Despite the odds: formation of the SARS-CoV-2 methylation complex.

Coronaviruses modify their single-stranded RNA genome with a methylated cap during replication to mimic the eukaryotic mRNAs. The capping process is initiated by several nonstructural proteins (nsp) encoded in the viral genome. The methylation is performed by two methyltransferases, nsp14 and nsp16, while nsp10 acts as a co-factor to both. Additionally, nsp14 carries an exonuclease domain which operates in the proofreading system during RNA replication of the viral genome. Both nsp14 and nsp16 were reported to independently bind nsp10, but the available structural information suggests that the concomitant interaction between these three proteins would be impossible due to steric clashes. Here, we show that nsp14, nsp10, and nsp16 can form a heterotrimer complex upon significant allosteric change. This interaction is expected to encourage the formation of mature capped viral mRNA, modulating nsp14's exonuclease activity, and protecting the viral RNA. Our findings show that nsp14 is amenable to allosteric regulation and may serve as a novel target for therapeutic approaches.

Selected Technical Aspects of Molecular Allergy Diagnostics.

Diagnosis of allergic diseases is a complex, multi-stage process. It often requires the use of various diagnostic tools. The in vitro diagnostics (IVD), which includes various laboratory tests, is one of the stages of this process. Standard laboratory tests include the measurement of the serum concentration of specific immunoglobulin E (sIgE) for selected allergens, full allergen extracts and/or single allergen components (molecules). The measurement of IgE sIgE to the allergen components is called molecular allergy diagnosis. During the standard laboratory diagnostic process, various models of immunochemical tests are used, which enable the measurement of sIgE for single allergens (one-parameter tests, singleplex) or IgE specific for many different allergens (multi-parameter tests, multiplex) in one test. Currently, there are many different test kits available, validated for IVD, which differ in the method type and allergen profile. The aim of the manuscript is to present various technical aspects related to modern allergy diagnostics, especially in the area of molecular allergy diagnostics.

The Two-Sided Experimental Model of ImmunoCAP Inhibition Test as a Useful Tool for the Examination of Allergens Cross-Reactivity on the Example of α-Gal and Mammalian Meat Sensitization-A Preliminary Study.

Cross-reactivity of allergens is the cause of various, sometimes unexpected, clinical reactions. There are no standard methods to investigate cross-reactivity. We present an experimental model of a two-sided inhibition test (IT) on ImmunoCAP membranes (CAP). We constructed the described model based on the known cross-allergy syndrome to red meat developing in people bitten by ticks (α-Gal syndrome; AGS). Some individuals who are bitten by ticks develop IgE antibodies specific to the carbohydrate determinant, galactose-α-1,3-galactose (α-Gal), present in the tick's saliva. These antibodies can cross-react with α-Gal molecules expressed on mammalian meat proteins. The well-known property of anti-α-Gal IgE antibodies binding by various sources of this allergen was used by us in the proposed model of the two-sided inhibition test on ImmunoCAP membranes. We expected that anti-α-Gal IgE antibodies bind allergens from mammalian meat and blocking them abolishes this reactivity, and the two-sided inhibition test model we proposed on ImmunoCAP membranes allowed us to observe such a relationship. We conducted the experiment three times on biological material from people with different clinical manifestations of allergy to α-Gal, each time obtaining similar results. In conclusion, the model of bilateral inhibition on ImmunoCAP membranes proposed by us seems to be an attractive, simple tool for direct testing of allergic cross-reactivity.

Assessment of RANTES, MIP4A, MMP7, MMP9, MMP14, TIMP 1, TIMP 2 and TIMP 3 concentration in the follicular fluid of patients undergoing in vitro fertilization/embryo transfer procedure.

