Adaptation of the sulfophosphovanillin method of analysis of total lipids for various biological objects as exemplified by <i>Drosophila melanogaster</i>

Lipid metabolism is crucial in physiology. In recent decades the model object Drosophila melanogaster has been actively used in the study of the fundamental issues of lipid metabolism and its disorders, including obesity, as well as in the search for therapeutic goals for the treatment of metabolic disorders in humans. Quick and accurate quantification of lipid content is an important step in solving these problems. For the first time the method of quantitative measurement of total lipids with the use of the sulfophosphovanillin (SPV) method was described by Zöllner and colleagues in 1962, and adapted for insects by Van Handel on females of the yellow fever mosquito Aedes aegypti. The advantages of this method compared to traditional gravimetric and chromatographic methods of analysis are the use of a small amount of biological material, lack of need for complex manipulations with the sample, its high sensitivity, reproducibility and simplicity of implementation with a minimum set of equipment. Here, a modification of the Van Handel protocol is described, which allows the method to be adapted for quantitative determination of total lipids for various organisms as exemplified a widely used model, D. melanogaster. To test the effectiveness of the modified method, we measured the content of total lipids in D. melanogaster females carrying hypomorphic mutations of the dilp6 and dfoxo insulin signaling pathway genes compared to the wild-type Canton-S line, and showed that dilp6 took part in the regulation of fat metabolism, while dfoxo did not. The results obtained emphasize the effectiveness of the colorimetric method with the use of SPV reaction and spectrophotometry for the quantitative analysis of total lipids

The neuroendocrine stress-response in insects: the history of the development of the concept

The neuroendocrine stress-response is an effective defense mechanism against adverse influences of various nature. This reaction is universal and appears in response to stimuli that are unusual not just for living and habitat of the species, but also for each population. Here we review a progressive change of theoretical concepts, approaches and methods of research in this scientific field: beginning with the development of the stress concept by H. Selye and up to the present day. In 1982 H. Selye defined stress as a combination of stereotypical phylogenetic programmed  reactions of the organism that are caused by any strong, superstrong or extreme influences and are followed by a reorganization of the organism’s adaptive forces. The stress-causing agent was named a stressor. In the dy namics of the complex of nonspecific protectiveadaptive reactions that respond to a stressful influence aimed at cultivating the organism’s resistance to any factor, it is possible to logically identify three stages (“the Selye triad”): 1) alarm state, 2) resistance state, 3) exhaustion state. The duration and the expression of each stage can vary depending on the nature and strength of the stressor agent, the species of the animal and the physiological state of the organism. The lack of a hypothalamic-pituitary-adrenocortical system in insects was considered a proof of their inability to develop a stress reaction of the warm-blooded animals’ type. Nevertheless, since the early 1980s, enough evidence of the development of stress reaction in insects has been gathered, which emphasizes the conservative nature of the stress reaction in mammals and insects. The similarity in the neurochemical and physiological changes in invertebrates and vertebrates in response to a stressful influence indicates that the response to a stressor is a complex of ancient mechanisms preserved in evolution. Insects present unique opportunities for experimentation, which can allow us to understand the basic mechanisms of stress reactions. In insect larvae the mechanism of stress reaction has been studied in detail. In this century, the main efforts of researchers are aimed at studying the mechanisms of stress reaction in imago and genetic control of its individual links. The study of stress reaction in insects has both theoretical importance, as it demonstrates the convergence of evolutionary pathways of adaptive transformations in such distant taxa as insects and mammals, and practical importance, since the patterns of this reaction’s mechanisms can be used in modeling hereditary or acquired human diseases, in developing breeding methods for economically valuable insects and in finding ways to fight insect pest

The transcription factor dFOXO controls the expression of insulin pathway genes and lipids content under heat stress in <i>Drosophila melanogaster</i>

The insulin/insulin-like growth factor signaling (IIS) pathway is one of the key elements in an organism’s response to unfavourable conditions. The deep homology of this pathway and its evolutionary conservative role in controlling the carbohydrate and lipid metabolism make it possible to use Drosophila melanogaster for studying its functioning. To identify the properties of interaction of two key IIS pathway components under heat stress in D. melanogaster (the forkhead box O transcription factor (dFOXO) and insulin-like peptide 6 (DILP6), which intermediates the dFOXO signal sent from the fat body to the insulin-producing cells of the brain where DILPs1–5 are synthesized), we analysed the expression of the genes dilp6, dfoxo and insulin-like receptor gene (dInR) in females of strains carrying the hypomorphic mutation dilp641 and hypofunctional mutation foxoBG01018. We found that neither mutation influenced dfoxo expression and its uprise under short-term heat stress, but both of them disrupted the stress response of the dilp6 and dInR genes. To reveal the role of identified disruptions in metabolism control and feeding behaviour, we analysed the effect of the dilp641 and foxoBG01018 mutations on total lipids content and capillary feeding intensity in imago under normal conditions and under short-term heat stress. Both mutations caused an increase in these parameters under normal conditions and prevented decrease in total lipids content following heat stress observed in the control strain. In mutants, feeding intensity was increased under normal conditions; and decreased following short-term heat stress in all studied strains for the first 24 h of observation, and in dilp641 strain, for 48 h. Thus, we may conclude that dFOXO takes part in regulating the IIS pathway response to heat stress as well as the changes in lipids content caused by heat stress, and this regulation is mediated by DILP6. At the same time, the feeding behaviour of imago might be controlled by dFOXO and DILP6 under normal conditions, but not under heat stress

