Toll-like receptor (TLR2, TLR4) polymorphisms and their influence on the incidence of urinary tract infections in children with and without urinary tract malformation.

Introduction: Toll-like receptors (TLRs) contribute to the innate immune system. They are an element of non-specific immunity, which enables organisms to react quickly to foreign antigens, without being previously exposed to them. TLRs are pattern recognition receptors. TLR gene polymorphisms are widely investigated in connection with various infections. The aims of the study were: to investigate the role of TLR2 and TLR4 polymorphisms in the course of urinary tract infections (UTIs); to test for differences in distribution of these polymorphisms between children with urinary tract malformations suffering from recurrent UTI (rUTI), children with malformations but without rUTI and healthy controls; to determine whether these polymorphisms predispose to rUTI; and to analyse how polymorphisms and urine neutrophil gelatinase-associated lipocalin (NGAL) and interleukin 8 (IL-8) concentrations affect one another. Material and methods: The group consisted of 133 children (1-18 years old), 68 female and 65 male. The group was divided into 4 subgroups: A (rUTI with urinary tract malformations), B (urinary tract malformations without rUTI), C (rUTI) and D (healthy controls). Polymorphisms were analysed using PCR-RFLP. IL-8 and NGAL urine concentrations were established using immunoenzymatic methods. Results: TLR2 Arg753Gln and TLR4 Arg299Gly appeared significantly more often among children with rUTI. No correlation between urine IL-8 and urine NGAL and polymorphisms was found. Urine NGAL concentration was significantly higher among children with urinary tract malformations. Conclusions: TLR2 Arg753Gln and TLR4 Asp299Gly may predispose to rUTI. Urine NGAL concentration suggests the presence of kidney tissue injury, of varying degrees, among children with urinary tract malformations.

CSF levels of myelin basic protein in pediatric patients with ventriculoperitoneal shunt infection.

INTRODUCTION: Hydrocephalus is a common disorder of the central nervous system (CNS) in the pediatric population. Surgical treatment options involve ventriculoperitoneal shunt (VPS) placement. VPS infection is the most common complication of surgically treated hydrocephalus in pediatric patients [1, 2],which may lead to neuronal damage. Myelin basic protein (MBP) has been proposed as a marker of neuronal injury in a variety of contexts, and MBP levels in the cerebrospinal fluid (CSF) may be used to assess the severity of neuronal damage [1, 3, 4]. Therefore, the aim of this study was to evaluate the CSF level of myelin basic protein (MBP) in a group of pediatric patients with VPS infection. MATERIAL AND METHODS: Thirty CSF samples were collected from pediatric patients with VPS infection. CSF levels of MBP were measured at three time points, marked by contamination detection, obtention of the first sterile CSF culture, and VPS shunt implantation. The collected data were compared with those of the control group composed of children with active congenital hydrocephalus and valid CSF values. RESULTS: The MBP level in the study group was higher than the corresponding control values in the second and third measurements. The highest MBP level was reached in the study group in the second and third measurements. CONCLUSIONS: The lack of normalization of MBP level in the CSF of children with shunt infection could be connected with ongoing brain damage. It takes longer than the normalization of CSF protein level and pleocytosis. The delay is associated with a prolonged reaction of the immunological system.

Vaccination in children with chronic severe neutropenia - review of recommendations and a practical approach.

While the management of childhood neutropenia associated with a modifiable factor should be appropriate for the primary cause, there are misconceptions regarding the management of severe congenital neutropenia, immune neutropenia and cases classified as "idiopathic". Antibiotic prophylaxis or granulocyte-colony stimulating factor (G-CSF) are prescribed by specialists in pediatric hematology or immunology, whereas immunization may be conducted by primary care physicians should clear recommendations by provided. There is a belief that severe neutropenia, as an immunodeficiency, is associated with compromised effectiveness and increased rate of complications of immunization. The immunization might be delayed or omitted, increasing the risk of unnecessary infection. We discuss the available data and recommendations regarding vaccination of children with chronic severe neutropenia. While there are virtually no studies addressing the safety and effectiveness of vaccination in neutropenia, expert opinions provide information on immunization policy in "phagocytic cells defects" or explicitly neutropenia. There are no contraindications for inactivated vaccines in neutropenia. Live bacterial vaccines are contraindicated. While in general the vaccination with live viral vaccines is encouraged, occasionally neutropenia might be associated with defects of adaptive immunity, which would preclude the administration of such vaccines. Although this should be easily phenotypically identified, we propose assessing immunoglobulin levels and performing a low-cost flow cytometry test for major lymphocyte subpopulations to exclude significant defects in adaptive immunity before administration of live viral vaccines to such patients. This can improve the adherence of patients' guardians and physicians to proposed vaccination policy and the professional and legal safety associated with the procedure.

