Changes in the Number of Residual γH2AX Foci in Ki-67-Positive and Ki-67-Negative Human Fibroblasts Irradiated with X-Rays in Doses of 2-10 Gy.

We studied changes in the number of residual γH2AX foci in cultured human fibroblasts with different expression of the cell proliferation marker protein Ki-67 24, 48, and 72 h after exposure to X-ray radiation in doses of 2-10 Gy. It was shown that, regardless of the expression of Ki-67, the number of residual γH2AX foci in irradiated cells linearly depends on the absorbed dose of X-ray radiation. However, the quantitative yield of residual γH2AX foci per unit of the absorbed dose in Ki-67+ cells 24 and 48 h after irradiation was higher than in Ki-67- cells by 1.8 and 2.0 times, respectively. In Ki-67- cells, the quantitative yield of residual γH2AX foci per unit of absorbed dose decreases by ~1.7 times with increasing the time after irradiation from 24 to 72 h. For the purposes of practical radiation biodosimetry, it can be recommended to quantify residual γH2AX foci in non-proliferating cells at least 72 h after irradiation.

[Real-world experience with netakimab in the treatment of spondyloarthritis].

AIM: To study the real-world efficacy and safety of netakimab in the treatment of ankylosing spondylitis (AS) and psoriatic arthritis (PsA). MATERIALS AND METHODS: The retrospective analysis included 23 patients (13 males; 56.5%) aged 23 to 73 years (median 42, interquartile range 28 to 52 years) with AS (n=12) or PsA (n=11) who received netakimab therapy from February 2021 to April 2023. Disease activity was assessed every 3-6 months based on the C-reactive protein (CRP) level for all patients according to the BASDAI and ASDAS-CRP indices for AS, DAPSA and PASI for PsA. These indicators were analyzed before therapy and at the last visit to assess the effectiveness of treatment. The results are presented as median (interquartile range). RESULTS: In all patients treated with netakimab (median duration of treatment 11 months), the CRP level decreased from 10.6 (3.1; 17.3) to 3.1 (1.9; 8.9) mg/L (absolute difference -7.5 mg/L, median relative reduction -60%; p=0.008), and the proportion of patients with elevated CRP decreased from 70 to 41%; p=0.039. In patients with AS (median duration of treatment 9 months), BASDAI score decreased from 5.8 (4.7; 6.5) to 3.0 (1.9; 3.8) points (absolute difference -2.8 points, median relative reduction of -45%; p=0.008) and ASDAS-CRP score decreased from 2.8 (1.9; 3.9) to 1.9 (1.7; 2.6) points (absolute difference -0.9 points, median relative reduction -21%; p=0.007). The proportion of patients with high AS activity (BASDAI≥4) decreased from 90% to 20% (p=0.031); however, there was no significant change in the CRP level (absolute difference -4.9 mg/L, median relative reduction -57%; p=0.110). In patients with PsA (median duration of treatment 18 months), the CRP level decreased from 12.0 (4.5; 17.3) to 3.3 (2.0; 7.8) mg/L (absolute difference -8.7 mg/L, median relative reduction -80%; p=0.041), the DAPSA score decreased from 23.0 (19.0; 30.5) to 6.3 (5.2; 13.5) points (absolute difference -16.7 points, median relative reduction -69%; p=0.018). Three (13%) patients reported mild to moderate adverse events. CONCLUSION: The obtained data confirm the effectiveness and safety of netakimab in treating AS and PsA in real-world practice.

Efficacy of secondary prevention and rehabilitation programs with distant support in patients with atrial fibrillation after intervention procedures: impact on psychological status.

