Verification of skin autofluorescence values by mass spectrometry in adolescents with type 1 diabetes: brief report.

BACKGROUND: Accumulation of advanced glycation end products (AGEs) in tissues is a major risk factor for diabetes-associated complications. Skin autofluorescence (SAF) values measured by a specific noninvasive approach (AGE Reader; DiagnOptics Technologies B.V., Gröningen, The Netherlands) reflect the overall AGE exposure in skin. SUBJECTS AND METHODS: In 16 adolescents with type 1 diabetes (age range, 11-18 years) we tested the association between SAF measured with an AGE Reader and the presence of glucuronic acid, 3-indoxyl sulfate, 3-hydroxybutyrate, phenol sulfate, and pentosidine in skin tissue determined with desorption electrospray ionization mass spectrometry (DESI-MS). These compounds are implicated in long-term diabetes complications. RESULTS: SAF values significantly correlated with levels of compounds measured by DESI-MS (r>0.9 and P<0.001 for each). CONCLUSIONS: The strong correlation between adolescents' SAF values measured with the AGE Reader and some glycation products measured with DESI-MS indicates that SAF values may be used as surrogate markers of skin exposure to glycemic end products in type 1 diabetes.

Lymphocyte subsets in pediatric migraine.

Aseptic inflammation due to activated immune cells has been implicated in the pathomechanism of migraine. We measured the prevalence of regulatory T cells (Tregs), along with that of CD4(+)/CD8(+) lymphocytes and their Th1/Th2 commitment in pediatric migraine. Children and adolescents suffering from migraine without aura, migraine with aura and hemiplegic migraine ictally (n = 53, 27, and 20, respectively), also interictally (n = 33) were recruited and compared to 24 healthy children. Our results indicated comparable prevalence of Tregs, CD4(+) and Th1/Th2 committed cells. CD8(+) prevalence was lower, and CD4(+)/CD8(+) ratio was higher in ictal phase irrespective of the subtype of migraine. No association between CD8(+) prevalence and gender, body weight, disease onset and attack duration in migraine subtypes was found. CD8(+) prevalence was normal in patients in interictal phase. These results suggest the absence of major systemic alteration of adaptive immunity in children and adolescents suffering from migraine; however, a transient decrease of CD8(+) prevalence during the ictal phase was detected irrespective of the subtype of migraine.

Role of nitric oxide (NO) metabolism and inflammatory mediators in childhood obesity.

OBJECTIVE AND DESIGN: The role of NO and adipocytokines in childhood obesity was studied, supposing that obesity provokes inflammation. Children were admitted to the pediatric clinic for a regular check up because of obesity. SUBJECTS: Obese (n = 79) and healthy (n = 12) children were selected and divided into subgroups according to their age, gender, glucose tolerance and nitric oxide synthase (NOS II) positivity. METHODS: Urine and blood nitrite plus nitrate, the expression of NOS II in white blood cells, serum adipocytokines and clinical characteristics were analyzed in each group. Significance was tested by unpaired two-tailed t test and by ANOVA. RESULTS: NOS II was only detected in the white blood cells of a subgroup (17/79) of obese children. Serum leptin and resistin concentrations were significantly higher, adiponectin was lower compared to healthy children. Significant correlations were observed between serum adiponectin and resistin levels (reciprocal, R (2) = 0.4), and between body mass index and serum leptin levels. CONCLUSIONS: NOS II expression in white blood cells was observed in a minority of patients. Low-grade inflammation in obese children was suggested by the increased resistin levels, particularly in NOS II-positive patients. Correlation between different adipocytokines was restricted for a few subgroups.

Synthesis of hepcidin derivatives in order to develop standards for immune adsorption method.

MeCN, acetonitrile; ECL, enhanced chemiluminescence; EDT, 1,2-ethanedithiole; HEPC12-A, rabbit anti-human hepcidin IgG, affinity purified; HEPC13-A, rabbit anti-mouse/human hepcidin IgG, affinity purified; HEPC61-P, human hepcidin-25 control/blocking synthetic peptide; HRP, horseradish peroxidase; IL-6, interleukin-6; KLH, keyhole limpet hemocyanin; LEAP, liver-expressed antimicrobial peptide; NEM, N-ethylmaleimide; NMP, N-methyl-pirrolidone; PBS, phosphate buffered saline; PVDF, polyvinylidene difluoride; SELDI-TOF-MS, surface-enhanced laser desorption ionization-time-of-flight mass spectrometry; TMB, tetramethylbenzidin; TNF-alpha, tumor necrosis factor-alpha.