Unexpected eosinophilia in children affected by hydrocephalus accompanied with shunt infection.

PURPOSE: The aim of the article is to describe an immunological reaction to shunt infection in children with hydrocephalus. The main cause of shunt infection involves methicillin resistant Staphylococcus epidermidis (Bhatia et al. Indian J Med Microbiol 35:120-123, 2017; Hayhurst et al. Childs Nerv Syst 24:557-562, 2008; Martínez-Lage et al. Childs Nerv Syst 26: 1795-1798, 2010; Simon et al. PLoS One, 2014; Snowden et al. PLoS One 8:e84089, 2013; Turgut et al. Pediatr Neurosurg 41:131-136, 2005), a bacterial strain which is responsible for the formation of biofilm on contaminated catheters (Snowden et al. PLoS One 8:e84089, 2013; Stevens et al. Br J of Neurosurg 26: 792-797, 2012). METHODS: The study group involved 30 children with congenital hydrocephalus after shunt system implantation, whose procedures were complicated by S. epidermidis implant infection. Thirty children with congenital hydrocephalus awaiting their first-time shunt implantation formed the control group. The level of eosinophils in peripheral blood was assessed in both groups. Cerebrospinal fluid (CSF) was examined for protein level, pleocytosis, interleukins, CCL26/Eotaxin-3, IL-5, IL-6, CCL11/Eotaxin-1, CCL3/MIP-1a, and MBP. Three measurements were performed in the study group. The first measurement was obtained at the time of shunt infection diagnosis, the second one at the time of the first sterile shunt, and the third one at the time of shunt reimplantation. In the control group, blood and CSF samples were taken once, at the time of shunt implantation. RESULTS: In the clinical material, the highest values of eosinophils in peripheral blood and CSF pleocytosis were observed in the second measurement. It was accompanied by an increase in the majority of analyzed CSF interleukins. CONCLUSION: CSF pleocytosis observed in the study group shortly after CSF sterilization is presumably related to an allergic reaction to Staphylococcus epidermidis, the causative agent of ventriculoperitoneal shunt infection.

Lectin pathway factors in patients suffering from juvenile idiopathic arthritis.

Both complement activation and certain infections (including those with Yersinia sp.) may contribute to the pathogenesis of juvenile idiopathic arthritis (JIA). We investigated factors specific for the lectin pathway of complement: mannose-binding lectin (MBL), ficolins and MBL-associated serine protease-2 (MASP-2), in 144 patients and 98 controls. One hundred and six patients had oligoarticular disease and 38 had polyarticular disease. In 51 patients (out of 133 tested), Yersinia-reactive antibodies were found (JIA Ye+ group). MBL deficiency was significantly more frequent in the JIA Ye+ group than in patients without Yersinia-reactive antibodies or in controls. Median serum ficolin-2 level was significantly lower (and proportion of values deemed ficolin-2 insufficient greater) in JIA patients irrespective of their Yersinia antibody status. The minority (C) allele at -64 of the FCN2 gene was less frequent among JIA patients than among control subjects. No differences were found in the frequency of FCN3 gene +1637delC or MASP2 +359 A>G mutations nor for median values of serum ficolin-1, ficolin-3 or MASP-2. However, high levels of serum ficolin-3 were under-represented in patients, in contrast to MBL. MBL, ficolin-1, ficolin-2, ficolin-3 and MASP-2 were also readily detectable in synovial fluid samples but at a considerably lower level than in serum. Our findings suggest a possible role for the lectin pathway in the pathogenesis of JIA, perhaps secondary to a role in host defence, and indicate that investigations on the specificity of lectin pathway recognition molecules towards specific infectious agents in JIA might be fruitful.

