The management of acute myocardial infarction in the Russian Federation: protocol for a study of patient pathways

Death rates from cardiovascular disease in Russia are among the highest in the world. In recent years, the Russian government has invested substantially in the healthcare system, with a particular focus on improving access to advanced technology, especially for acute myocardial infarction (AMI). This protocol describes a study to understand the management of AMI in different Russian regions, investigating the role of patient, clinical, and health system characteristic

Hospital stage of myocardial infarction treatment in 13 Regions of Russian Federation by results of the international research

Aim: To describe the characteristics of the patient with MI who is admitted to a hospital and to characterize the main diagnostic and treatment interventions in clini

Efficient chimeric mouse production using a novel embryonic stem cell line

Embryonic stem cells are commonly used for generation of transgenic mice. Embryonic stem cells could participate in the development of chimeric animals after injection into a blastocyst. Injection of genetically modified embryonic stem cells could lead to germ line transmission of a transgene or genomic modification in chimeric mice. Such founders are used to produce transgenic lines of mice. There are several projects dedicated to production of knock-out mouse lines (KOMP Repository, EUCOMM, Lexicon Genetics). Never-theless, there is a need for complex genome modifications, such as large deletions, reporter genes insertion into the 3’ gene regulatory sequence, or site-specific modifications of the genome. To do that, researchers need an embryonic stem cell line that is able to participate in chimeric animal formation even after prolonged culture in vitro. Several lines of mouse embryonic stem cells were produced in the Laboratory of Developmental Genetics of the Institute of Cytology and Genetics SB RAS. We tested DGES1 cell line (2n = 40, XY) (129S2/SvPasCrl genetic background) for chimeric mice production at the Center for Genetic Resources of Laboratory Animals at ICG SB RAS. Embryonic stem cells were injected into 136 blastocysts (B6D2F1 genetic background), which were transplanted into CD-1 mice. Among 66 progeny, 15 were chimeric, 4 of which were more than 80 % chimeric judged by coat color. All chimeras were males without developmental abnormalities. 10 of 15 males were fertile. Microsatellite analysis of the progeny of chimeric mice revealed embryonic stem cell line DGES1 contribution to the gamete formation. Thus, a novel DGES1 embryonic stem cell line could be efficiently used for transgenic mouse production using B6D2F1 blastocysts and CD-1 recipients

Immunological efficacy and tolerability of vaccination in patients with noncommunicable diseases

Aim. To study the efficacy and tolerability of Sputnik V vaccination in patients with noncommunicable diseases (NCDs) compared with healthy individuals.Material and methods. The retrospective analysis included data from 800 men and women aged 18 to 90 who were vaccinated from February to May 2021 at the National Medical Research Center for Therapy and Preventive Medicine. Sputnik V (Gam-COVID-Vac, Russia), consisting of two doses, was used as a vaccine. The mean age of the patients was 50,46±13,16 years. Patients were questioned in two stages: before vaccination and after the first dose of vaccination. The following clinical and paraclinical investigations were carried out: saturation assessment, measurement of temperature, blood pressure and heart rate at rest, pharynx examination, auscultation of the heart and respiratory organs. The blood concentration of antibodies (IgM and IgG) was determined after 21 days and 42 days. For analysis, patients were divided into three groups. The first group included healthy individuals (n=238, 29,8%), the second group — patients with one NCD (n=385, 48,1%), and the third group — patients with two or more NCDs (n=177, 22,1%).Results. The sex composition was comparable in all three groups, while the mean age differed. In the first group, the mean age was 43,93±10,80 years, while in the second and third groups — 50,51±12,55 and 59,12±12,37 years, respectively (p<0,001). Statistical analysis was performed with adjustment for age and sex. The median blood concentration of immunoglobulin G of vaccinated individuals of the cohort after 21 days was within 4,60 [2,00; 8,40]. A comparative analysis of the groups did not reveal significant differences. After 42 days, the median concentration of immunoglobulin G increased by 4 times, which is significant (p<0,001) and amounted to 16,20 [15,80; 16,40]. In the group of healthy individuals, the concentration of immunoglobulin G was significantly higher compared to groups of individuals with one, as well as two or more NCDs. Forty-two days after vaccination, the concentration of immunoglobulin M both in the cohort and in individual groups did not change significantly compared to the level after 21 days. In addition, 47%, 38% and 40% of participants from the first, second and third groups, respectively, reported some symptoms. A temperature increase on average was detected in 13% of vaccinated individuals. The largest number of persons with fever was registered in the group of healthy individuals — 19%, in the group of patients with one NCD — 14%, and in the group with two or more diseases, the temperature increase was the lowest and amounted to 3% (p<0,001). After the first dose, 18,6% of patients reported taking additional drugs as follows: first group — 23,9%, second group — 15,1%, third group — 19,2%. These are mainly analgesic, non-steroidal anti-inflammatory and antihistamine drugs.Conclusion. Vaccination with Sputnik V increases the immunoglobulin G level in the blood after 42 days by four times compared to after 21 days. This pattern is observed both in the group of healthy individuals and in patients with one, two or more chronic NCDs. Tolerability of vaccination in all groups was comparable and did not lead to visits to healthcare facilities

