Protein Synthesis Research Articles

New insights into the mechanisms of red blood cell enucleation: From basics to clinical applications

AbstractBackgroundRed blood cell (RBC) enucleation is a crucial step in the process of erythropoiesis. By removing the nucleus, RBCs gain greater flexibility, enabling them to traverse narrow capillaries with ease, thereby enhancing the efficiency of oxygen and carbon dioxide transport. This transformation underscores the intricate balance between cellular structure and function essential for maintaining homeostasis.TopicThis review delves into the multifaceted enucleation process, outlining its complex steps that encompass protein sorting, vesicle trafficking, cytoskeletal remodeling, and apoptosis regulation, while also exploring the potential of enhancing the enucleation rate of RBCs in vitro. We emphasize the intricate regulation of this process, which is orchestrated by multiple factors. This includes transcription factors that meticulously guide protein synthesis and sorting through the modulation of gene expression, as well as non‐coding RNAs that play a pivotal role in post‐transcriptional regulation during various stages of RBC enucleation. Additionally, macrophages participate in the enucleation process by engulfing and clearing the extruded nuclei, further ensuring the proper development of RBCs. Although many studies have deeply explored the molecular mechanisms of enucleation, the roles of apoptosis and anti‐apoptotic processes in RBC enucleation remain incompletely understood.ImplicationIn this review, we aim to comprehensively summarize the RBC enucleation process and explore the progress made in ex vivo RBC generation. In the future, a deeper understanding of the enucleation process could provide significant benefits to patients suffering from anemia and other related conditions.

INFLUENCE OF UROGENEITRAL INFECTION IN PREGNANT WOMEN ON THE UTERINE – FETOPLACENTARY COMPLEX

As reported by the World Health Organization (WHO), approximately 20 % of pregnant women globally are aff ected by an infection of the lower genital tract. Urogenital infection can result in complications during the onset and progression of pregnancy, as well as the postpartum period. These complications include placental dysfunction, premature birth, low birth weight, premature rupture of the fetal membranes, and postpartum endometritis. A marker of normal placental function is α2-fertility microglobulin (AFMG), the determination of which will refl ect the presence or absence of a uterine- fetoplacental complex violation during urogenital infection in pregnant women.The aim of the study is to reveal the infl uence of urogenital infection on the dysfunction of the uterine – fetoplacental complex in pregnant women.Materials and methods. A total of 75 pregnant women were examined, divided into two groups: the main group (MG) and the comparison group (CG). The MG consisted of 55 pregnant women, of whom 35 underwent local sanitation of the urogenital infection according to the identifi ed pathogen (Subgroup A) and 20 refused this procedure (Subgroup B). The CG consisted of 20 pregnant women without urogenital infection, as determined by bacterioscopic, bacteriological, and polymerase chain reaction (PCR) examinations. All pregnant women underwent determination of AFMG in blood serum via immunoenzymatic methodology. The statistical analysis was conducted using the MedStat software (serial number MS00019). The study was performed in accordance with the provisions of the Declaration of Helsinki. The study protocol was approved by the Local Ethics Committee of PFMU for all women who participated in this study. Statistical processing was performed using the MedStat program (serial number MS00019). The work is a part of the initiative research work of the Department of Obstetrics and Gynecology No. 1 of Poltava State Medical University «Pathogenetic role of endothelial dysfunction and genetic features in pathology during pregnancy and gynecological diseases» (state registration No. 0117U005253, term of implementation 2017-2023).Research results. The results of the study demonstrated that pregnant women with a urogenital infection had a history of chronic pyelonephritis 5.5 times more frequently (p = 0.03) and salpingo- oophoritis 10.2 times more frequently (p < 0.001), which may be a source of latent infection. The most prevalent infectious agents among pregnant women were: The most prevalent microorganisms were Ureaplasma urealyticum (21.4 %), Candida fungi (19.04 %), Gardnerella vaginalis (11.9 %), and Toxoplasma gondii (11.9 %). The level of AFMG in pregnant women with urogenital infection was observed to be 4.3 times lower than in pregnant women with CG (p < 0.001). The remediation of an infection of the lower genital tract during pregnancy has been observed to increase AFMG production by a factor of four (p < 0.001). The level of AFMG in pregnant women of CG was 2.2 times higher than in pregnant womenof Subgroup A (p < 0.004).Conclusions. It was demonstrated that urogenital infection in pregnant women has a considerable impact on the dysfunction of the utero- fetoplacental complex. It was also shown that local sanitation of the lower genital tract in these women, based on the identified pathogen, has a benefi cial eff ect on the placenta’s functioning. However, this approach does not fully address the issue of protein synthesis impairment.

