Disorders in the CMG helicase complex increase the proliferative capacity and delay chronological aging of budding yeast

The replication of DNA requires specialized and intricate machinery. This machinery is known as a replisome and is highly evolutionarily conserved, from simple unicellular organisms such as yeast to human cells. The replisome comprises multiple protein complexes responsible for various steps in the replication process. One crucial component of the replisome is the Cdc45-MCM-GINS (CMG) helicase complex, which unwinds double-stranded DNA and coordinates the assembly and function of other replisome components, including DNA polymerases. The genes encoding the CMG helicase components are essential for initiating DNA replication. In this study, we aimed to investigate how the absence of one copy of the CMG complex genes in heterozygous Saccharomyces cerevisiae cells impacts the cells' physiology and aging. Our data revealed that these cells exhibited a significant reduction in transcript levels for the respective CMG helicase complex proteins, as well as disruptions in the cell cycle, extended doubling times, and alterations in their biochemical profile. Notably, this study provided the first demonstration that cells heterozygous for genes encoding subunits of the CMG helicase exhibited a significantly increased reproductive potential and delayed chronological aging. Additionally, we observed a noteworthy correlation between RNA and polysaccharide levels in yeast and their reproductive potential, as well as a correlation between fatty acid levels and cell doubling times. Our findings also shed new light on the potential utility of yeast in investigating potential therapeutic targets for cancer treatment

Live while the DNA lasts. The role of autophagy in DNA loss and survival of diploid yeast cells during chronological aging

Aging is inevitable and affects all cell types, thus yeast cells are often used as a model in aging studies. There are two approaches to studying aging in yeast: replicative aging, which describes the proliferative potential of cells, and chronological aging, which is used for studying post-mitotic cells. While analyzing the chronological lifespan (CLS) of diploid Saccharomyces cerevisiae cells, we discovered a remarkable phenomenon: ploidy reduction during aging progression. To uncover the mechanism behind this unusual process we used yeast strains undergoing a CLS assay, looking for various aging parameters. Cell mortality, regrowth ability, autophagy induction and cellular DNA content measurements indicated that during the CLS assay, dying cells lost their DNA, and only diploids survived. We demonstrated that autophagy was responsible for the gradual loss of DNA. The nucleophagy marker activation at the start of the CLS experiment correlated with the significant drop in cell viability. The activation of piecemeal microautophagy of nucleus (PMN) markers appeared to accompany the chronological aging process until the end. Our findings emphasize the significance of maintaining at least one intact copy of the genome for the survival of post-mitotic diploid cells. During chronological aging, cellular components, including DNA, are exposed to increasing stress, leading to DNA damage and fragmentation in aging cells. We propose that PMN-dependent clearance of damaged DNA from the nucleus helps prevent genome rearrangements. However, as long as one copy of the genome can be rebuilt, cells can still survive

Depletion of the Origin Recognition Complex Subunits Delays Aging in Budding Yeast

Precise DNA replication is pivotal for ensuring the accurate inheritance of genetic information. To avoid genetic instability, each DNA fragment needs to be amplified only once per cell cycle. DNA replication in eukaryotes starts with the binding of the origin recognition complex (ORC) to the origins of DNA replication. The genes encoding ORC subunits have been conserved across eukaryotic evolution and are essential for the initiation of DNA replication. In this study, we conducted an extensive physiological and aging-dependent analysis of heterozygous cells lacking one copy of ORC genes in the BY4743 background. Cells with only one copy of the ORC genes showed a significant decrease in the level of ORC mRNA, a delay in the G1 phase of the cell cycle, and an extended doubling time. Here, we also show that the reducing the levels of Orc1-6 proteins significantly extends both the budding and average chronological lifespans. Heterozygous ORC/orcΔ and wild-type diploid cells easily undergo haploidization during chronological aging. This ploidy shift might be related to nutrient starvation or the inability to survive under stress conditions. A Raman spectroscopy analysis helped us to strengthen the hypothesis of the importance of lipid metabolism and homeostasis in aging

The impact of COVID-19 pandemic and distance learning on physical and mental health of Polish students

