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Abstract Intellectual disability and autistic features are associated with chromosome region 2q23.q23.2 duplication carrying LYPD6 and LYPD6B genes. Here, we analyzed LYPD6 and LYPD6B expression in patients with different neuropsychiatric disorders. Increased LYPD6 and LYPD6B expression was revealed in autism and other disorders. To study possible consequences of Lypd6 and Lypd6b overexpression in the brain, we used a mouse model with intracerebroventricular delivery of recombinant analogs of these proteins. A two-week infusion evoked significant memory impairment and acute stress. Both modulators downregulated hippocampal and amygdala dendritic spine density. No changes in synaptic plasticity were observed. Intracerebroventricular administration by both proteins downregulated hippocampal expression of Lypd6, Lypd6b, and α7 nicotinic acetylcholine receptor (nAChR). Similar to Lypd6, Lypd6b targeted different nAChR subtypes in the brain with preferential inhibition of α7- and α4β2-nAChRs. Thus, increased Lypd6 and Lypd6b level in the brain are linked to cholinergic system depression, neuronal atrophy, memory decline, and anxiety.

Three-finger proteins (TFPs), or Ly6/uPAR proteins, are characterized by the beta-structural LU domain containing three protruding “fingers” and stabilized by four conserved disulfide bonds. TFPs were initially characterized as snake alpha-neurotoxins, but later many studies showed their regulatory roles in different organisms. Despite a known expression of TFPs in vertebrates, they are poorly studied in other taxa. The presence of TFPs in starfish was previously shown, but their targets and functional role still remain unknown. Here, we analyzed expression, target, and possible function of the Lystar5 protein from the Asterias rubens starfish using bioinformatics, qPCR, and immunoassay. First, the presence of Lystar5 homologues in all classes of echinoderms was demonstrated. qPCR revealed that mRNA of Lystar5 and LyAr2 are expressed mainly in coelomocytes and coelomic epithelium of Asterias, while mRNA of other TFPs, LyAr3, LyAr4, and LyAr5, were also found in a starfish body wall. Using anti-Lystar5 serum from mice immunized by a recombinant Lystar5, we confirmed that this protein is expressed on the surface of coelomocytes and coelomic epithelium cells. According to ELISA, a recombinant analogue of Lystar5 bound to the membrane fraction of coelomocytes and coelomic epithelium but not to the body wall or starfish arm tip. Analysis by LC-MALDI MS/MS suggested integrin α-8-like protein expressed in the coelomocytes and coelomic epithelium as a target of Lystar5. Thus, our insights propose the important role of TFPs in regulation of starfish physiology and show prospects for their further research.

Making a correct genetically based diagnosis in patients with diseases associated with mitochondrial dysfunction can be challenging both genetically and clinically, as can further management of such patients on the basis of molecular-genetic data assessing the state of their mitochondria. In this opinion article, we propose a novel approach (which may result in a clinical protocol) to the use of a precise molecular-genetic tool in order to monitor the state of mitochondria (which reflects their function) during treatment of certain conditions, by means of not only signs and symptoms but also the molecular-genetic basis of the current condition. This is an example of application of personalized genomic medicine at the intersection of a person’s mitochondrial genome information and clinical care. Advantages of the proposed approach are its relatively low cost (compared to various types of sequencing), an ability to use samples with a low input amount of genetic material, and rapidness. When this approach receives positive outside reviews and gets an approval of experts in the field (in terms of the standards), it may then be picked up by other developers and introduced into clinical practice.

Three-finger proteins (TFPs), or Ly6/uPAR proteins, are characterized by the beta-structural LU domain containing three protruding "fingers" and stabilized by four conserved disulfide bonds. TFPs were initially characterized as snake alpha-neurotoxins, but later many studies showed their regulatory roles in different organisms. Despite a known expression of TFPs in vertebrates, they are poorly studied in other taxa. The presence of TFPs in starfish was previously shown, but their targets and functional role still remain unknown. Here, we analyzed expression, target, and possible function of the Lystar5 protein from the Asterias rubens starfish using bioinformatics, qPCR, and immunoassay. First, the presence of Lystar5 homologues in all classes of echinoderms was demonstrated. qPCR revealed that mRNA of Lystar5 and LyAr2 are expressed mainly in coelomocytes and coelomic epithelium of Asterias, while mRNA of other TFPs, LyAr3, LyAr4, and LyAr5, were also found in a starfish body wall. Using anti-Lystar5 serum from mice immunized by a recombinant Lystar5, we confirmed that this protein is expressed on the surface of coelomocytes and coelomic epithelium cells. According to ELISA, a recombinant analogue of Lystar5 bound to the membrane fraction of coelomocytes and coelomic epithelium but not to the body wall or starfish arm tip. Analysis by LC-MALDI MS/MS suggested integrin α-8-like protein expressed in the coelomocytes and coelomic epithelium as a target of Lystar5. Thus, our insights propose the important role of TFPs in regulation of starfish physiology and show prospects for their further research. [ABSTRACT FROM AUTHOR]

