Age features of morphological changes of the hematotesticular barrier in experimental diabetes mellitus.

Background. Diabetes mellitus is an acute medical and social problem in all parts of the world due to the constant increase in the incidence of the disease, severe complications, disability and high mortality. Hyperglycemia leads to oxidative stress and increased processes of formation of active oxygen species, which in turn make a significant contribution to the development of male infertility. Objective. Therefore, the aim of our study was to establish morphological changes in the vessels of the hemomicrocirculatory flow and testicular sustentocytes of adult rats with experimental streptozotocin diabetes mellitus (SDM). Methods. The material for the study were the testicles of 20 sexually mature 6-month-old rats (weighing 150-180 g). SDM in animals of the experimental group was simulated by a single intraperitoneal injection of streptozotocin (dissolved in 0.1 M citrate buffer solution with a pH of 4.5) at a dose of 6 mg per 100 g of mass. An equivalent dose of 0.1 M citrate buffer was injected intraperitoneally to animals of the control group. Histological, electron microscopic, biochemical, morphometric and statistical research methods were used. Results. It was found that in the early stages of SDM (28th day) on the background of hyperglycemia in the hemomicrocirculatory flow of the testes there is a spasm of the vessels of the afferent link, which is confirmed by a decrease in the lumen of arterioles and an increase in their Wagenworth index. On the 70th day of SDM on the background of elevated levels of glucose and glycosylated hemoglobin in the links of the hemomicrocirculatory flow of the testis there are signs of diabetic microangiopathy, manifested by: hemorheological disorders in micro-hemo-vessels, (erythrocyte sludge, adhesion of erythrocytes and platelets, microclasmatosis), decrease in the capacity of arterioles and capillaries (increase in the Wagenworth index, respectively by 1.8 and 1.9 times), microclasmatosis, thickening and proliferation of the basement membrane of capillaries. Against the background of diabetic microangiopathy, there is a decrease in the number of sustentocytes by 0.01 mm2 of testicular parenchyma by 1.8 times, compared with control indicators, the area of their profile increases by 2.2 times (in all cases p<0.05). The nucleolar areas probably do not change, which leads to an increase in the nuclear-cytoplasmic ratio by 2.9 times (p<0.05). Such morphometric changes of sustentocytes are caused by development of vacuolar hydropic dystrophies in them, and an apoptosis that is confirmed by data of histo-ultrastructural studies. Changes in sustentocytes against the background of the development of diabetic microangiopathy lead to a violation of the hematotesticular barrier and to dystrophic changes in the spermatogenic epithelium of the testis. Conclusion. Thus, on the 70th day of SDM in the hemomicrocirculatory flow of the testis, the development of diabetic microangiopathy is observed, which leads to a violation of the hematotesticular barrier, and as a consequence, to a violation of spermatogenesis.

Necrobiosis lipoidica: a rare clinical and pathomorphological case

According to modern scientific researches, necrobiosis lipoidica (NL) is a disease characterized by focal disorganization and lipid collagen dystrophy. It is believed that the basis of skin changes in this dermatosis is diabetic microangiopathy that is accompanied by sclerosis and obliteration of blood vessels, which leads to necrobiosis with subsequent deposition of lipids in the dermis. This pathology is registered relatively rarely, in 1 % of patients with diabetes mellitus (DM) on average. The combination of NL with DM, according to the literature data, ranges from 25 to 70 %; more often (in 40–60 % of cases) DM is preceded by NL, and in 10–25 % of cases they occur simultaneously. In addition, in 10–50 % of cases NL is diagnosed in people without concomitant diabetes. The variability of clinical, epidemiological features and the relatively low prevalence of this pathology is often the cause for misdiagnosis or late diagnosis. The described clinical case is typical in terms of the epidemiological data: sex, age, presence of DM. At the same time, it is rare in terms of the clinical picture: it is not classically diabetic by localization (symmetrical areas of the legs are typical), by appearance of necrobiosis areas — granulomatous type of necrobiosis in the form of granuloma annulare, by histological structure — area of chronic perivascular lymphoplasmocytic inflammation with the involvement of single giant cells, which required additional clinical and anamnestic data for an objective report of the pathologist. Biopsy in this case was used as a differential diagnosis between granuloma annulare and necrobiotic necrogranuloma. In addition, this method of diagnosis has played an additional therapeutic role. This case may have demonstrated the activation of the cellular and humoral immune response in the area of chronic inflammation in response to a mechanical damage and the resolution of inflammation with complete tissue repair.

