The aim of the study was to assess the effect of IFN-alpha and ribavirine combined therapy of chronic hepatitis C in children. The study comprised 37 children (22 boys and 15 girls) aged between 4 and 18 years (mean 12 years and 6 months) with chronic hepatitis C, diagnosed on the basis of serological, virological and histological criteria. The treatment included Intron A (Schering Plough) administered subcutaneously 3 M.U. 3 times a week and ribavirin (Rebetol-Schering Plough) orally 15 mg/kg body weight daily for the period of 12 months. In all children liver biopsy was performed before the beginning of treatment. Tests for the presence of HCV RNA in serum were performed after 12-weeks therapy (EVR), after 6-months therapy, immediately on its completion (ETR) and 6 months afterwards (SVR), after the end of treatment. In the course of applied treatment, ALT activity, hemoglobin levels, leukocyte and trombocyte counts in the peripheral blood were determined every 4 week. EVR was obtained in 25 children and in these chidren genetic material of HCV in serum of blood was not observed both, after 6-months therapy, as well after the end of treatment. In this group a sustained virological response (SVR) was observed in 21 children. In 12 children without EVR, the elimination of HCV RNA was not achieved in the course of further treatment. Conclusions: 1. Twelve-months of combined IFN-alpha and ribavirine therapy enables to obtain a sustained virological response in about 56% of the treated children. 2. The frequency of recurrences of HCV infection in children amounts to 16%. 3. During the combined IFN-alpha and ribavirine treatment numerous side effects were observed, which in individual cases were the reason for treatment modification.

The aim of this study was to assess the results of 12-month lamivudine therapy in children with chronic hepatitis B. MATERIAL AND METHODS: The study included 24 children (14 boys and 10 girls) aged from 7 to 18 years with chronic hepatitis B diagnosed according to current criteria. Lamivudine was administred in a daily dose of 3-5 mg/kg up to 100 mg for twelve months. The efficiency of treatment was estimated on the basis of absence of HBV DNA in blood serum, loss of HBeAg and normalization of ALT activity. RESULTS: Total virusological response and biochemical response were achieved in 6 children after twelve months of lamivudine therapy. In 4 children, absence of HBV DNA without the loss of HBeAg and without the normalization of ALT activity were observed. Fourteen children did not respond to 12-month of lamivudine therapy (the lack of HBV DNA elimination and the lack of HBeAg loss). None of the children showed elimination of HBsAg and production of HBs antibodies. CONCLUSIONS: The treatment is very well tolerated and does not cause undesired side effects.

Lyme borreliosis has become a serious diagnostic and therapeutic problem of modern medicine. The occurrence of the disease in Europe, and also in Poland, has recently increased. Diagnostic problems with Borrelia burgdorferi infection result from non-characteristic course of the disease in children, without the 1 st stage of the disease (erythema migrans), negative history forwards tick bite, lack of seasonal changes in occurrence and difficulties with interpretation of serological tests results. Direct laboratory investigations are also of little relevance.

The aim of this study was to evaluate the activity of NK cells in peripheral blood in children with chronic type C hepatitis, and to determine the correlation between the activity of NK cells and histopathological changes in the liver. The study included 25 children with chronic type C hepatitis. The control group consisted of 10 children without liver diseases in past medical history and normal activity of amino-transferases. In all children the activity of NK cells was evaluated in relation to leukaemic cells of erytroleukaemia K-562 without stimulation and after stimulation with IL-2 (in vitro), and the percentage and absolute count of NK cells in peripheral blood were determined by flow cytometry. RESULTS: In the studied group of children NK cells accounted for 11.68 +/- 6.73% of peripheral white blood cells, and their count was 241.08 +/- 128.56/ml. The cytotoxic activity of NK cells was 91.8 +/- 1.07% in solution E:T 25:1; and 6.72 +/- 3.68% in solution E:T 12.5:1. After stimulation with IL-2 it was 92.8 +/- 1.01% and 7.58 +/- 3.95%, respectively. The count of NK cells in serum and cytotoxic activity of NK cells in the studied group were not different from those of the control group. CONCLUSIONS: Key parameters of NK cells activity are not changed in children with chronic type C hepatitis.