[Evaluation of selected parameters of specific immunity in children with osteosarcoma at various stages of antitumour treatment].

UNLABELLED: Typical cells of specific immunity are lymphocytes. T cells may specifically recognize antigens using T Cell Receptors (TCRs). Antigen presentation by specialized cells is a necessary element of specific immunity. It leads to initiating an immune response. After antigen dependent activation T cells transform to memory and effector lymphocytes. Cells engaged directly in destruction of the tumour are CD8+cytotoxic T lymphocytes, CD4+ lymphocytes, Tγδ lymphocytes, NK, NKT cells and indirectly B lymphocytes. One of the main methods of osteosarcoma treatment in children is chemotherapy. The goal is to destroy the cancer cells by putting them in a state of apoptosis. All of the medications used as chemotherapy carry the risk of short-term and long-term problems including leukopenia, immune disorders such as immunodeficiency. THE AIM OF STUDY: was evaluation by flow cytometry selected elements of specific cellular immunity in children with osteosarcoma at various stages of antitumour treatment. MATERIALS AND METHODS: The study was performed on the group of 44 children with osteosarcoma, aged from 6 to 20 years (average 14.9 years; median 15.0 years). T and B lymphocytes and subpopulations: CD4+, CD8+, CD3+?HLA-DR+, CD3+γδ; NK, NKT cells were analyzed in peripheral blood with use of flow cytometry method with monoclonal antibodies. Examinations were performed before the therapy - in diagnostic period (examination I), after the neoadjuvant chemotherapy (examination II), 10-14 days after the surgery (examination III), 5 months after the surgery (after adjuvant chemotherapy, examination IV). RESULTS: The number of T and B lymphocytes was decreasing after each stage of cytostatic therapy, with the biggest differences for CD19+ cells (medians: I examination - 205.0; II exam. - 62.0; IV exam. - 24.0 cells/mL); in single cases the number of cells decreased even under 10/mL (norm 200-500 cells/mL). CONCLUSIONS: 1. In children and youth with osteosarcoma antineoplastic treatment contributes to the suppression of the immune system, decreasing definitely the number and percentage of B lymphocytes, T helper lymphocytes and NK cells. 2. Decreased number of CD3+, CD4+, CD19+ and NK lymphocytes during chemotherapy may contribute to the progression of neoplastic disease in the future, after treatment. 3. Evaluation of immunologic status in patients with osteosarcoma may be helpful in monitoring of antineoplastic therapy effectiveness, may prevent the formation of unfavourable clinical changes and may be the basis for correction of the cytostatic agents' administration.

Successful haploidentical PBSCT with subsequent T-cell addbacks in a boy with HyperIgM syndrome presenting as severe congenital neutropenia.

HIGM syndrome is a group of primary immunodeficiency disorders characterized by recurrent bacterial and opportunistic infections; it is also associated with normal to elevated serum IgM levels and a concomitant deficiency of IgG, IgA, and IgE. In this report, we give account of a boy with X-linked HIGM and a novel Y172C mutation within his CD40LG gene. He presented with severe neutropenia as the dominating symptom. His bone marrow showed maturation arrest at the promyelocyte/myelocyte stage, typical of congenital neutropenia. This boy suffered from life-threatening infections and required high doses of rhG-CSF, and a haploidentical PBSCT was also successfully performed, thus leading to reconstitution of CD40L expression on activated CD4+ T cells (as assessed with flow cytometry six months after the procedure). Two low-dose T-cell addbacks were required to re-establish full donor chimerism and clear CMV reactivation. The report demonstrates that in select cases, alternative donor allogeneic HSCT supported by DLI may be effective in correcting the defect in X-linked HIGM, and HSCT in HIGM children is not necessarily limited to matched sibling donor transplantation.

Correlation between serum interleukin-31 level and the severity of disease in children with atopic dermatitis.

