New Biosensor for Determination of Neuropilin-1 with Detection by Surface Plasmon Resonance Imaging.

Neuropilin-1 is transmembrane protein with soluble isoforms. It plays a pivotal role in both physiological and pathological processes. NRP-1 is involved in the immune response, formation of neuronal circuits, angiogenesis, survival and migration of cells. The specific SPRI biosensor for the determination of neuropilin-1 was constructed using mouse monoclonal antibody that captures unbound NRP-1 form body fluids. The biosensor exhibits linearity of the analytical signal between 0.01 and 2.5 ng/mL, average precision value 4.7% and recovery between 97% and 104%. The detection limit is 0.011 ng/mL, and the limit of quantification is 0.038 ng/mL. The biosensor was validated by parallel determination of NRP-1 in serum and saliva samples using the ELISA test, with good agreement of the results.

An Array SPRi Biosensor for the Determination of Follicle-Stimulating Hormone in Blood Plasma.

Follicle-stimulating hormone (FSH) regulates the development, growth, pubertal maturation and reproductive processes of the human body. The determination of serous FSH concentration is significant as an alternative to testicular biopsy in the case of boys suffering from cryptorchidism after orchidopexy, and as a means of determining the menopausal stage in women. The aim of this investigation is to develop a specific array surface plasmon resonance imaging (SPRi) biosensor for the determination of FSH in body liquids such as blood plasma, obtaining sufficient sensitivity to determine FSH at levels characteristic for that hormone in blood plasma, without any signal enhancement. The biosensor consists of a mouse monoclonal anti-FSH antibody attached to the gold surface of a chip via a cysteamine linker. Its linear response range is from 0.08 mIU mL-1 (LOQ) to 20 mIU mL-1, and well covers most of the range of FSH activities found in blood without dilution. The precision of measurement is between 3.2% and 13.1% for model samples, and between 3.7% and 5.6% for spiked plasma samples. Recoveries are in the range from 94% to 108%. The biosensor has good selectivity, and is validated by comparison with ECLE, with good agreement of the results.

Seroprevalance of anti-SARS-CoV-2 antibodies in the IgG class in hospitalized children in the north-eastern part of Poland in 2021.

Introduction: In children, SARS-CoV-2 infection may manifest with symptoms of fever, cough, muscle pain, diarrhea or waste smell and taste. However, in most cases it is mild or asymptomatic. Determination of the level of anti-SARS CoV-2 IgG antibodies in children enables to assess retrospectively the incidence of the SARS-CoV-2 infection among children. Aim of the study: The aim of the research was to analyze the results of anti-SARS-CoV-2 antibodies in the IgG class contained in children hospitalized in north-eastern Poland in 2021. Material and methods: The material for the study was venous blood collected once from children hospitalized in Children's University Hospital in Bialystok in 2021. The results obtained from 615 children aged from 1 month to 11 years were analyzed. Determination of the level of antibodies was performed using the automated SARS-COV-2 IgG II test on the VIDAS® analyzer. Results: The analysis of the results of the anti-SARS-CoV-2 IgG value showed that half of children (52%, n=319) had contact with SARS-CoV-2 virus of which approximately 15% (n=47) had documented infection with this virus. There was no difference in the frequency of infections among boys and girls, antibodies were detected in 51% and 53%, respectively. The highest percentage of positive results was observed in children aged 7 to 10 years, 62% (91/147). The least children with a positive result were in the group below 1 year of age, 24% (4/17). High values of the antibody index (>15.00) were most often found among younger children, aged 1-2 years. Conclusions: The presented results showed that infection with the SARS-CoV-2 virus does not depend on the sex of children but the value of anti-SARS-CoV-2 IgG antibodies decreased with the age of the child.

An Immunosensor for the Determination of Cortisol in Serum and Saliva by Array SPRi.