The hepatitis C virus (HCV) infection course and efficacy of treatment may be depended on HLA antigens. The aim of the study is attempt to define dependence between the course of HCV infection and efficacy its treatment and HLA A antigens in children and youth. PATIENTS AND METHODS: To the study included 61 patients (51 after treatment for HCV infection and 10 not treated). The average age was 13.77 years (range 5-18 years). Patients were divided to subgroups in depend on effect of the treatment: virusological and biochemical response. Antigens HLA A were molecularly typed on the low resolution method. To statistical analysis we applied the chi-square test. RESULTS: We demonstrated no statistical significant dependences between HCV infection course and efficacy of its treatment children and youth HLA A antigens. However we observed following tendencies: antigens HLA A *01 and HLA A *02 can be related to unprofitable course of infection and unsuccessful antiviral therapy; HLA A *03 can be favorable prognostic factor for HCV infection course and response to treatment; HLA A *24 can be related to mild course of infection and profitable response to treatment; HLA A *11 can be favorable prognostic factor for course of infection; HLA A *30 can be profitable for efficacy of treatment and HLA A *25 can be disadvantage for it. Probably study performed with larger group of patients could gain dependencies statistical significant. CONCLUSIONS: It is possible that course if HCV infection and efficacy of antiviral treatment are depended on HLA A antigens.

Haemophilus influenzae is a gram-negative bacteria. The capsular form of this bacteria, mainly type b, is responsible for severe bacterial meningitis. In the study course of Haemophilus influenzae meningitis in two children was presented. In one of these children the clinical course of meningitis was particularly serious. The child was unconscious, the generalized swellings, pneumonia, increase and damage of liver, diarrhoea, heavy anaemia as well as disorders of blood coagulation were observed. In second child the beginning of disease was not characteristic, what delayed the proper diagnosis. The duration of children treatment was from 16 to 18 days. The permanent damages of central nervous system were not observed in both children

Clinical picture of Borrelia burgdorferi infection has been presented in 89 children from Lodz region. The analysis showed significant domination of cases with non specific symptoms (41.6%) as: fever or headache and cases with affected central and peripheral nervous system (30.3%). Peripheral cranial nerves paralysis and symptoms of cerebrospinal meningitis dominated among children with neuroborreliosis. Unlike descriptions concerning adults, majority of the observed symptoms were changes characteristic for I stage of the disease. Dermatosis was found only in (19%) child and symptoms of arthritis in (9%) of them. Contact with tick was stated in 56.8% of the analysed children. Incidence of the disease occurred throughout the whole year, more frequently in summer and autumn months.

Recently, the incidence of fungal infections in children, including children with shunt-dependent hydrocephalus, has increased. The analysis comprised 8 children treated in the III Clinic of Pediatrics of ICZMP during the period of 12 months (12% of all infectious complications of the shunt system). The clinical picture of fungal infection included symptoms of shunt dysfunction: febrile conditions, vomiting, distress and loss of appetite. The most common pathogens isolated from the cerebro-spinal fluid were fungi from the Candida species. Mean value of pleocytosis in the cerebro-spinal fluid was 812 cell/microl, and mean protein concentration was 311 mg/dl. Treatment consisted of monotherapy with Diflucan, monotherapy with Ancotil or combined treatment with Ancotil and Amphotericine B. The drugs were administered intravenously and intraventricularly after removal of the shunt and application of external drainage. Sterility of cerebro-spinal fluid was obtained in the shortest time with the use of Ancotil. Prophylactic application of antifungal drugs decreases the frequency of infections in children with shunt-dependent hydrocephalus.

The aim of the study was an evaluation of the somatic development of children with chronic hepatitis B, C and HBs antigen positive and finding the correlation between the nutrition status, the duration of disease and the aminotransferase activity. The somatic development was examined on the basis of body height and weight measurement and calculation of body mass index and Cole's index. Lowered body height and weight was observed in 25% of the children with chronic HCV infection. In the group of children with chronic hepatitis B there was no body weight loss, but a lowered body height was observed in 9.1%. No correlation between the duration of the disease and the nutrition status in children with chronic hepatitis B and C was proved.

The importance of carbohydrate metabolism in the pathogenesis of chronic hepatitis C has been recognized in recent years. The aim of the study was to assess carbohydrate metabolism in children with chronic HCV-related hepatitis. MATERIAL AND METHODS: The study was comprised of 16 children with chronic hepatitis C and 16 healthy controls. In all the children anthropometric data and, after overnight fasting, serum levels of glucose and insulin were investigated and insulin resistance index HOMA IR was calculated. In the study group biopsy specimens were assessed. RESULTS: Macrovesicular steatosis in numerous hepatocytes was found in 3 children. Mean values of body mass, height, body mass index (BMI), levels of glucose, insulin and HOMA IR did not differ significantly between the two groups. We found correlation between serum levels of glucose and fibrosis (r=0.64, p<0.01) or staging (r= 0.7, p<0.004) in children with chronic hepatitis C. CONCLUSION: Neither obesity, which is recognized as a risk factor for diabetes mellitus and steatosis was found in children with HCV-related hepatitis. Correlation between serum levels of glucose and fibrosis or staging in children with chronic hepatitis C suggests the participation of carbohydrate metabolism in the pathogenesis of this disease and the risk, existing already in such young patients, of developing hepatic fibrosis with its consequences.

