Antibacterial and Antibiofilm Effects of Photodynamic Treatment with Curcuma L. and Trans-Cinnamaldehyde against Listeria monocytogenes.

Photodynamic inactivation (PDI) is a highly effective treatment that can eliminate harmful microorganisms in a variety of settings. This study explored the efficacy of a curcumin-rich extract, Curcuma L., (Cur)- and essential oil component, trans-cinnamaldehyde, (Ca)-mediated PDI against Listeria monocytogenes ATCC 15313 (Lm) including planktonic cells and established biofilms on silicone rubber (Si), polytetrafluoroethylene (PTFE), stainless steel 316 (SS), and polyethylene terephthalate (PET). Applying Ca- and Cur-mediated PDI resulted in planktonic cell reductions of 2.7 and 6.4 log CFU/cm2, respectively. Flow cytometric measurements (FCMs) coupled with CFDA/PI and TOTO®-1 staining evidenced that Ca- doubled and Cur-mediated PDI quadrupled the cell damage. Moreover, the enzymatic activity of Lm cells was considerably reduced by Cur-mediated PDI, indicating its superior efficacy. Photosensitization also affected Lm biofilms, but their reduction did not exceed 3.7 log CFU/cm2. Cur-mediated PDI effectively impaired cells on PET and PTFE, while Ca-mediated PDI caused no (TOTO®-1) or only slight (PI) cell damage, sparing the activity of cells. In turn, applying Ca-mediate PDI to Si largely diminished the enzymatic activity in Lm. SS contained 20% dead cells, suggesting that SS itself impacts Lm viability. In addition, the efficacy of Ca-mediated PDI was enhanced on the SS, leading to increased damage to the cells. The weakened viability of Lm on Si and SS could be linked to unfavorable interactions with the surfaces, resulting in a better effect of Ca against Lm. In conclusion, Cur demonstrated excellent photosensitizing properties against Lm in both planktonic and biofilm states. The efficacy of Ca was lower than that of Cur. However, Ca bears potent antibiofilm effects, which vary depending on the surface on which Lm resides. Therefore, this study may help identify more effective plant-based compounds to combat L. monocytogenes in an environmentally sustainable manner.

Occurrence and Characteristics of Staphylococcus aureus Strains along the Production Chain of Raw Milk Cheeses in Poland.

Cheeses produced from unpasteurized milk by traditional production methods may contain many groups of microorganisms, including Staphylococcus aureus. The aim of this study was to determine the occurrence of S. aureus in the artisanal cheese production chain from unpasteurized milk. We investigated the prevalence of S. aureus strains isolated from various stages of artisanal cheese of unpasteurized milk production from farms in the northeastern and southern parts of Poland and characterized them. Characterization included antimicrobial susceptibility by microbroth dilution and biofilm formation by in vitro assay. Among all strains, the presence of enterotoxigenic genes and genes involved with biofilm formation and antibiotic resistance were screened by PCR-based methods. A total of 180 samples were examined. A high percentage of strains were resistant to penicillin (54/58.1%) and tobramycin (32/34.4%). Some tested isolates also showed resistance to the macrolide class of antibiotics: azithromycin, clarithromycin, and erythromycin at 17/18.3%, 15/16.1%, and 21/22.6%, respectively. Among tested isolates, we also found phenotypic resistance to oxacillin (9/9.7%) and cefoxitin (12/12.9%). The blaZ gene encoding penicillin resistance was the most common gene encoding antibiotic resistance among the tested strains. All isolates showing phenotypic resistance to cefoxitin possessed the mecA gene. The study also evaluated the prevalence of biofilm-associated genes, with eno the most frequently associated gene. Eighty-nine out of 93 S. aureus isolates (95.7%) possessed at least one enterotoxin-encoding gene. The results of this study showed that production of raw milk cheeses may be a source of antibiotic resistance and virulent S. aureus. Our results suggest that artisanal cheese producers should better control production hygiene.

Are Omentin Rs2274907 and Vaspin Rs2236242 Gene Polymorphisms Related to Body Composition, Lipid Profile and Other Adipokines in Prepubertal Healthy Children?