Cortisol is a hormone which plays an essential role in the immune, endocrine, cardiovascular, renal and skeletal systems. Its level increases in response to stress, illness, injury or exhaustion, and it is therefore a significant diagnostic biomarker of stress. An immunosensor for the determination of cortisol by SPRi array was developed. The receptive part of the immunosensor is mouse monoclonal antibody against cortisol, immobilized via cysteamine linker. The optimum pH of the immunosensor is 7.4, and the optimum concentration of the antibody is 50 ng mL-1. The immunosensor is specific for cortisol, and its linear response ranges from 0.20 ng mL-1 (LOQ) to 8 ng mL-1. The precision of the determination was between 3.1% and 3.3%, and the recovery between 99% and 102%. The immunosensor was validated by simultaneous determination of cortisol in serum and saliva samples by a standard method, with good agreement between the results.

Two Biosensors for the Determination of VEGF-R2 in Plasma by Array SPRi.

Vascular endothelial growth factor receptor 2 (VEGF-R2) is a marker of angiogenesis and metastasis of cancer. Two biosensors for the determination of VEGF-R2 in plasma have been developed. One of them is based on a pure gold chip, and the other on a silver/gold bimetallic chip; both have the receptor, monoclonal rabbit antibody specific for human VEGF-R2, attached to the chip via a cysteamine linker. The biosensor with the gold chip exhibits linearity of the analytical signal between 0.03 and 2 ng/mL, a precision of 1.4% and recovery between 99% and 102%. The biosensor with the bimetallic chip exhibits linearity between 0.03 and 1 ng/mL, a precision of 2.2% and recovery between 99% and 103%. Both biosensors tolerate a 1:100 excess of VEGF, VEGF-R1 and VEGF-R3. Both biosensors were validated by parallel determination of VEGF-R2 in 27 different plasma samples using the ELISA immunosensor assay, with very good agreement of the results. Thermodynamic parameters of the interaction of VEGF-R2 with the antibody were determined by QCM (Quartz Crystal Microbalance) and SPRi (Surface Plasmon Resonance imaging) measurements.

Comparison of the interleukin 27 expression for the acute and chronic phase of mononucleosis in children.

BACKGROUND: The objective of the current study was to assess the serum level of interleukin 27 in children with mononucleosis and to compare the expression of this cytokine in the acute and chronic phase of the infection. METHODS: The level of IL-27 was determined using commercial enzyme-linked immunosorbent assay (ELISA) kits (Diaclone SAS, Besancon, France). Other laboratory findings were determined using routine laboratory methods. RESULTS: Serum level of IL-27 was found to be significantly higher in children with mononucleosis in comparison with healthy subjects (almost a 4-fold increase, 15.7 vs. 4.2 pg/mL, p < 0.001). It was also significantly higher in the acute phase compared to the chronic stage of the disease (more than a two-fold increase, 20.7 vs. 9.64 pg/mL, p < 0.001). This cytokine positively correlated with ALT, AST, LDH activity and WBC count (R = 0.498, p < 0.001; R = 0.586, p < 0.001; R = 0.170, p < 0.05, R = 0.329, p < 0.05, respectively) in the whole study, and only with AST activity in the chronic phase subgroup (R = 0.684, p < 0.05). CONCLUSION: In conclusion, this study shows that serum concentration of interleukin 27 in children with mononucleosis is increased, thus confirming the on-going inflammatory process. We also suggest that IL-27 can be a useful indicator to differentiate between the acute and chronic phase of the disease.

An SPRi Biosensor for Determination of the Ovarian Cancer Marker HE4 in Human Plasma.

Human epididymis protein 4 (HE4) is an ovarian cancer marker. Various cut-off values of the marker in blood are recommended, depending on the method used for its determination. An alternative biosensor for HE4 determination in blood plasma has been developed. It consists of rabbit polyclonal antibody against HE4, covalently attached to a gold chip via cysteamine linker. The biosensor is used with the non-fluidic array SPRi technique. The linear range of the analytical signal response was found to be 2-120 pM, and the biosensor can be used for the determination of the HE4 marker in the plasma of both healthy subjects and ovarian cancer patients after suitable dilution with a PBS buffer. Precision (6-10%) and recovery (101.8-103.5%) were found to be acceptable, and the LOD was equal to 2 pM. The biosensor was validated by the parallel determination of a series of plasma samples from ovarian cancer patients using the Elecsys HE4 test and the developed biosensor, with a good agreement of the results (a Pearson coefficient of 0.989). An example of the diagnostic application of the developed biosensor is given-the influence of ovarian tumor resection on the level of HE4 in blood serum.