OBJECTIVE: We have recently shown that terminal sialic acid residues are essential for alpha(1)-acid glycoprotein (AGP)-induced Ca(2+) mobilization in neutrophils. The aim of the present study was to establish the importance of sialic acid residues on AGP in modulating human neutrophil functions, with emphasis on the generation of reactive oxygen species (ROS). MATERIALS AND METHODS: ROS were measured by luminol-enhanced chemiluminescence in isolated human neutrophils. RESULTS: We found that AGP did not provoke ROS generation in resting or L-selectin presensitized neutrophils. Moreover, AGP did not affect the N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced ROS generation, but it slightly suppressed opsonized zymosan-induced responses. However, when the neutrophils were prestimulated with fMLP, the subsequent addition of AGP provoked a marked ROS response. Dose-response studies and time studies revealed that the ROS generating capacity of AGP was highest at a concentration of 0.05 mg/ml and when given 3-10 min after addition of fMLP. A desialylated form of AGP or pretreatment of neutrophils with 3'- and 6'-sialyllactose caused a substantially lower ROS response in neutrophils prestimulated with fMLP. CONCLUSIONS: Our data show that AGP can stimulate a second ROS response in fMLP preactivated neutrophils and that terminal sialic acid residues on AGP play a crucial role in this regard.

Background: It has been supposed that liver transplant recipients with hepatitis C virus infection have a higher incidence of infectious complications after transplantation. This study was designed to investigate whether neutrophil function is immediately affected by liver transplantation. Methods: Biochemical values, plasma levels of myeloperoxidase (MPO), hydrogen peroxide production of neutrophils and neutrophil-platelet complexes were analyzed in 32 patients who underwent liver transplantation and 20 healthy volunteers. Results: MPO levels were significantly increased 24 h after reperfusion. In post-hepatitic patients levels were significantly lower three d up to one wk post-transplant than in patients due to other liver diseases. One wk post-operatively the respiratory burst activity following N-formyl-methionyl-leucylphenylalanine (fMLP) or (tumor necrosis factor-alpha) TNF-alpha/fMLP stimulation was depressed in post-hepatitic recipients. Respiratory burst stimulated with phorbol 12-myristate 13-acetate in these patients was increased one wk after transplantation. One d after transplantation the neutrophil-platelet complexes decreased significantly throughout the post-operative period. Conclusions: The results of this study suggest a reduced post-operative neutrophil activation in liver transplant recipients suffering from post-hepatitic cirrhosis compared to cirrhosis due to other causes. We hypothesized that neutrophil dysfunction in those patients depends on the underlying disease with an increased susceptibility to bacterial or fungal infections.

The effect of glucomannan (GM), a natural polysaccharide isolated from the yeast Candida utilis, on reactive oxygen species (ROS) generation in human neutrophils in vitro and in rats with Mycobacterium butyricum induced adjuvant arthritis (AA) was tested by the luminol/isoluminol-enhanced chemiluminescence (CL) method. In vitro, GM (500mug/ml) significantly decreased spontaneous CL of human whole blood, while PMA (4beta-phorbol-12beta-myristate-alpha13acetate)-stimulated CL was decreased by GM in the concentrations of 100 and 500mug/ml. To specify the site of action of GM, its effect on extra- and intracellular ROS generation in isolated neutrophils was evaluated. GM significantly decreased spontaneous and PMA-stimulated CL and it was more effective extracellularly than intracellularly. In vivo experiments included healthy animals as controls, arthritic animals without any drug administration, and arthritic animals with GM administration (once daily in the oral dose of 15mg/kg, over a period of 28 days). On day 28, CL in whole blood, spleen and joint was monitored. Arthritic animals treated with GM showed decrease in spontaneous and PMA-stimulated CL of whole blood as well as CL of the joint, in comparison with untreated animals. The obtained findings demonstrated an antioxidant effect of GM in vitro and in rats with AA, which may be due to its free radical scavenger activity and to interaction with different receptors and/or modulation of postreceptor intracellular signalling pathways. The specific physicochemical parameters, such as structure of GM, its low molecular weight and good water solubility, play an important role in the above effects.

