Oligonucleotides Targeting DNA Repeats Downregulate Huntingtin Gene Expression in Huntington's Patient-Derived Neural Model System

Huntington's disease (HD) is one of the most common, dominantly inherited neurodegenerative disorders. It affects the striatum, cerebral cortex, and other subcortical structures leading to involuntary movement abnormalities, emotional disturbances, and cognitive impairments. HD is caused by a CAG•CTG trinucleotide-repeat expansion in exon 1 of the huntingtin (HTT) gene leading to the formation of mutant HTT (mtHTT) protein aggregates. Besides the toxicity of the mutated protein, there is also evidence that mtHTT transcripts contribute to the disease. Thus, the reduction of both mutated mRNA and protein would be most beneficial as a treatment. Previously, we designed a novel anti-gene oligonucleotide (AGO)-based strategy directly targeting the HTT trinucleotide-repeats in DNA and reported downregulation of mRNA and protein in HD patient fibroblasts. In this study, we differentiate HD patient-derived induced pluripotent stem cells to investigate the efficacy of the AGO, a DNA/Locked Nucleic Acid mixmer with phosphorothioate backbone, to modulate HTT transcription during neural in vitro development. For the first time, we demonstrate downregulation of HTT mRNA following both naked and magnetofected delivery into neural stem cells (NSCs) and show that neither emergence of neural rosette structures nor self-renewal of NSCs is compromised. Furthermore, the inhibition potency of both HTT mRNA and protein without off-target effects is confirmed in neurons. These results further validate an anti-gene approach for the treatment of HD

Oligonucleotide-Palladacycle Conjugates as Splice-Correcting Agents

2'-O-Methylribo phosphorothioate oligonucleotides incorporating cyclopalladated benzylamine conjugate groups at their 5'-termini have been prepared and their ability to hybridize with a designated target sequence was assessed by conventional UV melting experiments. The oligonucleotides were further examined in splice-switching experiments in human cervical cancer (HeLa Luc/705), human liver (HuH7_705), and human osteosarcoma (U-2 OS_705) reporter cell lines. Melting temperatures of duplexes formed by the modified oligonucleotides were approximately 5 degrees C lower than melting temperatures of the respective unmodified duplexes. The cyclopalladated oligonucleotides functioned as splice-correcting agents in the HeLa Luc/705 cell line somewhat more efficiently than their unmodified counterparts. Furthermore, the introduction of this chemical modification did not induce toxicity in cells. These results demonstrate the feasibility of using covalently metalated oligonucleotides as therapeutic agents

Aneurysmal Subarachnoid Hemorrhage

Aneurysmal subarachnoid hemorrhage (SAH) is a devastating neurological syndrome, which occurs at a rate of 3–25 per 100,000 population. Smoking and hypertension are the most important risk factors of subarachnoid hemorrhage. Rupture of cerebral aneurysm leads to rapid spread of blood into cerebrospinal fluid and subsequently leads to sudden increase of intracranial pressure and severe headache. Subarachnoid hemorrhage is associated with neurological (such as re‐bleeding and vasospasm) and systemic (such as myocardial injury and hyponatremia) complications that are causes of high mortality and morbidity. Although patients with poor‐grade subarachnoid hemorrhage are at higher risk of neurological and systemic complications, the early and aggressive management of this group of patient has decreased overall mortality by 17% in last 40 years. Early aneurysm repair, close monitoring in dedicated neurological intensive care unit, prevention, and aggressive management of medical and neurological complications are the most important strategies to improve outcome

Modulational instability in periodic quadratic nonlinear materials

We investigate the modulational instability of plane waves in quadratic nonlinear materials with linear and nonlinear quasi-phase-matching gratings. Exact Floquet calculations, confirmed by numerical simulations, show that the periodicity can drastically alter the gain spectrum but never completely removes the instability. The low-frequency part of the gain spectrum is accurately predicted by an averaged theory and disappears for certain gratings. The high-frequency part is related to the inherent gain of the homogeneous non-phase-matched material and is a consistent spectral feature.Comment: 4 pages, 7 figures corrected minor misprint