Introduction: Matrix metalloproteinases (MMP) degrade extracellular matrix. Some studies show that MMP9 concentration in follicular fluid (FF) may play a role in oocyte maturation and in vitro fertilization (IVF) success. The immunology of follicular fluid is still not fully understood. Aim: Assessment of RANTES, MIP4A, MMP7, MMP9, MMP14, TIMP 1, TIMP 2 and TIMP 3 concentration in the follicular fluid of patients undergoing in vitro fertilization/embryo transfer procedure. Material and methods: This case-control study included 20 randomly selected patients with a positive pregnancy (PPG) test and 20 patients with a negative pregnancy (NPG) test after IVF/ET. In FF obtained during oocyte retrieval, the concentrations of MIP4A, MMP7, MMP14, TIMP 1, TIMP 2, TIMP 3, RANTES, IL-12p40, and IL-17A were measured. Their effect on the characteristics of follicles, embryos, and the efficiency of IVF and ET were analysed. Results: There was no statistically significant relationship between a positive pregnancy test and the results of the immunoassay performed. The number of COC-1 correlates significantly and positively with RANTES (r = 0.34; p = 0.038) and IP-10 (r = 0.329; p = 0.038). MII correlates significantly and positively with RANTES (r = 0.341, p = 0.031). The number of top-quality embryos correlates significantly and positively with IL-17A (r = 0.451, p = 0.004) and TIMP 1 (r = 0.44, p = 0.005). Conclusions: The concentration of IL-17A and TIMP 1 may predict IVF/ET success. Further studies are required on the influence of the follicular fluid immunological environment on oocyte maturation and quality and, subsequently, embryo development.

Extended IgE profile of shrimp-sensitized patients based on Multiplex Examination ALEX2 - Allergy Explorer.

Introduction: The key to the correct diagnosis of shrimp allergy is a qualification to the most efficient diagnostic method and later interpretation of the result. To achieve this, it is necessary to apply a diagnostic strategy relevant to each patient's clinical situation and approach every case individually. Aim: In this study the allergen profile of shrimp-sensitized patients was analysed using ALEX2 Allergy Explorer. Material and methods: This study includes 50 adult patients with positive prick-by-prick tests with tiger shrimp bought from the local eco-market and an elevated concentration of IgE specific to the shrimp allergen extract (ImmunoCap). A total of 35 patients with negative skin prick tests with shrimp and not detectable sIgE shrimp in ImmunoCap were included in the control group. All patients had ALEX2 Allergy Explorer microarray test. Results: In the shrimp-sensitized group, 22 patients were sensitized to at least one allergen component of Penaeus monodon, 20 patients were sensitized to crab, and 20 were sensitized to lobster. Only 15 (30%) patients were sensitized to the Northern prawn (Pandalus borealis) allergen extract in ALEX2 and only 12 (24%) to Shrimp mix (Litopenaeus setiferus, Farfantepenaeus aztecus, Farfantepenaeus dourarum). Conclusions: Sensitization to shrimp tropomyosin in the research group was present only in 34% of cases. There may be other shrimp allergen components, not available in ALEX2, which are responsible for shrimp sensitization.

Nanohybrids of oxides nanoparticles-chitosan and their antimicrobial properties.

Growing international problem with pathogens acquiring resistance to antibiotics is the reason for the search for bactericidal substances against which microorganisms cannot become resistant. The aim of this study was to synthesize inorganic-organic nanohybrids and obtain materials with antimicrobial effects. Chitosan (CS) was deposited on nanocomposite carriers such as calcium oxide with titanium dioxide (CaO-TiO2), magnesium oxide with titanium dioxide (MgO-TiO2) and copper(II) oxide with titanium dioxide (CuO-TiO2). The efficiency of the process was examined at varying concentrations of chitosan and temperature. The parameters for nanohybrids synthesis were selected based on the highest amount of nano-chitosan deposited on the nanohybrids-for each carrier, the process conditions were as follows: chitosan solution at 5 g l-1and 20 °C. The materials were obtained using these parameters and were used for microbiological tests againstE. coliATCC 25922,S. aureusATCC 25923 andC. albicansATCC 10231. The growth inhibitory activity of the obtained materials was qualitatively defined. These results suggest that the synthesized nanohybrids and nanocomposites exhibit biostatic action. The material with the broadest effect was the CuO-TiO2-CS hybrid, which had biostatic properties against all tested strains at a minimal concentration of 1250µg ml-1. Further research is required to find eco-friendly, non-toxic, and more effective antimicrobials with a broad action to prevent the acquisition of resistance.