Distinct effects of CDK8 module subunits on cellular growth and proliferation in Drosophila.

The Mediator complex plays a pivotal role in facilitating RNA polymerase II-dependent transcription in eukaryotes. Within this complex, the CDK8 kinase module (CKM), comprising CDK8, Cyclin C (CycC), Med12, and Med13, serves as a dissociable subcomplex that modulates the activity of the small Mediator complex. Genetic studies in Drosophila have revealed distinct phenotypes associated with mutations in CKM subunits, but the underlying mechanisms have remained unclear. Using Drosophila as a model, we generated transgenic strains to individually or simultaneously deplete the four CKM subunits in all possible combinations, uncovering unique phenotypes in the eyes and wings. Depletion of CDK8-CycC enhanced E2F1 target gene expression and promoted cell-cycle progression, whereas Med12-Med13 depletion had no significant impact on these processes. Instead, depleting Med12-Med13 altered the expression of ribosomal protein genes and fibrillarin, reduced nascent protein synthesis, indicating a severe reduction in ribosome biogenesis and cellular growth compared to the loss of CDK8-CycC. These findings reveal distinct in vivo roles for CKM subunits, with Med12-Med13 disruption having a more pronounced effect on ribosome biogenesis and protein synthesis than CDK8-CycC loss.

Unveiling the Antibacterial Mechanism of Gold Nanoparticles by Analyzing Bacterial Metabolism at the Molecular Level.

The threat of drug-resistant bacteria is challenging, and it is urgent to explore new antibiotics. Gold nanoparticles (AuNPs) are known to be a group of promising antibacterial agents for replacing conventional antibiotics. Nevertheless, their antibacterial mechanism remains to be elucidated. Herein, we directly observed the interaction between antibacterial AuNPs and bacteria at the molecular level using neutral desorption extractive electrospray ionization mass spectrometry (ND-EESI-MS). We monitored and analyzed the dynamic changes of bacterial metabolites in real time after AuNP treatment. Ten substances representing 3 major metabolic pathways, including protein and nucleic acid synthesis, energy metabolism, and quorum sensing, were identified, indicating that AuNPs may exert antibacterial effects through multiple mechanisms influencing bacterial metabolism and communication. This study deepens the understanding of the antibacterial mechanism of AuNPs and is insightful for designing and screening new antibacterial agents.

Unlocking marine microbial treasures: new PBP2a-targeted antibiotics elicited by metals and enhanced by RSM-driven transcriptomics and chemoinformatics.

Elicitation through abiotic stress, including heavy metals, is a new natural product drug discovery technique. In this research, three compounds 1, 2, and 6, were achieved by triggering zinc and nickel on marine Sphingomonas sp. and Streptomyces sp., which were absent in normal culture. Compound 5 was obtained for the first time from marine bacteria. All compounds showed potent antibacterial activity against Staphylococcus aureus and bactericidal effect at 300µm, but 6 was more active. The potent compound 6 production was further enhanced through response surface methodology by optimizing the condition consisting of nickel 1mM ions, 20mg/L sucrose, 30mg/L salt and culture time 14days. Under these conditions, the SM-6 production was enhanced with a yield of 6.3mg/L, which was absent in the normal culture. Further transcriptome analysis of compound 6 unveiled its antibacterial activity on S. aureus by modulating heat shock protein genes, disrupting protein folding and synthesis, and perturbing cellular redox balance, leading to a comprehensive inhibition of normal bacterial growth. In addition, ADMET has shown that all compounds are safe for cardiac and hepatotoxicity. To determine the anti-bacterial mechanism, all compounds were docked with PBP2a and DNA gyrase enzyme, and TLR-4 protein for predicting vaccine construct, and the best docking score was achieved against PBP2a enzyme with the highest score of -10.2 for compound 6. In-silico cloning was carried out to ensure the expression of proteins generated and were cloned using S.aureus as a host. The simulation studies have shown that both SM-6-PBP2a and TLR-4-PBP2a complex are stable with the system. This study presents a new approach to anti-bacterial drug discovery from microorganisms through heavy metals triggering and enhancing the compound production through response surface methodology.