"Level Curvature" Distribution for Diffusive Aharonov-Bohm Systems: analytical results

We calculate analytically the distributions of "level curvatures" (LC) (the second derivatives of eigenvalues with respect to a magnetic flux) for a particle moving in a white-noise random potential. We find that the Zakrzewski-Delande conjecture is still valid even if the lowest weak localization corrections are taken into account. The ratio of mean level curvature modulus to mean dissipative conductance is proved to be universal and equal to $2\pi$ in agreement with available numerical data.Comment: 12 pages. Submitted to Phys.Rev.

Novel endosomolytic compounds enable highly potent delivery of antisense oligonucleotides

The therapeutic and research potentials of oligonucleotides (ONs) have been hampered in part by their inability to effectively escape endosomal compartments to reach their cytosolic and nuclear targets. Splice-switching ONs (SSOs) can be used with endosomolytic small molecule compounds to increase functional delivery. So far, development of these compounds has been hindered by a lack of high-resolution methods that can correlate SSO trafficking with SSO activity. Here we present in-depth characterization of two novel endosomolytic compounds by using a combination of microscopic and functional assays with high spatiotemporal resolution. This system allows the visualization of SSO trafficking, evaluation of endosomal membrane rupture, and quantitates SSO functional activity on a protein level in the presence of endosomolytic compounds. We confirm that the leakage of SSO into the cytosol occurs in parallel with the physical engorgement of LAMP1-positive late endosomes and lysosomes. We conclude that the new compounds interfere with SSO trafficking to the LAMP1-positive endosomal compartments while inducing endosomal membrane rupture and concurrent ON escape into the cytosol. The efficacy of these compounds advocates their use as novel, potent, and quick-acting transfection reagents for antisense ONs

Nonlinear localized waves in a periodic medium

We analyze the existence and stability of nonlinear localized waves in a periodic medium described by the Kronig-Penney model with a nonlinear defect. We demonstrate the existence of a novel type of stable nonlinear band-gap localized states, and also reveal an important physical mechanism of the oscillatory wave instabilities associated with the band-gap resonances.Comment: 4 pages, 5 figure

Impact of highly qualified personnels migration on economic and innovative development