Purpose/Aim: So far no research concerning the omentin-1 (ITLN1) rs2274907 and vaspin (SERPINA12) rs2236242 polymorphisms has been carried out in a healthy pediatric population. We analyzed associations of these polymorphisms with anthropometric parameters, lipid profile, as well as adiponectin, leptin and soluble leptin receptor (sOB-R) levels in prepubertal healthy children, to search for their possible role in the risk of obesity and obesity-related disorders.Materials and Methods: Frequencies of these polymorphisms were analyzed by the restriction fragment length polymorphism in 89 normal-weight children. The body composition was measured by dual-energy X-ray absorptiometry. Serum levels of adipokines were measured using ELISA methods.Results: We observed differences in values of HDL-cholesterol (p = 0.002) and triglycerides (p = 0.039) in children carrying different genotypes of the ITLN1 rs2274907 polymorphism. In children carrying different genotypes of the SERPINA12 rs2236242 polymorphism differences in BMI (p =0.025) and BMI Z-score (p = 0.01) values were found. Significant relations between anthropometric parameters and levels of HDL-cholesterol and triglycerides were associated with minor alleles of the studied polymorphisms. In addition, leptin/sOB-R ratio was related to HDL-cholesterol (p = 0.004) and triglycerides (p = 0.03) levels in children carrying minor allele of the SERPINA12 rs2236242 SNP.Conclusions: We suggest that both ITLN1 rs2274907 and SERPINA12 rs2236242 polymorphisms influence body composition and lipid profile in prepubertal healthy children. Relations between anthropometric parameters, lipid and adipokine levels may be associated with minor alleles of the studied polymorphisms. The possible role of these polymorphisms in the modulation of the risk of obesity and obesity-related disorders in the later life might be considered.

Cadmium Stress Reprograms ROS/RNS Homeostasis in Phytophthora infestans (Mont.) de Bary.

Heavy metal pollution causes many soils to become a toxic environment not only for plants, but also microorganisms; however, little is known how heavy metal contaminated environment affects metabolism of phytopathogens and their capability of infecting host plants. In this study the oomycete Phytophthora infestans (Mont.) de Bary, the most harmful pathogen of potato, growing under moderate cadmium stress (Cd, 5 mg/L) showed nitro-oxidative imbalance associated with an enhanced antioxidant response. Cadmium notably elevated the level of nitric oxide, superoxide and peroxynitrite that stimulated nitrative modifications within the RNA and DNA pools in the phytopathogen structures. In contrast, the protein pool undergoing nitration was diminished confirming that protein tyrosine nitration is a flexible element of the oomycete adaptive strategy to heavy metal stress. Finally, to verify whether Cd is able to modify P. infestans pathogenicity, a disease index and molecular assessment of disease progress were analysed indicating that Cd stress enhanced aggressiveness of vr P. infestans towards various potato cultivars. Taken together, Cd not only affected hyphal growth rate and caused biochemical changes in P. infestans structures, but accelerated the pathogenicity as well. The nitro-oxidative homeostasis imbalance underlies the phytopathogen adaptive strategy and survival in the heavy metal contaminated environment.

Enterococci from ready-to-eat food - horizontal gene transfer of antibiotic resistance genes and genotypic characterization by PCR melting profile.

BACKGROUND: The aim of this study was to evaluate the possibility of the horizontal transfer of genes encoding resistance to aminoglycosides (aac(6')-Ie-aph(2″)-Ia, aph(2″)-Ib, aph(2″)-Ic, aph(2″)-Id, ant(4')-Ia and ant(6')-Ia), tetracyclines (tetM, tetL, tetK, tetO and tetW), and macrolides (ermA, ermB, ermC, msrC, mefAB) in Enterococcus strains isolated from ready-to-eat dishes purchased in bars and restaurants in Olsztyn, Poland. RESULTS: It was found that 74% of tested strains were able to conjugal transfer at least one of the antibiotic resistance genes. Transfer of resistance to tetracyclines in strains was observed with a frequency ranging from 1.3 × 10-6 to 8.7 × 10-7 transconjugants/donor. The int gene and the tetM gene were transferred simultaneously, which indicated that a transposon of the Tn916/Tn1545 also participated in the conjugation process. The frequency of transferring genes of resistance to macrolides ranged from 3.2 × 10-6 to 2.4 × 10-8 transconjugants/donor. The ermB gene was transferred the most frequently. The frequency of acquisition of genes encoding aminoglycosides in strains isolated from food ranged from 1.7 × 10-6 to 3,2 × 10-8 transconjugants/donor. Transfer of the aac(6')-Ie-aph(2″) gene was the most frequent. In all reactions, the clonal character of transconjugants and recipients was confirmed by the polymerase chain reaction melting profile (PCR MP) method, which is an alternative to the pulsed field gel electrophoresis (PFGE) method. CONCLUSION: The findings of this study indicate that Enterococcus isolated from ready-to-eat food is able to horizontally transfer genes encoding various antibiotic resistance mechanisms. © 2018 Society of Chemical Industry.

The effect of weight loss on body composition, serum bone markers, and adipokines in prepubertal obese children after 1-year intervention.