Systematic genetic characterization of the human PKR kinase domain highlights its functional malleability to escape a poxvirus substrate mimic

Evolutionary arms races can arise at the contact surfaces between host and viral proteins, producing dynamic spaces in which genetic variants are continually pursued. However, the sampling of genetic variation must be balanced with the need to maintain protein function. A striking case is given by protein kinase R (PKR), a member of the mammalian innate immune system. PKR detects viral replication within the host cell and halts protein synthesis to prevent viral replication by phosphorylating eIF2α, a component of the translation initiation machinery. PKR is targeted by many viral antagonists, including poxvirus pseudosubstrate antagonists that mimic the natural substrate, eIF2α, and inhibit PKR activity. Remarkably, PKR has several rapidly evolving residues at this interface, suggesting it is engaging in an evolutionary arms race, despite the surface’s critical role in phosphorylating eIF2α. To systematically explore the evolutionary opportunities available at this dynamic interface, we generated and characterized a library of 426 SNP-accessible nonsynonymous variants of human PKR for their ability to escape inhibition by the model pseudosubstrate inhibitor K3, encoded by the vaccinia virus gene K3L. We identified key sites in the PKR kinase domain that harbor K3-resistant variants, as well as critical sites where variation leads to loss of function. We find K3-resistant variants are readily available throughout the interface and are enriched at sites under positive selection. Moreover, variants beneficial against K3 were also beneficial against an enhanced variant of K3, indicating resilience to viral adaptation. Overall, we find that the eIF2α-binding surface of PKR is highly malleable, potentiating its evolutionary ability to combat viral inhibition.

Systematic genetic characterization of the human PKR kinase domain highlights its functional malleability to escape a poxvirus substrate mimic.

Evolutionary arms races can arise at the contact surfaces between host and viral proteins, producing dynamic spaces in which genetic variants are continually pursued. However, the sampling of genetic variation must be balanced with the need to maintain protein function. A striking case is given by protein kinase R (PKR), a member of the mammalian innate immune system. PKR detects viral replication within the host cell and halts protein synthesis to prevent viral replication by phosphorylating eIF2α, a component of the translation initiation machinery. PKR is targeted by many viral antagonists, including poxvirus pseudosubstrate antagonists that mimic the natural substrate, eIF2α, and inhibit PKR activity. Remarkably, PKR has several rapidly evolving residues at this interface, suggesting it is engaging in an evolutionary arms race, despite the surface's critical role in phosphorylating eIF2α. To systematically explore the evolutionary opportunities available at this dynamic interface, we generated and characterized a library of 426 SNP-accessible nonsynonymous variants of human PKR for their ability to escape inhibition by the model pseudosubstrate inhibitor K3, encoded by the vaccinia virus gene K3L. We identified key sites in the PKR kinase domain that harbor K3-resistant variants, as well as critical sites where variation leads to loss of function. We find K3-resistant variants are readily available throughout the interface and are enriched at sites under positive selection. Moreover, variants beneficial against K3 were also beneficial against an enhanced variant of K3, indicating resilience to viral adaptation. Overall, we find that the eIF2α-binding surface of PKR is highly malleable, potentiating its evolutionary ability to combat viral inhibition.

Nutritional intervention to enhance recovery after arthroscopic knee surgery in adults: a randomized controlled pilot trial.

Essential amino acid (EAA) and omega-3 fatty acid ingestion independently attenuate leg skeletal muscle disuse atrophy in uninjured persons. However, no data exist regarding the effectiveness of combined EAA and omega-3 fatty acid ingestion to mitigate skeletal muscle disuse atrophy in response to anterior cruciate ligament reconstruction (ACLR) surgery. This pilot trial will explore the feasibility of recruitment and retention of ACLR outpatients from a single center across 18 months to consume either a combination of omega-3 fatty acids and EAAs, or a placebo control, for 4 weeks before and 2 weeks after surgery. Thirty adult (≥ 18 years old) ACLR outpatients will be recruited for this single center, double-blind, two-arm randomized controlled feasibility pilot trial. Participants will consume either 5 g⋅day-1 of omega-3 fatty acids (fish oil) and 40 g⋅day-1 of EAAs or 5 g⋅day-1 of a control fatty acid mixture (safflower oil) and 40 g⋅day-1 of non-essential amino acids (NEAAs). Fatty acid supplements will be consumed 4 weeks before and for 2 weeks after ACLR surgery, whereas the EAAs and NEAAs will be consumed 1 week before and for 2 weeks after ACLR surgery. The primary outcomes are feasibility of recruitment and retention, with the goal to recruit 30 outpatients across 18 months and retain 22 participants upon completion of the study protocol following 12 weeks of data collection. These results will be reported using descriptive statistics, along with reasons and timepoints for study dropout. Secondary exploratory outcomes will be reported using inferential statistics for purposes of hypothesis generation and elucidation of mechanistic targets for future work; no inferences to clinical efficacy will be made. These outcomes include integrated rates of skeletal muscle protein synthesis, skeletal muscle protein content and expression of translation factors, skeletal muscle and erythrocyte phospholipid composition, and measures of skeletal muscle mass, strength, and power. This work will set the foundation for a future randomized controlled trialpowered to detect an effect of EAA + omega-3 fatty acid intake on skeletal muscle size or function in response to ACLR surgery. ClinicalTrials.gov, NCT06233825. Registered 31 January 2024. https://clinicaltrials.gov/study/NCT06233825?term=NCT06233825&rank=1.

