Systematic investigation of organochlorine pesticides and polychlorinated biphenyls blood levels in Greek children from the Rhea birth cohort suggests historical exposure to DDT and through diet to DDE.

The blood levels of organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) have been thoroughly investigated in Greek children from the Rhea birth cohort study. This investigation aimed to assess exposure levels, explore their possible relationship with children's age and sex, and indicate potential sources of exposure. Exposure patterns and common sources of PCBs and OCPs were analyzed using bivariate and multivariate statistics. A total of 947 blood samples from study participants were analyzed for OCP and PCB exposure, with 375 samples collected at 4 years old, 239 at 6.5 years old, and 333 at 11 years old. Elevated levels of DDE were observed in 6.5-year-old children compared to corresponding levels in other European countries. Higher levels of DDE were found in 4-year-old children, with the lowest concentrations in the 11-year-old group. The DDT/DDE ratio was consistently less than 1 among all the examined subjects. These results indicate exposure to DDT and DDE both in utero and through breastfeeding and dietary intake. For the entire cohort population, the highest concentration was determined for PCB 28, followed by PCBs 138, 153, and 180. The sum of the six indicator PCBs implied low exposure levels for the majority of the cohort population. Spearman correlations revealed strong associations between PCBs and OCPs, while principal component analysis identified two different groupings of exposure. DDE exhibited a correlation with a series of PCBs (153, 156, 163, 180), indicating a combined OCP-PCB source, and an anticorrelation with others (52, 28, 101), implying a separate and competing source.

Monitoring of Polycyclic Aromatic Hydrocarbon Levels in Mussels (Mytilus galloprovincialis) from Aquaculture Farms in Central Macedonia Region, Greece, Using Gas Chromatography-Tandem Mass Spectrometry Method.

A new sensitive and selective gas chromatography tandem mass spectrometry (GC-MS/MS) method was developed for the analysis of 26 polycyclic aromatic hydrocarbons (PAHs), including 16 Environmental Protection Agency (EPA) and 15 + 1 European Union (EU) PAHs, in mussel samples from aquaculture farms in Thermaikos and Strymonian Gulf, Central Macedonia Region, in three sampling periods. Concentrations were found at moderate to low values at all sampling sites, without exceeding maximum levels set by EU. Low molecular weight PAHs were predominant in all samples. Seasonal variation of the concentrations was observed; values were slightly higher in the winter period. Use of diagnostic ratios for potential sources of PAHs showed both petrogenic and pyrolitic origin. In comparison to other related studies of mussels from the Mediterranean Sea, Greek mussels cultivated in the studied gulfs are low in contaminants due to minimal environmental pollution effects. Low concentrations of PAHs are in compliance with the low values of other POPs which were found in the mussels.

Infant dietary exposure to dioxins and dioxin-like compounds in Greece.

The dietary exposure of infants to polychlorinated dibenzo dioxins and furans (PCDD/Fs) and dioxin like polychlorinated biphenyls (dl-PCBs) is an issue of great social impact. We investigated for the first time the dietary intake of these compounds in infants living in Greece. We included in our study two age groups: 0-6 months, when infants are fed exclusively by human milk and/or formula milk, and 6 to 12 months, when solid food is introduced to nutrition. We took into consideration analytical results for PCDD/Fs and dl-PCBs concentrations in the most popular infant formulae in the Greek market, previous data for mother milk concentrations of PCDD/Fs and dl-PCBs from Greece, and finally analytical data for fat-containing food products from the Greek market. In the first study group, it was found than in infants exclusively fed by breast milk, the calculated sum of PCDD/Fs and dioxin-like PCBs (60.3-80.4 TEQ pg/kg body weight) was significantly higher than that of infants that consume a combination of human milk and formula (31.2-41.6 TEQ pg/kg body weight). In the second study group, separate daily intake estimations were performed for babies receiving human milk (estimated total daily intake 19.76-24.95 TEQ pg/kg body weight) and formula milk (estimated total daily intake 1.60-2.24 TEQ pg/kg body weight). The risks of this exposure should not be overestimated because nursing is restricted to a limited period of human life and besides, the potential consumption of higher levels of dioxin-like compounds is fully compensated by the significant benefits of breast-feeding.

Complexes of an alpha thymosin derivative with ¹85/¹87Re and (99m)Tc: structural analysis and initial biological evaluation.

Alpha thymosins are a family of immunostimulating peptides first isolated from thymus gland. In the present work, the structure of the ReO(V)³âº complex of an alpha 1 thymosin derivative containing the metal-chelating N,N-dimethylglycyl-L-seryl-L-cysteinyl group was studied with NMR, CD, and ESI. The analysis indicated the existence of two interconverting diastereomers depending on the orientation of the side chain of the chelated Ser syn- or anti- to the oxygen of the ReO(V)³âº core. The two diastereomers could be separated on HPLC under a slow gradient showing the ratio of syn/anti to be 3:2, in agreement with the NMR data. The conversion process was shown to involve the coordination of a water molecule to the ReO(V)³âº core through the incubation of the complex in ¹8O-enriched water and subsequent ESI analysis. HPLC analysis of the analogous radioactive (99m)TcO(V)³âº complex showed the formation of two isomers in the same syn/anti 3:2 ratio. Biodistribution studies of the (99m)TcO(V)³âº complex in Swiss albino mice with experimentally induced inflammation showed higher accumulated radioactivity in inflamed tissue compared to normal (ratio of inflamed/control tissue 3.9). (99m)Tc-labeled complexes of alpha thymosin derivatives are expected to facilitate research on alpha thymosins and accelerate exploitation of these peptides in immunotherapy protocols.