The application of soft materials, including gels, in different material and biological chemistry domains has gained momentum in the past few years. Environmental remediation, drug resistance, and delivery are significant concerns that are being investigated widely nowadays. In this work, we report the multifunctionality of a gelator molecule A2 (5,5′-((6-(phenylamino)-1,3,5-triazine-2,4-diyl)-bis-(azanediyl))-diisophthalic acid) in the gel state. The gelator molecule A2 was used to fabricate organogel and two metallogels using copper perchlorate and a Ru-based anticancer drug molecule, respectively. The organogel and Cu-metallogel show efficient adsorption capabilities of iodine in the vapor and solution phases. These iodine-loaded gel composites were utilized as efficient antibacterial agents against Escherichia coli and Helicobacter pylori (I10) bacterial strains. Additionally, owing to the pH-responsive nature of the gel material, the Ru-based anticancer drug-loaded metallogel, Ru-A2, was fabricated, and its activity in pH-responsive drug delivery to the cancerous cells was also investigated. [ABSTRACT FROM AUTHOR]

In this work, an atomistic-scale investigation of the phosphodiester P–O bond cleavage reaction by the enzyme ribonuclease A was carried out using computer simulation techniques. It is shown that during the reaction the network of non-covalent interactions in the active center of the ribonuclease changes significantly, while the role of these non-covalent interactions is different: coordination of the corresponding groups, electron density transfer, and ligand holding in the active center. It is shown that the process of proton transfer from Asp121 to His119 is the first stage of this reaction; at the same time, the hydrogen bond between the phosphate ligand and the imino group of Arg39 is broken, which, although keeping the ligand in the active center, does not allow the ligand to orient itself more conveniently for subsequent proton transfers. Furthermore, the key step of this reaction occurs: proton transfer with the participation of imidazole rings His12 and His119, in which the guiding role is played by several hydrogen bonds with the participation of Phe120, and the role of an electron density carrier is played by the pnictogen bond between the oxygen of the phosphate ligand and the pyridine-like nitrogen of the imidazole ring His119, which was detected for the first time. [ABSTRACT FROM AUTHOR]

Human adenoviruses (HAdVs) are common pathogens that are associated with a variety of diseases, including respiratory tract infections (RTIs). Without reliable, fast, and cost-effective detection methods for HAdVs, patients may be misdiagnosed and inappropriately treated. To address this problem, we have developed a multiplex loop-mediated isothermal amplification (LAMP) assay for the detection of the species Human adenovirus B (HAdV-B), Human adenovirus C (HAdV-C) and Human adenovirus E (HAdV-E) that cause RTIs. This multiplexing approach is based on the melting curve analysis of the amplicons with a specific melting temperature for each HAdV species. Without the need for typing of HAdVs, the LAMP results can be visually detected using colorimetric analysis. The assay reliably detects at least 375 copies of HAdV-B and -C and 750 copies of HAdV-E DNA per reaction in less than 35 min at 60 °C. The designed primers have no in silico cross-reactivity with other human respiratory pathogens. Validation on 331 nasal swab samples taken from patients with RTIs showed a 90–94% agreement rate with our in-house multiplex quantitative polymerase chain reaction (qPCR) method. Concordance between the quantitative and visual LAMP was 99%. The novel multiplexed LAMP could be an alternative to PCR for diagnostic purposes, saving personnel and equipment time, or could be used for point-of-care testing. [ABSTRACT FROM AUTHOR]

