Effect of different amino acid solutions on the oxidative stability of three different lipid emulsions in all-in-one admixtures.

BACKGROUND AND AIM: Lipid peroxidation in parenteral nutrition mixtures is still a challenge. We aimed to evaluate the effect of two different amino acid solutions used in different clinical situations on lipid peroxidation of three different lipid emulsions (Intralipid, ClinOleic, and SMOFlipid) in all-in-one admixtures during 24 h of simulated infusion. The selected amino acid solutions included one used in stable patients and one used in renal insufficiency (Aminomel10E and Nephrotect, respectively). METHODS: Eighteen all-in-one admixtures were prepared. The simulated infusion with light protection was conducted straight after the preparation for 24 h at room temperature. The lipid peroxidation process was evaluated in all-in-one admixtures and the original lipid emulsion by determining the malondialdehyde levels (high-performance liquid chromatography) and conjugated dienes and trienes (ultraviolet-visible spectrophotometry). RESULTS: Malondialdehyde in the original packaging was lower in SMOFlipid (9 µM) compared with Intralipid (27 µM, P = 0.0003) and ClinOleic (25 µM, P = 0.0001). During simulated infusion, ClinOleic showed a significantly lower rate of lipid peroxidation (26% decrease in aldehyde levels) in comparison with Intralipid and SMOFlipid (up to 39% and 31% increase in aldehyde levels, respectively) when the admixture was based on Aminomel10E. In admixtures based on Nephrotect, ClinOleic, and SMOFlipid showed better oxidative stability in comparison with Intralipid. Admixtures based on Nephrotect and Intralipid had higher levels of primary lipid peroxidation products than those based on ClinOleic (P = 0.030) or SMOFlipid (P = 0.071, not significant). CONCLUSIONS: Amino acid solutions influence the rate of lipid peroxidation. The observation should be confirmed in larger studies with different amino acid solutions.

Antioxidant balance in plasma of patients on home parenteral nutrition: A pilot study comparing three different lipid emulsions.

BACKGROUND & AIMS: Parenteral nutrition (PN) can supply all essential nutrients to a patient with gastrointestinal insufficiency. However, the sensitivity to lipid peroxidation might increase in those receiving PN, especially home parenteral nutrition (HPN). This study aimed to investigate whether PN affects the antioxidant balance of plasma of HPN patients without comorbidities and whether this balance is influenced by comorbidities and according to the type of lipid emulsion included in the PN. METHODS: Adult patients on HPN (n = 86) received one of three types of lipid emulsion (based on 1) soyabean oil, 2) olive and soyabean oil or 3) soyabean, coconut, olive and fish oil) in all-in-one mixtures; in addition healthy controls (n = 66) were studied as comparators. HPN patients were classified to the following subgroups: 1) patients without (n = 58) or with (n = 28) comorbidities 2) patients on Intralipid (GINTRA, n = 53), ClinOleic (GCLIN, n = 17) or SMOFlipid (GSMOFn = 16). The activities of total glutathione peroxidase (GSH-Px), selenium dependent glutathione peroxidase (Se-GSHPx) and glutathione S-transferase (GST) in plasma were determined spectrophotometrically. The antioxidant potential of plasma was determined using oxygen radical absorbance capacity (ORAC). The lipid peroxidation marker malondialdehyde (MDA) was analyzed with high performance liquid chromatography. RESULTS: MDA concentration was the highest in GINTRA and the lowest in GSMOF (p < 0.05). GSMOF also had the highest activity of GSH-Px. No differences in Se-GSHPx, GST and ORAC were observed among GINTRA, GCLIN and GSMOF. Comparing with healthy controls, significantly lower GST (p = 0.0293) and ORAC (p < 0.0001) were observed in the HPN patients. Among all measured parameters only the concentration of MDA was significantly higher in patients with comorbidities compared to those without them. Comorbidities did not influence MDA level in GINTRA and GSMOF being still the lowest in GSMOF (p = 0.0033). In contrast, significantly higher MDA level was observed for GCLIN in those with vs. without comorbidities (p = 0.0262). CONCLUSIONS: Patients on HPN have lower antioxidant defenses than healthy controls. The type of lipid emulsion used in HPN affects lipid peroxidation (even after taking into account comorbidities which often involve oxidative stress) being the highest in GINTRA and the lowest in GSMOF. Thus, to minimize the risk of oxidative stress, SMOFlipid can be considered in patients in HPN especially for those with comorbidities. ClinOleic can be considered in HPN patients without comorbidities. The observation should be confirmed in larger studies.