Experimental Research Models to Assess the Cross-Reactivity between Can f 5 and Human PSA-Two Different Perspectives.

The similarity in antigenic structures, including the degree of correspondence between the amino acid sequence and the spatial structure, is not always reflected in the actual cross-reactivity of allergens. Inhibition tests appear to be an invaluable tool for assessing potential cross-reactivity between allergens under natural conditions. In this publication, we present two experimental models of inhibition tests: solid phase (SP-IT) and liquid phase (LP-IT). As an exemplary research model, we used the cross-reactivity between human PSA and anti-Can f 5 IgE antibodies. We performed an SP-IT model using a microplate coated with human PSA. The LP-IT model was based on mixing anti-Can f 5 IgE positive serum with a material containing human PSA. Anti-Can f 5 IgE and PSA concentrations before and after inhibition were measured to evaluate inhibition effectiveness. The results of the performed experiments showed that both proposed models of inhibition tests are an effective tools for studying cross-reactive relationships between antigens. In the SP-IT, the concentration of anti-Can f 5 decreased by 21.6% and, in the LP-IT, it decreased by 34.51%. In turn, the PSA concentration in the SP-IT decreased by 11.25% and, in the LP-IT, it decreased by 15.49%. In conclusion, both the SP-IT and the LP-IT seem to be effective tools for assessing the actual cross-reactivity between different allergens.

Assessment of the post-SARS-CoV-2 vaccination response depending on the epidemiological status, demographic parameters and levels of selected cytokines in medical personnel.

Introduction: SARS-CoV-2 is responsible for causing the COVID-19 disease, which affected 174 million people worldwide. After vaccines were launched, the focus was on their effectiveness and the degree of their safety. Aim: The authors try to find factors that may affect the response to vaccination. Material and methods: The study was conducted in 47 adults (39 women and 8 men; age: 47.3 ±11.2). Participants were vaccinated with two doses of the Comirnaty mRNA vaccine. Each patient had a medical history taken and the concentration of specific sIgG antibodies against S1 protein and SARS-CoV-2 N protein, as well as of selected cytokines (IL-8, TGF-ß, IFN-γ) was determined before and 3 weeks after the first and second dose of the vaccine. Results: There were 18 convalescents among the respondents. A statistically significant increase in the concentration of specific sIgG S1 in subsequent determinations was observed. Higher levels of sIgG S1 were found after the first dose of the vaccine in COVID-19 convalescents. There was no statistically significant influence of age, body mass index and sex on the increase in the concentration of antibodies and the concentration of the determined cytokines. It was shown that the higher the initial TGF-ß concentration, the greater the increase in sIgG S1 after administration of the vaccine. Conclusions: Vaccination did not increase the levels of IL-8, IFN-ß and TGF-γ. A higher concentration of serum TGF-ß before vaccination correlated with the higher concentration of sIgG S1 antibodies after the first dose of the vaccine.

Synthesis and antimicrobial properties of CaCO3-nAg and nAg-CaCO3 nanocomposites.

The nanocomposites of nAg-CaCO3 and CaCO3-nAg with different mass content of silver nanoparticles were obtained in a two-stage continuous microwave process. The nanocomposites were characterized by XRD, TEM, UV-vis, and FTIR spectroscopy. The effects of microwave radiation, silver nanoparticles content in the composite and the molar ratio of tannic acid to silver on the size of salt crystallites were studied. The effectiveness of combining silver nanoparticles with CaCO3 was confirmed to exceed 99%. Depending on the input parameters applied, the size of salt crystals in nAg/CaCO3 nanocomposites ranged from 25 to 45 nm, while the size of salt crystals in CaCO3/nAg nanocomposites ranged from 26 to 41 nm. In addition, some of the compositions were tested for their antimicrobial activity on selected yeast and bacterial microorganisms. The prepared materials present a biocidal effect, which makes them suitable for use in antimicrobial preparations.