Introduction and aim. In Poland, on 25 March 2020, distance learning, which for many people was known only from literature, entered into force. As a result, education could no longer be provided in its current form and was therefore provided online. Therefore, the main aim of the presented research was an attempt to verify the pandemic and distance education impact on students psychophysical conditions and satisfaction. Material and methods. Some primary school students (grades 6th-8th), secondary school students from the Subcarpathian Voivodeship in Poland and also some university students from foreign countries completed an anonymous survey on the impact of the COVID-19 pandemic on their physical and mental condition as well as on some aspects of distance education. The research was conducted in December 2021 and 804 students (Primary and Secondary School) and 64 university students completed the survey. Results. Both the primary and the secondary school students claimed they were quite satisfied with distance education. The mostly approved aspect of online education was the fact that the time of learning was fitted to students’ needs. Difficulties in distance learning indicated by students were mainly connected with their mental health. Therefore, many students suffered from mental health and concentration problems, unwillingness to be active, apathy. The pandemic was also a stress factor for students and a reason why they were in a bad mood. The pandemic was the reason for an increased interest in natural sciences among 31.3% of primary school students, among 20.8% of secondary school students and among 17.3% of university students from foreign countries. Distance education influenced physical health of almost half of the respondents in both types of schools. The pandemic had an impact on mental health state of almost half of primary school students. 65% of secondary school students said that the pandemic influenced both their mental and physical health. In case of university students the pandemic did not influence their physical health in a significant way (65.4%). The study shows that primary school students used the Internet mainly to play games (almost 50% of the respondents), watch films, listen to music, do homework. Online lessons were only 20% of the answers. Conclusion. There has been a significant impact of COVID-19 on students in elementary school, high school, and university. This resulted in negative mental and physical effects. Respondents were stressed, depressed, and unmotivated. Approximately half of them engaged in physical activity very rarely or rarely. Even though they were satisfied with online learning, which was caused by the COVID-2019 pandemic, the greatest problem was mental concentration problems, lack of motivation, and social isolation

Links between Disease Severity, Bacterial Infections and Oxidative Stress in Cystic Fibrosis

Cystic fibrosis (CF) is one of the most common, yet fatal genetic diseases in Caucasians. The presence of a defective CF transmembrane conductance regulator and the massive neutrophils influx into the airways contribute to an imbalance in epithelial cell processes and extracellular fluids and lead to excessive production of reactive oxygen species and intensification of oxidative stress. The study included 16 controls and 42 participants with CF aged 10 to 38. The products of protein oxidation, total antioxidant capacity (TAC) and markers of lipid peroxidation were estimated in the serum of the subjects. Furthermore, we compared the level of oxidative stress in patients with CF according to the severity of disease and type of bacterial infection. Thiol groups and serum TAC decreased significantly in patients with CF (p p Pseudomonas aeruginosa had elevated 3-nitrotyrosine concentration levels (p Staphylococcus aureus noted a decrease in thiol groups (p P. aeruginosa or S. aureus had a slight effect on oxidative stress, while co-infection by two species did not affect the level of oxidative stress

Effects of Temperature on Lifespan of Drosophila melanogaster from Different Genetic Backgrounds: Links between Metabolic Rate and Longevity

Despite many studies of the aging process, questions about key factors ensuring longevity have not yet found clear answers. Temperature seems to be one of the most important factors regulating lifespan. However, the genetic background may also play a key role in determining longevity. The aim of this study was to investigate the relationship between the temperature, genetic background (fruit fly origin), and metabolic rate on lifespan. Experiments were performed with the use of the wild type Drosophila melanogaster fruit flies originating from Australia, Canada, and Benin and the reference OregonR strain. The metabolic rate of D. melanogaster was measured at 20 °C, 25 °C, and 28 °C in an isothermal calorimeter. We found a strong negative relationship between the total heat flow and longevity. A high metabolic rate leads to increased aging in males and females in all strains. Furthermore, our results showed that temperature has a significant effect on fecundity and body weight. We also showed the usefulness of the isothermal calorimetry method to study the effect of environmental stress conditions on the metabolic activity of insects. This may be particularly important for the forecasting of impact of global warming on metabolic activity and lifespan of various insects