NEW APPROACHES TO THE ORGANIZATION OF SKIN CANCER SCREENING

Skin cancer is the most common type of cancer in the Russian Federation. Despite the high incidence rate of skin cancer, there is no single standard for screening to identify this disease at an early stage, therefore, screening programs for detecting premalignant lesions and skin cancer at early stages are of great importance. The article describes the results of the action dedicated to the World Day against melanoma, held in Krasnoyarsk Regional Clinical Oncology Center named after A.I. Kryzhanovsky. Selection of participants was based on electronic applications, including photographs of skin lesions taken by the patients themselves. Out of the 343 people who applied, 143 (41.7 %) persons were selected for physician’s consultation. There were revealed 6 cases of melanoma, 13 basal cell tumors, and 1 lymphoproliferative disorders of the skin

Remote monitoring of adolescents with type 1 diabetes mellitus using a mobile application

Background: The prevalence of type 1 diabetes mellitus (T1DM) in childhood is increasing every year. Adolescence is the most challenging age for achieving optimal metabolic control of T1DM. Telemedicine has already been shown to be ­effective in children with the condition, but there are not enough studies in adolescents. The use of mobile apps may be associated with better glycemic control in patients with type 1 diabetes.Aims: To assess the effectiveness and safety of a model of medical care for adolescents with type 1 diabetes using remote counseling and a mobile application.Materials and methods: Were included adolescents aged ≥14 and 18 years with a T1DM duration&gt; 3 months, a glycated hemoglobin level (HbA1c)&gt; 7%. The duration of the study was 26 weeks. There were 3 face-to-face and at least 4 remote visits using a mobile application. All patients underwent standard examination and anthropometry, study of HbA1c, registration and analysis of indicators, assessment and correction of the treatment. The quality of life of adolescents was assessed at baseline and at the end of the study. Adolescents and physicians were interviewed about program evaluation.Results: 56 patients were included, 7 adolescents withdrew. HbA1c significantly decreased by the 12th week of the study (–0.3%; p = 0.005), by the end of the study the change in HbA1c was –0.5% (p &lt;0.001). There was an increase in the percentage of glucose measurements in the target range (+5.3 pp; p = 0.016) and a decrease in blood glucose variability (-3.1 pp; p =  0.015). There was a significant improvement in both the total assessment of the quality of life by patients (+2.9 points; p = 0.008) and individual components of its indicators: attitude to diabetes (+3.0 points; p = 0.049), attitude to treatment (+4.6 points; p = 0.010) and communication with others (+4.5 points; p = 0.015). The majority of doctors and patients assessed their participation in the study positively. The incidence of adverse events did not change significantly during the study from baseline.Conclusion: Remote counseling using a mobile app is a safe and effective approach for adolescents with T1DM in terms of glycemic control and quality of life, and provides convenience and speed of interaction

Behavioral risk factors and clinical course of cardiovascular diseases and other noncommunicable diseases during quarantine in various regions of Russia