Influence of Different Bacteria Strains Isolated from Septic Children on Release and Degradation of Extracellular Traps by Neutrophils from Healthy Adults.

Neutrophils are the first line of immune defense against pathogens. They use three major antimicrobial mechanisms: phagocytosis, degranulation, and release of neutrophil extracellular traps (NETs). NETs are structures which consist of nuclear DNA conjugated with antibacterial proteins. They are formed to entrap and kill pathogens. The aim of the study was to evaluate the influence of Escherichia coli (E. coli), Streptococcus pneumoniae (S. pneumoniae), Stenotrophomonas maltophilia (S. maltophilia), and Pseudomonas aeruginosa (P. aeruginosa), isolated from the peripheral blood of children with sepsis, on the release and degradation of NETs by neutrophils isolated from blood healthy adult subjects. Neutrophils were stimulated with the bacterial strains outlined above. The quantitative and qualitative analyses of NETs release were performed by fluorometric measurement and immunofluorescence, respectively. The ability of bacteria to degrade NETs was studied qualitatively. Oxidative burst was assessed by flow cytometry. Histone H3 citrullination was evaluated by Western blot. We found that NETs were formed only when neutrophils were incubated with S. pneumoniae. E. coli, P. aeruginosa, and S. maltophilia did not induce the release of the NETs. P. aeruginosa, S. pneumoniae, and E. coli induced the production of reactive oxygen species (ROS) by neutrophils. Two studied bacterial strains (S. pneumoniae and E. coli) were able to degrade NETs. However, none of the strains induced the citrullination of histone H3. We conclude that the ability of bacteria to induce and degrade NETs depends on the specific bacterial strain.

Role of Chlamydia pneumoniae in the pathogenesis of hypertrophy and adenoid tissue inflammation in children.

OBJECTIVE: A tropism to epithelial cells and lymphocytes, an inhibition of apoptosis in host cells, an ability to occurrence in persistent form resistant to antibiotic treatment are the features of Chlamydia pneumoniae, which can have connection with chronic inflammation of an adenoid tissue and adenoid hypertrophy. This study aimed to (1) detect the C. pneumoniae in an adenoid in children undergoing adenoidectomy, (2) estimate a connection between C. pneumoniae occurrence and the size of adenoid, (3) demonstration in which of adenoid cells C. pneumoniae occurs most often. MATERIAL AND METHODS: The examined group consisted of 200 children aged from 2 to 16 years (mean age 6,4) undergoing adenoidectomy. In all children during qualification for adenoidectomy a fiberoscopic examination of the nasopharynx was performed. A part of removed adenoid tissue was analysed by real-time PCR for C. pneumoniae. Adenoids from children with positive PCR examination and from 10 children with negative PCR examination were examined using immunohistochemistry (IHC). RESULTS: C. pneumoniae in the adenoid was present in 5,5% children. Positive results were obtained most frequently (24,14%, 7/29) in the eldest group (10-16 years). A statistical analysis demonstrated the correlation between C. pneumoniae occurrence in an adenoid tissue and the size of adenoid. In immunohistochemistry C. pneumoniae was found the most frequently in lymphocytes and in epithelial cells. CONCLUSIONS: A presence of C. pneumoniae in lymphocytes and epithelial cells of the adenoid first of all in older children with adenoid hypertrophy confirms the participation of this bacteria in adenoid pathology.

Chlamydia pneumoniae and typical bacteria occurrence in adenoid in children qualified for adenoidectomy.