Morphophysiological alterations caused by insertional mutagenesis of contactin 5 (Cntn5) gene in transgenic mice

Transgenesis has become a routine for modern biological studies. The most popular method for producing transgenic animals–pronuclear microinjection–frequently leads to host gene disruption due to a random transgene integration. In this paper, we report our analysis of morphophysiological parameters of the transgenic mouse line GM9, in which a transgene designed for milk-specific expression of the human granulocyte-macrophage colony-stimulating factor (GM-CSF) gene was integrated into the intron of the Contactin 5 gene (Cntn5). We studied Cntn5 expression with RT-PCR and discovered that its expression in the brain, the primary organ of Cntn5 activity, was unperturbed. However, transgenic animals had less Cntn5 transcripts in other tissues such as the kidney and heart. In addition, we observed a decreased amount of splice variants of Cntn5 exons that flank the transgene integration site. These data suggest that the transgene integration event might affect proper Cntn5 splicing in some tissues. Publications exist that imply that some polymorphisms in the Cntn5 gene are associated with obesity and arterial hypertension in humans. We evaluated core parameters of lipid metabolism and heart activity in mice homozygous and heterozygous for Cntn5 mutation using wild- type animals as control. Our results uncovered that homozygous mutant mice have lower body weight than controls and that it is caused by slower accumulation of fat tissue. Cntn5 mutants also exhibit abnormalities in blood circulation: homozygous Cntn5 mutants are characterized by a higher blood pressure and heart beat rate, as well as faster blood flow in the tail vessels. Heterozygous animals showed intermediate results for all of these parameters

Body composition as an indicator of metabolic changes in mice obtained by <i>in vitro</i> fertilization

To identify body systems subject to epigenetic transformation during in vitro fertilization (IVF), comparative morphological and functional studies were performed on sexually mature offspring of outbred CD1 mice, specific-pathogen-free (SPF), obtained by IVF (experiment) and natural conception (control). The studies included assessment of age-related changes in body weight and composition, energy intake and expenditure, and glucose homeostasis. To level the effects caused by the different number of newborns in the control and in the experiment, the size of the fed litters was halved in the control females. Males obtained using the IVF procedure were superior in body weight compared to control males in all age groups. As was shown by analysis of variance with experiment/control factors, gender, age (7, 10 and 20 weeks), the IVF procedure had a statistically significant and unidirectional effect on body composition. At the same time, IVF offspring outperformed control individuals in relative fat content, but were behind in terms of lean mass. The effect of the interaction of factors was not statistically significant. IVF offspring of both sexes had higher fat to lean mass ratios (FLR). Since adipose tissue contributes significantly less to total energy intake compared to muscle, the main component of lean mass, it is not surprising that at the same level of IVF locomotor activity offspring consumed less food than controls. When converted to one gram of body weight, this difference reached 19 %. One of the consequences of reduced utilization of IVF energy substrates by offspring is a decrease in their tolerance to glucose loading. The integral criterion for the effectiveness of restoring the initial glucose level is the area under the curve (AUC), the value of which was 2.5 (males) and 3.2 (females) times higher in IVF offspring compared to the corresponding control. Thus, the totality of our original and literature data shows an increase in the risk of metabolic disorders in IVF offspring, which is confirmed by epidemiological studies of a relatively young cohort of people born using assisted reproductive technologies