INTRODUCTION: Atopic dermatitis (AD) is a chronic inflammatory skin disease, characterized by intense itch, typical localization and a specific image of skin lesions. Pathogenesis of pruritus in AD is not fully understood, but recent studies emphasize the role of interleukin-31 (IL-31). This relatively recently described cytokine is considered to be a potential mediator inducing pruritus in AD. AIM: To assess the correlation of serum IL-31 level and the disease severity in children with AD. MATERIAL AND METHODS: Twenty-five children (16 girls and 9 boys) with AD aged from 4 months to 17 years (mean age: 4.2 years) were enrolled in the study. Disease severity in children with AD was assessed using the SCORAD (Severity SCORing of Atopic Dermatitis) index. Serum IL-31 levels were measured using ELISA with standard kits from EIAab R&D Systems. RESULTS: Serum IL-31 level was significantly higher in AD children than in healthy children. There was no statistic correlation between serum IL-31 level and the disease severity or itch intensity. CONCLUSIONS: The disease severity and itch intensity do not correlate with serum IL-31 level in children with atopic dermatitis.

Role of Lung Ultrasonography (LUS) as a Tool for Evaluating Children with Pediatric Inflammatory Multisystem Syndrome Temporally Associated with SARS-CoV-2 (PIMS-TS).

BACKGROUND: Pediatric inflammatory multisystem syndrome temporally associated with SARS-CoV-2 (PIMS-TS) is a novel entity. The inflammatory process involves the circulatory, digestive, respiratory, and central nervous systems, as well as the skin. Making a diagnosis requires extensive differential diagnoses, including lung imaging. The aim of our study was to retrospectively assess the pathologies found in lung ultrasound (LUS) in children diagnosed with PIMS-TS and to evaluate the usefulness of the examination in diagnostics and monitoring. METHODS: The study group consisted of 43 children diagnosed with PIMS-TS, in whom LUS was performed at least three times, including on admission to hospital, on discharge, and 3 months after disease onset. RESULTS: Pneumonia (mild to severe) was diagnosed in 91% of the patients based on the ultrasound image; the same number had at least one pathology, including consolidations, atelectasis, pleural effusion, and interstitial or interstitial-alveolar syndrome. By the time of discharge, the inflammatory changes had completely regressed in 19% of the children and partially in 81%. After 3 months, no pathologies were detected in the entire study group. CONCLUSION: LUS is a useful tool for diagnosing and monitoring children with PIMS-TS. Inflammatory lesions of the lungs resolve completely when the generalized inflammatory process subsides.

Comparison of COVID-19 and RSV Infection Courses in Infants and Children under 36 Months Hospitalized in Paediatric Department in Fall and Winter Season 2021/2022.

BACKGROUND: The study aimed to determine the differences between COVID-19 and Respiratory syncytial virus (RSV) infections in young children hospitalized in the pediatric department. METHODS: This retrospective study included 52 children with COVID-19 and 43 children with RSV infection younger than 36 months hospitalized in a pediatric department between September 2021 and March 2022. Clinical and laboratory findings, methods of treatment and hospitalization length were compared. RESULTS: In the RSV group, significantly higher rates of cough (93.2% vs. 38.5%), rhinitis (83.7% vs. 50%), dyspnea (83.7% vs. 21.1%), crackles (69.8% vs. 5.8%) and wheezes (72.1% vs. 9.6%) were observed. The COVID-19 group had significantly higher rates of fever (80.8% vs. 37.2%) and seizures (13.5% vs. 0%). Patients with RSV infection had significantly higher rates of bronchodilator therapy (88.37% vs. 5.77%) and oxygen therapy (48.8% vs. 7.7%) and required a longer hospital stay (8 vs. 3 days). In admission, the majority of the patients from both groups were not treated with antibiotics, but because of clinical deterioration and suspected bacterial co-infections, antibiotics were administered significantly more frequently in the RSV group (30.2% vs. 9.6%). CONCLUSIONS: RSV infection in infants and small children had a more severe course than COVID-19 infection. RSV infection was associated with a longer hospitalization period and required more elaborate treatment.

Biochemical and cardiovascular predictors of PIMS-TS risk in children after COVID-19 recovery: preliminary results of the LATE-COVID-Kids Study.