OBJECTIVE: The ability to occur in a persistent form and to inhibit apoptosis in host cells are the features of Chlamydia pneumoniae which may be associated with adenoid hypertrophy. This study aimed to (1) demonstrate the presence of C. pneumoniae DNA in the adenoid in children qualified for adenoidectomy, (2) evaluate the correlation between the presence of C. pneumoniae in the adenoid and the child's age, the size of adenoid, the incidence of recurrent respiratory tract infections, (3) demonstrate the co-occurrence of C. pneumoniae and typical bacteria in the adenoid tissue. METHODS: A group of 200 children aged from 2 to 16 years (mean age 6.4) undergoing adenoidectomy from February 2010 to May 2011 were enrolled to the study. Adenoid tissue removed during planned adenoidectomy was analyzed for the presence of C. pneumoniae by real-time PCR and for the presence of typical bacteria by bacteriological culture. Statistical analysis was conducted using SPSS PASW Statistics 18 software. RESULTS: C. pneumoniae was detected in the adenoid in 5.5% of the 200 children. Positive results were obtained most frequently (24.1%, 7/29) in the eldest group (10-16 years). In PCR positive children the degree of choanal narrowing by an adenoid tended to be larger than in PCR negative children. The occurrence of C. pneumoniae in the adenoid was not associated with a presence of a specific typical bacterium. CONCLUSIONS: The presence of C. pneumoniae in an adenoid was most frequently detected in the children of 10-16 years qualified for adenoidectomy.

Chronic rhinosinusitis in children--bacteriological analysis in terms of cytological examination.

INTRODUCTION: The mechanisms of inflammatory response occurring in chronic rhinosinusitis in children are multifactorial. Besides the history and a physical examination, amongst diagnostic tools there are cytological and bacteriological examinations. OBJECTIVES: 1 Determining the nature of the bacterial flora present in the nasal cavities and paranasal sinuses in children with chronic rhinosinusitis amongst patients of The Department of Paediatric Otolaryngology,Warsaw Medical University. 2 Determining the relation between bacterial strains and cytological examination of nasal mucosa in children with chronic rhinosinusitis. MATERIALS AND METHODS: The study group included 64 patients with chronic rhinosinusitis without polyps. The control group included 30 randomly chosen children. Diagnostic tests performed in both groups were: middle meatal culture and cytological examination from the inferior nasal concha and middle meatus. Statistical analysis was accomplished with Statistica 8.0. CONCLUSIONS: Damage to the respiratory epithelial surface is understood as damage to the innate immune barrier, and repeated antibiotic therapy with the subsequent repopulation of the epithelium accidentally by various bacteria can become responsible for the pathogenic effect of bacteria in chronic rhinosinusitis.

Antibodies against N-homocysteinylated proteins and their determinants in patients on long-term hemodialysis.

INTRODUCTION: The factors that determine the level of antibodies against N-homocysteinylated (N-Hcy) proteins have not been established so far. The clinical significance of these antibodies and their effect on cardiovascular (CV) risk in patients with end-stage renal disease (ESRD) are still unknown. OBJECTIVES: The aim of this study was to identify the factors that determine the level of antibodies against N-Hcyalbumin and N-Hcy hemoglobin in patients on long-term hemodialysis (HD). PATIENTS AND METHODS: The study involved 247 subjects on long-term HD (110 women, 137 men; age range, 23-89 years) and 60 controls matched for age, sex, and CV risk factors (serum creatinine level <140 micromol/l). Serum antibodies against N-Hcyalbumin and N-Hcyhemoglobin were determined using an in-house enzyme-linked immunosorbent assay. Total homocysteine (tHcy), folate, and 8-isoprostaglandin F2alpha (8-iso-PGF(2alpha)) were also measured. RESULTS: Patients on HD had higher serum levels of anti-N-Hcy-albumin (absorbancy at 490 nm: 0.56 [0.49-0.623] vs. 0.259 [0.198-0.338], P <0.0001) and anti-N-Hcy-hemoglobin antibodies (0.659 [0.597-0.723] vs. 0.379 [0.289-0.442], P <0.0001) as compared with controls. The level of both antibodies correlated with tHcy (r = 0.56, P <0.0001 and r = 0.67, P <0.0001, respectively), 8-iso-PGF(2alpha) (r = 0.48, P <0.0001 and r = 0.63, P <0.0001, respectively), and folate (r = -0.18, P = 0.0054 and r = -0.38, P <0.0001, respectively), but not with HD duration, the initial cause of ESRD, and CV comorbidity. CONCLUSIONS: In HD patients, tHcy is an independent predictor of antibodies against N-Hcy proteins. Folate and 8-iso-PGF(2alpha) concentrations were not independently associated with the levels of both antibodies.