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.

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.

[Assessment of selected bone metabolism marker concentrations in children with cystic fibrosis]. / Analiza stezen wybranych parametrów metabolizmu kostnego u dxieci chorych na mukowiscydoze.

AIM: The aim of this study was to assess bone formation and resorption processes and bone metabolism regulators, such as osteoprotegerin and fetuin-A in children with cystic fibrosis. MATERIAL AND METHODS: We examined 45 children with cystic fibrosis aged 5-13 years treated at the Institute of Mother and Child in Warsaw. The control group consisted of 35 healthy children in the same synage range without any diseases which may influence bone metabolism. We determined serum calcium and phosphate levels by colorimetric methods, vitamin D3 by the chemiluminiscence method and bone metabolism markers (osteocalcin, 5b isoenzyme of tartrate-resistant acid phosphatase, osteoprotegerin, fetuin-A) by immunoenzymatic methods. RESULTS: Mean serum concentrations of calcium and phosphate in the studied children were within the reference ranges. However, the level of 25-hydroxyvitamin D3 was significantly lower in patients with cystic fibrosis compared to the controls (19.3±7.6 vs 25.2±8.9 ng/ml, p<0.01). In cystic fibrosis children we observed a statistically significant lower concentration of osteocalcin (81.9±28.9 vs 97.9±28.6 ng/ ml, p<0.01) and similar activity of 5b isoenzyme of tartrate-resistant acid phosphatase (12.5±2.9 vs 13.4±3.5 U/L) as compared to healthy peers. Mean serum concentration of osteoprotegerin in patients with CF was significantly lower than in the control children (4.1±0.98 vs 4.59±0.86 pmol/l, p<0.05). Serum concentration of fetuin-A was comparable in both groups of children. CONCLUSIONS: In children with cystic fibrosis changes in the profile of bone metabolism markers were observed. Even patients with CF who are clinically stable and supplemented with vitamins are at risk of osteopenia and osteoporosis in their later life. Therefore, they should be under a comprehensive medical and nutritional care in order to achieve their optimal peak bone mass.

The effect of vegetarian diet on selected essential nutrients in children.

INTRODUCTION: Vegetarian diets are considered to promote health and reduce the risk of some chronic diseases. It is also known that restriction or exclusion of animal foods may result in low intake of essential nutrients. The aim of the presented study was to assess the intake and serum status of vitamin B12, folate, vitamins A, E and D, as well as concentrations of homocysteine, total antioxidant status and iron balance in Polish vegetarian children. MATERIAL AND METHODS: The study included 50 children, aged 5-11 who had been referred to the Institute of Mother and Child for dietary consultation. From those, 32 were vegetarians (aged 6.5±4.2 years) and 18 omnivores (aged 7.9±2.7 years). Dietary constituents were analyzed using the nutritional programme Dietetyk2®. Folate and vitamin B12 were determined with a chemiluminescence immunoassay, total homocysteine with a fluorescence polarization immunoassay and TAS (total antioxidant status) by colorimetric method. Vitamin A and E in serum were determined by the high-pressure liquid chromatography method (HPLC) and vitamin D by immunoenzymatic assay (ELISA). Concentrations of iron, ferritin, transferrin and total iron-binding capacity (TIBC) in serum were determined by commercially available kits. RESULTS: In vegetarian children daily intake of vitamin B12 (1.6 eg) was in the recommended range, that of folate (195 eg) and vitamin A (1245 eg) higher, but vitamin E slightly lower (6.6 eg) and three-fold lower vitamin D (1.1 eg) than references allowance. Serum concentrations of vitamin B12 (548 pg/ml), folate (12.8 ng/ml), vitamin A (1.2 emol/L), vitamin E (15.6 emol/l) were within physiological range, but that of vitamin D (13.7 eg/L) was only half of the lowest limit of the reference value. In vegetarian children in comparison to omnivorous similar levels of homocysteine (6.13 emol/L vs 5.45 emol/L) and vitamin A (1,17 emol/L vs 1.32 emol/L) were observed. Lower (p<0.05) values of vitamin E (15.6 emol/L vs 18.4 emol/L) and TAS (1.21 mmol/L vs 1.30 mmol/L; p<0.0001) were found. Concentrations of iron markers were in physiological range. CONCLUSION: Obtained results indicated that intakes of vitamin B12 and folic acid from vegetarian diets are sufficient to maintain serum concentrations of both homocysteine and iron in the range observed in omnivorous children. High consumption of vitamin A and low vitamin E only slightly affected their serum values. Significantly lower concentration of serum vitamin E in vegetarian children in comparison to nonvegetarians may be reflected with statistically significant lowering of total antioxidant status. Insufficient intake of vitamin D and its low serum concentration should be under close monitoring in vegetarian children. In order to prevent vitamin D deficiency appropriate age-dependent supplementation should be considered.