Pregnancy of women with type 1 diabetes mellitus – the eff ect of preconception care on perinatal results. Ten years of experience

Introduction: Despite the ever-improving medical care, pregnancies of women with type 1 diabetes mellitus (T1DM) are at increased risk of complications for both mother and child. Optimal compensation of diabetes before and during pregnancy is an essential protective factor reducing the risk of congenital malformations, pregnancy loss, and other complications. The pregnancy of women with T1DM should be planned, ideally at a time of optimal diabetes compensation. Target glycated hemoglobin (HbA1c) values until the range of 42–48 mmol/mol should be achieved at least three months before pregnancy. Our work aimed to evaluate the perinatal results of pregnancies in women with T1DM and the eff ect of preconception counseling and adequate T1DM compensation before pregnancy on perinatal outcomes. Methods and results: Retrospective analysis of pregnancy and perinatal outcomes of women with T1DM were followed up at the Department of Gynecology and Obstetrics, General University Hospital in Prague and First Faculty of Medicine, Charles University between 2008 to 2018. A total of 221 women with T1DM were included in the analysis. Adequate (HbA1c ≤ 48 mmol/mol at least 3 months before conception) and inadequate diabetes compensation at the beginning of the pregnancy had 59 (26.7%) and 162 (72.3%) women, respectively. Pregnancies of women with adequate diabetes compensation were more often planned (55.9 vs. 24.7%; P < 0.0001), had a lower incidence of any form of diabetic microangiopathy (13.6 vs. 37.7%; P = 0.001), pre-eclampsia (0 vs. 6.8%; P = 0.036), better compensation of diabetes during pregnancy (mean HbA1c during pregnancy 39.9 ± 6.7 vs. 49.9 ± 12.2; P < 0.0001), and their pregnancy was less often terminated from medical indication (congenital malformation of the fetus or decompensation of T1DM) (0 vs. 7.4%; P = 0.032). Pregnancies of women with adequate diabetes compensation before conception were less often complicated by fetal macrosomia (birth weight > 95th percentile; 22.0 vs. 35.8%; P = 0.027). Conclusion: The pregnancy of women with T1DM is burdened by a number of perinatal and neonatal complications. In the study group, most women with T1DM became pregnant unintentionally at a time of inadequate diabetes compensation. Women who achieved adequate diabetes compensation before pregnancy had a lower incidence of perinatal complications. Therefore, it is advised that women with T1DM should plan their pregnancy, attend preconception and antenatal care, and give birth in perinatal centers, which provide coordinated care from diabetologists, gynecologists, obstetricians, and neonatologists. Key words: preconceptual counseling – type 1 diabetes mellitus – perinatal outcomes

Dissecting the Pathogenesis of Diabetic Retinopathy Based on the Biological ceRNA Network and Genome Variation Disturbance

Background. Diabetic retinopathy (DR) is the most important manifestation of diabetic microangiopathy. It is essential to explore the gene regulatory relationship and genomic variation disturbance of biological networks in DR progression. Methods. In this study, we constructed a comprehensive lncRNA-mRNA ceRNA network of DR procession (CLMN) and explored its topological characteristics. Results. Modular and functional analysis indicated that the organization of CLMN performed fundamental and specific functions in diabetes and DR pathology. The differential expression of hub ceRNA nodes and positive correlation reveals the highly connected ceRNA regulation and important roles in the regulating of DR pathology. A large proportion of SNPs in the TFBS, DHS, and enhancer regions of lncRNAs will affect lncRNA transcription and further cause expression variation. Some SNPs were found to disrupt the lncRNA functional elements such as miRNA target binding sites. These results indicate the complex nature of genotypic effects in the disturbing of CLMN and further contribute to gene expression variation and different disease phenotypes. Conclusion. The identification of individual genomic variations and analysis of biological network disturbance by these genomic variations will help provide more personalized treatment plans and promote the development of precision medicine for DR.