Ser128Arg E-selectin and Thr715Pro P-selectin polymorphisms and severe preeclampsia.

OBJECTIVE: To investigate the association of functional E and P selectin polymorphisms with severe preeclampsia. STUDY DESIGN: P-selectin Thr715Pro and E-selectin Serl28Arg polymorphism were determined with the polymerase chain reaction in 126 women with severe preeclampsia and in 106 healthy pregnant women. Genotype distribution and allele prevalence of both groups were compared. Regression analysis was used to analyze the association between disease characteristics and genotype. RESULTS: Genotype distributions and the allele prevalence were similar in the tested populations. Furthermore, the genotypes investigated had no impact on the risk of eclampsia of the hemolysis, elevated liver enzymes and low platelets (HELLP) syndrome. Multiple linear regression analysis identified the P-selectin 715Pro allele as an independent determinant of the time of onset of hypertension (p = 0.034) along with maternal age, smoking and body mass index before pregnancy. CONCLUSION: The polymorphisms tested are not associated with the risk of severe preeclampsia.

Association between estrogen receptor alpha (ESR1) gene polymorphisms and severe preeclampsia.

Associations have been reported between estrogen receptor alpha (ESR1) gene polymorphisms and various pathological conditions, including cardiovascular diseases. Our aim was to investigate whether two polymorphisms of the ESR1 gene (ESR1 c.454 -397T>C: PvuII restriction site and c.454 -351A>G: XbaI restriction site) are associated with preeclampsia. In a case-control study, we analyzed blood samples from 119 severely preeclamptic patients and 103 normotensive, healthy pregnant women using the polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) method. All of the women were Caucasian. There was no association between severe preeclampsia and the PvuII and XbaI ESR1 gene polymorphisms separately. However, with the simultaneous carriage of both polymorphisms, the TT/AA genotype combination was significantly more frequent in severely preeclamptic patients than in healthy control subjects (24.4% vs. 9.7%, p=0.003), whereas the TT/AG combination was significantly less frequent in the severely preeclamptic group than in the control group (5.0% vs. 18.4%, p=0.002). According to the haplotype estimation, the homozygous T-A haplotype carriers had an increased risk of severe preeclampsia independent of maternal age, prepregnancy BMI, primiparity and smoking status (adjusted odds ratio [OR]: 4.36, 95% confidence interval [CI]: 1.65-11.53). The GG genotype of the XbaI polymorphism was associated with a lower risk of fetal growth restriction in patients with severe preeclampsia (OR: 0.23, 95% CI: 0.07-0.73). In conclusion, the homozygous T-A haplotype carriers of ESR1 PvuII and XbaI polymorphisms showed an increased risk of severe preeclampsia. In addition, the GG genotype of the XbaI polymorphism decreased the risk of fetal growth restriction in severely preeclamptic patients.

Selectin polymorphisms and perinatal morbidity in low-birthweight infants.

BACKGROUND: Studies have shown an association between altered expression of selectins and premature birth, early sepsis and bronchopulmonary dysplasia. AIM: To investigate the possible link between functional polymorphisms of the E-, P- and L-selectin genes and perinatal morbidity. METHODS: We compared the genotype distribution of the E-selectin Ser128Arg, P-selectin Thr715Pro and L-selectin Pro213Ser polymorphisms in 125 low-birthweight singleton infants with those of 156 healthy term neonates. We also analysed the association of genotype with risk of sepsis and bronchopulmonary dysplasia. RESULTS: We found no association between E-selectin or P-selectin polymorphisms and premature birth, nor did we find any association between E-selectin or P-selectin and early postnatal sepsis or bronchopulmonary dysplasia. Carriers of the 213Ser L-selectin allele were found to be more prevalent in low-birthweight infants, particularly in those with bronchopulmonary dysplasia. We found no association between the L-selectin polymorphism and early postnatal sepsis. CONCLUSION: Our results underline the importance of L-selectin in perinatal pathology, but further studies are needed to evaluate the alteration of L-selectin levels in carriers of the 213Ser allele and their possible contribution to premature birth and bronchopulmonary dysplasia.