Purpose/Aim: The influence of weight loss on bone turnover and bone quality in childhood remains controversial, but it may implicate interactions between adiposity and bone metabolism. Therefore, we studied the relationships between bone markers and adipokines during weight loss in obese children. MATERIALS AND METHODS: We determined serum leptin, soluble leptin receptor, adiponectin, bone alkaline phosphatase (BALP), C-terminal telopeptide of type I collagen (CTX-I), osteocalcin (OC), carboxylated-OC (Gla-OC), undercarboxylated-OC (Glu-OC), sclerostin, body composition, and bone mineral density (BMD) in 40 obese prepubertal children before and after therapy. The control group, matched for sex and age, consisted of 40 non-obese children. RESULTS: We found that values of the total body less head-bone mineral content (TBLH-BMC) and TBLH-BMD were significantly increased, but TBLH-BMD Z-score was decreased by 25% (p = 0.002) in obese children with weight loss after therapy. We observed increases of CTX-I to OC ratio (p = 0.009), and Gla-OC concentrations (p = 0.049). Changes in TBLH-BMD Z-score in patients were positively correlated with changes in BMI Z-score (p = 0.001), percentage of fat mass (p = 0.004), and BALP activity (p = 0.01). Changes in BALP activity were negatively correlated (p < 0.001) with changes in adiponectin concentrations, while changes in sclerostin levels were positively correlated (p = 0.001) with leptin changes. CONCLUSIONS: We suggest that alterations in adipokines metabolism were associated with a lower rate of bone mineral accrual as a result of decreased bone formation rather than increased bone resorption. The lower rate of bone mass accrual in weight losing children may be an effect of reduced BALP levels related to increase in adiponectin levels.

The levels of bone alkaline phosphatase (BALP) and soluble epidermal growth factor receptor-2 (ECD/HER-2) in pediatric patients with osteosarcoma during clinical treatment.

OBJECTIVE: Aim: The aim of this study was to assess the usefulness of bone-specific alkaline phosphatase (BALP) and the extracelluar domain of human epidermal growth factor receptor 2 (ECD/HER-2) measurements in pediatric patients with osteosarcoma as prospective prognostic and predictive markers for monitoring the treatment and early detection of disease recurrence. PATIENTS AND METHODS: Material and methods: We studied 22 patients (5 girls, 17 boys) aged 7-20 years with osteosarcoma (OS) treated at the Institute of Mother and Child in Warsaw. All the patients were evaluated for the serum levels of BALP and ECD/HER-2 before treatment, during pre- and postoperative chemotherapy and after the completion of treatment. Healthy children (n=22) were the reference group. The levels of BALP and ECD/HER-2 were measured using immunoenzymatic methods. RESULTS: Results: The values of BALP and ECD/HER-2 proteins were higher (p<0.01; p<0.05, respectively) in patients with osteosarcoma at the time of diagnosis compared with the control group. The values of both markers significantly decreased during chemotherapy in most patients with remission. In contrast to ECD/HER-2, the value of BALP after therapy was higher in patients with progression than with remission (p<0.001). CONCLUSION: Conclusions: Our results demonstrate the different pattern of BALP and ECD/HER-2 proteins during clinical treatment in patients with osteosarcoma. Higher values of BALP may characterize the progression of the disease and unfavourable prognosis. Further longitudinal studies are necessary to confirm the prognostic values of BALP and ECD/HER-2 proteins in this group of patients.

Relations between oxidized low-density lipoproteins and fat-soluble vitamin concentrations in obese children - preliminary study.

INTRODUCTION: Although lipid peroxidation products are formed during normal cell metabolism, they appear mostly in pathological conditions via producing an excess of free radicals that can react with unsaturated fatty acids, in particular low-density lipoprotein (LDL). The aim of this study was to investigate the relations between oxidized LDL (oxLDL), the fat-soluble vitamin status and the anthropometric parameters in prepubertal obese children. MATERIAL AND METHODS: Thirty-two obese (SDS-BMI >2) and 25 non-obese children (SDS-BMI <-1+1>) were included in the study. The concentration of oxLDL was determined in the serum by the ELISA assay. Vitamin A and E were measured by the high-pressure liquid chromatography method. Total cholesterol, LDL- and HDL-cholesterol, and triglyceride levels were determined by enzymatic methods. RESULTS: The concentrations of oxLDL and vitamin A were higher in obese children than in normalweight controls by about 50% (p=0.01) and 40% (p=0.001), respectively. In obese children the significant positive correlation was found between oxLDL and vitamin A concentrations (<0.05). In addition, oxLDL correlated positively with BMI values (<0.05) and the amount of fat mass (kg) (<0.02) in these patients. Concentrations of vitamins A and E correlated with the level of total cholesterol (<0.05; <0.01, respectively).Moreover, a positive correlation between vitamin E and LDL-cholesterol was observed (<0.05). CONCLUSIONS: Our preliminary study shows that oxLDL starts early during the prepubertal period and may precede atherosclerotic lesions. We suggest there is an occurrence of relationships between vitamin A and oxidized LDL in prepubertal obese children. Vitamin A and E concentrations are also associated with dyslipidemia in these patients.