Vascular endothelial growth factor and transforming growth factor ß in hypertrophic adenoids in children suffering from otitis media with effusion.

OBJECTIVES: The study objective was to assess the levels of VEGF-A and TGF-ß cytokines in the children with adenoid hypertrophy concomitant with exudative otitis media (OME) and in children with adenoid hypertrophy (HA) alone. METHODS: The study material consisted of hypertrophic adenoids removed during adenoidectomy from 39 children (20 girls and 19 boys), aged 2-7 years suffering from OME. The reference group included 41 children (19 girls and 22 boys), aged from 3 to 9 years with adenoid hypertrophy. The levels of VEGF-A and TGF-ß were determined in supernatants obtained from phytohemagglutinin-stimulated cell cultures of the adenoids using a commercial enzyme-linked immunosorbent assay kit. RESULTS: The median VEGF-A and mean TGF-ß concentrations in the study group were significantly higher than those in the reference group (503 pg/mL versus 201 pg/mL, P < 0.001 and 224 pg/mL versus 132 pg/mL, P < 0.001, respectively). ROC analysis revealed that the area under the curve (AUC) for VEGF-A was 0.952 with diagnostic sensitivity and specificity of 95%, whereas for TGF-ß it was 0.902 with 60% sensitivity and the same specificity as for VEGF-A. There was no significant difference between the AUC for VEGF-A and TGF-ß (P = 0.573). CONCLUSIONS: The changes in the levels of VEGF-A and TGF-ß may indicate bacterial pathogen as one of the causes of exudative otitis media in children. Determination of VEGF-A and TGF-ß could be used as additional and objective tests to confirm the clinical diagnosis.

The influence of different intensity of treatment on hormonal markers of gonadal function in acute lymphoblastic leukemia survivors.

Anti-cancer treatment in children can deteriorate gonadal function and affect future fertility. We analyzed the hormonal markers of gonadal function in adolescent leukemia survivors, treated in childhood with different levels of aggressiveness. We analyzed hormone levels in 69 adolescents and young adults, leukemia survivors stratified into standard (SR), intermediate (IR), and high (HR) risk groups, and in 80 healthy controls (38 men) at a similar age. We assessed follicular stimulating hormone (FSH), luteinizing hormone (LH), and inhibin B in the whole group, testosterone in males, and E2 and anti-Müllerian hormone (AMH) in females. Males classified into HR group presented, in comparison to control, higher levels of FSH, LH, lower inhibin B, and normal testosterone, whereas in SR and IR group, the hormonal values were comparable to the control. In females, in all risk groups, the levels of FSH, LH, E2, and inhibin B were comparable with the control, but the mean AMH levels were slightly lowered. We did not observe the effect of prophylactic cranial irradiation (12 or 18 Gy) or the time of treatment (before vs. during puberty) on hormone levels. In females, a positive correlation was found between the time interval after the end of treatment and AMH levels. Male leukemia survivors having undergone more intensive chemotherapy show the symptoms of disturbed spermatogenesis and need to be followed-up in the future. Women, irrespective of the risk group, can develop the signs of preterm ovarian insufficiency. They should be informed about the impact of the treatment on gonadal function.

Use of the Sysmex XT-4000i hematology analyzer in the differentiation of cerebrospinal fluid cells in children.

BACKGROUND: Routine analysis of pleocytosis and cellular composition of cerebrospinal fluid (CSF) is carried out with a phase-contrast microscope. The use of hematological analyzers seems to be an alternative to the manual method. The aim of the study was to assess the usefulness of the automated technique for counting and differentiating CSF cells in children. METHODS: The study group consisted of 59 children (28 girls and 31 boys) aged from 4 to 17 years suffering from viral and bacterial meningitis. Children were divided into three subgroups according to CSF cell count: 1st group had a pleocytosis of 6-50 cells/µL, 2nd group-51-100 cells/µL, and 3rd group->100 cells/µL. A reference group involved 32 children (17 girls and 15 boys) aged from 2 to 18 years with a normal range of 0-5 cells/µL. Examination of CSF was performed in parallel by two different method, manual and automated. RESULTS: The analysis of pleocytosis revealed that the values obtained by the manual method were statistically significantly lower in relation to the values obtained by automated technique in subgroups I and II. The number of mononuclear and polymorphonuclear cells in subgroups I, II, and III determined by both manual and automated methods was comparable. CONCLUSION: We conclude that automated method cannot fully replace the previously used manual method and some of the dubious cases, such as samples with low pleocytosis rates or abnormal cells indicated by the analyzer, will still require microscopic examination.