Introduction: Alzheimer's disease is increasingly becoming a cause of early disability and death. Attempts to create a pharmacotherapeutic agent that provides an effective result in treating this pathology have so far been unsuccessful. This article presents the results of experimental studies aimed at creating a more effective treatment of Alzheimer's disease. Material and Methods: The object of the study was polyisoprenoids isolated from Siberian fir (Abies sibirica Ledeb.). Experiments were conducted on 108 outbred adult male mice from the CD1 (cluster of differentiation 1) stock, weighing 28-30 g (at 5 weeks old). The effect of the sum of polyisoprenoids on learning and memory was studied in a dose range of 5, 20, 50, 100, 200, and 500 mg/kg. Gliatilin was used as a reference drug at a dose of 90 mg/kg. The experimental model of cognitive dysfunction was created by chronic (for 20 days) intraperitoneal injection of scopolamine at a dose of 2 mg/kg. Cognitive dysfunction was assessed by changes in the indicators of conditioned passive avoidance reflex (CPAR). Results and Discussion: The results of the experiments showed that chronic administration of scopolamine caused a pronounced decrease in the reproducibility of CPAR. Simultaneously, the studied substance at doses of 20 and 50 mg/kg restored the reproducibility of the reflex to a level close to the values of intact control. Gliatilin showed a similar effect. The authors believe that cholinergic receptors are involved in the development of the anti-amnestic effect. Conclusion: The presented study demonstrates the discovery of a new agent with anti-amnestic action based on the sum of polyprenols from Siberian Fir (Abies sibirica Ledeb.), family Pinaceae. This agent can be used for the treatment of neurodegenerative pathologies, particularly Alzheimer's disease. [ABSTRACT FROM AUTHOR]

Reverse transcriptases (RTs) are a family of enzymes that synthesize DNA using an RNA template and are involved in retrovirus propagation and telomere lengthening. In vitro, RTs are widely applied in various methods, including RNA-seq, RT-PCR, and RT-LAMP. Thermostable RTs from bacterial group II introns are promising tools for biotechnology due to their higher thermostability, fidelity, and processivity compared to commonly used M-MuLV RT and its mutants. However, the diversity of group II intron-encoded RTs is still understudied. In this work, we biochemically characterized a novel RT from a thermophilic bacterium, Anoxybacillus flavithermus, which was isolated from a hot spring in New Zealand and has an optimal growth temperature of around 60 °C. The cloned RT, named Afl RT, retained approximately 40% of the specific activity after a 45 min incubation at 50 °C. The optimal pH was 8.5, the optimal temperature was between 45 and 50 °C, and Mn2+ ions were found to be an optimal cofactor. The processivity analysis with MS2 phage gRNA (3569 b) demonstrated that Afl RT elongated fully up to 36% of the template molecules. In reverse transcription and RT-qLAMP, the enzyme allowed up to 10 copies of MS2 phage genomic RNA to be detected per reaction. Thus, Afl RT holds great potential for a variety of practical applications that require the use of thermostable and processive RTs. [ABSTRACT FROM AUTHOR]

LAMP (Loop-mediated isothermal amplification) is a popular method for the molecular diagnostics of numerous pathogens, specifically useful for point-of-care testing. However, the efficacy and sensitivity of LAMP still need to be maximised for the best performance in clinical settings. Adding a novel fourth primer pair is a promising way to accelerate the LAMP speed. Here, we report PI primers that are part of inner primers and can be used in LAMP without a specific design. PI primers were tested in quantitative LAMP detecting SARS-CoV-2 and MS2. The new primers have increased the speed and sensitivity of quantitative LAMP, RT-LAMP, and duplex LAMP with artificial templates and RNA samples from nasal swabs. Adding PI primers could become a valuable option for LAMP optimisation, especially when a desirable LAMP target is a highly variable DNA sequence with a few conservative sites for primers. [ABSTRACT FROM AUTHOR]