Experimentally-driven mathematical model to understand the effects of matrix deprivation in breast cancer metastasis.

Normal epithelial cells receive proper signals for growth and survival from attachment to the underlying extracellular matrix (ECM). They perceive detachment from the ECM as a stress and die - a phenomenon termed as 'anoikis'. However, metastatic cancer cells acquire anoikis-resistance and circulate through the blood and lymphatics to seed metastasis. Under normal (adherent) growth conditions, the serine-threonine protein kinase Akt stimulates protein synthesis and cell growth, maintaining an anabolic state in the cancer cell. In contrast, previously we showed that the stress due to matrix deprivation is sensed by yet another serine-threonine kinase, AMP-activated protein kinase (AMPK), that inhibits anabolic pathways while promoting catabolic processes. We illustrated a switch from Akthigh/AMPKlow in adherent condition to AMPKhigh/Aktlow in matrix-detached condition, with consequent metabolic switching from an anabolic to a catabolic state, which aids cancer cell stress-survival. In this study, we utilized these experimental data and developed a deterministic ordinary differential equation (ODE)-based mechanistic mathematical model to mimic attachment-detachment signaling network. To do so, we used the framework of insulin-glucagon signaling with consequent metabolic shifts to capture the pathophysiology of matrix-deprived state in breast cancer cells. Using the developed metastatic breast cancer signaling (MBCS) model, we identified perturbation of several signaling proteins such as IRS, PI3K, PKC, GLUT1, IP3, DAG, PKA, cAMP, and PDE3 upon matrix deprivation. Further, in silico molecular perturbations revealed that several feedback/crosstalks like DAG to PKC, PKC to IRS, S6K1 to IRS, cAMP to PKA, and AMPK to Akt are essential for the metabolic switching in matrix-deprived cancer cells. AMPK knockdown simulations identified a crucial role for AMPK in maintaining these adaptive changes. Thus, this mathematical framework provides insights on attachment-detachment signaling with metabolic adaptations that promote cancer metastasis.

TUDY OF CHRONIC TOXICITY OF AN IMPROVED A-SILICONE IMPRESSION MATERIAL PRODUCED IN UKRAINE

The objective of this study is to evaluate the potential toxic effects of an improved A-silicone impression material with decorative properties on the organs and mucous membranes of laboratory animals. Materials and Methods The experiment was conducted on white laboratory rats aged 9 months and weighing 230–285 g. Findings indicate that prolonged contact (30 days) with the material does not result in significant changes in the function of major organ systems in these laboratory rats. Results. A single exposure of laboratory rats to A-silicone impression material with disinfectant properties did not cause significant changes in the physiological state of the animals. Weights of internal organs, including the liver, spleen, heart, and adrenal glands, remained within normal limits, indicating that the material had no toxic effects. The analysis of the peripheral blood composition did not reveal any abnormalities in the number of red blood cells, leukocytes, hemoglobin and other cellular components. The functional state of the liver and kidneys, including the activity of the enzymes AlаT and AсаT, levels of total protein, albumin, creatinine and urea, remained stable and did not change under the influence of the material. There was also no effect on carbohydrate metabolism. No abnormalities in protein synthesis and nitrogen excretion functions were detected, which confirms the safety of the material during the 30-day observation. Conclusion. It was established that this material in the tested dose does not affect the enzyme-synthetic function of the liver and does not exhibit cytolytic effects. The material was found to have no toxic impact on liver function with prolonged use, nor does it cause statistically significant deviations in the measured indicators between the control and experimental groups. The absence of sugar, ketones, and protein in the urine indicates no adverse effect on the renal nitrogen-excreting and saluretic functions, as well as on the vital organs of the rats.