Introduction: We aimed to characterize biochemical and cardiovascular predictors of the paediatric inflammatory multisystem syndrome temporally associated with SARS-CoV-2 (PIMS-TS) risk based on the data from the LATE-COVID-Kids study. Methods: 148 consecutive COVID-19 convalescents hospitalized for the clinical evaluation after the acute phase of COVID-19 were classified into two groups related to symptoms: 33 children finally diagnosed with PIMS-TS and 115 children without PIMS-TS (control group). Results: PIMS-TS children were significantly younger (6.79 ±4.57 vs. 9.10 ±4.94 years). After adjustment, in comparison to those without, PIMS-TS children had a higher level of antithrombin III (111 ±9.30 vs. 105 ±11.4), higher heart rate (HR)/min (100 (89.0-111) vs. 90 (79.7-100)) and sinus rhythm (p = 0.03) but lower PQ interval (p = 0.02) on admission to hospital. The lymphocytes (absolute count and percentage) were significantly higher in children with PIMS-TS, and the opposite results were obtained for IgA and neutrophils. Furthermore, children with PIMS-TS had a higher level of thyroid stimulating hormone (2.76 (2.16-4.18) vs. 2.36 (1.73-2.83)) and red cell distribution width (p < 0.005) compared to those without. Conclusions: It is the first data on the possible predictors of PIMS-TS risk in the Long-COVID period. These results need to be further validated to next create the PIMS SCORE algorithm, which might enable the effective prediction of children with the risk of PIMS-TS occurrence after COVID-19 recovery.

COVID-19 in unvaccinated patients with inborn errors of immunity-polish experience.

At the beginning of the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) pandemic, patients with inborn errors of immunity (IEI) appeared to be particularly vulnerable to a severe course of the disease. It quickly turned out that only some IEI groups are associated with a high risk of severe infection. However, data on the course of Coronavirus Disease 2019 (COVID-19) in patients with IEI are still insufficient, especially in children; hence, further analyses are required. The retrospective study included 155 unvaccinated people with IEI: 105 children and 50 adults (67.7% and 32.3%, respectively). Male patients dominated in the study group (94 people, 60.6%). At least two comorbidities were found in 50 patients (32.3%), significantly more often in adults (56% vs. 21%). Adult patients presented significantly more COVID-19 symptoms. Asymptomatic and mildly symptomatic course of COVID-19 was demonstrated in 74.8% of the entire group, significantly more often in children (88.6% vs. 46%). Moderate and severe courses dominated in adults (54% vs. 11.4%). Systemic antibiotic therapy was used the most frequently, especially in adults (60% vs. 14.3%). COVID-19-specific therapy was used almost exclusively in adults. In the whole group, complications occurred in 14.2% of patients, significantly more often in adults (30% vs. 6.7%). In the pediatric group, there were two cases (1.9%) of multisystem inflammatory syndrome in children. Deaths were reported only in the adult population and accounted for 3.9% of the entire study group. The death rate for all adults was 12%, 15.4% for adults diagnosed with common variable immunodeficiency, 12.5% for those with X-linked agammaglobulinemia, and 21.4% for patients with comorbidity. The results of our study imply that vaccinations against COVID-19 should be recommended both for children and adults with IEI. Postexposure prophylaxis and early antiviral and anti-SARS-CoV-2 antibody-based therapies should be considered in adults with IEI, especially in those with severe humoral immune deficiencies and comorbidity.

Mannan-binding lectin deficiency in pediatric patients with inflammatory bowel disease.

OBJECTIVE: The incidence of inflammatory bowel disease (IBD) in Europe has increased significantly. At least a fourth of patients are children. Mannan-binding lectin (MBL) is believed to be an important component of innate immunity, acting as an opsonin and activator of the lectin pathway (LP) of complement. The data relating any of the LP factors to IBD are sparse and contradictory and were obtained mainly from adult patients. The aim of this study was to investigate the possible role of MBL in Crohn's disease (CD) and ulcerative colitis (UC) in children. METHODS: MBL2 gene single nucleotide polymorphisms (PCR-RFLP) and MBL concentrations (ELISA) were determined. RESULTS: The frequency of MBL2 gene variants responsible for MBL deficiency (LXPA/O and O/O) is significantly higher in CD patients compared with controls or children with UC. A relatively high frequency of the codon 52 mutation (D allele) was noted in these patients. Practically no difference was found between UC and control (C) groups. Similarly, the average MBL levels as well as the number of MBL-deficient (MBL concentrations < 150 ng/ml) individuals differed between CD patients and controls or children suffering from UC. Again, there was no difference between UC and C groups. CONCLUSIONS: These data suggest that MBL deficiency may be associated with CD but not with UC in pediatric patients. The possible role of MBL in IBD requires confirmation in larger series and further investigation of the mechanisms involved.