Diagnostic Usefulness of Insulin-Like Growth Factor 1 and Insulin-Like Growth Factor Binding Protein 3 in Children with Suspected Pituitary Dwarfism.

BACKGROUND: Pituitary dwarfism (also known as short stature) is a medical condition in which the pituitary gland does not produce enough growth hormone (GH). To confirm the diagnosis of growth hormone deficiency the overnight profile of GH secretion and GH provocative tests are usually performed; however, due to wide GH fluctuations throughout the day and night and the invasiveness of stimulation tests, their clinical utility is limited. Therefore, screening for IGF-1 (insulin-like growth factor 1) and IGFBP-3 (insulin-like growth factor binding protein type 3) is proposed, suggesting that these tests provide a more accurate reflection of the mean plasma GH level, although the results of these tests are still problematic. In this context, the aim of this study was to assess the diagnostic usefulness of IGF-1 and IGFBP-3 in children with suspected pituitary dwarfism. METHODS: Studies were carried out in 127 children with abnormal growth and low spontaneous 24-hour plasma GH profiles and abnormal results of GH stimulation tests. Fasting serum IGF-1 and IGFBP-3 were determined by chemiluminescent quantitative measurement using the IMMULITE 1000 IGF-1 and IGFBP-3 kits (Siemens Healthcare Diagnostics, United Kingdom) on the IMMULITE 1000 analyzer (Siemens Healthcare Diagnostics, USA). Results were compared to the normal range by children's age. RESULTS: Mean serum IGF-1 concentrations were within the lower normal range (41.7% cases), and 58.3% results were below the normal reference range in the study group. The average serum IGFBP-3 levels were within the lower normal range. CONCLUSIONS: We conclude that IGF-1 test can be a useful tool in the diagnosis of pituitary dwarfism in children suspected of this condition, but due to relatively poor sensitivity the testing cannot be performed alone, but in combination with other tests. The IGFBP-3 test is not useful for the diagnosis of this disease.

Comparison of the Diagnostic Power of Serum IL-6, IL-8 and TNF-α for the Idiopathic Anterior Uveitis in Children.

BACKGROUND: Uveitis is the inflammation of the uvea that often occurs in children. There are many causes of disease, but some of them do not have any reasons and are then called idiopathic uveitis. Cytokines play an important role in the regulation of the immune response. Determination of cytokine profiles could contribute to the explanation of the etiology of uveitis and could serve to evaluate the inflammation intensity as well as be helpful in the early diagnosis this disease. The purpose of this study was to determine the serum level of selected inflammatory cytokines, such as interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor α (TNF-α) and to compare their diagnostic power as a markers of idiopathic anterior uveitis in children. METHODS: The study was carried out on 28 children diagnosed with idiopathic anterior uveitis. The reference group comprised 30 healthy children. Serum IL-6, IL-8, and TNF-α concentrations were measured with specific enzyme-linked immunoassay (ELISA) methods. RESULTS: The mean values of IL-6, IL-8, and TNF-α in the children with idiopathic anterior uveitis were significantly higher than those found in the reference group. The highest sensitivity, specificity, accuracy, positive and negative predictive value, and likelihood ratio of a positive test result were achieved for IL-8. There was a significant difference between the area under the curve for IL-6 and IL-8. CONCLUSIONS: Increased serum concentrations of interleukin IL-6, IL-8, and TNF-α may suggest that these cytokines induce inflammatory changes in the ocular surface. Analysis of cytokine levels showed that IL-8 has the highest diagnostic power and is the best marker for diagnosis of idiopathic anterior uveitis in children.

[Vitamin D deficiency in psoriatic patients in north-east of Poland].