MiR-124-3p Suppresses the Dysfunction of High Glucose-Stimulated Endothelial Cells by Targeting G3BP2

Background: Diabetic retinopathy (DR) is the most important manifestation of diabetic microangiopathy. MicroRNAs (miRNAs), members of non-coding RNAs, have been frequently reported to regulate various diseases including DR. MiR-124-3p is involved in DR based on bioinformatics. The current study aimed to investigate the role of miR-124-3p in high glucose (HG)-treated human retinal microvascular endothelial cells (HRMECs), an in vitro model of DR.Methods: Bioinformatics analysis was applied to reveal the targets downstream miR-124-3p. A series of assays including CCK-8, luciferase reporter, western blot, and tube formation assays were used to explore the function and mechanism of miR-124-3p in HG-stimulated HRMECs.Results: We found out that miR-124-3p was downregulated in HG-stimulated HRMECs. Functionally, miR-124-3p overexpression restrained the HG-induced cell injury of HRMECs. Mechanistically, we predicted 5 potential target mRNAs of miR-124-3p. G3BP stress granule assembly factor 2 (G3BP2) was validated to bind with miR-124-3p. Rescue assays showed that miR-124-3p suppressed cell injury of HG-stimulated HRMECs through G3BP2. In addition, miR-124-3p regulated the p38MAPK signaling pathway by G3BP2, and G3BP2 promoted injury of HG-treated HRMECs through the activation of the p38MAPK signaling pathway.Conclusion: MiR-124-3p suppressed the dysfunctions of HG-treated HRMECs by targeting G3BP2 and activating the p38MAPK signaling. This new discovery provided a potential biomarker for DR treatment.

Hepatic Hyaline Arteriolosclerosis in Patients with Chronic Hyperglycemia

Introduction/Objective Hyaline arteriolosclerosis is a common feature of diabetic microangiopathy. It results from hyperglycemia-induced endothelial cell dysfunction and can be found in many sites including the retina, kidney, and skin. This histologic finding is important because studies have shown that in the kidney it is an independent risk factor for cardiovascular complications. To our knowledge, an association between hyperglycemia and hyaline arteriolosclerosis in the liver has not be studied. We aimed to investigate whether chronic hyperglycemia is associated with this histologic finding in liver arterioles. Methods/Case Report The first 50 liver biopsies performed at our institution starting January 1, 2020 were scanned into a digital pathology system (Philips, Cambridge, MA). For each biopsy, the patient’s highest recorded hemoglobin A1c (HbA1c) up to one year from the date of biopsy was recorded. The biopsies were then grouped into two groups: those from patients with a HbA1c greater than 6.5%, and those with a HbA1c value less than 6.5 %. Next, the digested periodic acid Schiff (DPAS) intensity of the arteriolar vessels in each biopsy was graded 0 to 3 by a single pathology resident who was blinded to its corresponding HbA1c group. Grade 0 to 1 was considered negative staining. Grade 2 to 3 was considered positive staining and indicative of hyaline arteriolosclerosis. By the end of the data collection, each case had been given a staining category and a HbA1c group. A 2 x 2 contingency table was constructed. Results (if a Case Study enter NA) 37 of 50 patients had a recorded HbA1c meeting the study criteria. Of these 37 patients, 11 had an HbA1c greater than 6.5 % while 26 had an HbA1c less than 6.5%. Seven of the 11 biopsies from patients with an elevated HbA1c showed positive staining. Six of the 26 biopsies from patients with an HbA1c less than 6.5% showed positive staining. Conclusion We hypothesized that patients living with hyperglycemia may be more likely to exhibit hepatic hyaline arteriolosclerosis. A chi-squared test of independence was performed to examine the association between a liver biopsy’s HbA1c group and staining category. The relation between these variables was significant at p = &lt; .05, χ 2 (2, N= 36) = 5.57, p = .0182, indicating liver biopsies with a HbA1c greater than 6.5 % are more likely to have this histologic finding. Future studies are needed to characterize diabetic hepatopathy, and its relatedness to macrovascular complications.