Reproductive effects of the tumor necrosis factor (TNF) deficiency in mice

TNF is a multifunctional cytokine that, at physiological concentrations, maintains the balance between apoptosis and survival of male germ cells and, at higher concentrations, has adverse effects on various stages of the reproductive process. Although ant-cytokine therapies have been used in millions of patients, the consequences of cytokine deficiency for reproductive functions are poorly understood and need attention. In this work, we have studied behavioral interactions between males and females, spermatogenesis, male fertility, and embryonic developmental characteristics of the progeny in TNFα knockout mice (TNF-/-). We have demonstrated that TNF is involved in the regulation of sexual behavior, spermatogenesis, pre- and postimplantation development. Complete TNF deficiency led to decreased reproductive efficiency: a lower number of viable embryos were observed in TNF-/- mice than in wild-type mice. The decrease in fertility was caused by preimplantation embryo loss in TNF-/- mice. Preimplantation loss in females might be caused by asospermia in TNF-/- males. Additionally, the sensitivity of reproductive functions to female stimuli was different between TNF-/- mice and wild-type mice, while interactions with females increased the concentrations of sper­matozoids in both TNF-/- and wild-type mice. Still higher levels were observed in knockout animals, which led to increase in the number of immature spermatozoids in epididymides

Magnetic resonance spectroscopy of hippocampal and striatal neurometabolites in experimental PTSD rat modeling

The spectrum of the metabolites in the dorsal region of the hippocampus and striatum was studied using the method of 1H magnetic resonance spectroscopy at experimental modeling of the posttraumatic stress disorder syndrome (PTSD) in rats. PTSD was reproduced by exposure of the cat cue to rats daily along 10 day by 10 minutes at once. The anxiety level of animals was estimated 12 days later after the end of the experimental series of stress. Based on the anxiety index, the rats were divided into 3 phenotypes. The animals with an anxiety index &gt; 0.8 (group 1) had lower plasma corticosterone compared with rats form the control group. In animals with an anxiety index in the range 0.7–0.8 (group 2), an elevated corticosterone level was noted. The rats with an anxiety index &lt; 0.7 (group 3) had a lower plasma corticosterone level compared with animals from the control group. Rats of group 2 were characterized by an increased level of GABA in the hippocampus compared with controls. In the remaining groups, the percentages of GABA in the hippocampus and striatum did not differ significantly from the control. The distribution of NAA differed form that of GABA. The highest level of NAA was found in the striatum for rats from group 1, whereas NAA in animals form groups 1 or 3 did not differ from the control. The NAA level in the hippocampus was similar between all groups, including the control. The results obtained indicate that multiple exposures to psychological stress associated with the sense of proximity of a natural enemy in some animals cause an anxiolytic reaction. These animals are characterized by a stable corticosterone level and a stable level of neurometabolites in the studied structures of the brain. For rats with the highest level of anxiety, a lowered level of corticosterone with a constant level of neurometabolites in the hippocampus and striatum is characteristic. And only in rats with an intermediate level of anxiety, synchronization was observed between the increase in plasma corticosterone and the increase in hippocampal GABA content. The results obtained are in good agreement with the ideas of the protective action of glucocorticoids under PTSD manifested in  restraining violations of the psycho-physiological status. The mate rials allow the neurobiological mechanisms of the protective action of glucocorticoids to be detailed

Hospital Stage of Myocardial Infarction Treatment in 13 Regions of Russian Federation by Results of the International Research

Aim. To describe the characteristics of the patient with MI who is admitted to a hospital and to characterize the main diagnostic and treatment interventions in clinic. Material and methods. This study is observational and the part of big international project. It includes a representative sample of patients with MI admitted to 16 clinics in 13 regions of Russian Federation (Arkhangelsk region, Belgorod region, Bryansk region, Tver region, Saratov region, Rostov region, Samara region, the Republic of Tatarstan, Perm region, Tyumen region, Khanty-Mansiysk Autonomous district, Kemerovo region, Altai region). Patients were selected at random from among those experiencing a MI that were alive on the next morning after hospitalization. Enrollment took place from June 2015 to August 2016. Results. Of 1,128 patients included in the study, 872 were male (77.3%) and 256 females. 21.4% of patients had a previous MI, 8.3% had undergone PCI, and 2.2% CABG. Turning to cardiovascular risk factors, 46.2% of patients smoked prior to hospitalization, 34.6% were obese and 52.1% had a high cholesterol level. Only 40.0% of patients had no contact with the health care system within 12 months before the MI. Every fourth patient (25.1%) had undergone dispensarisation within 12 months before MI, women significantly often than men (33.5% and 22.6%, p < 0.001). Initial revascularization was performed in 73.2% of patients, PCI was the initial revascularization attempt in 49.4% of patients, and PCI with stenting in 46.7%. Conclusion. Patient with a MI in Russian clinics is likely to have had a history of cardiovascular disease, and to have regular contact with the health care system within 12 months before the development of cardiovascular event. We demonstrated the high rates of appropriate MI treatment, without significant gender and age difference (except for thrombolysis), however, there is a reserve for increasing the proportion of patients who are undergoing revascularization