ADIPOQ -11377C>G Polymorphism Increases the Risk of Adipokine Abnormalities and Child Obesity Regardless of Dietary Intake.

OBJECTIVE: The aim of the present study was to verify whether selected functional single nucleotide polymorphisms in LEP, LEPR, and ADIPOQ loci are associated with the development of obesity and serum levels of the respective adipokines in prepubertal white children with obesity. METHODS: Frequencies of -2548G>A LEP (rs7799039), Q223R (rs1137101) and K656N (rs8129183) LEPR, and -11377C>G (rs266729) and -11426A>G (rs16861194) ADIPOQ polymorphisms were analyzed by restriction fragment length polymorphism in 101 obese (standard deviation score [SDS]-body mass index [BMI] >2) and 67 normal-weight (SDS-BMI <- 1 + 1 >) children. Serum adipokine concentrations were measured using the enzyme-linked immunosorbent assay method. RESULTS: The GC/GG genotypes of -11377C>G ADIPOQ polymorphism were associated with a higher risk of obesity (P = 0.022, odds ratio 2.08 [95% confidence interval 1.11-3.90]). Individuals carrying the GG genotype had a higher leptin/total adiponectin ratio by 25% than CC homozygotes (P trend = 0.05). In the multivariate linear regression model, we found differences among particular genotypes of this polymorphism in concentrations of high molecular weight (HMW) adiponectin (P trend = 0.043) and HMW/total adiponectin ratio (P trend = 0.048), with the lowest values in GG homozygotes. Positive correlations between SDS-BMI and dietary reference intake percentage were observed in individuals homozygous for allele C (r = 0.403, P = 0.01) and CG heterozygotes (r = 0.428, P = 0.004). No significant correlations between both parameters were found in the GG homozygotes. CONCLUSIONS: Among the analyzed polymorphisms, only -11377C>G ADIPOQ single nucleotide polymorphism was associated with obesity during the prepubertal period. Adipokine abnormalities coexisting with the lack of relations between SDS-BMI and dietary intake may predict a higher risk of future obesity-related disorders in obese children carrying the GG genotype than in those with other genotypes.

[Serum concentrations of visfatin and oxidative stress markers in tobacco smoking pregnant women].

Tobacco smoking during pregnancy increases the risk of severity of oxidative stress, which can affect adipokines expression in adipose tissue. The aim of the study was to assess the effects of smoking on the level of visfatin and selected markers of oxidative stress (ox-LDL- oxidized low density lipoprotein, TOC - total oxidant capacity, TAC - total antioxidant capacity) and the association between them in pregnant women. Material and Methods: The research material was the venous blood of healthy women admitted to given birth in the Department of Obstetrics and Gynecology Institute of Mother and Child in the years 2015 - 2016. Based on an interview and determination of serum cotinine, women were divided into following groups: smoking (n=41) and non-smoking group (n=43). Concentrations of biochemical parameters were determined by enzyme immunoassay using commercial kits. Results: In serum of smoking pregnant women concentrations of visfatin, ox-LDL, and TOC were significantly higher while level of TAC was lower than observed in nonsmoking group (p<0.001). We found important association between number of cigarettes daily consumed and serum level of visfatin (r=0.43; p<0.01), ox-LDL (r=0.36; p<0.05), and TOC (r=0.39; p<0.05). Multivariate regression analysis showed a positive correlation between concentration of visfatin and level of ox-LDL as well as TOC both in the smoking women (ox- LDL: ß=0.381; TOC: ß=0.388; p<0.05), and in tobacco abstinent group (ox- LDL: ß= 0.470; TOC: ß=0.326; p<0.05). Conclusions: These results indicate that smoking during pregnancy adversely affect the oxidant/ antioxidant balance. The association between visfatin concentrations and markers of oxidative stress may suggest prooxidant tendency of this adipokine.

Influence of exogenous leptin on redox homeostasis in neutrophils and lymphocytes cultured in synovial fluid isolated from patients with rheumatoid arthritis.