In this study, we report the synthesis and full characterization of five novel ruthenium(II) cymene complexes with the general formula [Ru(cym)(L')Cl], featuring N,O- and N,N-coordinating pyrazolone-based hydrazone ligands. We have characterized these complexes using single X-ray crystallography, Fourier-transform infrared spectroscopy (FT-IR), Nuclear magnetic resonance (NMR), elemental analysis, and Electrospray Ionization Mass Spectroscopy (ESI-MS). Crystallographic analysis confirmed that all of the complexes have a similar type of half-sandwich, pseudo-octahedral "three-legged piano-stool" geometry where the cymene moiety displays the typical η 6 -coordination mode and the hydrazone ligands coordinate to the Ru(II) center in a bidentate fashion. These complexes, with multiple catalytic sites, demonstrated high efficiency in catalyzing one-pot cascade deacetalization-Knoevenagel condensation reactions under mild conditions, achieving up to 92% of final product yield at 75 °C after 4 h of reaction time under solvent-free condition. Additionally, DFT calculations provided insight into the catalytic mechanism, suggesting a pathway driven by metal-ligand cooperation, assisted by the basic oxygen site of the pyrazolone ring and by the weakly acidic character of the NNH proton of the hydrazone group.

Introduction: The template method (TM), pioneered by Ozaki for aortic valve neocuspidization (AVNeo), has been widely adopted for aortic valve replacement, though it requires specialized instruments. This study introduces a novel formula method (FM), which uses the diameter of the aortic valve fibrous ring (AV-D) to determine the dimensions of the neocusps to be trimmed from autologous without the need for templates, potentially reducing costs and complexity. We aimed to compare the clinical outcomes of the FM with the established TM in patients undergoing AVNeo., Methods: A retrospective and prospective study was conducted on 31 patients who underwent isolated AVNeo between January 21, 2019 and December 15, 2022. Patients were divided into two groups: FM (n = 17) and TM (n = 14). The formula for the cusp free margin horizontal length is L1 = AV-D + 10 mm, cusp height is H = AV-D, cusp suture margin is L2 is a parabola that joins L1 and H, and cusp wings to be secured to aortic sinus = 3mm. The primary endpoints were major adverse valve-related events, including cardiac death, reoperation, and infective endocarditis. Secondary endpoints included significant aortic regurgitation, peak pressure gradients, aortic valve area, and New York Heart Association (NYHA) functional class. Intraoperative times, early postoperative outcomes, and mid-term hemodynamic performance were evaluated for both techniques., Results: Both the FM and TM demonstrated comparable intraoperative and postoperative outcomes. The cardiopulmonary bypass time, myocardial ischemia time, and blood loss were similar between the groups. Mid-term outcomes also showed no significant differences in valve function or hemodynamic parameters, with both groups exhibiting substantial reverse left ventricular remodeling. The FM group had a peak pressure gradient of 14.1 ± 4.3 mmHg compared to 18.4 ± 12.0 mmHg in the TM group (p = 0.219). The aortic valve area was 2.43 ± 0.3 cm² in the FM and 2.4 ± 0.2 cm² in the TM (p = 0.890). No significant differences were observed in freedom from reoperation or adverse events., Conclusion:  Both techniques showed excellent mid-term hemodynamic performance and comparable intraoperative and postoperative outcomes. The FM for AVNeo provides a cost-effective and practical alternative to the TM, offering similar clinical outcomes without the need for expensive templates; it has the potential to improve the accessibility of AVNeo, particularly in resource-limited settings. However, further research with larger cohorts and long-term follow-up is needed to fully assess the durability and long-term benefits of the FM., Competing Interests: Human subjects: Consent for treatment and open access publication was obtained or waived by all participants in this study. I.M. Sechenov First Moscow State Medical University IRB issued approval N/A. The study was conducted in compliance with the ethical requirements of the Ministry of Health of the Russian Federation Order No. 922n, dated November 15, 2012, "On the approval of the Procedure for providing medical care to the adult population in the field of 'surgery'." The study was also approved by the Institutional Review Board of I.M. Sechenov First Moscow State Medical University. Patients were informed of the procedure and were required to sign informed consent for surgery. Individual written consent to publish their study data was also obtained. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Komarov et al.)