Diagnostic capabilities of different methods of laser doppler flowmetry spectral indexes assessment in patients with diabetic microangiopathy

The article contains the results of a study of two different methods for calculating the spectral parameters of laser Doppler flowmetry in patients with diabetic microangiopathy caused by type 2 diabetes mellitus (main group) and those with excluded diabetes mellitus (control group). Spectral indices were calculated using either average or maximum amplitudes of the frequency ranges. When comparing the contribution of respiratory and pulse fluxmotions using average amplitudes, there were significant (p < 0.05) differences between the main and control groups. On the contrary, when using the maximum amplitudes, no significant differences were noted (p > 0.05). Also, significant correlations were found between the contributions of respiratory and pulse fluxmotions and the estimated glomerular filtration rate in the main group, using both calculation methods. These studies indicate the feasibility of using a technique based on the analysis of average amplitudes to increase the specificity of laser Doppler flowmetry as a method for diagnosing diabetic microangiopathy.

Thiamine and diabetes: back to the future?

The first reports of a link between thiamine and diabetes date back to the 1940s. Some years later, a role for thiamine deficiency in diabetic neuropathy became evident, and some pilot studies evaluated the putative effects of thiamine supplementation. However, the administration of thiamine and its lipophilic derivative benfotiamine for the treatment of this complication gained consensus only at the end of the ‘90 s. The first evidence of the beneficial effects of thiamine on microvascular cells involved in diabetic complications dates to 1996: from then on, several papers based on in vitro and animal models have addressed the potential use of this vitamin in counteracting diabetic microangiopathy. A few pilot studies in humans reported beneficial effects of thiamine administration on diabetic nephropathy, but, despite all promising proofs-of-concept, the possible role of thiamine in counteracting development or progression of retinopathy has not been addressed until now. Thiamine is a water-soluble vitamin, rapidly expelled from the body, with no issues of over-dosage or accumulation; unfortunately, it is non-patentable, and neither industry nor independent donors are interested in investing in large-scale randomized controlled clinical trials to investigate its potential in diabetes and its complications. Consequently, science will not be able to disprove a promising hypothesis and, more importantly, diabetic people remain deprived of a possible way to ameliorate their condition.

Age features of morphological changes of vessels of testicular hemomicrocirculatory flow in streptozotocin diabetes mellitus

Medico-social problem of diabetes is caused by early disability and mortality of patients due to specific complications of micro- and macroangiopathies. Therefore, the aim of our study was to establish morphological changes in vessels of the hemomicrocirculatory flow of the testes of immature rats with experimental streptozotocin diabetes mellitus (SDM). The material for the study were the testicles of 20 two-month-old immature (weighing 65-95 g) white outbred male rats, which were divided equally into 2 groups: experimental and control ones. SDM in animals of the experimental group was simulated by a single intraperitoneal injection of streptozotocin (dissolved in 0.1 M citrate buffer solution with a pH of 4.5) at a dose of 7 mg per 100 g of mass. The control group of animals received intraperitoneally an equivalent dose of 0.1 M citrate buffer. Histological, electron microscopic, biochemical, morphometric and statistical research methods were used. It was found that in the early stages of SDM (14th day) on the background of hyperglycemia in the hemomicrocirculatory flow of the testes there is a spasm of the vessels of the afferent link, which is confirmed by a decrease in the area of arterioles lumen and an increase in their VI. On the 56th day of SDM, on the background of elevated levels of glucose and glycosylated hemoglobin in the links of the hemorrhagic circulatory flow of the testes there are initial signs of diabetic microangiopathy, manifested by: hemorheological disorders in micro-hemo-vessels (erythrocyte sludges, adhesion of erythrocytes and platelets, microclasmatosis), decreased capacity of arterioles and capillaries (increase in VI, respectively by 1.2 and 1.9 times), microclasmatosis, thickening and proliferation of the basement membrane of capillaries. Thus, on the 56th day of SDM in the hemomicrocirculatory flow of the testes, the development of diabetic microangiopathy is observed, which leads to the disruption of the blood-testis barrier, and as a consequence, to a violation of spermatogenesis.