Background: Recent studies point out the important role of vitamin D in the pathogenesis of many autoimmune diseases, in between psoriasis. Vitamin D regulates function of the dendritic cells, proliferation and maturation of the keratynocytes and lymphocytes T. The aim of the study was to evaluate serum vitamin D (25OHD) concentrations in psoriatic patients in the north-east of Poland and their comparison in the summer and winter period of the year. We also evaluated relationship between 25OHD concentration and disease severity, prevalence of psoriatic comorbidities and laboratory results. Material and Methods: 25OHD serum concentration was evaluated by electrochemiluminescent method in 115 patients with exacerbated plaque type psoriasis. 39 patients were evaluated in the summer and 79 in the winter time. 28 patients from winter group were treated with narrow band ultraviolet B radiation (NB-UVB). The results were compared with 38 healthy persons. Results: 25OHD serum concentrations of psoriatic patients were significantly lower than in the control group (p=0,0003). Vitamin D deficiency was diagnosed in 66% of patients in the summer time and in 63% in winter time; in the control group: 24% during the summer and 71% during the winter. After UVB phototherapy we observed reduction of skin lesions, measured as a significant decline in the Psoriasis Area and Severity Index (PASI) (p<0.001). The serum 25OHD concentration increased (p<0.001). After the treatment 65% of the psoriatic patients reached normal range of 25OHD concentration, 35% of patients were still vitamin D insufficient. We demonstrate positive correlation between increase in 25OHD and number of NB-UVB phototherapy sessions (r=0.38). Conclusions: We have observed vitamin D deficiency both in psoriatic patients and in the control group. Among psoriatic patients the praevalence of deficiency were higher than in the control group, especially during the summer months. Frequent vitamin D defficiency in the groups studied indicates the need of for its supplementation. The 25OHD serum concentrations increased after phototherapy with UVB.

Circulating Levels of Soluble α-Klotho and FGF23 in Childhood Cancer Survivors: Lack of Association with Nephro- and Cardiotoxicity-A Preliminary Study.

Background/Objectives: The survival rate among pediatric cancer patients has reached 80%; however, these childhood cancer survivors (CCSs) are at a heightened risk of developing chronic conditions in adulthood, particularly kidney and cardiovascular diseases. The aims of this study were to assess the serum α-Klotho and FGF23 levels in CCSs and to determine their association with nephro- and cardiotoxicity. Methods: This study evaluated a cohort of 66 CCSs who remained in continuous remission, with a mean follow-up of 8.41 ± 3.76 years. Results: The results of this study revealed that CCSs exhibited significantly higher levels of soluble α-Klotho compared to healthy peers (1331.4 ± 735.5 pg/mL vs. 566.43 ± 157.7 pg/mL, p < 0.0001), while no significant difference was observed in their FGF23 levels. Within the participant cohort, eight individuals (12%) demonstrated a reduced estimated glomerular filtration rate (eGFR) below 90 mL/min/1.73 m2. The relationship between treatment with abdominal radiotherapy and reduced eGFR was confirmed (p < 0.05). No correlations were found between potential treatment-related risk factors, such as chemotherapy or radiation therapy, serum levels of α-Klotho and FGF23, and nephro- and cardiotoxicity. Conclusions: In conclusion, this preliminary cross-sectional study revealed elevated levels of α-Klotho among childhood cancer survivors but did not establish a direct association with anticancer treatment. The significance of elevated α-Klotho protein levels among CCSs warrants further investigation.

Vitamin D Deficiency and Carotid Media-Intima Thickness in Childhood Cancer Survivors.

Childhood cancer survivors (CCS) are predisposed to developing numerous late effects of anticancer treatment later in life. The existing literature suggests that vitamin D deficiency (VDD) may influence cardiovascular abnormalities and metabolic diseases. The objectives of this study were to investigate the prevalence of VDD among childhood cancer survivors and examine the association of vitamin D deficiency and carotid intima-media thickness (IMT). The study comprised 111 childhood cancer survivors (62 males, 49 females) with a median follow-up time of 6.14 years. Vitamin D status was determined by measuring serum 25(OH)D levels using the automatic immunoenzymatic method. Ultrasonography of the common carotid artery (CCA), the carotid bulb, and the proximal part of the internal carotid artery (ICA) was conducted. Vitamin D deficiency (<20 ng/mL) was detected in 69.4% of CCS. A higher parathormone level and increased BMI were observed among VDD survivors. No effects of type of diagnosis, radiotherapy or hematopoietic stem cell transplantation on vitamin D status were observed. Our findings reveal that survivors with VDD exhibited significantly greater thickness in the CCA and carotid bulb. In conclusion, the results of our study of childhood cancer survivors demonstrate that vitamin D deficiency is prevalent in up to 70% of individuals. We did not confirm the hypothesis that factors related to anticancer treatment used during childhood contributed to the higher prevalence of VDD. Additionally, we did not verify the contribution of vitamin D deficiency to the increase in IMT thickness.