Introduction Type A aortic dissection (TAAD) is a life-threatening condition that often leads to cerebral malperfusion (CM), a severe complication that can result in permanent neurological damage. Traditionally, a cardiopulmonary bypass (CPB) with selective antegrade cerebral perfusion (ACP) is employed during aortic arch reconstruction to protect cerebral circulation. However, the use of CPB carries inherent higher risks, including embolic events, hypothermia, and interrupted cerebral perfusion, especially in patients with CM. This study evaluates an innovative off-pump extra-corporeal ACP technique using an axillo-axillary shunt to provide uninterrupted bihemispheric cerebral perfusion during branch-first stage total aortic arch replacement (BF-TAR) for TAAD with CM; the shunt depends on cardiac contradiction to transfuse blood from the donor axillary artery to the recipient axillary artery, which then flows to through the carotid and vertebral arteries to the brain. Methods Between 2021 and 2023, 18 patients with TAAD complicated by CM underwent BF-TAR; the novel axillo-axillary shunt technique was employed for ACP because of the risks of ischemic neurologic injury. Outcomes measured included operative mortality, neurological complications, cardiopulmonary bypass times (measured after completion of the branch-first stage), and overall morbidity. Results The axillo-axillary shunt provided stable, continuous ACP in all patients. No new permanent neurological deficits were observed. Five (27.8%) patients experienced transient neurological symptoms such as blurred vision, dizziness, and confusion, which resolved within 48 hours. Operative mortality was 5.6% (1 patient), and minor complications included transitory lower limbs ischemia in 3 patients (16.7%) and deep sternal wound infection in 1 patient (5.6%). All transitory complications were managed by "watchful waiting". The mean CPB time was 145.3 ± 48.6 minutes, while the mean cross-clamp time was 100.6 ± 17.4 minutes, which was better than the average of 227 ± 32 minutes and 147 ± 23 minutes reported in other studies. Postoperative imaging confirmed well-reconstructed aortic arches with no residual malperfusion or graft-related complications. Conclusion The off-pump axillo-axillary shunt technique provides a safe and effective method for maintaining continuous bihemispheric cerebral perfusion during total aortic arch replacement in patients with TAAD complicated by CM. This approach minimizes the risks associated with CPB, including embolic events and interrupted cerebral perfusion while achieving favorable neurological and surgical outcomes. Further studies with larger cohorts and longer follow-ups are warranted to validate the long-term benefits of this innovative technique., Competing Interests: Human subjects: Consent was obtained or waived by all participants in this study. IRB of Bakulev Scientific Center of Cardiovascular Surgery · Moscow, RUS issued approval Protocol №: 4 (15.12.2021). The Institutional Review Board approved this short study after initial tests showed it was a viable and effective approach to cerebral perfusion with improved hemodynamic parameters as compared to conventional measures. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Elias et al.)

Making a correct genetically based diagnosis in patients with diseases associated with mitochondrial dysfunction can be challenging both genetically and clinically, as can further management of such patients on the basis of molecular-genetic data assessing the state of their mitochondria. In this opinion article, we propose a novel approach (which may result in a clinical protocol) to the use of a precise molecular-genetic tool in order to monitor the state of mitochondria (which reflects their function) during treatment of certain conditions, by means of not only signs and symptoms but also the molecular-genetic basis of the current condition. This is an example of application of personalized genomic medicine at the intersection of a person's mitochondrial genome information and clinical care. Advantages of the proposed approach are its relatively low cost (compared to various types of sequencing), an ability to use samples with a low input amount of genetic material, and rapidness. When this approach receives positive outside reviews and gets an approval of experts in the field (in terms of the standards), it may then be picked up by other developers and introduced into clinical practice., Competing Interests: The authors declare no conflict of interest., (© 2024 The Author(s). Published by IMR Press.)

Objectives: Veterans experience high rates of fatal and non-fatal firearm injuries. This risk may be compounded among Veterans who are rural-residing, aging, and/or experiencing cognitive decline or dementia. Firearm safety discussions are not broadly implemented across Department of Veterans Affairs (VA) healthcare settings due, in part, to concerns of causing Veterans to disengage from care. This study examines perceptions about firearm safety discussions to inform healthcare-based harm-reduction efforts., Methods: We conducted interviews with 34 Veterans (median age 70) and 22 clinicians from four VA facilities that treat high rates of rural patients with firearm-related injuries., Results: Most Veterans accepted the idea of universal firearm safety discussions at the VA. Some reported they might not be forthright in such discussions, but raising the topic would not stop them from engaging with VA care. Veterans and clinicians unanimously endorsed firearm safety discussions for older patients experiencing cognitive decline or dementia., Conclusions: VA patients and clinicians are amenable to firearm safety discussions during healthcare visits and especially endorse the need for such discussions among high-risk populations., Clinical Implications: Universal firearm safety discussions could be incorporated into standard VA practice, particularly for Veterans experiencing cognitive decline or dementia, without risking Veteran disengagement from care.