Circulating Leptin, Adiponectin, and Tumor Necrosis Factor-Alpha in Patients Undergoing Surgery Due to Colorectal Cancer.

BACKGROUND: Adipocytokines have been proposed as factors mediating associations between obesity and inflammation in patients with colorectal cancer (CRC). Thus, the aim of this study was to determine the clinical relationships between blood concentrations of leptin (LEP), adiponectin (ADP), and tumor necrosis factor alpha (TNF-alpha) and the outcomes measured in patients with CRC undergoing surgery. PATIENTS AND METHODS: History, body composition, and blood concentrations of LEP, ADP, and TNF-alpha were determined in 107 patients undergoing surgery due to CRC. The patients were followed up for 619.72 ± 371.65 days. RESULTS: Compared to patients with stage II CRC, individuals with clinical stage I CRC had significantly lower ADP and higher TNF-alpha blood concentrations. We found significant correlations between the clinical stage of CRC (early vs. localized vs. metastatic) and the following: crude blood ADP concentration (R = 0.25; p = 0.015), ADP-to-TNF-alpha ratio (R = 0.31; p = 0.002), and ADP when indexed to body surface area (R = 0.25; p = 0.008) and to fat mass (R = 0.25; p = 0.016). The risk of death during the long-term follow-up period was independently related to the clinical stage of CRC, impairment of the patient's functional status, and higher blood carcinoembryonic antigen concentration. In Kaplan-Meier survival analysis, patients with blood LEP concentrations adjusted to a visceral adipose tissue score of ≥0.47 had a significantly better likelihood of surviving than their counterparts. CONCLUSIONS: In patients with CRC undergoing surgery, blood ADP and TNF-alpha concentrations were associated with the clinical stage of the cancer, likelihood of radical tumor excision, occurrence of nonsurgical postoperative complications, and long-term survival, which suggests the role of dysregulation in the endocrine function of adipose tissue in response to the neoplasmatic process.

Function of Follicular Cytokines: Roles Played during Maturation, Development and Implantation of Embryo.

A balance within the immune system is necessary for the proper development of ovarian follicles. Numerous cytokines were detected in follicular fluid, the role of which in reproductive physiology seems crucial. They influence the development and maturation of the follicle, ovulation, and corpus luteum formation, as well as embryo implantation and maintenance of pregnancy. The analysis of follicular fluid requires its collection by puncturing of the ovary, which is usually executed in connection with various gynaecological procedures. When interpreting such test results, clinical indications for a given procedure and the method of patient preparation should be taken into account. This review revealed the results of currently available studies on the concentration of pro-inflammatory cytokines in follicular fluid in various forms of infertility. Additionally, it presented cytokines, whose concentration has a significant impact on the size of ovarian follicles, their number, the effectiveness of in vitro fertilisation, development of the embryo, and chances of correct implantation. Despite the many recent publications, the knowledge of follicular fluid immunology in the context of reproductive pathology is superficial and further research is required to extensively understand the roles of individual cytokines in reproductive pathology. In the future, this knowledge may enable patients' individual qualifications to individual methods of infertility treatment, as well as the possible adjustment of the treatment regimen to the patient's immune profile.

Assessment of serum concentrations of matrix metalloproteinase 1, matrix metalloproteinase 2 and tissue inhibitors of metalloproteinases 1 in atopic dermatitis in correlation with disease severity and epidermal barrier parameters.

