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Authors = Alexander Blazhev ORCID = 0000-0003-0092-9488

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17 pages, 1279 KiB  
Article
Unlocking the Uterine Code: Microbiota, Immune Cells, and Therapy for Recurrent Reproductive Failure
by Svetla Blazheva, Svetlana Pachkova, Tatyana Bodurska, Petar Ivanov, Alexander Blazhev, Tzvetan Lukanov and Emiliana Konova
Microorganisms 2024, 12(3), 547; https://doi.org/10.3390/microorganisms12030547 - 9 Mar 2024
Cited by 7 | Viewed by 3097
Abstract
The uterine microbiota has been the subject of increasing study, but its interaction with the local immune system remains unclear. Successful embryo implantation relies on endometrial receptivity, which is pivotal for immunological tolerance to fetal antigens and precise regulation of inflammatory mediators. Emerging [...] Read more.
The uterine microbiota has been the subject of increasing study, but its interaction with the local immune system remains unclear. Successful embryo implantation relies on endometrial receptivity, which is pivotal for immunological tolerance to fetal antigens and precise regulation of inflammatory mediators. Emerging data suggest a dynamic interplay between endometrial microflora and the immune system, making dysbiosis a potential determinant of pregnancy outcomes. Imbalances in the regulation of immune cells in the endometrium and decidua have been associated with infertility, miscarriage, and obstetric complications. A thorough comprehension of the immune system in the female reproductive tract shows potential for improving women’s health and pregnancy outcomes. The objective of this study was to evaluate the patterns of endometrial microbiota in patients with recurrent implantation failure (RIF) and recurrent pregnancy loss (RPL) and to explore their implications for endometrial immune cells and chronic endometritis (CE). Immune cells in biopsies from 107 RIF and 93 RPL patients were examined using flow cytometry. The endometrial microbial composition was analyzed using real-time polymerase chain reaction (RT-PCR). The research uncovered disrupted endometrial microbiota in most women with RIF and RPL, which was often associated with significant effects on lymphocytes, T cells, and uNK cells. Full article
(This article belongs to the Special Issue Vaginal Microbiome in Women's Health)
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12 pages, 919 KiB  
Article
Early Diagnostic Markers of Late-Onset Neonatal Sepsis
by Preslava Gatseva, Alexander Blazhev, Zarko Yordanov and Victoria Atanasova
Pediatr. Rep. 2023, 15(3), 548-559; https://doi.org/10.3390/pediatric15030050 - 20 Sep 2023
Cited by 5 | Viewed by 3790
Abstract
Objective: Early diagnosis of nosocomial infections in newborns is a great challenge, because in the initial phase of systemic infection, clinical symptoms are often non-specific, and routinely used hematological markers are not sufficiently informative. The aim of this study was to determine the [...] Read more.
Objective: Early diagnosis of nosocomial infections in newborns is a great challenge, because in the initial phase of systemic infection, clinical symptoms are often non-specific, and routinely used hematological markers are not sufficiently informative. The aim of this study was to determine the potential of early inflammatory markers to diagnose late-onset neonatal sepsis—procalcitonin (PCT), interleukin 6 (IL-6), interleukin 8 (IL-8) and endocan (ESM-1). Material and methods: A prospective clinical–epidemiological study was conducted in a third-level NICU in Pleven, Bulgaria. Patients with suspected late-onset sepsis and healthy controls were tested. A sandwich ELISA method was used to measure the serum concentrations of biomarkers. Results: Sixty newborns were included, of which 35% symptomatic and infected, 33.3% symptomatic but uninfected and 31.7% asymptomatic controls. The mean values of PCT, IL-6, I/T index and PLT differ significantly in the three groups. For ESM-1, IL-8 and CRP, the difference was statistically insignificant. The best sensitivity (78%) and negative predictive value (84%) was found for IL-6. The combinations of PCT + IL-6 and PCT + IL-6+ I/T+ PLT showed very good diagnostic potential. Conclusion: The introduction into the routine practice of indicators such as PCT and IL-6 may provide an opportunity to promptly optimize the diagnostic and therapeutic approach to LOS. Full article
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10 pages, 1252 KiB  
Article
Anaplasma and Ehrlichia Species in Ixodidae Ticks Collected from Two Regions of Bulgaria
by Iskren Stanilov, Alexander Blazhev and Lyuba Miteva
Microorganisms 2023, 11(3), 594; https://doi.org/10.3390/microorganisms11030594 - 26 Feb 2023
Cited by 6 | Viewed by 3347
Abstract
The aim of the study was to determine prevalence of Anaplasmataceae-infected ticks in the Black Sea Coast and the Pleven regions of Bulgaria. A total of 350 ticks from different tick species were collected. Two hundred fifty-five ticks were removed from dogs [...] Read more.