Aim      To evaluate the effectivity of secondary prevention/rehabilitation programs with remote support for the psychological condition of patients with paroxysmal atrial fibrillation (AF) following interventional procedures (radiofrequency catheter ablation and cryoablation).Material and methods  This prospective, controlled, randomized clinical study was performed in three parallel groups. Each group consisted of 45 patients with AF after interventional procedures. In groups 1 and 2, secondary prevention/rehabilitation programs with remote support were performed, including a single individual in-hospital counseling (on risk factors of AF and their control and on major aspects of the disease, treatment and prevention of complications) and three months of remote support (by phone in group 1 and by e-mail in group 2). Patients of group 3 (control group) received standard recommendations at discharge from the hospital. The psychological status was evaluated using the Hospital Anxiety and Depression Scale, the PHQ-9 questionnaire, the Spielberg-Hanin scale for reactive and personal anxiety, and the visual analogue scale for stress assessment. The follow-up duration was 12 months.Results At the end of the follow-up period, the proportion of patients with anxiety symptoms considerably decreased in both intervention groups (р<0.001 for each group) and was significantly less than in the control group (р<0.001 for both comparisons). Also, in intervention group 1, the proportion of patients with clinically pronounced anxiety symptoms was significantly decreased. For 12 months of follow-up, the severity of depressive symptoms significantly decreased in all three groups. However, in both intervention groups, this decrease was significantly greater than in the control group (р<0.001 for group 1 and р=0.020 for group 2). In both intervention groups at 12 months, the stress level was significantly reduced whereas in the control group, it remained practically unchanged. The greatest (50% on average) decrease in the stress level was observed in intervention group 2.Conclusion      Secondary prevention and rehabilitation programs with remote support during a 12-month follow-up resulted in improvement of the psychological status in patients with AF after interventional procedures.

[The prevalence of anemia and its associations with other geriatric syndromes in subjects over 65 years old: data of Russian epidemiological study EVKALIPT].

BACKGROUND: A low hemoglobin level in older adults impairs cognitive ability and functional status and associates with risk of falls and fractures, sarcopenia, malnutrition, depression, frailty, and decreased autonomy. Epidemiological data on the anemia prevalence in the geriatric population in our country is not available. AIM: To assess the prevalence of anemia and analyze its associations with geriatric syndromes (GS) in subjects aged 65 years. MATERIALS AND METHODS: 4308 subjects (30% of men) aged 65107 years, living in 11 regions of the Russian Federation, were examined and divided into age groups (6574 years, 7584 years and 85 years). All the participants underwent a comprehensive geriatric assessment and determined hemoglobin level. RESULTS: The anemia prevalence in older adults was 23.9%. It has been shown that with an increase in age per 1 year, the risk of anemia detection increases by 4%. The incidence of anemia was higher in males than females (28.1% versus 22.1%; p0.001). In most cases, anemia was mild. The results of a comprehensive geriatric assessment show that patients with anemia had lower hand grip force, Barthel Index, the sum of points on Lawton instrumental activities of daily living scale, Mini Nutritional Assessment scale, the Mini-Cog test and higher the sum of points on the Geriatric Depression Scale (GDS-15) and the Age Is No Barrier scale. Patients with anemia were more likely to use hearing aids, absorbent underwear, and assistive devices during movement. Patients with anemia had a higher incidence of all GS, except for orthostatic hypotension and chronic pain syndrome. The presence of GS is associated with an increased risk of anemia by 1.33.4 times. CONCLUSION: EVKALIPT study obtained domestic data on the prevalence of anemia in older patients and examined its associations with other GS.

[Epithelial protective therapy in comorbid diseases. Practical Guidelines for Physicians].

This document was produced with the support of the National Medical Association for the Study of Comorbidities (NASС). In 2021 the first multidisciplinary National Consensus on the pathophysiological and clinical aspects of Increased Epithelial Permeability Syndrome was published. The proposed guidelines are developed on the basis of this Consensus, by the same team of experts. Twenty-eight Practical Guidelines for Physicians statements were adopted by the Expert Council using the "delphic" method. Such main groups of epithelial protective drugs as proton pump inhibitors, bismuth drugs and probiotics are discussed in these Guidelines from the positions of evidence-based medicine. The clinical and pharmacological characteristics of such a universal epithelial protector as rebamipide, acting at the preepithelial, epithelial and subepithelial levels, throughout gastrointestinal tract, are presented in detail.

[The Drosophila CG9890 Protein is Involved in the Regulation of Ecdysone-Dependent Transcription].