Family history of early-onset cardiovascular disorders is associated with a higher risk of severe preeclampsia.

AIM: The aim was to evaluate familial early-onset cardiovascular disorders as potential risk factors for severe preeclampsia. STUDY DESIGN: A case-control study was carried out by interviewing 162 primiparous severely preeclamptic women and 521 primiparous healthy control patients after delivery to determine the frequency of cardiovascular disorders (chronic hypertension, myocardial infarction, stroke) developed before the age of 50 among their parents. The chi2-test was utilized to estimate odds ratios (OR) and 95% confidence intervals (95% CI). The association was adjusted for pre-pregnancy body mass index, maternal age, and smoking habits before pregnancy using logistic regression analysis. RESULTS: Maternal and paternal early-onset chronic hypertension (adjusted OR: 3.84, 95% CI: 2.25-6.54; and adjusted OR: 3.26, 95% CI: 1.76-6.05) as well as paternal early-onset myocardial infarction (adjusted OR: 3.33; 95% CI: 1.51-7.32) were independent risk factors for severe preeclampsia. Early-onset stroke affected only the fathers of severely preeclamptic patients. Among the severely preeclamptic patients a positive family history of cardiovascular disorders developed before the age of 50 increased the risk of early-onset preeclampsia (developing before the 32nd gestational week) by 5.05-fold (95% CI: 3.08-8.31) compared with the control group. CONCLUSION: Our results suggest that the presence of familial early-onset cardiovascular disorders is a predisposing factor for severe preeclampsia.

Lack of association between insulin-like growth factor I receptor G(+3174)A polymorphism and retinopathy of prematurity.

BACKGROUND: Retinopathy of prematurity (ROP) is a potentially blinding eye disorder that affects premature infants. Insulin-like growth factor I (IGF-I) has been established as being necessary for vascular growth in the neonatal retina. Low IGF-I levels during the early postnatal days have been found to be predictive for ROP. The effects of IGF-I are mediated through IGF-I receptors (IGF-IR), which, in turn, suppress IGF-I production. The G(+3174)A polymorphism of the IGF-IR gene has been shown to be associated with low IGF-I levels. We tested the association of this IGF-IR polymorphism with ROP. METHODS: We enrolled in the ROP group those infants (n=108) who had been treated with laser or cryo therapy due to ROP stage 2+ or 3 (n=91) or had a ROP stage 4 or 5 (n=17) (ROP group). The median gestational age of these infants was 28 weeks (range 24-35 weeks) and birth weight was 970 g (range 630 to 2,000 g). The distribution of IGFR-1 G(+3174)A genotype in the ROP group was compared to that in 120 gestational age-matched infants with ROP stage 1 or 2 not requiring intervention [gestational age 30 (range 24-37) weeks, birth weight 1,235 (640-1,960) g] (LBW group) and 164 term newborns [gestational age 39 (range 35-42) weeks, birth weight 3,450 (2,500-4,350) g] (term group). Genotyping was done using PCR-RFLP methods. RESULT: The prevalence of IGF-IR G(+3174)A polymorphism was the same in the ROP group, the LBW group and the term group, showing no association between this single-nucleotide polymorphism (SNP) and ROP. CONCLUSION: Our results do not support the hypothesis that the carrier state of IGF-IR G(+3174)A polymorphism has an impact on the risk of ROP in infants. A possible cause for the lack of association is that the rapid nutritional and metabolic changes during postnatal life have a greater effect on IGF-I levels than this SNP does.

Genetic polymorphisms for vascular endothelial growth factor in perinatal complications.

Low birth weight (LBW) infants have increased susceptibility to perinatal complications. An immature and impaired vascular system may possibly participate in these complications. There is evidence that supports the notion that vascular endothelial growth factor (VEGF), which is an essential regulator of embryonic angiogenesis, plays a central role in the pathogenesis of perinatal complications. We aimed to test whether functional genetic polymorphisms of VEGF are associated with the risk of preterm birth or perinatal morbidity. We enrolled 128 LBW infants (< or = 1500 grams). VEGF T-460C, VEGF C-2578A and VEGF G+405C polymorphisms were determined by real-time PCR or PCR-RFLP, respectively. Their genotypes were compared with VEGF genotypes of 200 healthy, term neonates. The prevalence of the VEGF+405 C allele was higher in LBW infants than in healthy, term neonates (OR [95% CI]: 1.29 [1.01-1.65]). Carrier state for the VEGF -2578A allele was an independent risk factor for enterocolitis necrotisans (NEC) (adjusted OR [95% CI]: 2.77 [1.00-7.65]). The carrier state for the VEGF -2578AA genotype was associated with a decreased risk of acute renal failure (ARF) (adjusted OR [95% CI]: 0.2 [0.05-0.78]). These results suggest that VEGF G+405C polymorphism might be associated with a higher risk of preterm birth and that VEGF C-2578A polymorphism may participate in the development of perinatal complications such as NEC and ARF.