The aim of the study was to determine prevalence of Anaplasmataceae-infected ticks in the Black Sea Coast and the Pleven regions of Bulgaria. A total of 350 ticks from different tick species were collected. Two hundred fifty-five ticks were removed from dogs in the Black Sea Coast region, and 95 Ixodes ricinus ticks were collected by flagging vegetation with a white flannel cloth in two areas in the region of Pleven. After the DNA isolation of the ticks, a genus-specific polymerase chain reaction (PCR) was performed to identify Anaplasmataceae. Second PCRs were performed with species-specific primers to identify Ehrlichia canis (E. canis) and Anaplasma phagocytophilum (A. phagocytophilum). The results showed that 26.9% of the Ixodes ricinus ticks were infected with Anaplasmataceae in the Black Sea Coast region and 36.8% in the Pleven region. The infection with E. canis was detected in 35.7% and A. phagocytophilum in 25.0% of positive ticks from the Black Sea Coast region. In the Pleven region, 22.9% of ticks were positive for E. canis, while 42.9% were positive for A. phagocytophilum. The molecular identification of E. canis in ticks collected from Bulgaria was performed for the first time. In conclusion, the present study revealed a higher prevalence of ticks infected with Anaplasmataceae, particularly A. phagocytophilum, in the Pleven region than in the Black Sea Coast region. Full article
(This article belongs to the Special Issue Advanced Research on Ticks and Tick-Borne Diseases)
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9 pages, 893 KiB  
Article
Elevated IgG and IgM Autoantibodies to Advanced Glycation End Products of Vascular Elastin in Hypertensive Patients with Type 2 Diabetes: Relevance to Disease Initiation and Progression
by Krasimir Kostov and Alexander Blazhev
Pathophysiology 2022, 29(3), 426-434; https://doi.org/10.3390/pathophysiology29030034 - 1 Aug 2022
Cited by 5 | Viewed by 3487
Abstract
The increased glycation of elastin is an important factor in vascular changes in diabetes. Using the ELISA method, we determined serum levels of IgM and IgG autoantibodies to advanced glycation end products of vascular elastin (anti-AGE EL IgM and anti-AGE EL IgG) in [...] Read more.
The increased glycation of elastin is an important factor in vascular changes in diabetes. Using the ELISA method, we determined serum levels of IgM and IgG autoantibodies to advanced glycation end products of vascular elastin (anti-AGE EL IgM and anti-AGE EL IgG) in 59 hypertensive patients with type 2 diabetes (T2D) and 20 healthy controls. Serum levels of matrix metalloproteinases-2 and -9 (MMP-2 and MMP-9) and the C-reactive protein (CRP) were also determined. The levels of anti-AGE EL IgM antibodies in the T2D group were similar to those in the control group, while those of anti-AGE EL IgG antibodies were significantly higher (p = 0.017). Significant positive correlations were found between the levels of anti-AGE EL IgM antibodies and MMP-2 (r = 0.322; p = 0.013) and between the levels of anti-AGE EL IgG antibodies and CRP (r = 0.265; p = 0.042). Our study showed that elevated anti-AGE EL IgG antibody levels may be an indicator of the enhanced AGE-modification and inflammatory-mediated destruction of vascular elastin in hypertensive patients with T2D. Anti-AGE EL IgM antibodies may reflect changes in vascular MMP-2 activity, and their elevated levels may be a sign of early vascular damage. Full article
(This article belongs to the Collection Feature Papers in Pathophysiology)
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12 pages, 1420 KiB  
Article
Prevalence of Borrelia burgdorferi Sensu Lato in Ixodes ricinus Ticks Collected from Kaylaka Park in Pleven, Bulgaria
by Alexander Blazhev, Iskren Stanilov, Lyuba Dineva Miteva, Milena Atanasova, Svetla Blazheva and Spaska Stanilova
Microorganisms 2022, 10(4), 772; https://doi.org/10.3390/microorganisms10040772 - 3 Apr 2022
Cited by 6 | Viewed by 3646
Abstract
We aimed to determine the presence and distribution of Borrelia burgdorferi sensu lato (s.l.) in Ixodes ricinus ticks collected from urbanized and wild areas in Kaylaka Park (Bulgaria). A total of 546 ticks were collected over three years (2017–2019). The presence of Borrelia [...] Read more.