Previously we showed that the CG9890 protein, which has zinc finger domains, interacts with ENY2-containing complexes and is localized mainly on the promoters of active genes. The CG9890 protein is involved in the regulation of the expression of some of the genes on the promoters of which it is located, and among these genes there are genes for the ecdysone cascade. In this work, the role of the CG9890 protein in the regulation of ecdysone-dependent inducible transcription was studied. For this, 12 ecdysone-dependent genes on the promoters of which the CG9890 protein is localized were identified. Their activation was studied after the addition of 20-hydroxyecdysone to cells, both in normal conditions and after RNA interference of CG9890. The expression of ecdysone-dependent genes is significantly increased in response to the treatment of cells with ecdysone, in contrast to the control genes. Moreover, in the cell line after RNA interference CG9890, the transcription of 8 out of 12 genes was significantly higher than in the control line. Thus, the CG9890 protein is involved in the regulation of transcription of ecdysone-dependent genes, and, in most cases, acts as a repressor.

[Healthy life expectancy of people over age 65: results of the Russian epidemiological study EVCALIPT.]

Population health is an important indicator of the general well-being of the population, and it has it's practical significance, as it determines the costs of providing care, social and medical assistance for the elderly. The study presents an assessment of healthy life expectancy indicators of people over age 65 based on the results of the Russian epidemiological study EVCALIPT and a comparison of this results with data from other surveys in Russia and European countries.

Increased Yield of Residual γH2AX Foci in p53-Deficient Human Lung Carcinoma Cells Exposed to Subpicosecond Beams of Accelerated Electrons.

We studied quantitative yield of residual (24 h post-irradiation) phosphorylated histone (γH2AX) foci as a marker of DNA double strand breaks in wild-type A549 and p53-deficient H1299 human lung carcinoma cells after exposure to subpicosecond (energy 4 MeV, pulse duration 400 fsec, peak dose rate during the pulse 16 GGy/s) and quasi-continuous (energy 3.6 MeV) beams of accelerated electrons in a dose range of 0.5-10.0 Gy. The efficiency of pulse irradiation in A549 and H1299 cells assessed by the yield of residual foci was higher than the efficiency of quasi-continuous exposure by 1.8 and 5.3 times, respectively. Significant differences in quantitative yield of residual γH2AX foci between wild-type and p53-deficient cell lines were observed only after exposure to subpicosecond, but not quasi-continuous beams of accelerated electrons.

Changes in the Number of Mesenchymal Stem Cells in Bone Marrow at Different Periods after In Vivo Exposure of the Bone Marrow to Local Infrared Laser Radiation.

We determined optimal parameters of bone marrow (BM) irradiation in vivo for rapid increase in the number of mesenchymal stem cells (MSC) at the initial stages of the culturing without changing the karyotype, polyploidy, which are observed at higher passages. Such an increase is necessary to achieve the required number of cells at the initial passages for subsequent transplantation into the body. It was shown that after irradiation with λ=0.97 µm, the maximum and similar increase in the content of BM MSC in comparison with the control (by 2.4 times) was observed on day 2 in the irradiated and contralateral tibia. An insignificant difference in the number of BM MSC for the irradiated and contralateral tibia remained at all terms after irradiation, with a general decrease in the number of BM MSC up to 21 days. After laser irradiation with λ=1.56 µm, the maximum number of BM MSC was also observed on day 2. In this case, the concentration of these cells in the irradiated and contralateral limbs exceeded the control by 3.1 and 1.7 times, respectively. With increasing the time after exposure, the number of BM MSC in both limbs showed the same tendency to a decrease as after irradiation at λ=0.97 µm.

[Asymptomatic bacteriuria in pregnant women the normal condition of healthy women urine].

OBJECTIVE: to study the dynamics of the microbiota of a freshly excreted middle portion of urine in primigravida at different times of all three trimesters of pregnancy. MATERIALS AND METHODS: A single-center prospective observational cohort study was conducted with a consecutive enrollment of 30 women at different gestational ages: I - 8-12 weeks, II - 22-24 weeks, III - 30-32 weeks. A midstream specimen of morning vesical urine was taken for the study, then it was cultivated using nutrient media for aerobic and anaerobic microorganisms under appropriate conditions. Statistical analysis was performed using SPSS ver.26 (IBM SPSS Inc., Chicago, IL, USA). RESULTS: The freshly released middle portion of urine in all 30 observations in the I-III trimesters contains aerobic-anaerobic associations of microorganisms. Coagulase-negative staphylococci, Enterococcus spp., Corynebacterium spp., Lactobacillus spp., Eubacterium spp. prevail in the urine during pregnancy. The E. coli, Candida spp. detection frequency decreases by the third trimester, but Lactobacillus spp. detection frequency rises. Significant differences in the detection frequency were found only in Propionibacterium spp. and Lactobacillus spp. The average level of bacteriuria in most cases is 102-103 CFU/ml with significant differences only in E. faecium, Lactobacillus, Propionibacterium spp. in the III trimester. DISCUSSION: The study of urine at different times of all three trimesters of pregnancy refutes the previous ideas about asymptomatic bacteriuria. The urine microbiota in primigravida during pregnancy has wide spectrum and quite stable until delivery. Such bacteriuria can be considered asymptomatic, but it is a consequence of a healthy state and it is not a disease or its predictor. CONCLUSION: The term asymptomatic bacteriuria is not correct in the context of risk factor of urinary tract infection in pregnant women.