Aim. To assess the behavioral risk factors and the clinical course of cardiovascular diseases (CVDs) and other noncommunicable diseases (NCDs) during quarantine in various regions of Russia.Material and methods. This multicenter cohort cross-sectional study included 205 men and women from 6 Russian cities. Further, 4 of them (Saransk, Nizhny Novgorod, Penza, Ulyanovsk) were combined into one group — the Volga region. The study included men and women aged 30-69 years with one or more NCDs (hypertension, coronary artery disease with or without myocardial infarction, type 2 diabetes, chronic obstructive pulmonary disease/asthma and cancer in patients receiving chemotherapy and/or radiation therapy) who were self-isolated during coronavirus disease 2019 (COVID-19) pandemic. For all patients, a questionnaire was used, which included socio-demographic parameters, behavioral risk factors, status of the underlying disease, incidence of COVID-19 and its complications. Self-assessment of the state of health was carried out using the European Quality of Life Questionnaire.Results. In every third Muscovite, the intensity of physical activity decreased, and in the groups of patients from Omsk and the Volga region, it was 45% and 43%, respectively. An increase in meal frequency and an impairment of eating habits in Moscow and Omsk was noted in 18,2% and 18,7% of participants, while in Volga region subjects, these parameters were 2 times higher (42,4%). At the same time, no significant changes of alcohol consumption and smoking was revealed in the cohorts. Hypertensive crises during a pandemic were noted in all three subgroups, but more of them were recorded in the Volga region — in every third patient (p&lt; 0,05 compared to Moscow), in the Omsk group — in every fourth patient, and among Muscovites — no more than 5%. Clinical deterioration in patients with angina was noted in 15% of cases, while the smallest number was noted in Omsk subjects (5,3%), three times less than in other subgroups. Changes in intensity and regimen of hypoglycemic therapy were noted in patients from Omsk, while 30% of them (p&lt; 0,05 compared with the Volga region) increased the doses of medications taken. Chronic obstructive pulmonary disease was registered in the group with the largest number of Volga region patients — 14,1% (p&lt; 0,05 compared to Omsk), while 17% of patients in this group increased the dose of drugs. Any cancer was recorded in 13,6% of Muscovites, while in the other two groups — about 5%. The largest number of patients from the Volga region noted a health decline over the past year (30,8%), while every fifth patient from Omsk (19,6%) and 13,6% of Muscovites reported health changes.Conclusion. During quarantine and self-isolation, changes in dietary habits and physical activity decline were noted among patients with NCDs, while alcohol consumption and smoking remained practically unchanged. The change in clinical status was characterized by an increase in hypertensive crisis incidence, an increase in doses of antihypertensive and hypoglycemic medication. Depending on the region, the health decline was noted by 13-31% of patients with NCDs

The era of Belousov Yuri: continuing the tradition. To the 80th birthday

The article is dedicated to the outstanding scientist, doctor, teacher, public figure, corresponding member of the Russian Academy of Sciences, professor, doctor of medical sciences Belousov Yuri, who would have turned 80 on September 23, 2022. The presented materials make it possible to recall the main milestones in the biography of this remarkable person. The article is based on materials collected by relatives, friends, colleagues and students of Yuri Belousov

C-peptide levels and the prevalence of islets autoantibodies in children with type 1 diabetes mellitus with different duration of the disease

BACKGROUND: Type 1 diabetes mellitus (T1DM) is characterized by the development of absolute insulin deficiency. In some patients residual insulin secretion may persist for a long time. C-peptide is a widely used to measure the pancreatic beta cells function, in clinical practice and in research studies.AIM: To assess C-peptide levels and presence of islets autoantibodies (Ab) in children with different duration of T1DM and to identify factors associated with the preserved secretion.MATERIALS AND METHODS: Single-center cross-sectional study including data from 703 cases of children with T1DM, examined in the Endocrinology Research Center in 2016-2020, who was underwent a study of C-peptide levels and was positive for one or more islets antibodies (ZnT8, IA-2, GAD, ICA, IAA). There were 3 groups of patients: 1st — T1DM duration &lt; 1 year, 2nd — from 1 to 5 years, 3rd — &gt; 5 years.RESULTS: The median of the fasting C-peptide level in the 1st group was 0.6 ng/ml [0.27; 1.09]; in the 2nd group — 0.2 ng/ml [0.01; 0.8]; in the 3rd group — 0.01 ng/ml [0.01; 0.037]. The preserved secretion of C-peptide (&gt; 0.6 ng/ml) was determined in 51.4% in the 1st group, in 31.4% — in the 2nd group and in 11.4% in the 3rd group. In patients with obesity during the first year from the T1DM diagnosis C-peptide levels above 1.1 ng/ml was determined significantly more often (65.2%), as well as at the period of 1 to 5 years of the disease (35.7%), compared with children with normal BMI (18.5% and 14.5%, respectively) or overweight (15.7% and 19%, respectively), p &lt;0.01. A negative correlation was found between C-peptide levels and the duration of T1DM (r = -0.489, p = 0.000), the daily dose of insulin (r = -0.637, p = 0.000), a positive association was found with the age of diagnosis of T1DM (r = 0.547, p = 0.000). The frequency of the presence of one type of islets autoantibodies in all groups was 29.5%, 2 types — 33.6%, 3 and more types — 36.9%. The titer of IA-2, ZnT8 decreased with the disease duration (p &lt;0.05 and p &lt;0.01, respectively), while the titer of ICA increased (p &lt;0.01). No associations between the types, number, antibodies titer and C-peptide levels, age of disease manifestation were found.CONCLUSION: C-peptide levels in children with T1DM in groups with older age at diagnosis and with obesity is significantly higher for the first 5 years of the disease. The study have shown the titer of IA-2, ZnT8 is decreasing with the disease duration, in turn, the frequency of detection of ICA increases. No association between C-peptide levels and the type, number and titer of antibodies were found