Changes that occur with age in the opsonin-independent oxidative activity of peripheral phagocytes in whole blood were examined by means of luminol chemiluminescence (LCL). The chemiluminescence was registered simultaneously by non-stimulated and stimulated cells and the age-related alterations of total and extracellular generation of reactive oxygen species (ROS) were studied using model systems. It was found that the rate of phagocyte activation by the glass surface of the measuring chambers, assessed by the time of the peak appearance after the start of LCL response, increased. However, the maximum oxidative activity and the integral oxidative capacity of the cells during adhesion, evaluated by the maximum LCL intensity and the area under the LCL curve, respectively, declined. No age-dependence of formyl-methionyl-leucyl-phenylalanine (fMLP)-stimulated oxidative cellular activity for total ROS generation was detected. The maximum oxidative activity and the integral oxidative capacity of peripheral phagocytes to generate extracellular superoxide in response to fMLP was decreased. The likely causes for the observed alterations in phagocyte function are discussed and an analysis of the obtained results is given on the background of the contradictory data published on phagocyte oxidative activity age-related changes. Copyright (c) 2009 John Wiley & Sons, Ltd.

The human promyelocytic leukemia HL 60 and PLB 985 cell lines can differentiate into terminally mature neutrophil-like cells via dimethyl sulfoxide (DMSO) induction. In this study the luminol-dependent chemiluminescence (LCL) of both neutrophil-like cells was analayzed and compared in response to phorbol myristate acetate (PMA) and opsonized zymosan (OZ) stimulants. It was shown that, like human blood neutrophils, both neutrophil-like cells expressed high levels of CD11b, but unlike human blood neutrophils these cells almost lack LCL-detectable intracellular oxidase activity. By studying the pattern of activation to OZ and PMA and priming with GM-CSF, we concluded that there is no difference between the percentage of differentiation and function of DMSO-induced HL 60 and PLB 985. However, the LCL capacity (area under the curve) of DMSO induced PLB 985 cells was higher than that of HL 60 cells in response to both PMA and OZ, which implies a higher capacity to generate reactive oxygen species in PLB 985 cells. Copyright (c) 2009 John Wiley & Sons, Ltd.

Production of reactive oxygen species (ROS) by macrophages from blood monocytes of healthy donors (MP(N)) and patients with IHD (MP(IHD)) before, during, and after their incubation with low-density lipoprotein (LDL) isolated from the blood plasma of healthy donors (LDL(N)) and patients with a high cholesterol level (LDL(H)) was estimated by the method of luminol-dependent and stimulated by opsonized zymosan (OZ) or phorbol-12-myristate-13-acetate (PMA) chemiluminescence (CL). Intrinsic luminol-dependent, and zymosan- or PMA-stimulated chemiluminescence of MP(IHD) have exceeded the same types of chemiluminescence of MP(N) by factors of 1.4, 1.8, 2.7, and 1.6, respectively (p < 0.05-0.01). The effect of zymosan on MP(N) and MP(IHD) was stronger than that of TPA by factors of 4.3 and 3.2, respectively, but manifested itself 2.5-3.0 times slower. LDL(N) and LDL(H) incubated with MP(N) during 15-60 min increased ROS production by a factor of 1.4 and 2.5 respectively, but influenced ROS production by MP(IHD)(as estimated by luminol-dependent chemiluminescence). Effects LDL(N) and LDL(H) on MP(IHD) were not detected at all. Repeated increase in zymosan-stimulated CL of MP(N) was also observed after their 15-180 min preincubation with LDL(N) and LDL(H) which followed after taking out LDL, washing MP(N) and adding Hanks' solution with opsonized zymosan. This increase was also stronger after MP(N) incubation with LDL(H) than after MP(N) incubation with LDL(N), and no increase was observed in experiments with MP(IHD). Thus, the results obtained by a chemiluminescent method showed that fresh macrophages from the blood of patients with IHD had higher ROS production than macrophages from healthy donors. LDL(N) and LDL(H) could exhibit primary and secondary (after preincubation) stimulating effect on CL in MP(N); but had no effect on MP(IHD). An analysis of macrophage chemiluminescence is a sensitive test for evaluation the degree of macrophage's stimulation and it may be effectively used for the dynamic control for treatment effectiveness in clinics.