OBJECTIVES: Leptin is an adipose cells derived hormone that regulates energy homeostasis within the body. Energy metabolism of immune cells influences their activity within numerous pathological states, but the effect of leptin on these cells in unclear. On the one hand, it was observed that leptin induces neutrophils chemotaxis and modulates phagocytosis. On the other hand, neutrophils exposed to leptin did not display detectable Ca(2+) ions mobilization or ß2-integrin upregulation. In this study, we investigated the effect of leptin on the redox homeostasis in lymphocytes and neutrophils. MATERIAL AND METHODS: Neutrophils and lymphocytes were isolated by density-gradient centrifugation of blood from healthy volunteers. Cells were cultured with or without leptin (100 ng/ml for lymphocytes and 500 ng/ml for neutrophils) or with or without synovial fluid (85%) for 0-72 h. Culture media were not changed during incubation. Cells were homogenized and homogenate was frozen until laboratory measurements. Redox homeostasis was assessed by the reduced glutathione (GSH) vs. oxidized glutathione (GSSG) ratio and membrane lipid peroxidation evaluation. RESULTS: Lymphocytes cultured with leptin and synovial fluid showed a significant increase of the GSSG level. The GSSG/GSH ratio increased by 184 ±37%. In neutrophils incubated in a similar environment, the GSSG/GSH ratio increased by just 21 ±7%, and the effect was observed irrespectively of whether they were exposed to leptin or synovial fluid or both together. Neither leptin nor synovial fluid influenced lipid peroxidation in neutrophils, but in lymphocytes leptin intensified lipid peroxidation. CONCLUSIONS: Leptin altered the lymphocytes, but not neutrophils redox state. Because firstly neutrophils are anaerobic cells and have just a few mitochondria and secondly lymphocytes have typical aerobic metabolism, the divergence of our data supports the hypothesis that leptin induces oxidative stress by modulation of mitochondria.

Body composition parameters and adipokines levels in relation to bone mineral density in patients with malignant bone tumors after treatment.

BACKGROUND: Recent data support the concept that adipokines, which are secreted by fat cells, are important modulators of bone metabolism. The aim of this study was to assess the relationships between body composition parameters, adipokines (leptin, adiponectin) and bone mineral density (BMD) in patients with malignant bone tumors after anticancer therapy. PROCEDURE: The study included 35 patients (median age 14.8 years) with diagnosed malignant bone tumors treated according to obligatory protocols. Total fat mass, lean mass, bone mineral content (BMC) and BMD measurements were performed after treatment completion by dual energy-ray absorptiometry (DXA). Serum concentrations of leptin, leptin receptor and adiponectin were determined using immunoenzymatic assays. The control group consisted of 28 healthy children (median age 14.3 years). RESULTS: Patients with bone tumor after chemotherapy had significantly higher fat mass (P < 0.01), increased ratio of fat mass/lean mass (P < 0.001), and a decrease (P < 0.001) in total body and lumbar spine BMD compared with controls. We observed higher serum leptin concentration (P < 0.01) and lower soluble leptin receptor (P < 0.001) and adiponectin (P = 0.01) in patients than in controls. The ratios of leptin/leptin receptor and leptin/adiponectin were about three-fold higher in patients compared with the control group (P = 0.001). We found a significant positive correlation between BMD and body composition and a negative correlation between BMD and adiponectin in the patients group. CONCLUSIONS: Changes in body composition parameters coexisting with disturbed adipokine levels, especially higher levels of leptin and lower levels of adiponectin, might be associated with bone status in patients treated for malignant bone tumors.

[Adipokines, body composition and bone mineral density in underweight children]. / Adipokiny, parametry skladu ciala i gestosc mineralna kosci u dzieci z niedoborem masy ciala.

UNLABELLED: One of the important factors affecting bone health is body weight. Underweight children are predisposed to disturbances in bone metabolism, which may result in osteopenia and osteoporosis in later life. AIM: The aim of the study was to assess the relationship between adipokines, bone metabolism, and anthropometric parameters in underweight prepubertal children. MATERIALS AND METHODS: The study included 60 children aged 5-10 years. Among them, there were: 30 underweight children (BMI z-score ≤-1) and 30 normal-weight children (BMI z-score <-1 + 1 >). Body composition (fat mass, lean body mass, bone mass) and bone mineral density examination were performed by densitometry. Serum concentrations of bone metabolism markers and adipokines were determined by immunoenzymatic methods. RESULTS: In underweight children we observed significantly lower fat mass (p<0.0001), lean mass (p<0.001), bone mineral content (p<0.01) and bone mineral density both the total body (p<0.01) as well as lumbar spine L2-L4 (p<0.05) compared with normal-weight children. In the group of underweight children, serum concentration of bone resorption marker (CTX) was significantly higher than in normal-weight children (2.006±0.649 vs. 1.624±0.492 ng/ml, p<0.05), with no differences in the concentrations of osteocalcin and sclerostin between studied groups. The ratio of adipokines (leptin/adiponectin) was approximately 2-fold lower in underweight than in normal-weight subjects. In underweight children we observed positive correlations between concentrations of sclerostin and bone turnover markers (OC, CTX) and between adiponectin and CTX. However, there was no correlation between fat mass and leptin concentration in this group of children. CONCLUSIONS: Low body weight in prepubertal period is related with an alteration in the adipokines profile and bone metabolism markers, resulting in a decrease in bone mineral density.