The use of Flash glucose monitoring in children with type 1 diabetes mellitus in real clinical practice

BACKGROUND: In 2018, a Frestyle Libre flash glucose monitoring system (FGM) appeared in Russia and became a potential alternative to the traditional CGM. Studies carried out to date have shown the advantages of FGM over SMBG, but only a few of them relate to real clinical practice, especially in children with type 1 diabetes.OBJECTIVE: To evaluate the efficacy of FGM in children with T1DM in relation to glycemic control indicators, the occurrence of severe hypoglycemia and diabetic ketoacidosis, as well as the satisfaction of patients and their parents with the use of FGM.MATERIALS AND METHODS: Single-center, prospective, observational cohort study. Children 4–18 years old with T1DM and HbA1c level less than 10.0% were invited to participate in the study on intensified insulin therapy (by MDI or CSII). The duration of the patient’s participation in the study was 6 months. At baseline and every 3 months thereafter, face-to-face consultations were conducted with an assessment of the general condition, HbA1c study, an assessment of glycemic indicators, progress in relation to glycemic control targets and correction of the therapy. A total of 228 patients (110 boys and 118 girls) who met the inclusion criteria were included in the study. The median age was 11.2 (8.6–14.7) years, the duration of type 1 diabetes was 3.8 (2–7.1), 136 patients received insulin therapy by CSII for 1.3 (0.8–2.6) years.RESULTS: In the general group of patients, 3 and 6 months after the start of FGM use, the HbA1c values decreased statistically significantly by 0.2%. In addition, the number of children with HbA1c &lt;7.5% increased by 6.1 and 4.9% at 3 and 6 months, respectively, but these changes were not statistically significant. The number of cases of DKA when using FGM decreased by 74%, and the number of cases of severe hypoglycemia by 83%, thus the number of episodes decreased by 4 and 6 times, respectively. Patients and / or their parents rated the ease of use and their experience with FGM on a scale from 0 (strongly agree) to 4 (strongly disagree). The majority of children and parents positively (0 or 1) assessed the convenience of installing and wearing the sensor (72.7–98.2%) using the FGM system in general (75.0–96.4%) and in comparison with the SMBG glucometer (92.3–98.2%).CONCLUSION: The installation and use of FGM is convenient and comfortable for the vast majority of children and parents, while compared to SMBG, the use of FGM is more convenient and simpler, and glucose measurement is much faster and less painful

The role of specific pancreatic antibodies in the differential diagnosis of complete clinical and laboratory remission of type 1 diabetes mellitus and MODY in children

BACKGROUND: T1D is characterized by autoimmune destruction of pancreatic β-cells, which develops due to genetic and environmental risk factors. Shortly after initiating the treatment with insulin, 80% of children with T1D may require smaller doses of insulin and develop clinical and laboratory remission of the disease so called «honeymoon». The issue of whether there is a need of differential diagnosis between autoimmune DM and non-immune forms of DM raises in cases of preclinical diagnosis of T1D and laboratory remission for more than 6 months.AIM: To study the clinical, immunological, genetic characteristics of T1D remission phase and MODY in children, to determine the diagnostic criteria for T1D and MODY in children.MATERIALS AND METHODS: A single-centre, cross sectional noncontrolled comparative study of two independent cohorts. Data of 150 children examined in the Endocrinology Research Center (January 2016–June 2021). First cohort included patients with complete clinical and laboratory remission of T1D (n=36), second cohort included patients with MODY, confirmed by genetic study (n=114).RESULTS: The median age of diabetes manifestation was significantly higher in patients with T1D — 11.25 years [8.33; 13.78] than in patients with MODY — 7.5 years [4.6; 12.2] (p=0.004). In patients with T1D remission the level of glycated hemoglobin was 6.0% [5.6; 6.4], in group with MODY — 6.5% [6.2; 6.7] (p&lt;0.001). Patients with monogenic diabetes had impaired fasting glucose — 6.27 mmol/l [5.38; 6.72], while patients with remission phase had normoglycemia — 5.12 mmol/l [4.17; 5.87]. The oral glucose tolerance test was perform to all patients, two-hour glucose level did not significantly differ in two groups (p=0.08). A strong family history of diabetes in patients with MODY registered more often (93% vs. 66.7%). A positive autoantibody titer detected more often in patients with remission of T1D (77.8%) than in patients with MODY (11.4%). In addition, no more than 1 type of autoantibodies was detected in patients with MODY.CONCLUSION: Antibodies ZnT8 and IA2 showed the greatest significance for the differential diagnosis of T1D and MODY in cases with long absents of insulin requirement in children with diabetes mellitus. Genetic test is recommended in seronegative cases. If only one type of AT is detected, specialist should decide on the need to do diagnostic genetic test based on a comprehensive analysis of the patient’s clinic characteristics, including family history, manifestation and blood glucose levels