Nanopore-Based Sequencing of the Full-Length Transcriptome of Male and Female Cleavage-Stage Embryos of the Chinese Mitten Crab (Eriocheir sinensis)

The cleavage stage plays a crucial role in embryo development, characterized by a swift surge in cell proliferation alongside the accurate genetic material transmission to offspring. To delve into the characteristics of sex development during the cleavage stage of embryos, we generated the full-length transcriptome of Eriocheir sinensis male and female cleavage-stage embryos using Oxford Nanopore Technologies (ONT). Notably, this investigation represents the first sequencing effort distinguishing between genders in E. sinensis embryos. In the transcriptome structure analysis, male and female cleavage-stage embryos, while not clustered, exhibited a comparable frequency of alternative splicing (AS) occurrences. We also successfully identified 2875 transcription factors (TFs). The quantitative analysis showed the top 150 genes, in which the highly expressed genes in male embryos predominantly related to protein synthesis and metabolism. Further investigation unveiled 500 differentially expressed genes (DEGs), of which 7 male-biased ribosomal protein genes (RPGs) were particularly noteworthy and further confirmed. These analyses suggest that there may be a more active protein synthesis process in male E. sinensis cleavage-stage embryos. Furthermore, among the 2875 identified TFs, we predicted that 18 TFs could regulate the differentially expressed RPGs, with most TFs belonging to the zf-C2H2 and Homeobox families, which are crucial for embryonic development. During the cleavage stage of E. sinensis, the differential RPGs between genders were intricately linked to energy metabolism. We proposed that these RPGs exert regulatory effects on gene expression in E. sinensis, thereby regulating the difference of development between male and females. Our research sheds light on the developmental mechanisms of E. sinensis during the embryo stage and establishes a groundwork for a deeper understanding of sex development in E. sinensis. The results also provide comprehensive full-length transcriptome data for future gene expression and genetic studies in E. sinensis.

Respiratory Outcomes of Interrupted Modulator Therapies in Children With Cystic Fibrosis.

Cystic fibrosis (CF) is a multisystemic disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, resulting in defective synthesis or function of the CFTR protein. Historically, CF treatment focused on managing symptoms and complications. Fortunately, modulator drugs are now available to directly target the defective CFTR protein. However, in some countries, such as Turkey, these drugs are not covered by social insurance. Consequently, many CF patients face barriers to accessing modulatory therapies or must interrupt their treatment. This study demonstrates the impact of interrupting modulator therapy on pulmonary function, emphasizing the need for uninterrupted continuous treatment. In this study, 39 CF patients receiving elexacaftor-tezacaftor-ivacaftor (ETI) at our clinic were retrospectively analyzed. Among the patients, 18 experienced one or more interruptions, ranging from 15 to 210 days during ETI treatment. We analyzed pulmonary function test results from 27 interruption periods. At the beginning of the interruption, the mean percent predicted FEV1 (ppFEV1) was 69.59% ± 25.87%, which decreased to 64.96% ± 24.52% by the end of the interruption. There was a significant decrease with a mean change of 4.62 ± 8.49 (p = 0.008). However, no significant correlation was found between the interruption duration and FEV1 change. Our results demonstrate that pulmonary functions are adversely affected by interruption periods, regardless of their duration. Even short interruptions have a significant impact on pulmonary functions. This underscores the need for uninterrupted continuation of modulatory treatment and for improved policies to ensure equitable access to treatment.

Bone loss with aging is independent of gut microbiome in mice

Emerging evidence suggests a significant role of gut microbiome in bone health. Aging is well recognized as a crucial factor influencing the gut microbiome. In this study, we investigated whether age-dependent microbial change contributes to age-related bone loss in CB6F1 mice. The bone phenotype of 24-month-old germ-free (GF) mice was indistinguishable compared to their littermates colonized by fecal transplant at 1-month-old. Moreover, bone loss from 3 to 24-month-old was comparable between GF and specific pathogen-free (SPF) mice. Thus, GF mice were not protected from age-related bone loss. 16S rRNA gene sequencing of fecal samples from 3-month and 24-month-old SPF males indicated an age-dependent microbial shift with an alteration in energy and nutrient metabolism potential. An integrative analysis of 16S predicted metagenome function and LC-MS fecal metabolome revealed an enrichment of protein and amino acid biosynthesis pathways in aged mice. Microbial S-adenosyl methionine metabolism was increased in the aged mice, which has previously been associated with the host aging process. Collectively, aging caused microbial taxonomic and functional alteration in mice. To demonstrate the functional importance of young and old microbiome to bone, we colonized GF mice with fecal microbiome from 3-month or 24-month-old SPF donor mice for 1 and 8 months. The effect of microbial colonization on bone phenotypes was independent of the microbiome donors’ age. In conclusion, our study indicates age-related bone loss occurs independent of gut microbiome.