INTRODUCTION: Matrix metalloproteinases (MMPs) are a group of proteolytic enzymes, conditioning the integrity of skin cells, however, their role in the inflammatory process of atopic dermatitis (AD) and the direct effect on the epidermal barrier parameters remain unexplained. AIM: To assess MMP-1, MMP-2, tissue inhibitors of metalloproteinases (TIMP)-1 concentrations in blood serum in the context of transepidermal water loss (TEWL) and stratum corneum hydration in AD. Moreover, serum levels of MMPs and TIMP-1 were analysed in relation to the Eczema Area and Severity Index (EASI). MATERIAL AND METHODS: Forty-three AD patients and 22 control group subjects have been investigated. Serum concentrations of MMP-1, MMP-2, and TIMP-1 have been evaluated with ELISA. TEWL and stratum corneum hydration have been assessed with a TM300 Tewameter and a CM825 Corneometer. Skin lesions in patients with AD have been evaluated with the Eczema Area and Severity Index. RESULTS: MMP-1 and MMP-2 serum concentrations were significantly higher in the AD group. The results of TIMP-1 serum concentration were similar for both groups. The correlation between the serum concentration and the EASI was demonstrated only for MMP-2 for patients with severe and moderate AD. Patients with AD and TIMP-1 serum concentration greater than MMP-1 presented lower TEWL and higher epidermal hydration. CONCLUSIONS: The results of this study warrant further investigation. The predominance of TIMP-1 over MMP-1 in blood serum can potentially limit TEWL and maintain the proper water content of the epidermis. Future work is necessary to establish how reliable the role of MMP-2 concentration is as an indicator of the severity of AD.

Evaluation of the effect of the interleukin-25 serum concentration on the intensity of the symptoms of atopic dermatitis and epidermal barrier.

INTRODUCTION: Interleukin 25 (IL-25) is a cytokine with proinflammatory and anti-inflammatory effects, and its biological function of reciprocal epidermal hyperplasia and of inhibiting the filaggrin synthesis points to an essential role connecting the inflammatory process with damage to the epidermal barrier in the course of atopic dermatitis (AD). AIM: To assess the IL-25 in serum concentration in AD patients and to analyse its possible correlation with the disease intensity and selected epidermal barrier parameters such as transepidermal water loss (TEWL). MATERIAL AND METHODS: The study involved 43 patients with AD and 22 healthy volunteers. The IL-25 concentration was measured using the ELISA method. The intensity of disease symptoms was investigated using W-AZS and EASI indicators. The epidermal barrier was evaluated using a Tewameter TM300 and Corneometer CM825. RESULTS: The concentration of IL-25 in serum was higher in the study group than in the control group. IL-25 serum concentration correlates with W-AZS/EASI in patients with a severe and moderate course of AD. The concentration of IL-25 affects the TEWL within the affected, evaluated skin surface. CONCLUSIONS: An elevated IL-25 concentration in serum is characteristic for patients with moderate and severe AD intensity. The IL-25 concentration in serum correlates with TEWL and with the moisture level in the affected area. However, further studies are necessary to determine the role played by IL-25 in the course of the disease and how it affects the functional parameters of the epidermal barrier.

Omalizumab improves forced expiratory volume in 1 second in patients with severe asthma.

INTRODUCTION: Asthma is a multiphenotypic disease, and therapeutic managenment in patients with severe asthma is particulary difficult, with conventional treatment of severe asthma showing poor efficacy. AIM: To analyse forced expiratory volume in 1 s (FEV1) following the adminstration of omalizumab. MATERIAL AND METHODS: Six patinents (mean age: 50 ±12.6) with severe, uncontrolled asthma according to the GINA guidelines were enrolled in the study. RESULTS: Treatment with omalizumab increased in all subjects FEV1 by 17.28 ±13.4% after months and 18.57 ±13.4% after 12 months of treatment. CONCLUSIONS: These results provides further evidence that therapy with omalizumab improves spiromtric parameters in severe asthma.

Insulin Signaling in Bupivacaine-induced Cardiac Toxicity: Sensitization during Recovery and Potentiation by Lipid Emulsion.