[Umbilical cord serum hepcidin levels and maternal smoking during pregnancy]. / Stezenia hepcydyny we krwi pepowinowej dzieci matek palacych tyton w przebiegu ciazy.

Anemia during pregnancy is a risk factor of disturbance in pre- and postnatal child's development. Hepcidin plays the key role in iron metabolism, as protein participating in the regulation of intestinal absorption of this element and its release from macrophages, and transport across the placenta. Maternal smoking during pregnancy can result in disturbances of iron homeostasis leading to a subclinical deficiency. The depletion of maternal iron can cause fetal hypoxia condition and decreased expression of hepcidin. The aim of the study was to evaluate the effect of smoking on the levels of hepcidin and erythropoietin (as an indicator of hypoxia) and their relationships in umbilical cord blood. The research material was the umbilical cord blood of 50 newborns born in the Department of Obstetrics and Gynecology (Institute of Mother and Child in Warsaw) in the years 2013-2014. Based on an interview and determination of cotinine in the blood of mothers, newborns were divided into following groups: children of smoking mothers (n=20) and children of tobacco abstinent mothers (n=30). Hepcidin and erythropoietin concentrations were determined by enzyme immunoassay using commercial kits (DRG, Germany). It has been shown that hepcidin concentrations were significantly lower in children of smoking mothers than in the group of tobacco abstinent (37.5 ng/mL vs 45.1 ng/mL, p<0.001). However, the level of erythropoietin was higher in children of smoking mothers than in children of non-smoking women (p<0.001). A negative correlation between the levels of hepcidin and erythropoietin (r = -0.41, p<0.05) and number of smoked cigarettes (r = -0.43, p<0.05) was observed. These results indicate that smoking during pregnancy significantly affects hepcidin levels in children born at term. Decrease of hepcidin concentration coexisting with high level of erythropoletin in umbilical cord blood in children of smoking pregnant women may be the cause of subclinical deficiency of iron in the newborn.

The effect of leptin on the respiratory burst of human neutrophils cultured in synovial fluid.

OBJECTIVES: Leptin is a hormone responsible for nutritional status and immune competence coordination. In rheumatoid arthritis (RA) increased leptin levels were observed in both serum and synovial fluid. Its influence on development of the disease still remains unclear. So far, research on leptin's influence on the emission of reactive oxygen intermediates (ROI) measured with chemiluminescence (CL) has provided unclear and contradictory results. In this study, we evaluated the influence of leptin on oxidative activity of neutrophils isolated from blood of healthy volunteers and cultured in different amounts of synovial fluid (SF) from patients with RA. MATERIAL AND METHODS: Neutrophils' oxidative metabolism was measured by two types of CL. The first one, luminol-dependent CL (CL-lum), allows one to determine phagocytic activity and the level of ROI generated in a myeloperoxidase-dependent manner. The second method used was lucigenin-dependent CL (CL-luc), which monitors ROI production dependent on the NADPH oxidase enzyme complex located in the cell membranes of neutrophils and enables one to determine the scope of extracellular ROI emission. RESULTS: Neutrophils stimulated by opsonized zymosan show a decrease in the level of CL-lum, proportional to the increasing concentration of both SF and serum collected from healthy donors. The observed effect of decreased CL-lum may, therefore, be dependent on the physical conditions (viscosity of fluids used). None of these experiments showed any effect of leptin on the level of CL-lum. CONCLUSIONS: The present study showed that leptin does not affect the level of any of the CL types in inactive neutrophils incubated in normal serum, and it does not affect the level of oxidative activity in resting neutrophils incubated with SF. However, leptin influences extracellular ROI emission (measured by CL-luc). Leptin reduces extracellular emission of ROI, and this effect is dependent on concentration and duration of exposure to leptin.

[Assessment of anthropometric parameters and serum leptin and fetuin-A levels in children and adolescents with osteosarcoma after anticancer treatment]. / Ocena parametrów antropometrycznych oraz stezen leptyny i fetuiny-A u dzieci i mlodziezy po zakonczeniu leczenia miesaka kosciopochodnego.