We aimed to determine the presence and distribution of Borrelia burgdorferi sensu lato (s.l.) in Ixodes ricinus ticks collected from urbanized and wild areas in Kaylaka Park (Bulgaria). A total of 546 ticks were collected over three years (2017–2019). The presence of Borrelia in 334 of the collected I. ricinus was detected by dark-field microscopy (DFM) and two nested PCRs (nPCR) targeting the borrelial 5S-23S rRNA intergenic spacer and Flagellin B (FlaB) gene. DFM was performed on a total of 215 ticks, of which 86 (40%) were positive. PCR was performed on 153 of the ticks. In total, 42.5% of the 5S-23S rRNA intergenic spacer and 49% of FlaB were positive. Considering as positive any single tick in which Borrelia sp. was detected regardless of the used method, the infection rate reached 37% (10/27) in the nymphs and 48.5% (149/307) in the adults (48.7% (77/158) females, 48.3% (72/149) males). The incidence of B. burgdorferi infection in I. ricinus did not differ statistically significantly between female, male, and nymph. This study provides evidence that Lyme disease spirochetes are present in various regions of Kaylaka Park with extremely high prevalence in their vectors. Full article
(This article belongs to the Special Issue Epidemiology, Surveillance and Prevention of Tick-Borne Diseases)
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7 pages, 3202 KiB  
Article
Changes in Serum Levels of Matrix Metalloproteinase-1 and Tissue Inhibitor of Metalloproteinases-1 in Patients with Essential Hypertension
by Krasimir Kostov and Alexander Blazhev
Bioengineering 2022, 9(3), 119; https://doi.org/10.3390/bioengineering9030119 - 15 Mar 2022
Cited by 12 | Viewed by 2677
Abstract
Hypertension (HTN) is a leading risk factor for cardiovascular (CV) disease. Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) are thought to be actively involved in the remodeling of the CV extracellular matrix (ECM) during hypertensive damage. Therefore, in this study, we aimed [...] Read more.
Hypertension (HTN) is a leading risk factor for cardiovascular (CV) disease. Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) are thought to be actively involved in the remodeling of the CV extracellular matrix (ECM) during hypertensive damage. Therefore, in this study, we aimed to investigate serum levels of MMP-1 and TIMP-1 in patients with essential HTN and compare them with those of normotensive individuals. We measured serum concentrations of MMP-1 and TIMP-1 in 60 patients with HTN and 20 healthy controls using an ELISA. The obtained results showed that in patients with HTN, the mean levels of MMP-1 (1.82 ± 0.9 ng/mL) were significantly higher (p = 0.03) than the mean levels in the control group (1.19 ± 0.7 ng/mL). The levels of TIMP-1 in patients with essential HTN (0.44 ± 0.1 ng/mL) were also significantly higher (p = 0.005) than those in the control group (0.33 ± 0.1 ng/mL). In HTN, elevated serum MMP-1 levels may be associated with increased collagen degradation in the CV ECM, whereas elevated TIMP-1 levels may favor its accumulation and the development of pathological remodeling and fibrosis of the heart and arterial vessels. Full article
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7 pages, 562 KiB  
Article
Circulating Levels of Endothelin-1 and Big Endothelin-1 in Patients with Essential Hypertension
by Krasimir Kostov and Alexander Blazhev
Pathophysiology 2021, 28(4), 489-495; https://doi.org/10.3390/pathophysiology28040031 - 25 Oct 2021
Cited by 12 | Viewed by 3711
Abstract
The role of endothelin-1 (ET-1) in the pathogenesis of hypertension (HTN) is not clearly established. There is evidence that its circulating levels are elevated in some forms of experimental and human HTN, but this was not a consistent finding. Based on these controversial [...] Read more.