Anti-Müllerian hormone levels in serum 1 year after unilateral orchiopexy.

The aim of this study was to determine the serum anti-Müllerian hormone (AMH levels) in a group of boys with cryptorchidism before and 1 year after surgery. Additionally, the effect of orchiopexy, based on the testicular position and morphology, was evaluated. Fifty boys who were 1-4 years old (median, 2.4 years) with unilateral cryptorchidism were evaluated before and 1 year after they underwent orchiopexy. Prior to the procedure, all of the patients had undergone karyotyping to exclude chromosomal abnormalities. Blood samples were collected while obtaining blood for standard laboratory tests routinely performed just before and 12 months after surgery. Twelve months after the orchiopexy, in most cases surgically treated testes were in the middle or lower pole of the scrotum. The descended glands were of comparable size and density in comparison with the testes on the opposite side. In the treated children, serum AMH levels measured at 12 months after orchiopexy were similar to preoperative basal levels. The authors found that AMH levels in children with unilateral undescended testes do not change significantly 1 year after surgical procedure. We still do not have trustworthy factors to predict fertility in children affected with cryptorchidism. We observed an upward trend in AMH concentration 1 year after orchiopexy, but it was statistically insignificant. Therefore, we plan further measurements of AMH in boys after orchiopexy to assess the influence of this procedure on gonadal function.

Assessment of gonadal function in boys and adolescents at the diagnosis of neoplastic disease.

OBJECTIVE: We assessed the gonadal function in boys with a newly diagnosed neoplastic disease prior to chemotherapy. Eighty-four boys (48 prepubertal and 36 pubertal) were evaluated, including 50 with acute lymphoblastic leukemia (ALL) or non-Hodgkin lymphoma (NHL), 10 with Hodgkin lymphoma (HL), and 24 with solid tumors. The control group consisted of 24 healthy prepubertal and 24 pubertal boys. The levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), inhibin B, and testosterone were determined, and testicular volumes were measured. RESULTS: Patients in prepuberty and early puberty (Tanner stages 1-3) diagnosed with ALL/NHL or solid tumor presented normal serum reproductive hormone levels, whereas in ALL/NHL patients in Tanner stages 4-5, the mean values of inhibin B were significantly lower (45.18 +/- 33.85 vs. 153.57 +/- 71.44 ng/L, p = 0.0027). In patients with HL in Tanner stages 4-5, a statistically significant lower mean inhibin B level (100.44 +/- 67.45 versus 153.57 +/-71.44 ng/L, p = 0.0027), higher mean FSH level (6.3 +/- 3.6 versus 4.6 +/- 2.2 mIU/mL, p = 0.05), and higher mean LH level (5.9 +/- 4.0 versus 3.6 +/- 1.8 mIU/mL, p = 0.05) were observed. No statistically significant differences were noted in assessed hormones in patients with solid tumors, independently of Tanner stage. CONCLUSION: Our analysis indicates that adolescents with ALL/NHL and HL prior to treatment, exhibit reduced levels of inhibin B, which indirectly suggests the possibility of spermatogenesis dysfunction.

Increased percentage of T cells with the expression of CD127 and CD132 in hypertrophic adenoid in children with otitis media with effusion.