[Effectiveness and tolerance of antibiotics in pediatrics patients]. / Skutecznosc i tolerancja antybiotyków w pediatrii.

Antibiotics represent the most widely prescribed therapeutic agents. Preschool children being most exposed to antibiotic drugs, especially in the community setting. Unnecessary antibiotic prescribing remains the cardinal contributing factor to the development of antibiotic resistance. Early antibiotic exposure, especially to broad-spectrum antibiotics, may suppress the developing immune system and produce a reduced anti-allergic response. Early antibiotic use in children is associated with an increased risk for asthma.

[Intravenously applied immunoglobulin in systemic, autoimmune and vasculitis diseases in children]. / Preparaty immunoglobulin w leczeniu chorób ukladowych o podlozu autoimmunizacyjnym i zapaleniach naczyn u dzieci.

Intravenous immunoglobulin (IVIG) have been widely used in clinical practice for more than 35 years. Their specificity and diversity and their relative safety make them potent therapy in antibody deficiencies, certain infections and several autoimmune and inflammatory disorders. IVIG is a biological drug that is used routinely for idiopathic thrombocytopenic purpura, Kawasaki disease, and Guillain-Barré syndrome. Therapeutic approaches for autoimmune diseases are primarily based on suppressive measures that down regulate an over productive immune system. IVIG efficacy has been established in many clinical trials for various autoimmune diseases, vasculitis and neuroimmunological, dermatological and hematological disorders. Despite the evidence of efficacy, there are no generally accepted therapeutic guidelines, the dosage and timing of IVIG therapy, and questions of costs/benefits ratio still remain insufficiently documented and multicentric controlled clinical trials are necessary. Most available evidence for a benefit for IVIG in children comes from uncontrolled open series or case reports.

Nomograms of the fetal thymus for clinical practice.

INTRODUCTION: The fetal thymus may be visualized using ultrasonography (USG) and is typically located in the mediastinum. In the past years, the size of the fetal thymus has served not only as a marker of genetic or heart defects but also as a predictive factor for intrauterine growth restriction, premature birth, preeclampsia, chorioamnionitis or even neonatal sepsis. MATERIAL AND METHODS: A total of 410 fetuses were qualified for the study. Fetuses with heart defects were excluded from the study. The fetal thymus was evaluated with ultrasonography between the 14th and 40th week of gestation. After obtaining a standard transverse view encompassing the three great vessels, thymus measurements were attempted, i.e. maximal transverse diameter, circumference and surface area. Linear regression was used for statistical analysis, yielding 3 models, each with a different dependent variable. The confidence interval for each model was set at 80% to aid the comparison with centile grid growth charts for neonates and children. The test was regarded as statistically significant when p < 0.05. RESULTS: From a total of 410 fetuses the thymus transverse diameter, circumference and area were successfully measured in 410, 320 and 330 cases, respectively. The probabilities are lower than 0.0005 for each model, which means that each model is quite statistically significant. CONCLUSIONS: The coverage of healthy thymus nomograms in the fetal population may be the basis for the identification of fetuses at risk of hypoplasia or thymic hyperplasia, which seems particularly important from the point of view of the detection of potential inborn immunological disorders.

[Mannan-binding lectin (MBL) in duodenal ulcer and gastritis]. / Lektyna wiazaca mannan (MBL) w chorobie wrzodowej dwunastnicy i zapaleniu blony sluzowej zoladka.