The aim of our study was to characterize the priming effect of extracellular nucleotides on reactive oxygen species (ROS) production induced by formyl-methionyl-leucyl-phenylalanine (fMLP), interleukin-8 (IL-8), leukotriene B4 (LTB4), and platelet activating factor (PAF). Also, we investigated the roles played by different protein kinase C (PKC) isoforms in nucleotide-induced priming. ROS production was determined by an isoluminol-based assay. MATERIAL AND METHOD: Nucleotide-induced priming was concentration- and time-dependent. The concentration of UTP able to cause maximal priming was 10 microM. When UTP (10 microM) was administered prior the agonist, the increase of the amplitude of the response reached the maximum at 1 minute of preincubation with the nucleotide. RESULTS: Calcium depletion of neutrophil caused significant inhibition of ROS production induced by all agonists tested, but did not affect the priming effect of the nucleotides. We tested the effect of several PKC inhibitors on the nucleotide-induced priming. GF 109203X (5 microM), an inhibitor of all neutrophil's PKC isoforms, or RO 31-8220 (5uM), an inhibitor of classical and novel PKC isoforms, abolished the responses induced by fMLP (10 nM) IL-8 (10 nM), LTB4 (100 nM) or PAF (100 nM). Go 6976 (100 nM), a selective inhibitor of classical PKC isoforms, had no effect on nucleotide-induced priming, suggesting that activation of these PKC isoforms does not play a role in the priming effect. Rottlerin (5 microM), a PKC delta inhibitor, almost abolished the effect of fMLP in the absence or in the presence of UTP, indicating that PKC delta is essential for the fMLP-induced effect; rottlerin also caused inhibition of ROS production induced by IL-8, LTB4 or PAF, however the priming effect of UTP was not affected for these chemoattractants. Our data suggest that classical PKC isoforms do not play a role in chemoattractant-induced ROS production. CONCLUSION: Although fMLP induced effect appears to be highly dependent on PKC delta activation, other chemoattractants are able to cause ROS production through PKC delta-independent mechanisms.

It has been known that administration of antibiotics may lead to excessive release of bacterial endotoxins and complicate clinical course of patients with Gram-negative infections. This concern may also apply to phages. Endotoxin may in turn activate neutrophils to produce reactive oxygen species (ROS) that are believed to play an important role in the pathogenesis of multiple organ dysfunction in the course of sepsis. We showed that a purified T4 phage preparation with low-endotoxin content could significantly diminish the luminol-dependent chemiluminescence (CL) of peripheral blood polymorphonuclear leukocytes (PMNs) both stimulated by lipopolysaccharides (LPSs) isolated from different Escherichia coli strains. This effect was also observed for live bacteria used for PMNs stimulation and was independent of bacterial susceptibility for T4-mediated lysis. Our data suggest, that phage-mediated inhibition of LPS- or bacteria-stimulated ROS production by PMNs may be attributed not only to phage-PMNs interactions, but also to phage-LPS interactions and bacterial lysis (in case of homologous phage). Interestingly, the T4 preparation did not influence ROS formation by PMNs stimulated with PMA. This suggests that the observed phenomena are also dependent upon the nature of activator. Bacteriophage-mediated inhibition of ROS formation by cells exposed to endotoxin provides new evidence for possible interactions between phages and mammalian cells. It helps in understanding the role of phages in our environment and may also be of important clinical significance.

To determine the relation between neutrophil function and the clinical characteristics of systemic lupus erythematosus (SLE), the superoxide anion ([Formula: see text]) production by neutrophils, mediated by FcgammaR and FcgammaR/CR cooperation, was studied in 64 SLE patients classified according to their prevalent clinical manifestations. Three clinically distinct patterns were designated:(1) manifestations associated with the occurrence of cytotoxic antibodies (SLE-I group);(2) manifestations associated with circulating immune complexes (IC; SLE-II group), and (3) manifestations associated with IC and cytotoxic antibodies (SLE-III group).[Formula: see text] production was evaluated by a lucigenin-dependent chemiluminescent assay in neutrophils stimulated with IC-IgG opsonized or not with complement. No difference in [Formula: see text] production was observed when neutrophil responses from healthy controls were compared to the unclassified patients. However, when the SLE patient groups were considered, the following differences were observed:(1) SLE-I neutrophils showed lower [Formula: see text] production mediated by the IgG receptor (FcgammaR) with the cooperation of complement receptors (FcgammaR/CR) than observed in the SLE-II, SLE-III, and healthy groups;(2) neutrophils from the SLE-II group showed a decreased [Formula: see text] production mediated by FcgammaR/CR compared to the SLE-III group,(3) SLE-III neutrophils produced more [Formula: see text] than neutrophils from the SLE-II and control groups, and (4) CR showed inefficiency in mediating the [Formula: see text] production by neutrophils from the SLE-I group. Comparative experiments on the kinetics of chemiluminescence (CL; T (max) and CL(max)) disclosed differences only for the SLE-I group. Taken together, these results suggest that differences in oxidative metabolism of neutrophils mediated by FcgammaR/CR may reflect an acquired characteristic of disease associated with distinct clinical manifestations.