BACKGROUND: The impact of local anesthetics on the regulation of glucose homeostasis by protein kinase B (Akt) and 5'-adenosine monophosphate-activated protein kinase (AMPK) is unclear but important because of the implications for both local anesthetic toxicity and its reversal by IV lipid emulsion (ILE). METHODS: Sprague-Dawley rats received 10 mg/kg bupivacaine over 20 s followed by nothing or 10 ml/kg ILE (or ILE without bupivacaine). At key time points, heart and kidney were excised. Glycogen content and phosphorylation levels of Akt, p70 s6 kinase, s6, insulin receptor substrate-1, glycogen synthase kinase-3ß, AMPK, acetyl-CoA carboxylase, and tuberous sclerosis 2 were quantified. Three animals received Wortmannin to irreversibly inhibit phosphoinositide-3-kinase (Pi3k) signaling. Isolated heart studies were conducted with bupivacaine and LY294002-a reversible Pi3K inhibitor. RESULTS: Bupivacaine cardiotoxicity rapidly dephosphorylated Akt at S473 to 63 ± 5% of baseline and phosphorylated AMPK to 151 ± 19%. AMPK activation inhibited targets downstream of mammalian target of rapamycin complex 1 via tuberous sclerosis 2. Feedback dephosphorylation of IRS1 to 31 ± 8% of baseline sensitized Akt signaling in hearts resulting in hyperphosphorylation of Akt at T308 and glycogen synthase kinase-3ß to 390 ± 64% and 293 ± 50% of baseline, respectively. Glycogen accumulated to 142 ± 7% of baseline. Irreversible inhibition of Pi3k upstream of Akt exacerbated bupivacaine cardiotoxicity, whereas pretreating with a reversible inhibitor delayed the onset of toxicity. ILE rapidly phosphorylated Akt at S473 and T308 to 150 ± 23% and 167 ± 10% of baseline, respectively, but did not interfere with AMPK or targets of mammalian target of rapamycin complex 1. CONCLUSION: Glucose handling by Akt and AMPK is integral to recovery from bupivacaine cardiotoxicity and modulation of these pathways by ILE contributes to lipid resuscitation.

Intraosseous lipid emulsion: an effective alternative to IV delivery in emergency situations.

OBJECTIVE: To determine whether intraosseous infusion of a lipid emulsion reverses cardiac pharmacotoxicity in anaesthetized rats. DESIGN: Prospective, randomized animal study. SETTING: Academic research laboratory. SUBJECTS: Adult, male Sprague-Dawley rats. INTERVENTIONS: We assigned 25 male Sprague-Dawley rats into four groups: intraosseous lipid emulsion, intraosseous saline, IV lipid emulsion, and sham/null. Rats were anesthetized with 1.5% isoflurane and 95% oxygen. The left internal carotid artery and both internal jugular veins were cannulated and a flow probe was placed on the right carotid artery. Subsequently, in animals assigned to the intraosseous groups, the greater trochanter of the left proximal femur was exposed and the intraosseous space was cannulated. After surgical recovery, bupivacaine (10 mg/kg) was injected IV over 20 seconds followed 10 seconds later by treatment with one of the following: intraosseous lipid-emulsion (10 mL/kg over 180 s), intraosseous saline (10 mL/kg over 180 s), IV lipid-emulsion (10 mL/kg over 90 s), or no treatment (sham/null). MEASUREMENTS AND MAIN RESULTS: Electrocardiogram, aortic blood pressure, and carotid blood flow were recorded continuously. Rats treated with intraosseous lipid emulsion experienced a significantly faster recovery of hemodynamic variables (return of 50% flow; median [CI]: 160 s [105-263 s]) than did rats treated with saline (471 s [283-611 s]; p < 0.05) or animals with no treatment (415 s [340-539 s], p < 0.05), but at a similar rate to animals treated with IV lipid emulsion (176 s [152-217 s], p = not significant). All groups experienced persistent negative chronotropic effects. A compensatory increase in systemic arterial pressure was observed in rats treated with lipid emulsion. CONCLUSION: These proof-of-principle data indicate that intraosseous infusion of lipid emulsion rapidly reverses bupivacaine-induced cardiac toxicity in rats. Further studies are warranted to optimize this novel route of lipid emulsion injection in emergency situations when intravascular access is not secured.