UNLABELLED: H. pylori is considered to be a major etiological agent of chronic gastritis. Mannan-binding lectin (MBL) is one of the key factors of the innate immunity. It belongs to collectins binding to saccharides on the surface of a variety of pathogenic microorganisms, H. pylori including. MBL after coating the bacteria is recognized by the receptor on macrophages, facilitates phagocytosis by direct opsonization and activates complement via the lectin pathway which protects the host against infection. Single nucleotide polymorphisms of MBL2 gene affect both MBL concentration and biological activity. AIM: Evaluation of the correlation between the occurrence of MBL2 gene mutations, MBL concentration in serum, and prevalence of duodenal ulcer and chronic gastritis with or without H. pylori infection. MATERIAL AND METHODS: The study comprised 174 children aged 6-17 years (mean 13.1 x 4.2), among them 74 children with H. pylori infection-associated chronic gastritis (group Hp+), 41 with H. pylori infection excluded (group Hp-), and 11 children with duodenal ulcer (DU). The control group consisted of 46 healthy children without gastritis, endoscopically checked. Determination of serum MBL concentration was performed using ELISA. Polymorphisms of the promoter region as well as mutations of exon 1 MBL2 gene were identified with polymerase chain reaction methods. H. pylori was detected with both urease test and histopathology Giemsa staining. RESULTS: MBL serum concentration was significantly lower in children with both Hp+ (1809 ng/ml) and Hp-gastritis (1439 ng/ml) when compared to the control group (2545 ng/ml) (p < 0.05 and p < 0.01, respectively). The same trend (however the difference was insignificant) was observed in the case of DU group (1643 ng/ml). The frequency of MBL2 gene mutations did not differ between any group of patients as compared to controls. CONCLUSIONS: No influence of MBL2 genotype on the incidence of duodenal ulcer or gastritis was found. MBL serum concentrations are lower in chronic gastritis patients (with or without H. pylori infection) when compared to the age-matched healthy individuals.

[Alterations of metalloproteinases and their inhibitors concentrations in peripheral blood in children with congenital heart disease undergoing cardiac surgery with cardiopulmonary bypass]. / Zmiany stezen metaloproteinaz i ich inhibitorów w krwi obwodowej dzieci z wrodzona wada serca poddanych zabiegowi kardiochirurgicznemu z uzyciem krazenia pozaustrojowego.

Cardiac surgery with cardiopulmonary bypass (CPB) in children with congenital heart disease induces neutrophil activation, degranulation and systemic inflammatory response. Matrix metalloproteinase-9 (MMP-9) and matrix metalloproteinase-2 (MMP-2) are enzymes involved in degranulation and leukocyte extravasation. These are secreted as a pro-enzyme in response to several inflammatory mediators and are inhibited by tissue inhibitor of metalloproteinase-1 (TIMP-1) and tissue inhibitor of metalloproteinase-2 (TIMP-2). To explore metalloproteinase activation during cardiac surgery we investigated MMP-9, MMP-2, TIMP-1 and TIPM-2 levels in young children during and after surgery. We measured the dynamics of these enzyme concentrations in peripheral blood. Additionally we measured CD11b and CD66b molecule expression on neutrophils. These investigations were carried out in 39 children, aged 5-38 months who were undergoing cardiacsurgery with cardiopulmonary bypass (CPB). Serum concentrations of MMPs and their inhibitors, CD11b and CD66b expression on neutrophils were sequentially measured before induction of anesthesia, at the initiation of CPB, after 30 minutes of CPB, at the end of CPB, 4 and 48 hours after CPB. MMP-9 concentration increased at the end of CPB and remained elevated for a period of 48 hours. The concentration of MMP-9 detected at the end of CPB positively correlated with time of CPB (r=0.68, p=0.0045). TIMP-1 concentration decreased significantly after 30 minutes of CPB, remained lowered to the end of CPB, and returned to the start of CPB level after 48 hours. CD11b and CD66b expression on neutrophils increased at the initiation of CPB. Our data confirm that MMPs play an important role in inflammatory complications after cardiac surgery in children. These findings suggest that kinetics of MMPs concentrations in serum after cardiac surgery appear to depend on many factors. We demonstrated the link between CPB duration and the MMP-9 concentration. Future studies will determine whether inhibition of MMPs activity diminishes morbidity in children after cardiac surgery.

Proteolytic and cytokine balance abnormalities in children with congenital heart disease undergoing cardiac surgery with cardiopulmonary bypass.