Integrated Profiling Identifies Long-Term Molecular Consequences of Prenatal Dexamethasone Treatment in the Rat Brain-Potential Triggers of Depressive Phenotype and Cognitive Impairment.

Prenatal excess of glucocorticoids (GCs) is considered to be one of the highly impacting factors contributing to depression development. Although GCs are crucial for normal fetal development and their administration (mainly dexamethasone, DEX) is a life-saving procedure for those at risk of preterm delivery, exposure to excess levels of GCs during pregnancy can yield detrimental consequences. Therefore, we aimed to systematically investigate the brain molecular alterations triggered by prenatal DEX administration. We used a rat model of depression based on prenatal exposure to DEX and performed integrative multi-level methylomic, transcriptomic, and proteomic analyses of adult rats' brains (i.e., frontal cortex (FCx) and hippocampus (Hp)) to identify the outcomes of DEX action. Each of the investigated levels was significantly affected by DEX in the long-term manner. Particularly, we found 200 CpG islands to be differentially methylated in the FCx and 200 in the Hp of prenatally DEX-treated rats. Global transcriptomic analysis uncovered differential expression of transcripts mostly in FCx (271) and 1 in Hp, while proteomic study identified 146 differentially expressed proteins in FCx and 123 in Hp. Among the identified enriched molecular networks, we found altered pathways involved in synaptic plasticity (i.e., cAMP, calcium, and Wnt signaling pathways or tight junctions and adhesion molecules), which may contribute to cognitive impairment, observed in DEX-treated animals. Moreover, in the FCx, DEX administration in the prenatal period downregulates the expression of ribosome protein genes associated both with large and small ribosomal subunit assembly which can lead to a global decrease in translation and protein synthesis processes and, indirectly, alterations in the neurotransmission process.

PATHOPHYSIOLOGICAL AND STRUCTURAL PECULIARITIES OF CARDIAC FIBROSIS DEVELOPMENT IN ATRIAL FIBRILLATION

Introduction. In recent years, the study of the pathogenesis of atrial fibrillation (AF) and its complications resulting from this condition has drawn substantial attention from cardiologists. A comprehensive understanding of this issue is essential for investigating the structural and functional disturbances occurring in the heart during AF. These data are also important for developing novel therapies aimed at preventing the onset and progression of cardiac fibrosis. The aim of this study is to identify the primary pathways that activate cardiac connective tissue cells involved in fibrosogenesis, to examine the structural characteristics of this pathological process, and to investigate the role of fibroblasts in initiating and progressing cardiac fibrosis. Materials and Methods. The primary data for this article were collected through an in-depth analysis of recent scientific literature, with a focus on studies published in the last 5–7 years. Results. A detailed review of primary sources reveals that this cardiological issue has only been partially explored. Findings indicate a close link between the pathophysiological and structural mechanisms of AF and numerous resulting complications, with cardiac fibrosis being a primary consequence. Two major forms of fibrous damage to atrial cardiomyocytes play a key role in the development of cardiac fibrosis associated with AF. This includes both reactive and reparative types of cardiac fibrosis, which develop concurrently in atrial fibrillation. These structural changes lead not only to damage and loss of cardiomyocytes but also to pathological remodeling of fibroblasts, the intercellular matrix, and the atria and ventricles. Disruptions in bioelectrical potential conduction are also observed. Activated fibroblasts play a central role in initiating cardiac fibrosis, with key stages of protein synthesis involved in atrial fibrosis now better understood. Specific intracellular signaling pathways, which present potential therapeutic targets for preventing cardiac fibrosis in AF treatment, have been identified. Conclusion. Atrial fibrillation and cardiac fibrosis are interdependent, with each potentially accelerating the progression of the other. These pathological processes are underpinned by significant structural and functional disruptions within cardiac cells and the intercellular matrix. Fibroblasts, myofibroblasts, and intensive fibrous tissue formation within the matrix are pivotal in the development of cardiac fibrosis. A preventive strategy targeting early intervention in cardiac fibrosis shows the greatest therapeutic promise.

