS-CM treated with HERG-blocking drugs recapitulated the LQT2 phenotype. In LQT3 iPS-CM, expressing NaV1.5-N406K, APs and [Ca2+]i transients were markedly prolonged. AP prolongation was sensitive to tetrodotoxin and to inhibiting Na+-Ca2+ exchange. These results suggest that LQTS mutations act partly on cytosolic Ca2+ cycling, potentially providing a basis for functionally targeted interventions regardless of the specific mutation site.

Spinocerebellar ataxia type 3 is caused by a polyglutamine expansion in the ataxin-3 protein, resulting in gain of toxic function of the mutant protein. The expanded glutamine stretch in the protein is the result of a CAG triplet repeat expansion in the penultimate exon of the ATXN3 gene. Several gene silencing approaches to reduce mutant ataxin-3 toxicity in this disease aim to lower ataxin-3 protein levels, but since this protein is involved in deubiquitination and proteasomal protein degradation, its long-term silencing might not be desirable. Here, we propose a novel protein modification approach to reduce mutant ataxin-3 toxicity by removing the toxic polyglutamine repeat from the ataxin-3 protein through antisense oligonucleotide-mediated exon skipping while maintaining important wild type functions of the protein. In vitro studies showed that exon skipping did not negatively impact the ubiquitin binding capacity of ataxin-3. Our in vivo studies showed no toxic properties of the novel truncated ataxin-3 protein. These results suggest that exon skipping may be a novel therapeutic approach to reduce polyglutamine-induced toxicity in spinocerebellar ataxia type 3.

Patterns of left atrial (far-field signals) or coronary sinus (CS) muscle (near-field) have been defined in CS recordings. The purpose of this study was to define the activation patterns from the coronary sinus in patients with anterior and posterior type of atrioventricular nodal reentrant tachycardia (AVNRT) circuits. Methods and Results: This retrospective study involved a total of 149 patients with 155 episodes of AVNRT which were divided into 3 patterns. In the anterior pattern (123 tachycardias), the atrial deflection from the His bundle electrogram preceded that from the proximal CS electrogram. In the posterior pattern (23 tachycardias), the proximal CS electrogram (CSp) was recorded earlier than the His bundle atrial electrogram. In the left atrial pattern (9 tachycardias), activation of distal CS sites preceded both proximal CS and atrial activation from the His bundle electrogram. A decapolar catheter with a 5-mm interelectrode distance was used for CS recording. The CS electrograms were analyzed to determine the total signal duration as well as the duration of the initial component. An initial slow wave was defined as a duration exceeding 10 ms. 1) The duration of the initial component in patients with the anterior pattern was longer than in those with the posterior pattern in CSp (7.3 ± 3.1 ms vs. 4.5 ± 2.0 ms), CS7-8 (7.4 ± 2.9 ms vs. 3.8 ± 1.5 ms), CS5-6 (7.3 ms ± 3.3 ms vs. 4.4 ms ± 2.5 ms) and CS3-4 (6.7 ms ± 2.4 ms vs. 4.5 ms ± 2.0 ms) (p < 0.01). 2) Similarly the total electrogram duration in the CS was longer in patients with an anterior compared to a posterior pattern in CSp (38.3 ms ± 10.1 ms vs. 26.8 ± 6.1 ms), CS7-8 (31.8 ms ±6.6 ms vs. 27.2 ms ± 6.0 ms), CS5-6 (31.3 ms ± 6.8 ms vs. 26.5 ms ± 4.9 ms), and CS3-4 (30.0 ms ± 6.3 ms vs. 25.0 ms ± 5.1 ms) (p < 0.01). 3) The percentage of tachycardias showing an initial slow wave followed by rapid activation was higher for anterior pattern patients compared with posterior pattern patients in CSp (62% vs. 13%), CS7-8 (79% vs. 4%), CS5-6 (72% vs. 4%), CS3-4 (54% vs. 9%) and distal CS (47% vs. 0%) (p < 0.01). Conclusions: The pattern of an initial slow wave followed by a rapid wave in the CS was characteristic of an anterior AVNRT circuit and is explained by the initial involvement of far field left atrial components. In contrast, the predominant early rapid waves in the posterior AVNRT circuit are compatible with early CS activation from the right atrium.

Background Left atrial appendage (LAA) exclusion has been performed in patients with atrial fibrillation (AF) to prevent thrombus formation and subsequent cardioembolic events. Left atrial electrical remodeling is a recognized factor in the recurrence of AF. The effects of LAA exclusion on P‐wave characteristics and left atrial electrical remodeling have not been well described. The purpose of this study was to evaluate the effect of LAA ligation on P‐wave morphology in patients with AF. Methods and Results Fifteen patients who were in sinus rhythm during the LAA ligation procedure were included in the study. We evaluated the P‐wave characteristics, including P‐wave duration, P‐wave amplitude, PQ interval, and P‐wave dispersion, before and after ligation. Eleven patients had paroxysmal AF and 4 patients had persistent AF (12 male patients and 3 female patients). P‐wave duration immediately after ligation was significantly shorter compared with baseline in all limb leads except lead aVR (P

Supraventricular tachycardia (SVT) with ventriculoatrial (VA) block can represent a diagnostic challenge. We present a case of SVT where His-His interval shortening was repeatedly observed during episodes of VA block. This novel observation is more diagnostically suggestive of atrioventricular nodal re-entrant tachycardia, as opposed to orthodromic re-entry using a nodofascicular or nodoventricular pathway where a constant His-His is recorded during episodes of VA block. (Level of Difficulty: Intermediate.).