[Influence of vegetarian diet on serum values of homocysteine and total antioxidant status in children]. / Wplyw stosowania diety wegetarianskiej na stezenie homocysteiny oraz calkowita aktywnosc przeciwutleniajaca w surowicy u dzieci.

UNLABELLED: The vegetarian diet may play a preventive role in the development of chronic diseases such as coronary heart and cardiovascular disease. However increase of homocysteine (Hcy) concentration in peoples avoiding animal products may contribute to an increased atherosclerotic risk in these subjects. Recent evidence has suggested that role of hyperhomocysteinemia in atherogenesis is associated with process of autooxidation, which can promote the production of hydroxyl radicals, resulting in oxidative modification of low density lipoprotein and endothelium injury. The oxidant-antioxidant imbalance depends not only on the amount of enhanced free oxygen species but also insufficiency of antioxidant defence system. Total antioxidant status (TAS) expresses capacity for scavenging of free radicals minimizes oxidative damage. The aim of this study was to asses concentrations of homocysteine and total antioxidant status in serum of children on vegetarian and omnivorous diet. We also studied levels of vitamin A (retinol) and vitamin E (alpha-tocopherol) particular components of TAS. MATERIAL AND METHODS: The study included 35 children, aged 5-16 who had been referred to Institute of Mother and Child for dietary consultation. From those, 17 were lacto-ovo-vegetarians and 18 omnivores. Dietary constituents were analyzed using the nutritional programme Dietetyk2 and completed with supplementation data. Concentration of homocysteine was estimated in serum with fluorescence polarization immunoassay and TAS by colorimetric method. Levels of vitamin A and E were determined using high-pressure liquid chromatography (HPLC). RESULTS: The mean concentration of homocysteine was similar in both studied groups (vegetarians: 6.13 +/- 1.01 micromol/l vs. omnivores: 5.45 +/- 0.98 micromol/l). In vegetarian children serum level of TAS was significantly lower (1.21 +/- 0.06 mmol/I) as compared to those in non-vegetarian ones (1.30 +/- 0.05 mmol/l, p < 0.0001), but remained within the physiological range (1.16-1.40 mmol/l). The mean concentrations of vitamins A and E in serum of vegetarians were lower than observed in omnivores group, but only in case of alpha-tocopherol the difference was statistically significant (vegetarians: 15.58 +/- 4.12 vs. omnivores: 18.41 +/- 3.10; p < 0.05). CONCLUSION: In studied group of children on lacto-ovo-vegetarian diet, serum concentrations of homocysteine are normal and daily food intake of antioxidants and their level in blood are sufficient for preserving adequate total antioxidant status.

[Effect of vitamin D supplementation on serum 25-hydroxyvitamin D and bone turnover markers concentrations in vegetarian children]. / Stezenia 25-hydroksywitaminy D oraz markerow obrotu kostnego w surowicy krwi dzieci na diecie wegetarianskiej suplementowanej preparatem wapniowo-witaminowym.