MECHANISMS OF ACTION OF PROTEIN SHAKES IN NORMALIZING METABOLIC PROCESSES, THEIR ROLE IN IMPROVING GLYCEMIC CONTROL IN PATIENTS WITH TYPE 2 DIABETES, AND IN PREVENTING CARDIOVASCULAR DISEASES

This review article delves into the significance of protein shakes, particularly whey protein, in various health and fitness contexts. The primary focus is on their biochemical properties, metabolic benefits, and their role in supporting muscle mass, weight loss, and metabolic health. The article synthesizes findings from multiple studies to highlight the potential of protein shakes in improving glycemic control, reducing inflammation, and enhancing overall health, particularly for individuals with type 2 diabetes, obesity, and sarcopenia. The review also explores the effectiveness of protein shakes in athletic performance and recovery, as well as their impact on glucose metabolism and homeostasis. It underscores the necessity for further research to optimize the use and dosage of protein shakes for diverse populations to fully harness their benefits. Whey proteins are rich in essential amino acids, particularly leucine, which stimulates muscle protein synthesis via the mTOR pathway. This makes them highly effective in muscle building and recovery. Whey protein consumption enhances glycemic control by increasing insulin secretion and improving insulin sensitivity. Studies have shown significant reductions in fasting glucose levels and HbA1c in patients with type 2 diabetes. Protein shakes aid in weight loss by promoting satiety and increasing thermogenesis. They help maintain muscle mass during weight loss, which is crucial for sustaining metabolic rate. Clinical trials have demonstrated their efficacy in reducing body fat and improving metabolic markers. Whey protein, combined with resistance training, significantly improves muscle mass and strength in older adults, helping to prevent sarcopenia. This is essential for maintaining physical functionality and reducing the risk of falls and fractures. In the rehabilitation of patients with severe obesity, protein shakes are effective in reducing body weight, improving metabolic health, and maintaining muscle mass. They play a crucial role in comprehensive rehabilitation programs that include diet and physical exercise. Protein shakes support metabolic adaptations by preserving muscle mass and enhancing thermogenesis, which helps maintain a high metabolic rate and prevent weight regain. Whey proteins enhance muscle mass and strength, speed up recovery after training, and boost endurance. They also support the immune system and reduce oxidative stress, contributing to better overall athletic performance . Proteins shakes aid in regulating blood glucose levels and improving insulin sensitivity, which is crucial for preventing and managing diabetes. Their antioxidant and anti-inflammatory properties further enhance metabolic health.

The miR3367-lncRNA67-GhCYP724B module regulates male sterility by modulating brassinosteroid biosynthesis and interacting with Aorf27 in Gossypium hirsutum.

Cytoplasmic male sterile (CMS) lines play a crucial role in utilization of heterosis in crop plants. However, the mechanism underlying the manipulation of male sterility in cotton by long non-coding RNA (lncRNA) and brassinosteroids (BRs) remains elusive. Here, using an integrative approach combining lncRNA transcriptomic profiles with virus-induced gene silencing experiments, we identify a flower bud-specific lncRNA in the maintainer line 2074B, lncRNA67, negatively modulating with male sterility in upland cotton (Gossypium hirsutum). lncRNA67 positively regulates cytochrome P274B (GhCYP724B), which acted as an eTM (endogenous target mimic) for miR3367. The suppression of GhCYP724B induced symptoms of BR deficiency and male semi-sterility in upland cotton as well as in tobacco, which resulted from a reduction in the endogenous BR contents. GhCYP724B regulates BRs synthesis by interacting with GhDIM and GhCYP90B, two BRs biosynthesis proteins. Additionally, GhCYP724B suppressed a unique chimeric open reading frame (Aorf27) in 2074A mitochondrial genome. Ectopic expression of Aorf27 in yeast inhibited cellular growth, and over expression of Aorf27 in tobacco showed male sterility. Overall, the results proved that the miR3367-lncRNA67-GhCYP724B module positively regulates male sterility by modulating BRs biosynthesis. The findings uncovered the function of lncRNA67-GhCYP724B in male sterility, providing a new mechanism for understanding male sterility in upland cotton.