BACKGROUND: Cardiac surgery with cardiopulmonary bypass (CPB) in children with congenital heart disease induces a systemic inflammatory response. This inflammatory response is thought to be produced by exposing patients to proinflammatory factors. AIM: To explore the role of cytokines and proteolytic enzymes in inflammatory complications after cardiac surgery in children. METHODS: We investigated the dynamics of concentrations of IL-6, IL-8 and IL-10, and metalloproteinases (MMPs) MMP-2 and MMP-9, and their inhibitors - tissue inhibitors of metalloproteinases (TIMPs) TIMP-1 and TIMP-2. These investigations were carried out in 28 children, aged 4-34 months, who underwent a cardiac operation with CPB. Serum concentrations of proteins were sequentially measured before induction of anaesthesia, at the initiation of CPB, after 30 minutes of CPB, at the end of CPB, and 4 and 48 hours after CPB. RESULTS: The serum levels of IL-6 increased dramatically 4 hours after CPB compared with the level before anaesthesia (141.83+/-25.49 vs. 10.68+/-5.01 ng/ml, p=0.00004) and correlated with duration of CPB (r=0.74, p=0.00028). The serum levels of IL-8 increased 4 hours after CPB compared with the level before anaesthesia (267.1+/-41.3 vs. 8.5+/-6.3 ng/ml, p=0.00002). A significant increase of IL-10 concentration at the end of surgery and 4 hours after CPB was detected (95.12+/-23.57 vs. 10.34+/-6.45 ng/ml, p=0.000004 and 59.41+/-21.4 vs. 10.34+/-6.45 ng/ml, p=0.00004, respectively ). The MMP-9 concentration increased at the end of CPB and remained elevated for a period of 48 hours (44.40+/-13.95 vs. 19.53+/-7.58, p=0.000004 and 38.97+/-10.76 vs. 19.53+/-7.58, p=0.00004, respectively). The concentration of MMP-9 detected at the end of CPB positively correlated with duration of CPB (r=0.68, p=0.0045). The TIMP-1 concentration decreased significantly after 30 minutes of CPB, and remained lowered to the end of CPB (respectively 52.68+/-17.72 vs. 83.29+/-17.06 ng/ml, p=0.000006 and 34.94+/-10.58 vs. 83.29+/-17.06 ng/ml, p=0.00004,respectively). CONCLUSIONS: Cardiac surgery causes an increase of IL-6 and IL-8 concentrations in peripheral blood 4 hours after CPB termination. The concentration of anti-inflammatory IL-10 cytokine increases immediately after the end of CPB. We showed an increase of the MMP-2 and MMP-9 concentrations during and after CPB and simultaneous decrease of TIMP-1 inhibitor. We demonstrated a link between CPB duration and IL-6 and MMP-9 concentrations.

[Comparable evaluation of usefulness of traditional methods and polymerase chain reaction technique in detection of Helicobacter pylori infection]. / Porównawcza ocena przydatnosci tradycyjnych metod i techniki lancuchowej reakcji polimerazy w wykrywaniu zakaienia Helicobacter pylori.

UNLABELLED: H. pylori is a well recognized pathogenic factor of chronic gastritis and class I carcinogen. Specimens are collected on gastroscopy performed due to clinical symptoms suggesting upper alimentary tract organic disease for pathomorphological examinations and for the estimation of the presence of H. pylori infection in urease test, histological examination in Giemsa staining method or in a culture. The aim of the project was to compare the detection of H. pylori in gastric mucosa in children by conventional method and polymerase chain reaction technique (PCR). MATERIAL AND METHODS: The study comprised 137 children and adolescents, aged 5-18 years (mean 13.2 years), in whom basing on the evaluation of the level of serum anti-H.pylori antibodies (ELISA), urease test and histological examination (HE), H. pylori infection was diagnosed or excluded. H. pylori genes: ureA, vacA, cagA were identified in gastric mucosa specimens with PCR method. RESULTS: On the grounds of standard methods H. pylori was detected in 86 (62.8%) and excluded in 51 (37.2%) children. UreA gene was detected in gastric mucosa in 82 (95.3%) children with H. pylori (Hp+) and in 8 (15.7%) without H. pylori (Hp-) infection (p < 0.001). CagA H. pylori gene was identified in 21 (24.4%) children from Hp+ group and in 1 (2%) from Hp- group (p < 0.001), and vacA gene in 23 (26.7%) and 6 (11.8%) (NS), respectively. H. pylori strains type I (cagA+/vacA+) were found in gastric mucosa of 15 (17.4%) Hp+ patients. CONCLUSIONS. Testing of H. pylori genes (particularly ureA gene) with PCR technique in mucosa bioptates enables to diagnose H. pylori infection even in patients with the infection not detected with standard methods.