Gender dependent association between perinatal morbidity and estrogen receptor-alpha Pvull polymorphism.

AIMS: Assuming the importance of estrogen in perinatal physiology, we tested the association of an estrogen receptor-alpha (ER-alpha) gene Pvull pP polymorphism with perinatal morbidity in premature infants. METHODS: The ER-alpha Pp genotype was determined in 69 low-birth weight (LBW) boys and 72 LBW girls, 86 term boys and 81 term girls. The association between risk factors, genotype, gender and perinatal morbidity was tested with binary logistic regression analysis. RESULTS: Boys carrying "p" allele were at lower risk for necrotizing enterocolitis (OR [95% Cl]: 0.24 [0.07-0.83]) and patent ductus arteriosus (OR [95% Cl]: 0.24 [0.05-0.97]). The carrier state of the "p" allele was associated with a 34-h shorter period of oxygen supplementation on average (P=0.0018). Boys with pp genotype were at greater risk for intraventricular hemorrhage (OR [95% Cl]: 4.39 [1.15-16.82]). No association between ER-alpha Pvull polymorphism and morbidity was present in girls. CONCLUSIONS: Since homozygocity for any Pvull alleles (i.e. having PP or pp genotype) increases the risk for at least one of the most common perinatal complications, it is likely that the heterozygous carrier state of Pvull genotypes has a protective effect, which is gender-dependent.

The association of the carrier state of the tumor necrosis factor-alpha (TNFalpha) -308A allele with the duration of oxygen supplementation in preterm neonates.

BACKGROUND: High levels of inflammatory cytokines lead to lung damage in premature newborns. We investigated whether single nucleotide polymorphisms (SNP) of innate immunity cytokine genes influence the length of oxygen supplementation. METHODS: We genotyped 123 very low birth weight (VLBW) infants for the tumour necrosis factor (TNF)- alpha G(-308)A, interleukin (IL)-1beta C(3954)T, IL-6 G(-174)C and IL-10 G(-1082)A SNPs. Genomic DNA was isolated from remnant dried blood samples from the neonates. We tested the association between SNPs and ventilation characteristics using a stepwise multiple regression analysis model. RESULTS: The carrier state of the TNF-alpha G(-308)A allele was associated with a 40-hour longer period of mechanical ventilation (p=0.004) and, on average, an additional 36 hours of oxygen supplementation (p=0.0008). The association was significant after its adjustment for perinatal risk factors for lung damage. CONCLUSIONS: The TNF-alpha G(308)A genotype - which is associated with increased TNF-alpha levels - might influence the supplemental oxygen requirement of VLBW infants.

[Hepcidin, the negative regulator of iron absorbtion]. / Hepcidin, a vasháztartás negatív irányú regulátora.

The task of the iron regulatory system is to balance between iron-stores and iron-consumption. Recently the predominant negative regulator of iron absorbtion in the small intestine and iron release from macrophages has been discovered. That is the hepcidin. Hepcidin, the 25-amino acid peptide produced by the hepatocytes, may be a new mediator of innate immunity and the long-sought iron regulatory hormone. Hepcidin is an acute phase peptide, its production is increased in inflammation and in iron overload. According to evidence obtained in mouse models hepcidin decreases the iron absorbtion in the small intestine and inhibits iron release from macrophage and iron transport across placenta. Hereby the hepcidin decreases the plasma iron level. The exact patomechanism of hepcidin is not clarified and its receptor is still unknown. The discovery of hepcidin and its role in iron metabolism could lead to new diagnostic possibilities on the field of hemochromatosis, other iron-regulatory diseases and anaemia of inflammation.