Oxidative Stress Markers in Urine and Serum of Patients with Bladder Cancer

Oxidative stress is defined as an imbalanced state of the production of reactive oxygen species and antioxidant capacity that causes oxidative damage to biomolecules, leading to cell injury and finally death. Oxidative stress mediates the development and progression of several cancer diseases, including bladder cancer. The aim of our study was to determine markers of levels of the oxidative stress in serum and urine in the same patients in parallel in serum and urine. Furthermore, we tried to estimate the associations between oxidative stress markers and the type of cancer, its clinical stage and grade, as the well as correlations between serum and urinary markers in patients with bladder cancer. Sixty-one bladder cancer and 50 healthy volunteers as a control group were included. We determined the serum and urine levels of advanced oxidation protein products (AOPP), Amadori products, total antioxidant capacity, total oxidant status (TOS), oxidative status index (OSI), and malondialdehyde. We confirm that almost all markers are elevated in serum and urine from patients with bladder cancer than from healthy subjects. Moreover, we did not find differences in the level of oxidative stress markers and the type of tumor, its clinical stage, and grade. We noted correlations between serum and urinary biomarkers, in particular TOS and OSI. Our results clearly indicate the participation of oxidative stress in the development of bladder cancer

Oxidative Stress Markers in Urine and Serum of Patients with Bladder Cancer

Oxidative stress is defined as an imbalanced state of the production of reactive oxygen species and antioxidant capacity that causes oxidative damage to biomolecules, leading to cell injury and finally death. Oxidative stress mediates the development and progression of several cancer diseases, including bladder cancer. The aim of our study was to determine markers of levels of the oxidative stress in serum and urine in the same patients in parallel in serum and urine. Furthermore, we tried to estimate the associations between oxidative stress markers and the type of cancer, its clinical stage and grade, as the well as correlations between serum and urinary markers in patients with bladder cancer. Sixty-one bladder cancer and 50 healthy volunteers as a control group were included. We determined the serum and urine levels of advanced oxidation protein products (AOPP), Amadori products, total antioxidant capacity, total oxidant status (TOS), oxidative status index (OSI), and malondialdehyde. We confirm that almost all markers are elevated in serum and urine from patients with bladder cancer than from healthy subjects. Moreover, we did not find differences in the level of oxidative stress markers and the type of tumor, its clinical stage, and grade. We noted correlations between serum and urinary biomarkers, in particular TOS and OSI. Our results clearly indicate the participation of oxidative stress in the development of bladder cancer

The Role of Oxidative Stress in Atopic Dermatitis and Chronic Urticaria

Atopic dermatitis (AD) and chronic urticaria (CU) are common skin diseases with an increasing prevalence and pathogenesis that are not fully understood. Emerging evidence suggests that oxidative stress plays a role in AD and CU. The aim of the single-center cross-sectional study was to compare markers of oxidative stress in 21 patients with AD, and 19 CU patients. The products of protein oxidation, total antioxidant capacity (TAC), and markers of lipid peroxidation were estimated in the serum. AD patients had a higher level of advanced protein oxidation products and a lower level of thiol groups than healthy participants. However, CU patients had statistically higher levels of AOPP and 3-nitrotyrosine than healthy subjects. The level of thiol groups and serum TAC decreased significantly in patients with CU. There was no difference in serum concentration of lipid peroxidation products, Amadori products, ratio of reduced to oxidized glutathione, and ability of albumin to binding cobalt between AD or CU patients compared to healthy subjects. We found a moderate positive significant correlation between AOPP and age in patients with AD. In patients with CU, TAC was negatively correlated with age. These results may shed light on the etiopathogenesis of AD or CU, and confirm an oxidative burden in these patients. Furthermore, our study could be useful in developing new therapeutic methods that include using antioxidants in dermatological diseases

Ribosomal Protein uL11 as a Regulator of Metabolic Circuits Related to Aging and Cell Cycle

Aging is a biological phenomenon common to all living organisms. It is thought that the rate of aging is influenced by diverse factors, in many cases related to the control of energy metabolism, i.e., the so-called pro-longevity effects of starvation. Translation, regarded as the main energy consumption process, lies at the center of interest, as it has a significant impact on the longevity phenomenon. It has been shown that perturbations in the translational apparatus may lead to a lower rate of aging. Therefore, the main aim of this study was to investigate aging in relation to the protein biosynthesis circuit, taking into account the uL11 ribosomal protein as a vital ribosomal element. To this end, we used set of yeast mutants with deleted single uL11A or uL11B genes and a double disruptant uL11AB mutant. We applied an integrated approach analyzing a broad range of biological parameters of yeast mutant cells, especially the longevity phenomenon, supplemented with biochemical and high throughput transcriptomic and metobolomic approaches. The analysis showed that the longevity phenomenon is not fully related to the commonly considered energy restriction effect, thus the slow-down of translation does not represent the sole source of aging. Additionally, we showed that uL11 can be classified as a moonlighting protein with extra-ribosomal function having cell-cycle regulatory potential