UNLABELLED: Cancer and the use of a comprehensive anti-cancer treatment are unfavorable factors, which have a significant impact on bone mass accumulation, bone mineralization and consequently the occurrence of osteoporosis. Bone turnover is regulated by complex mechanisms, among which an important role play OPG/RANK/RANKL signaling pathway, adipokines, and fetuin-A. The aim of the study was to evaluate bone mineral density and concentrations of leptin and fetuin-A in patients with osteosarcoma after anti-cancer treatment. MATERIALS AND METHODS: The study included 50 children and adolescents aged 10-21 years. The study group consisted of 25 patients with osteosarcoma and 25 healthy counterparts as a control group. The examination was conducted 2 months after the last course of postoperative chemotherapy and included densitometric measurements: bone mineral content (BMC), bone mineral density (BMD), fat mass, lean mass and biochemical measurements: serum concentrations of calcium, magnesium, phosphate, 25-hydroksyvitamin D, alkaline phosphatase, leptin and fetuin-A. Concentrations of leptin and fetuin-A were determined by immunoenzymatic methods. RESULTS: In patients with osteosarcoma after anti-cancer treatment, we observed significantly reduced bone mineral content, bone mineral density and lean body mass compared with the healthy children (p < 0.05, p < 0.01, p < 0.05, respectively). Mean values of z-score of the whole body BMD and z-score of the lumbar BMD L1-L4 were significantly lower in patients than in the controls (p < 0.001). The serum concentrations of phosphate, magnesium, and alkaline phosphatase in both studied groups were similar, while calcium was significantly lower (p < 0.05) in patients than in the healthy children. The concentration of 25-hydroxyvitamin D was about two-fold lower, while leptin approximately 2.5-fold higher in patients than in the controls. The mean value of fetuin-A was similar in both studied groups. Statistically significant positive correlations between body composition parameters and the values of BMD, as well as between anthropometric parameters and leptin and fetuin-A were observed. CONCLUSION: The deficit in bone mass observed in patients with malignant bone tumors after anti-cancer treatment might be the result of decreased serum calcium and vitamin D concentrations. The observed correlation between anthropometric and biochemical parameters may indicate the link between bone and adipose tissue metabolism.

Assessment of Biochemical Bone Turnover Markers in Polish Healthy Children and Adolescents.

BACKGROUND: Assessing bone turnover in paediatric populations is crucial for understanding the physiological changes occurring during skeletal development and identifying potential abnormalities. The objective of this study was to assess osteocalcin (OC), bone alkaline phosphatase (BALP), and C-terminal telopeptide of type I collagen (CTX-I) levels reflecting bone formation and resorption for age and sex in Polish healthy children and adolescents. MATERIALS AND METHODS: A total of 355 healthy normal-weight children and adolescents (46.5% girls) aged 1-18 years old were recruited. Total body less head (TBLH) and spine L1-L4 were used in children to assess bone mineral density (BMD) by dual-energy X-ray absorptiometry (DXA). Bone marker concentrations were determined by immunoenzymatic methods. RESULTS: Bone marker levels in girls and boys started with higher values in the first year of life and subsequently decreased until reaching a nadir during the prepubertal period. The pubertal peak values of bone markers were reached at 11-13 years old in boys and at 9-11 years old in girls. After puberty, the adolescents showed a gradual decline in bone marker concentrations to the values observed in adults. We found positive correlations between OC level and TBLH-BMD (r = 0.329, p = 0.002), TBLH-BMD Z-score (r = 0.245, p = 0.023), and L1-L4 BMD (r = 0.280, p = 0.009) in the prepubertal group. CONCLUSIONS: We showed serum levels of bone turnover markers-BALP, OC, and CTX-I-in relation to age and sex in healthy Polish children and adolescents. The age intervals of these markers for girls and boys aged 1-18 years old may be clinically useful in the assessment of bone metabolism in individuals with skeletal disorders.

[The effect of tobacco smoking on serum concentration of selected angiogenic factors and somatomedin C in pregnant women and umbilical cord blood]. / Wplyw palenia tytoniu na stezenia wybranych czynników angiogennych oraz somatomedyny C w surowicy krwi kobiet ciezarnych i krwi pepowinowej.