The role of endothelin-1 (ET-1) in the pathogenesis of hypertension (HTN) is not clearly established. There is evidence that its circulating levels are elevated in some forms of experimental and human HTN, but this was not a consistent finding. Based on these controversial data, we tested serum levels of ET-1 and Big ET-1 (the precursor of ET-1) in patients with essential HTN, comparing the results with those of healthy normotensive controls. The levels of ET-1 and Big ET-1 were measured by ELISA. Our results in patients with essential HTN showed that the mean levels of ET-1 (5.01 ± 2.1 pg/mL) were significantly higher (F = 6.34, p = 0.0144) than the mean levels in the control group (3.2 ± 1.0 pg/mL). The levels of Big ET-1 in patients with essential HTN (0.377 ± 0.1 pmol/L) were similar to those in the control group (0.378 ± 0.07 pmol/L) and did not differ significantly (F = 0.00, p = 0.9531). These data suggest that ET-1, but not Big ET-1, may play an important role in the pathogenesis of primary HTN. Full article
(This article belongs to the Special Issue Vasodilators: New Insights)
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13 pages, 955 KiB  
Article
Estimation of Ixodes ricinus (Acari: Ixodidae) Populations of Kaylaka Park in the Town of Pleven, Bulgaria
by Alexander Blazhev, Milena Atanasova, Krasimir Kostov, Tsetsa Doychinova, Svetla Blazheva and Milena Karcheva
Insects 2021, 12(9), 808; https://doi.org/10.3390/insects12090808 - 9 Sep 2021
Cited by 4 | Viewed by 2872
Abstract
(1) Background: Ticks are vectors of a large number of pathogenic microorganisms, which cause serious diseases in both humans and animals. Kaylaka Park is located in northern Bulgaria close to the city of Pleven. Part of the park is urbanized and visited daily [...] Read more.
(1) Background: Ticks are vectors of a large number of pathogenic microorganisms, which cause serious diseases in both humans and animals. Kaylaka Park is located in northern Bulgaria close to the city of Pleven. Part of the park is urbanized and visited daily by many citizens. The aim of our study was to determine the presence and distribution of hard ticks in the park area by surveying and comparing four urbanized with four wild areas. (2) Methods: Ticks were collected by flagging from 2016 to 2020 during the spring–summer season (March–July). Air temperature, relative humidity, collection time and flagging area were measured during the campaign. (3) Results: A total of 622 ticks were collected: 285 females (46%), 272 (44%) males and 64 (10%) nymphs. All were identified as Ixodes ricinus. Wild areas showed statistically significant higher values of ticks collected per minute (p = 0.009) and nymph densities (p = 0.003) compared to urbanized sampling sites. Other densities indices did not have a significant difference between urban and wild areas. Highest numbers of Ixodes ticks were collected at a temperature of 20 °C and at 60% relative humidity. The active questing began in March, peaked in end of April and declined in June. (4) Conclusions: In the present study, we found that ecological factors in the Kaylaka Park area are favourable for the development and distribution of tick populations. The results give us reason to consider that there is a high risk to visitors from tick bites in the Kaylaka Park area. Full article
(This article belongs to the Special Issue Ecology of Ticks and Their Control)
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10 pages, 589 KiB  
Article
Serum Anti-Collagen IV IgM and IgG Antibodies as Indicators of Low Vascular Turnover of Collagen IV in Patients with Long-Term Complications of Type 2 Diabetes
by Krasimir Kostov and Alexander Blazhev
Diagnostics 2021, 11(5), 900; https://doi.org/10.3390/diagnostics11050900 - 19 May 2021
Cited by 3 | Viewed by 3343
Abstract
Thickening of the vascular basement membrane (BM) is a fundamental structural change in the small blood vessels in diabetes. Collagen type IV (CIV) is a major component of the BMs, and monitoring the turnover of this protein in type 2 diabetes (T2D) can [...] Read more.
Thickening of the vascular basement membrane (BM) is a fundamental structural change in the small blood vessels in diabetes. Collagen type IV (CIV) is a major component of the BMs, and monitoring the turnover of this protein in type 2 diabetes (T2D) can provide important information about the mechanisms of vascular damage. The aim of the study was through the use of non-invasive biomarkers of CIV (autoantibodies, derivative peptides, and immune complexes) to investigate vascular turnover of CIV in patients with long-term complications of T2D. We measured serum levels of these biomarkers in 59 T2D patients with micro- and/or macrovascular complications and 20 healthy controls using an ELISA. Matrix metalloproteinases-2 and -9 (MMP-2 and MMP-9) were also tested. In the T2D group, significantly lower levels of CIV markers and significantly higher levels of MMP-2 and MMP-9 were found compared to controls. A significant positive correlation was found between IgM antibody levels against CIV and MMP-2. These findings suggest that vascular metabolism of CIV is decreased in T2D with long-term complications and show that a positive linear relationship exists between MMP-2 levels and CIV turnover in the vascular wall. Full article
(This article belongs to the Special Issue Biomarkers of Vascular Diseases 2.0)
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15 pages, 1180 KiB  
Article
Use of Glycated Hemoglobin (A1c) as a Biomarker for Vascular Risk in Type 2 Diabetes: Its Relationship with Matrix Metalloproteinases-2, -9 and the Metabolism of Collagen IV and Elastin
by Krasimir Kostov and Alexander Blazhev
Medicina 2020, 56(5), 231; https://doi.org/10.3390/medicina56050231 - 11 May 2020
Cited by 19 | Viewed by 5086
Abstract
Background and objectives: HbA1c measurements may be useful not only in optimizing glycemic control but also as a tool for managing overall vascular risk in patients with diabetes. In the present study, we investigate the clinical significance of HbA1c as a biomarker [...] Read more.