DNA Damage in Splenocytes of Mice Exposed to Secondary Radiation Created by 650 MeV Protons Bombarding a Concrete Shielding Barrier.

The proportion of splenocytes with a high level of DNA double-strand breaks was determined in mice exposed to primary and secondary radiation created by bombarding of a concrete barrier (thickness 20, 40, and 80 cm) by 650 MeV protons. The proportion of splenocytes with a high level of DNA double-strand breaks was assessed by flow cytometric analysis of γH2AX+ and TUNEL+ cells. It is shown that concrete barrier can significantly reduce primary proton radiation; the severity of negative biological effects in mice irradiated in the center of the proton beam decreased with increasing the thickness of this barrier. However, the spectrum of secondary radiation changes significantly with increasing the barrier thickness from 20 to 80 cm and the distance from central axis of the beam from 0 to 20 cm, and the proportion of the neutron component increases, which also causes negative biological effects manifesting in a significant (p<0.05) increase in the percentage of splenocytes with a high level of DNA damage in mice irradiated at a distance of 20 cm from the center of the proton beam and receiving relatively low doses (0.10-0.17 Gy).

Colony-Forming Ability and Residual Foci of DNA Repair Proteins in Human Lung Fibroblasts Irradiated with Subpicosecond Beams of Accelerated Electrons.

We performed a comparative study of the colony-forming ability and the number of residual foci of DNA repair proteins in cultured human lung fibroblasts (MRC-5 cell line) after exposure to subpicosecond beams of accelerated electrons with an energy of 3.6 MeV and quasi-continuous radiation (accelerated electrons with an energy of 4 MeV and X-rays). The yield of damages causing reproductive cell death after pulsed subpicosecond radiation exposure was higher by ~1.8 times than after quasi-continuous radiation exposure. The quantitative yield of residual γH2AX foci (phosphorylated H2AX histone, a protein marker of DNA double breaks) in cells irradiated with subpicosecond beams of accelerated electrons was shown to be ~2.0- 2.5-fold higher than in cells irradiated with quasi-continuous beams of accelerated electrons.

[The effect of physical health and functional status on cognitive functions in women aged 55-64 years old.]

Cognitive impairment is one of the most common causes of reduced quality of life in older people. The aim of the study to evaluate impact of functional status, physical health and cognitive functions in women aged 55-64 years old. The study included 250 women aged 55-64 years (mean age 59,3±2,9 years). Socio-demographic and economic factors, functional and cognitive status were analyzed, the presence and prevalence of geriatric syndromes and their association with social, demographic and economic characteristics, risk factors for the development of chronic non-communicable diseases were studied. The prevalence of cognitive impairments was independently associated with education level, low income of patients, diabetes mellitus, glomerular filtration rate, and vertebral artery diameter. In this case, the level of education was a protective factor.

Chromatin Modifiers in Transcriptional Regulation: New Findings and Prospects.

Histone-modifying and remodeling complexes are considered the main coregulators that affect transcription by changing the chromatin structure. Coordinated action by these complexes is essential for the transcriptional activation of any eukaryotic gene. In this review, we discuss current trends in the study of histone modifiers and chromatin remodelers, including the functional impact of transcriptional proteins/ complexes i.e., "pioneers"; remodeling and modification of non-histone proteins by transcriptional complexes; the supplementary functions of the non-catalytic subunits of remodelers, and the participation of histone modifiers in the "pause" of RNA polymerase II. The review also includes a scheme illustrating the mechanisms of recruitment of the main classes of remodelers and chromatin modifiers to various sites in the genome and their functional activities.