The use of inhibition assay in Api g 7 suspected allergy in a female patient with anaphylaxis: A case report.

The symptoms of celery allergy are mainly presented as oral allergy symptom, but there are several case reports of patients who experienced anaphylaxis. Defensin (Api g 7), as a novel allergen in celery root, was described in 2022 r. The female patient had a history of several episodes of dyspnea and cough, associated with ingestion of spice mixes containing dried celery. Up to the point of hospitalization, there were no objective tests, either sIgE or skin prick tests, that would confirm celery sensitization. During hospitalization, patient had a positive double-blind placebo-controlled food challenge with cooked celery. The patient was sensitized to mugwort defensin Art v 1. An inhibition assay with celery allergen extract was performed to prove cross-sensitization between Art v 1 and celery allergen responsible for symptoms in the patient. In conclusion, Api g 7 is an important celery allergen that can be responsible for severe reactions. Its cross-reactivity with Art v 1 is characteristic. Negative diagnostic tests with celery do not exclude Api g 7 sensitization.

Diagnostics of Allergy to Furry Animals-Possibilities in 2024.

Diagnosing allergies is not always easy. Sometimes the symptoms reported by the patient do not match the results of diagnostic tests. Diagnostics of IgE-dependent allergy, including allergy to furry animals, can be based on two basic strategies, "BOTTOM-UP" and "TOP-DOWN", and almost all available allergy tests can be used ranging from skin tests, through specific IgE, to molecular panels and challenge tests. Currently, most information regarding the diagnosis and incidence of allergies to furry animals focuses almost exclusively on the two most common pets-dogs and cats. This is certainly due to the fact that allergy to cats and/or dogs is a common phenomenon, has a huge impact on quality of life, and is a challenge for doctors of many specialties. However, the growing number of owners of other pet species means that diagnostic methods must keep up with the changing needs of patients. Further research in these groups will certainly provide new facts and facilitate clinical decision-making when working with allergic patients.

Blood type ABO and the cytokine profile of follicular fluid in women undergoing IVF/ET.

OBJECTIVES: ABO blood type was hypothesised to be related to a number of infertility processes. There is still an open debate on ABO blood group's incompatibility and infertility. It was associated with ovarian reserve in women with subfertility. There is still not enough information on the influence of blood type and the immunology of follicular fluid (FF). MATERIAL AND METHODS: 78 patients were selected, who underwent in vitro fertilization (IVF) between April 2021 and January 2022. FF samples from each individual patient were taken on the day of ovarian puncture and stored at -80°C until immunological assessment. Concentration of chosen interleukins - IL-1α, IL-2, IL-4, IL-5, IL-6, IL-8 IL-10, IL-15, IL-1ß, IL-18, IFN, LIF, TNFα, GCSF and PIBF-1 were measured using commercially available ELISA kits. RESULTS: All assessed cytokines were present in the FF of exanimated patients. The concentration was compared to the blood type ABO of all women undergoing in vitro fertilization. No statistical relevance was found between blood type ABO and the concentration of GCSF, PIBF1, LIF, IL-15, IL-5, IL-8, IL-1 alfa, IL-1 beta, INF gamma, IL-2HS, IL-4HS, IL-6HS, IL-10HS in the FF obtained during ovarian puncture (p > 0,05). There was no statistically significant correlation between blood type ABO and the quality of embryo, and the positive pregnancy test in patients undergoing IVF/ET. CONCLUSIONS: The blood type ABO does not influence the wide cytokine profile of FF obtained during ovarian puncture in women with infertility of different origin, as well as embryo quality and pregnancy rate.