Clinical and genetic analysis of a patient with Ataxia and vitamin E deficiency due to homozygous variant of TTPA gene

To explore the clinical phenotype and genetic characteristics of a patient with Ataxia and vitamin E deficiency syndrome (AVED) due to a variant of TTPA gene. A patient diagnosed with AVED (proband), intended for assisted reproductive technology for pregnancy in Zhongnan Hospital of Wuhan University in July 2023, was selected as research subject. Clinical data of the proband were collected, and 2 mL of peripheral venous blood samples were collected from the proband and her father and siblings for serum vitamin E level testing. Whole exome sequencing (WES) was carried out. Pathogenic variants were selected based on American public archive of interpretations of clinically relevant variants (ClinVar). Sanger sequencing was performed to validate the candidate variants detected by WES. Pathogenicity of variants was classified based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), and the impact of variants was analyzed using multiple bioinformatics tools including SIFT, Mutation Taster, CADD, and SpliceAI. Information on the protein domains was obtained from ClinVar and dbSNP databases, and a hotspot map for the variants of protein-coding region was constructed. This study was approved by the Medical Ethics Committee of Zhongnan Hospital of Wuhan University (No. 2023068K). The proband has a significantly low serum level of vitamin E (5.186 μg/mL), while her father and siblings were normal. WES revealed that she has harbored a homozygous missense c.2T>A (p.0?) variant of the TTPA gene, for which her father and younger sister were heterozygous carriers. Based on the guidelines from the ACMG, the missense c.2T>A (p.0?) variant of the TTPA gene was classified as pathogenic (PVS1+PM2+PM3). Multiple bioinformatics tools had predicted this variant to be located in the initiation codon region and may lead to abnormal synthesis of the TTPA protein, indicating it was deleterious. The hotspot map based on ClinVar and dbSNP databases showed an even distribution of variants across 5 structural domains of the TTPA protein, with high conservation of the first amino acid residue across various species. The homozygous c.2T>A (p.0?) variant of the TTPA gene probably underlie the AVED in the proband. Above discovery has enriched the mutational spectrum of AVED and provided a basis for the diagnosis, genetic counseling, and assisted reproductive strategies for this family.

Molecular mechanism analysis of a family with hereditary coagulation FXI deficiency caused by compound heterozygous mutations.

The purpose of this study was to determine the molecular basis of a Chinese family with factor XI (FXI) deficiency. The qRT-PCR was used to detect the transcription of F11 mRNA in transfected cells. ELISAs and western blot were used to detect the expression of FXI protein in culture media and lysates. Genetic analysis revealed that the proband carried a heterozygous nonsense mutation c.1107C>A (p.Tyr351stop) in exon 10 and a heterozygous missense mutation c.1562A>G (p.Tyr503Cys) in exon 13. The expression study revealed that p.Tyr351stop mutation resulted in the degradation of F11 mRNA. The p.Tyr503Cys mutation, however, had no effects on biosynthesis and secretion of FXI protein, but it had affected the catalytic activity of FXI. The inherited FXI deficiency of this family is related to nonsense mutation p.Tyr351stop and missense mutation p.Tyr503Cys.

The Interplay Between the MYC Oncogene and Ribosomal Proteins in Osteosarcoma Onset and Progression: Potential Mechanisms and Indication of Candidate Therapeutic Targets

High-grade osteosarcoma (OS) is the most common primary bone tumor mainly affecting children and young adults. First-line treatment consists of neo-adjuvant chemotherapy with doxorubicin, cisplatin, and methotrexate and surgery. The mean long-term survival rate for localized disease at diagnosis is 65–70%, dropping down to 20% when metastases are present at diagnosis. Therefore, curing OS is a clinical challenge, particularly for patients that do not respond to standard treatments. MYC has frequently been reported to be involved in the pathogenesis of OS and its high expression may be associated with drug resistance and patients’ worse prognosis. Moreover, MYC is a master regulator of ribosomal proteins (RPs) synthesis and ribosome biogenesis (RiBi), which is often up-regulated in human tumors. In recent years, RPs have been recognized not only for their traditional role in ribosome assembly but also for their extra-ribosomal functions, many of which are linked to the onset and progression of cancer. In this review we focus on the role and possible interplay of MYC and RPs expression in association with drug resistance and worse prognosis in OS and discuss therapeutic options that target de-regulated MYC, RiBi, or RPs, which are already clinically available or under evaluation in clinical trials.