[The prevalence of osteoarthritis and its association with geriatric syndromes in people over 65: data from the Russian epidemiological study EVKALIPT].

BACKGROUND: Osteoarthritis (OA) in elderly and senile patients is not only common, but also one of the main diseases affecting the duration of active life, its quality, the appearance of addictions and loss of autonomy. Data on the relationship between OA and geriatric syndromes (GS) in our country are extremely scarce. AIM: To estimate the prevalence of OA and to analyze its associations with HS in persons aged 65 years. MATERIALS AND METHODS: The study included 4308 people (30% of men) aged 65 to 107 years, living in 11 regions of Russia. The patients were divided into 2 groups: with OA (n=2464) and without OA (n=1821). All patients underwent a comprehensive geriatric assessment. RESULTS: The prevalence of OA was 57.6%. With age, the frequency of OA increased significantly. According to the results of a comprehensive geriatric assessment, patients with OA had lower walking speed, the sum of points on the Bartel, Lawton scales and a short battery of physical functioning tests and higher the sum of points on the geriatric scale of depression and the age is not a hindrance scale. Patients with OA rated the quality of life and health status lower and higher the intensity of pain syndrome. Patients with OA were more likely to use any assistive device, with the exception of a wheelchair. In patients with OA, the most common HS were chronic pain syndrome (92%), senile asthenia syndrome (64%), basic (66%) and instrumental (56%) dependence in everyday life, cognitive impairment (62%), probable depression (51%) and urinary incontinence (50%). Univariate regression analysis showed that OA is associated with a 1.23.0-fold increase in the risk of a number of GS and a 28% decrease in the risk of malnutrition. CONCLUSION: OA is widespread in the elderly population. The presence of OA is associated with a number of GS associated with loss of autonomy.

Functional Features of Hemocoagulation in rats with Experimentally formed Arterial Hypertension in Conditions of Increased Motor Activity

In rats with experimentally formed arterial hypertension, lipid perxidation in the plasma, amplification of blood chotting mechanisms with a decrease in anticoagulation and fibrinolysis was noted. Regular forced jogging provided the experimental rats with a positive dynamic of all the indicators considered. Thus, with increased muscular activity, the level of acyl hydro-peroxides of plasma decreased in rats with arterial hypertension formed due to the enhancement of its antioxidant activity. In addition, with the increase in muscle activity in experimental rats, normalization of clotting factor activity, indices of general coagulation tests, antithrobin III activity and protein C was achieved. This was accompanied by a normalization of the level of plasminogen, a2-antiplasmin and spontaneous euglobulin lysis time. In rats with formed arterial hipertension with stgandard physical activity, the initial violations of the measured parameters were completely preserved (AU)

The Development of Reporter System for the Investigation of Molecular Mechanisms of Ecdysone Response.

To study the mechanisms of transcriptional regulation, a convenient experimental approach is to use the artificial chimeric constructs carrying the regulatory elements of interest. In the present work, we describe the creation and characterization of a novel genetic construct that makes it possible to study the transcriptional regulation of the early-late gene of the ecdysone cascade. Using the data of genome-wide experiments, we have isolated the main regulatory region of the hr4 gene, which was successfully used to create a chimeric reporter construct expressing a fluorescent protein upon the treatment with the ecdysone hormone. This reporter system can be used to study the mechanisms of the ecdysone response, both in cell culture and in tissues, at various stages of the Drosophila development.

Biodistribution and Tumors MRI Contrast Enhancement of Magnetic Nanocubes, Nanoclusters, and Nanorods in Multiple Mice Models.