While Drs Wolff, Parkinson, and White fully described the syndrome in 1930, prior case reports had described the essentials. Over the ensuing century this syndrome has captivated the interest of anatomists, clinical cardiologists, and cardiac surgeons. Stanley Kent described lateral muscular connections over the atrioventricular (AV) groove which he felt were the normal AV connections. The normal AV connections were, however, clearly described by His and Tawara. True right-sided AV connections were initially described by Wood et al., while Öhnell first described left free wall pathways. David Scherf is thought to be the first to describe our current understanding of the pathogenesis of the WPW syndrome in terms of a re-entrant circuit involving both the AV node–His axis as well as the accessory pathway. This hypothesis was not universally accepted, and many theories were applied to explain the clinical findings. The basics of our understanding were established by the brilliant work of Pick, Langendorf, and Katz who by using careful deductive analysis of ECGs were able to define the basic pathophysiological processes. Subsequently, Wellens and Durrer applied invasive electrical stimulation to the heart in order to confirm the pathophysiological processes. Sealy and his colleagues at Duke University Medical Center were the first to successfully surgically divide an accessory pathway and ushered in the modern era of therapy for these patients. Morady and Scheinman were the first to successfully ablate an accessory pathway (posteroseptal) using high-energy direct-current shocks. Subsequently Jackman, Kuck, Morady, and a number of groups proved the remarkable safety and efficiency of catheter ablation for pathways in all locations using radiofrequency energy. More recently, Gollob et al. first described the gene responsible for a familial form of WPW. The current ability to cure patients with WPW is due to the splendid contributions of individuals from diverse disciplines throughout the world.

Ventricular arrhythmias in patients with ARVD/C are common. Differentiation between idiopathic ventricular tachycardia and arrhythmogenic right ventricular dysplasia is of utmost importance. Baseline sinus rhythm electrocardiography as well as electrocardiographic differences during ventricular arrhythmias (VT or PVCs) can be helpful in differentiating the two disease states. The pathological fibrofatty myocyte replacement in ARVD/C as well as ventricular arrhythmia origin are likely responsible for these differences.

A limiting factor in brain research still is the difficulty to evaluate in vivo the role of the increasing number of proteins implicated in neuronal processes. We discuss here the potential of antisense-mediated RNA targeting approaches. We mainly focus on those that manipulate splicing (exon skipping and exon inclusion), but will also briefly discuss mRNA targeting. Classic knockdown of expression by mRNA targeting is only one possible application of antisense oligonucleotides (AON) in the control of gene function. Exon skipping and inclusion are based on the interference of AONs with splicing of pre-mRNAs. These are powerful, specific and particularly versatile techniques, which can be used to circumvent pathogenic mutations, shift splice variant expression, knock down proteins, or to create molecular models using in-frame deletions. Pre-mRNA targeting is currently used both as a research tool, e.g. in models for motor neuron disease, and in clinical trials for Duchenne muscular dystrophy and amyotrophic lateral sclerosis.AONs are particularly promising in relation to brain research, as the modified AONs are taken up extremely fast in neurons and glial cells with a long residence, and without the need for viral vectors or other delivery tools, once inside the blood brain barrier. In this review we cover 1. The principles of antisense-mediated techniques, chemistry and efficacy.2. The pros and cons of AON approaches in the brain compared to other techniques of interfering with gene function, such as transgenesis and short hairpin RNAs, in terms of specificity of the manipulation, spatial and temporal control over gene expression, toxicity and delivery issues.3. The potential applications for Neuroscience. We conclude that there is good evidence from animal studies that the CNS can be successfully targeted, but the potential of the diverse AON-based approaches appears to be under-recognized.

To date there are 9 known diseases caused by an expanded polyglutamine repeat, with the most prevalent being Huntington's disease. Huntington's disease is a progressive autosomal dominant neurodegenerative disorder for which currently no therapy is available. It is caused by a CAG repeat expansion in the HTT gene, which results in an expansion of a glutamine stretch at the N-terminal end of the huntingtin protein. This polyglutamine expansion plays a central role in the disease and results in the accumulation of cytoplasmic and nuclear aggregates. Here, we make use of modified 2'-O-methyl phosphorothioate (CUG)n triplet-repeat antisense oligonucleotides to effectively reduce mutant huntingtin transcript and protein levels in patient-derived Huntington's disease fibroblasts and lymphoblasts. The most effective antisense oligonucleotide, (CUG)(7), also reduced mutant ataxin-1 and ataxin-3 mRNA levels in spinocerebellar ataxia 1 and 3, respectively, and atrophin-1 in dentatorubral-pallidoluysian atrophy patient derived fibroblasts. This antisense oligonucleotide is not only a promising therapeutic tool to reduce mutant huntingtin levels in Huntington's disease but our results in spinocerebellar ataxia and dentatorubral-pallidoluysian atrophy cells suggest that this could also be applicable to other polyglutamine expansion disorders as well.