Лібералізація та раціоналізація є основними факторами, що сприяють модифікації міграційних процесів та зростанню відповідних міграційних показників. До факторів, що визначають глобальний міграційний рух, також можна віднести розширення та придбання маси технологій, спрямованих на зменшення відчуття дистанції, які дозволяють швидко та оперативно змінювати місця перебування, не змінюючи умов та принципів роботи. , контактні аудиторії тощо. Саме в цьому контексті актуальним стає фокус досліджень міграційних процесів загалом і тих сфер, де невеликі зміни в яких потенційно можуть мати значний вплив на економічну ситуацію та подальший розвиток, у тому числі інноваційний. . Гіпотеза дослідження полягає в тому, що міграція висококваліфікованих кадрів здійснює комплекс впливів на економічний та інноваційний розвиток країни-реципієнта та країни-донора. Досягти поставленої мети дозволяють застосовані аналітичні методи, порівняльний аналіз, класифікація. Автори згрупували основні показники впливу міграційних процесів на країни виїзду (країни-донори) та виїзди (країни-реципієнти) за економічною та інноваційною складовою. Аналіз представляє результати в якісному та кількісному вираженні. Проаналізовано ІТ-ринок, наведено показники його розвитку та визначено основні тенденції, у тому числі у порівнянні зі світовими. Описано особливості міграційних процесів фахівців даної галузі. Як наслідки, пропонується включити міграцію висококваліфікованої робочої сили до показників оцінки Country Innovation Index.Liberalization and rationalization are the main factors that favor the modification of migration processes and the growth of relevant migration indicators. Factors that determine the global migration movement can also be attributed to the expansion and acquisition of a mass of technologies that aim at reducing the sense of distance, which allow you to quickly and quickly change locations of stay, without changing the conditions and principles of work, contact audiences, etc. It is in this context that the focus of research on migration processes in general, and those areas where small changes in which, can potentially have a significant impact on the economic situation and further development, including innovation, becomes relevant. The hypothesis of investigation is the migration of highly skilled personnel exerts a set of influences on the economic and innovative development of the recipient country and the donor country. Applied analytical methods, comparative analysis, classification, allow to achieve the aim. The authors grouped the main indicators of the migration processes impact on the countries of departure (donor countries) and those leaving (recipient countries) in terms of economic and innovation component. The analysis presents the results in qualitative and quantitative terms. The IT market is analyzed, indicators of its development are given, and major trends are identified, including in comparison with the world ones. The peculiarities of migration processes of specialists in this field are described. As implications, it is proposed to include the migration of highly skilled workforce in the assessment indicators Country Innovation Index.Liberalization and rationalization are the main factors that favor the modification of migration processes and the growth of relevant migration indicators. Factors that determine the global migration movement can also be attributed to the expansion and acquisition of a mass of technologies that aim at reducing the sense of distance, which allow you to quickly and quickly change locations of stay, without changing the conditions and principles of work, contact audiences, etc. It is in this context that the focus of research on migration processes in general, and those areas where small changes in which, can potentially have a significant impact on the economic situation and further development, including innovation, becomes relevant. The hypothesis of investigation is the migration of highly skilled personnel exerts a set of influences on the economic and innovative development of the recipient country and the donor country. Applied analytical methods, comparative analysis, classification, allow to achieve the aim. The authors grouped the main indicators of the migration processes impact on the countries of departure (donor countries) and those leaving (recipient countries) in terms of economic and innovation component. The analysis presents the results in qualitative and quantitative terms. The IT market is analyzed, indicators of its development are given, and major trends are identified, including in comparison with the world ones. The peculiarities of migration processes of specialists in this field are described. As implications, it is proposed to include the migration of highly skilled workforce in the assessment indicators Country Innovation Index

Physiological and pathophysiological homeostasis of astroglial channel proteins by Nedd4-2

Nedd4-2 is an E3 ubiquitin ligase, missense mutation of which is related to familial epilepsy, indicating its critical role in regulating neuronal network activity. However, Nedd4-2 substrates involved in neuronal network function have yet to be identified. Using mouse lines lacking Nedd4-1 and Nedd4-2, we identified astrocytic channel proteins inwardly rectifying K+ channel 4.1 (Kir4.1) and Connexin43 as Nedd4-2 substrates. We found that the expression of Kir4.1 and Connexin43 is increased upon conditional deletion of Nedd4-2 in astrocytes, leading to an elevation of astrocytic membrane ion permeability and gap junction activity, with a consequent reduction of γ-oscillatory neuronal network activity. Interestingly, our biochemical data demonstrate that missense mutations found in familial epileptic patients produce gain-of-function of Nedd4-2 gene product. Our data reveal a process of coordinated astrocytic ion channel proteostasis that controls astrocyte function and astrocyte-dependent neuronal network activity, and elucidate a potential mechanism by which aberrant Nedd4-2 function leads to epilepsy

Idiopathic pulmonary fibrosis: Best practice in monitoring and managing a relentless fibrotic disease

Idiopathic pulmonary fibrosis (IPF) is a fibrosing interstitial lung disease that is, by definition, progressive. Progression of IPF is reflected by a decline in lung function, worsening of dyspnea and exercise capacity, and deterioration in health-related quality of life. In the short term, the course of disease for an individual patient is impossible to predict. A period of relative stability in forced vital capacity (FVC) does not mean that FVC will remain stable in the near future. Frequent monitoring using multiple assessments, not limited to pulmonary function tests, is important to evaluate disease progression in individual patients and ensure that patients are offered appropriate care. Optimal management of IPF requires a multidimensional approach, including both pharmacological therapy to slow decline in lung function and supportive care to preserve patients' quality of life