The objective of this study was to evaluate the effect of cigarette smoking on concentration of selected angiogenic factors (vascular endothelial growth factor VEGF, placenta growth factor PIGF) and somatomedin C (insulin-like growth factor-I) in blood of mothers and umbilical cord blood. The correlations between studied biochemical parameters and markers of estimated intensity of cigarette smoking as well as birth weight were also determined. Fifty healthy pregnant women were divided into two groups: smoking and tobacco abstinent group according to serum cotinine concentration. The current smokers were defined as those who had smoked 5 cigarettes per day for 2 years before conception and continued smoking during pregnancy. In the group of smoking mothers the mean serum concentration of cotinine was 91.6 microg/L and correlated positively with number of cigarettes daily consumed (r = 0.58, p < 0.01) as well as with time of smoking before conception (r = 0.40, p < 0.05). The mean serum concentration of PIGF in III trimester of pregnancy was significantly higher in the group of smokers than in non-smoking ones (p < 0.0001) and correlated with serum cotinine concentration (r = 0.41, p < 0.05) and number of cigarettes daily consumed (r = 0.58, p < 0.01). The concentration of serum VEGF was similar in both studied group. The mean serum level of IGF-I was significantly lower in group of smokers than in non-smokers in the I and III trimester of gestation (p < 0.01). Also in umbilical cord blood of smoking newborn the concentration of IGF-I was lower by 20% than in nonsmoking group (p < 0.05). We observed negative correlation between number of cigarettes daily consumed and serum level of IGF-I in blood of mothers as well as in blood of their children (I trimester: r = -0.43, p < 0.05; III trimester: r = -0.70, p < 0.001; umbilical cord blood: r = -0.45, p < 0.05). In both studied groups there were a positive correlation between birth weight and concentrations of IGF-I in blood of mothers and umbilical cord blood (group of smokers: mothers r = 0.43, p < 0.05, cord blood r = 0.50, p < 0.01; group of tobacco abstinent: mothers r = 0.51, p < 0.01, cord blood r = 0.41, p < 0.05). The birth weight of the smoking mothers' infants was lower by about 400 g (p < 0.01) and their birth body length by 1.5 cm (p < 0.05) and negatively correlated with number of cigarettes smoked per day (r = -0.55; p < 0.005). Our results indicate, that tobacco smoking during pregnancy increased serum PIGF levels in the final stages of gestation and has no effect on the concentration of VEGF, which may lead to an increase of trophoblast proliferation and uteroplacental dysfunction. Lower than in tobacco abstinent levels of IGF-I in serum of smoking mothers and in umbilical cord blood and their close relationship with birth weight, may suggests a direct effect of this factor on birth anthropometric parameters.

Identification of the Enterotoxigenic Potential of Staphylococcus spp. from Raw Milk and Raw Milk Cheeses.

This study aimed to genotypic and phenotypic analyses of the enterotoxigenic potential of Staphylococcus spp. isolated from raw milk and raw milk cheeses. The presence of genes encoding staphylococcal enterotoxins (SEs), including the classical enterotoxins (sea-see), non-classical enterotoxins (seg-seu), exfoliative toxins (eta-etd) and toxic shock syndrome toxin-1 (tst-1) were investigated. Isolates positive for classical enterotoxin genes were then tested by SET-RPLA methods for toxin expression. Out of 75 Staphylococcus spp. (19 Staphylococcus aureus and 56 CoNS) isolates from raw milk (49/65.3%) and raw milk cheese samples (26/34.7%), the presence of enterotoxin genes was confirmed in 73 (97.3%) of them. Only one isolate from cheese sample (1.3%) was able to produce enterotoxin (SED). The presence of up to eight different genes encoding enterotoxins was determined simultaneously in the staphylococcal genome. The most common toxin gene combination was sek, eta present in fourteen isolates (18.7%). The tst-1 gene was present in each of the analyzed isolates from cheese samples (26/34.7%). Non-classical enterotoxins were much more frequently identified in the genome of staphylococcal isolates than classical SEs. The current research also showed that genes tagged in S. aureus were also identified in CoNS, and the total number of different genes detected in CoNS was seven times higher than in S. aureus. The obtained results indicate that, in many cases, the presence of a gene in Staphylococcus spp. is not synonymous with the ability of enterotoxins production. The differences in the number of isolates with genes encoding SEs and enterotoxin production may be mainly due to the limit of detection of the toxin production method used. This indicates the need to use high specificity and sensitivity methods for detecting enterotoxin in future studies.

Biofilm Formation Capacity and Presence of Virulence Determinants among Enterococcus Species from Milk and Raw Milk Cheeses.

Bacterial biofilm is one of the major hazards facing the food industry. Biofilm-forming ability is one of the most important virulence properties of enterococci. The genus Enterococcus includes pathogenic, spoilage, and pro-technological bacteria. The presence of enterococci in milk and dairy products is usually associated with inadequate hygiene practices. The study examined the isolates' capacity for biofilm formation and identification of the genetic determinants of its formation among 85 Enterococcus strains isolated from raw milk (n = 49) and soft-ripened cheeses made from unpasteurized milk (n = 36). E. faecalis and E. faecium were the dominant species. The obtained results showed that 41.4% isolates from milk and 50.0% isolates from cheeses were able to form biofilm. All of the isolates analyzed had at least one of the studied genes. As regards the isolates from raw milk, the most prevalent gene was the gelE (85.6%), followed by the asa1 (66.7%). None of the isolates from cheeses showed the presence of cylA and sprE. The most prevalent gene among the strains from this source was the epbC (94.4%), followed by the gelE (88.9%). In isolates from both sources, the presence of proteins from the Fsr group was noted the least frequently. Nevertheless, results showed that were no significant differences between the biofilm-producing Enterococcus spp. and non-biofilm-producing isolates in term of occurrences of tested virulence genes. The ability to produce a biofilm by enterococci isolated from raw milk or ready-to-eat products emphasizes the need for continuous monitoring of the mechanisms of microbial adhesion.