PURPOSE: Vitamin D plays a key role in bone mineralization by regulating calcium and phosphate metabolism. Deficiency of this vitamin may lead to disturbances in bone metabolism as well as to osteopenia and osteoporosis. AIM: 1. Assessment of daily intake of calcium and vitamin D in children on vegetarian diet. 2. Measurement of serum 25-hydroxyvitamin D and biochemical bone turnover markers levels in vegetarian children supplemented with calcium and vitamin D, before and after the intervention. MATERIAL AND METHODS: We examined 35 prepubertal children on vegetarian diet who were divided into two groups: group A (without supplementation) and group B in which the children received daily 400 IU vitamin D3 and 500 mg calcium. Total serum calcium, phosphate, 25-hydroxyvitamin D and bone turnover markers were measured at start and end of the intervention. RESULTS: Concentration of calcium and phosphate in serum of all tested children were within physiological range. The mean value of 25-hydroxyvitamin D at start of supplementation was low in both the vitamin D-treated group and in the controls (about 19 ng/ml). After 4-months supplementation, the vitamin D level was significantly higher in the vitamin D-treated group (38.3+/-8.5 ng/ml) than in the control group (13.7+/-5.3 ng/ml) (p<0.001). No statistically significant differences in bone formation (osteocalcin) and resorption (carboxy-terminal telopeptide of type I collagen, isoenzyme 5b of tartrate-resistant acid phosphatase) markers were observed. CONCLUSIONS: The presented results indicate that in vegetarian children the daily dietary intake of vitamin D in the winter months is not sufficient to maintain adequate serum vitamin D concentration. Vitamin D daily supplementation increases vitamin D concentration in these individuals to the required levels.

[Oxidative DNA damage in mothers of children with isolated orofacial clefts]. / Oksydacyjne uszkodzenia DNA u matek dzieci z izolowanymi rozszczepami twarzoczaszki.

Recently, there is increasing evidence suggesting that oxidative stress may contribute to birth defects. 8-Hydroxy-2'-deoxyguanosine (8-OHdG) is considered to be a sensitive biomarker of oxidative DNA damage. It would seem important to establish whether levels of 8-OHdG and antioxidants are different in mothers of children with orofacial clefts comparing to women who gave birth to healthy children. The present study was carried out to investigate this point. Total plasma antioxidant status (TAS), serum 8-OHdG, plasma alpha-tocopherol and retinol were determined in 27 healthy mothers of children with isolated orofacial clefts (cleft lip - 11, cleft lip and palate - 9, cleft palate - 7; mean age - 26.6+/-3.5 years; mean time after delivery - 6.5+/-8.4 months) and 14 control mothers of children without birth defect (mean age - 26.4+/-3.0 years, mean time after delivery - 5.7+/-5.8 months). 8-OHdG concentrations were significantly increased in mothers of children with clefts compared to controls (median - 4.7 ng/mL; range: 0.5-8.6 ng/mL vs. median 1.9 ng/mL; range: 0.1-3.2 ng/ mL; p<0.001 in Wilcoxon test). TAS (median - 1.03 micromol/L; range 0.87 - 1.26 micromol/L vs. median - 1.13 micromol/L; range: 1.03- 1.26 micromol/L; p<0,05) and alpha-tocopherol level (median - 15.8 micromol/L; range: 11.3-26.8 micromol/L vs. median - 20.6 micromol/L; range: 13.8-26.5 micromol/L; p<0,05) were significantly reduced in mothers of affected children. There were no differences in vitamin A levels between the study group and controls. The ratio of alpha-tocopherol to plasma total cholesterol was lower in women who gave birth to children with clefts compared to controls (median - 3.16 micromol/mmol vs. median - 3.7 micromol/ mmol; p>0.05). No correlation was found between TAS, the vitamins and 8-OHdG. We conclude that depletion of antioxidant systems as well as increased free radical generation are likely to be involved in the pathophysiology of orofacial clefts in humans. The results indicate the necessity of futher studies to establish whether peri-conceptional antioxidant supplementation will prevent these serious defects.