Magnetic resonance imaging (MRI) is a powerful technique for tumor diagnostics. Iron oxide nanoparticles (IONPs) are safe and biocompatible tools that can be used for further enhancing MR tumor contrasting. Although numerous IONPs have been proposed as MRI contrast agents, low delivery rates to tumor site limit its application. IONPs accumulation in malignancies depends on both IONPs characteristics and tumor properties. In the current paper, three differently shaped Pluronic F-127-modified IONPs (nanocubes, nanoclusters, and nanorods) were compared side by side in three murine tumor models (4T1 breast cancer, B16 melanoma, and CT26 colon cancer). Orthotopic B16 tumors demonstrated more efficient IONPs uptake than heterotopic implants. Magnetic nanocubes (MNCb) had the highest r2-relaxivity in vitro (300 mM-1·s-1) compared with magnetic nanoclusters (MNCl, 104 mM-1·s-1) and magnetic nanorods (MNRd, 51 mM-1·s-1). As measured by atomic emission spectroscopy, MNCb also demonstrated better delivery efficiency to tumors (3.79% ID) than MNCl (2.94% ID) and MNRd (1.21% ID). Nevertheless, MNCl overperformed its counterparts in tumor imaging, providing contrast enhancement in 96% of studied malignancies, whereas MNCb and MNRd were detected by MRI in 73% and 63% of tumors, respectively. Maximum MR contrasting efficiency for MNCb and MNCl was around 6-24 hours after systemic administration, whereas for MNRd maximum contrast enhancement was found within first 30 minutes upon treatment. Presumably, MNRd poor MRI performance was due to low r2-relaxivity and rapid clearance by lungs (17.3% ID) immediately after injection. MNCb and MNCl were mainly captured by the liver and spleen without significant accumulation in the lungs, kidneys, and heart. High biocompatibility and profound accumulation in tumor tissues make MNCb and MNCl the promising platforms for MRI-based tumor diagnostics and drug delivery.

Cognitive Tagging of Neurons: CREMediated Genetic Labeling and Characterization of the Cells Involved in Learning and Memory.

In this study, we describe use of Cre-mediated recombination to obtain a permanent genetic labeling of the brain neuronal networks activated during a new experience in animals. This method utilizes bitransgenic Fos-Cre-eGFP mice in which a green fluorescent protein is expressed upon tamoxifen-induced Cre-recombination only in the cells where immediate early gene c-fos expression takes place due to the new experience. We used the classical fear conditioning model to show that ex vivo microscopy of the eGFP protein in Fos-Cre-eGFP mice enables mapping of the neurons of the various brain regions that undergo Cre-recombination during acquisition of a new experience. We exposed the animals to the new environment in brief sessions and demonstrated that double immunohistochemical staining enables a characterization of the types of neocortical and hippocampal neurons that undergo experience-dependent Cre-recombination. Notably, Fos-Cre-eGFP labeled cells appeared to belong to excitatory pyramidal neurons rather than to various types of inhibitory neurons. We also showed that a combination of genetic Cre-eGFP labeling with immunohistochemical staining of the endogenous c-Fos protein allows one to identify and compare the neuronal populations that are activated during two different episodes of new experiences in the same animal. This new approach can be used in a wide spectrum of tasks that require imaging and a comparative analysis of cognitive neuronal networks.

[Social, Demographic and Behavioral Factors and Their Impact on 5-year Survival in Subjects Aged Over 75 Years in Moscow Population].

OBJECTIVE: to study the impact of social, demographic and behavioral factors on 5-year survival rate in persons older than 75 years living in Moscow and the Moscow region. MATERIALS AND METHODS: We included in this prospective observational study 283 patients (25% men, age 75-98, mean 86.8±4.3 years) after stay at the Russian gerontological clinical research center. To study social and behavioral factors we recorded anthropometric parameters and used questionnaires. Duration of follow-up was 5 years. Endpoint was death from any cause. RESULTS: One hundred five patients (37.1%) died during the follow-up (median 3.8 years). Survival analysis using the Kaplan-Meier method revealed a number of parameters that affected 5-year survival: age, defcient and excessive body weight, consumption of less than 1 liter of water per day, start of the night sleep after midnight, nightime sleep duration ≥10 h, level of physical activity and history of blood donation. At multivariate analysis adjusted for age and sex the following factors were independently associated with death during follow-up: weight defciency (hazard ratio [HR] 7.36, 95% confdence interval [CI] 2.15-25.26, p=0.001), second degree obesity (HR 0.47, 95% CI 0.25-0.87, p=0.016), start of night sleep after midnight (HR 2.53, 95% CI 1.32-4.85, p=0.005), nightime sleep duration ≥10 h (HR 3.89, 95% CI 1.77-8.59, p=0.001), and history of blood donation (HR 0.24, 95% CI 0.09-0.62, p=0.003). CONCLUSION: In people older than 75 years late sleep initiation and long duration of night sleep, and weight loss increase the risk of death in the next 5 by years, 2.5, 3.9 and 7.4 times, respectively, while overweight and history of blood donation are associated with 53% and 76% reduction of the risk of death.