The hypertrophic adenoid may promote chronic suppurative otitis media in children as it fulfills its immune function. The number of lymphocytes in the adenoid and their cooperation in the immune response depend of on their proliferation and migration to the effector sites. Interleukin 7 (IL-7) is essential for the normal development and function lymphocytes. IL-7 plays pivotal role for activation and proliferation of T and B cells. The heterodimeric interleukin-7 receptor (IL-7R) is composed of the IL-7Rα (127) and the common cytokine receptor γc (CD132). The aim of this study was to evaluate the percentage of lymphocytes T (CD4(+) and CD8(+)) with IL-7R (CD127 and CD132) expression in hypertrophic adenoid in children suffering with otitis media with effusion for a duration of 3 months. Adenoid excised due to hypertrophy with or without chronic otitis media with effusion was used as study material. CD4(+) CD127(+), CD4(+)132(+), CD8(+)CD127(+) and CD8(+)CD132(+) cell subpopulations were identified using monoclonal antibodies and flow cytometry. The percentage of CD4(+) and CD8(+) T cells with CD127 receptor expression in hypertrophic adenoid of children with otitis media with effusion was statistically significantly higher than in hypertrophic adenoid group. The percentage of CD4(+) T cells with CD132 expression in the study group was statistically significantly higher than in the reference group. The percentage of CD8(+) T cells with CD132(+) expression was not statistically different in both groups. The increased percentage of T lymphocytes with IL-7R expression (CD127 and CD132) in hypertrophic adenoid seems to influence the quantity of lymphocytes and upset the immunological function of tonsils which can influence the course of otitis media with effusion.

The Relationship between Selected Inflammation and Oxidative Stress Biomarkers and Carotid Intima-Media Thickness (IMT) Value in Youth with Type 1 Diabetes Co-Existing with Early Microvascular Complications.

Recent years have confirmed the importance of oxidative stress and biomarkers of inflammation in estimating the risk of cardiovascular disease (CVD) and explaining not fully understood pathogenesis of diabetic macroangiopathy. We aimed to analyze the relation between the intima-media thickness (IMT) of common carotid arteries and the occurrence of classical cardiovascular risk factors, together with the newly proposed biomarkers of CVD risk (high-sensitivity C-reactive protein (hsCRP), myeloperoxidase (MPO), adiponectin, N-terminal-pro B-type natriuretic peptide (NT-proBNP) and vitamin D) in youth with type 1 diabetes (T1D) recognized in screening tests to present early stages of microvascular complications (VC). The study group consisted of 50 adolescents and young adults with T1D, mean age 17.1 years (10-26 age range), including 20 patients with VC (+) and 30 VC (-). The control group (Control) consisted of 22 healthy volunteers, mean age 16.5 years (11-26 age range). In the VC (+) patients, we found a significantly higher concentration of HbA1c, lipid levels, hsCRP and NT-proBNP. BMI and blood pressure values were highest in the VC (+) group. Higher levels of MPO and lower levels of vitamin D were found in both diabetic groups vs. Control. IMT in VC (+) patients was significantly higher and correlated positively with HbA1c, hsCRP, NT-pro-BNP and negatively with vitamin D levels. In conclusion, youth with T1D and VC (+) present many abnormalities in the classical and new CVD biomarkers. hsCRP and MPO seem to be the most important markers for estimating the risk of macroangiopathy. NT-proBNP may present a possible marker of early myocardial injury in this population.

Two Biosensors for the Determination of Interleukin-6 in Blood Plasma by Array SPRi.

Interleukin-6 (IL-6) is a biomarker of inflammation, the advanced stage of COVID-19, and several cancers, including ovarian cancer. Two biosensors for the determination of IL-6 in blood plasma by array SPRi have been developed. One of these biosensors consists of the mouse monoclonal anti-IL-6 antibody as the receptor immobilized via the cysteamine linker. The second contains galiellalactone as the receptor, being an inhibitor specific for IL-6, immobilized via octadecanethiol (ODM) as the linker. Both biosensors are specific for IL-6. The biosensor with the antibody as the receptor gives a linear analytical response between 3 (LOQ) and 20 pg mL-1 and has a precision between 8% and 9.8% and recovery between 97% and 107%, depending on the IL-6 concentration. The biosensor with galiellalactone as the receptor gives a linear analytical response between 1.1 (LOQ) and 20 pg mL-1, and has a precision between 3.5% and 9.3% and recovery between 101% and 105%, depending on IL-6 concentration. Both biosensors were validated. Changes in IL-6 concentration in blood plasma before and after resection of ovarian tumor and endometrial cyst, as determined by the two developed biosensors, are given as an example of a real clinical application.