Background and objectives: HbA1c measurements may be useful not only in optimizing glycemic control but also as a tool for managing overall vascular risk in patients with diabetes. In the present study, we investigate the clinical significance of HbA1c as a biomarker for hyperglycemia-induced vascular damages in type 2 diabetes (T2D) based on the levels of matrix metalloproteinases-2, -9 (MMP-2, MMP-9), anti-collagen IV (ACIV), and anti-elastin (AE) antibodies (Abs) IgM, IgG, and IgA, and CIV-derived peptides (CIV-DP) reflecting collagen and elastin turnover in the vascular wall. The aim is to show the relationship of hyperglycemia with changes in the levels of vascular markers and the dynamics of this relationship at different degrees of glycemic control reported by HbA1c levels. Materials and Methods: To monitor elastin and collagen IV metabolism, we measured serum levels of these immunological markers in 59 patients with T2D and 20 healthy control subjects with an ELISA. Results: MMP-2, MMP-9, and the AEAbs IgA levels were significantly higher in diabetic patients than in control subjects, whereas those of the AEAbs IgM, ACIVAbs IgM, and CIV-DP were significantly lower. MMP-9 levels were significantly lower at HbA1c values >7.5%. Conclusions: A set of three tested markers (MMP-2, MMP-9, and AEAbs IgA) showed that vascular damages from preceding long-term hyperglycemia begin to dominate at HbA1c values ≥7.5%, which is the likely cut-point to predict increased vascular risk. Full article
(This article belongs to the Special Issue Interdisciplinary Medicine)
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13 pages, 640 KiB  
Article
Serum Concentrations of Endothelin-1 and Matrix Metalloproteinases-2, -9 in Pre-Hypertensive and Hypertensive Patients with Type 2 Diabetes
by Krasimir Kostov, Alexander Blazhev, Milena Atanasova and Anelia Dimitrova
Int. J. Mol. Sci. 2016, 17(8), 1182; https://doi.org/10.3390/ijms17081182 - 1 Aug 2016
Cited by 27 | Viewed by 7028
Abstract
Endothelin-1 (ET-1) is one of the most potent vasoconstrictors known to date. While its plasma or serum concentrations are elevated in some forms of experimental and human hypertension, this is not a consistent finding in all forms of hypertension. Matrix metalloproteinases -2 and [...] Read more.
Endothelin-1 (ET-1) is one of the most potent vasoconstrictors known to date. While its plasma or serum concentrations are elevated in some forms of experimental and human hypertension, this is not a consistent finding in all forms of hypertension. Matrix metalloproteinases -2 and -9 (MMP-2 and MMP-9), which degrade collagen type IV of the vascular basement membrane, are responsible for vascular remodeling, inflammation, and atherosclerotic complications, including in type 2 diabetes (T2D). In our study, we compared concentrations of ET-1, MMP-2, and MMP-9 in pre-hypertensive (PHTN) and hypertensive (HTN) T2D patients with those of healthy normotensive controls (N). ET-1, MMP-2, and MMP-9 were measured by ELISA. Concentrations of ET-1 in PHTN and N were very similar, while those in HTN were significantly higher. Concentrations of MMP-2 and MMP-9 in PHTN and HTN were also significantly higher compared to N. An interesting result in our study is that concentrations of MMP-2 and MMP-9 in HTN were lower compared to PHTN. In conclusion, we showed that increased production of ET-1 in patients with T2D can lead to long-lasting increases in blood pressure (BP) and clinical manifestation of hypertension. We also demonstrated that increased levels of MMP-2 and MMP-9 in pre-hypertensive and hypertensive patients with T2D mainly reflect the early vascular changes in extracellular matrix (ECM) turnover. Full article
(This article belongs to the Special Issue Liquid Biopsy for Clinical Application)
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