Placental leucine aminopeptidase efficiently generates mature antigenic peptides in vitro but in patterns distinct from endoplasmic reticulum aminopeptidase 1.

All three members of the oxytocinase subfamily of M1 aminopeptidases, endoplasmic reticulum aminopeptidase 1 (ERAP1), ERAP2, and placental leucine aminopeptidase (PLAP), also known as insulin-regulated aminopeptidase, have been implicated in the generation of MHC class I-presented peptides. ERAP1 and 2 trim peptides in the endoplasmic reticulum for direct presentation, whereas PLAP has been recently implicated in cross-presentation. The best characterized member of the family, ERAP1, has unique enzymatic properties that fit well with its role in Ag processing. ERAP1 can trim a large variety of long peptide sequences and efficiently accumulate mature antigenic epitopes of 8-9 aa long. In this study, we evaluate the ability of PLAP to process antigenic peptide precursors in vitro and compare it with ERAP1. We find that, similar to ERAP1, PLAP can trim a variety of long peptide sequences efficiently and, in most cases, accumulates appreciable amounts of correct length mature antigenic epitope. Again, similar to ERAP1, PLAP continued trimming some of the epitopes tested and accumulated smaller products effectively destroying the epitope. However, the intermediate accumulation properties of ERAP1 and PLAP are distinct and epitope dependent, suggesting that these two enzymes may impose different selective pressures on epitope generation. Overall, although PLAP has the necessary enzymatic properties to participate in generating or destroying MHC class I-presented peptides, its trimming behavior is distinct from that of ERAP1, something that supports a separate role for these two enzymes in Ag processing.

Levels of perfluorooctanesulfonate (PFOS) and perfluorooctanoate (PFOA) in blood samples from different groups of adults living in Greece.

The production of perfluorinated compounds which finds application in a wide variety of consumer products has led to their accumulation in the environment. This fact, in combination with their chemical properties of amphiphilicity and chemical stability is the cause of their occurrence in human tissues, which according to reports is widespread, not only in occupationally exposed workers, but also in the general population. In this study, we analyzed 182 blood samples from three different groups of adults living in Greece, in order to evaluate blood levels of two environmentally most common perfluorinated compounds, perfluorooctanesulfonate (PFOS) and perfluorooctanoate (PFOA). The samples examined were from a group of individuals living in Athens, a group of inhabitants of the semi-urban and rural area of Argolida, and a group of cancer patients from the St. Savas Anticancer Hospital in Athens. The latter group was chosen in order to examine the possible relation of PFOS and PFOA levels with cancer incidence in the general population. The analytical results showed the presence of PFOS and PFOA in all samples examined, at levels similar to those reported from other European countries. There was no significant difference between the three groups however there was a significant difference between the levels of men and women in all groups examined. No correlation was found between age and PFOS and PFOA levels. This is the first study of this kind to be conducted in samples from Greece.

Determination of PCDD/F, dioxin-like PCB and PAH levels in olive and olive oil samples from areas affected by the fires in summer 2007 in Greece.

During the summer of 2007, a series of massive forest fires broke out in several areas across Greece. The main sources of PCDD/Fs and dioxin-like PCBs in Greece are considered to be the uncontrolled combustion of municipal solid waste in open landfills and accidental fires in forest, rural and industrial areas. Combustion may also lead to the formation of PAHs, which are fat soluble substances of considerable toxicity. The objective of this study was to investigate PCDD/F, dioxin-like PCB and PAH contamination of olive oil produced in fire-affected areas. Olive oil is a very significant agricultural product of Greece. Samples for this study were collected from all affected oil producing areas after the fire and 1year later. PCDD/Fs, dioxin-like PCBs and PAHs were at normal levels in all samples analysed.

Detection of interactions of the beta-amyloid peptide with small molecules employing transferred NOEs.

The interaction of pineal hormone melatonin, the histological dye thioflavin T, and the olive tree polyphenol oleuropein, with the 28 amino acid residue N-terminal fragment of the beta-amyloid peptide (beta-AP) of Alzheimer's disease, [beta-AP(1-28)], was detected in solution through the observation of transferred NOEs (trNOEs) in 1D and 2D NOE spectroscopy (NOESY) experiments. The trNOE method is applied for the first time in the detection of interactions of soluble beta-AP(1-28) with small molecules and may provide a means of screening for the identification of possible inhibitors of the formation of neurotoxic beta-AP assemblies.

A HER-2/neu peptide admixed with PLA microspheres induces a Th1-biased immune response in mice.

The elimination of cancer cells requires strong cellular immune responses, and these responses are induced by the activation of Th1 lymphocytes. In this work, the possibility of inducing a Th1 type of immune response in vivo by mixing a HER-2/neu synthetic CTL (cytotoxic T lymphocyte) peptide [HER-2/neu (789-797)], with poly-lactide (PLA) microspheres was investigated. Various formulations of the peptide were administered to HLA-A2.1 transgenic (HHD) mice. Cellular experiments, assessing proliferation and cytokine determination in splenocyte culture supernatants, were carried out in order to evaluate the type of immune response to the antigen. The in vivo administration of the peptide antigen admixed with the PLA microspheres induced a potent immune response which was comparable to that induced by the combination of the antigen in complete Freund's adjuvant (CFA). Furthermore, the cytokine profile produced by the T lymphocytes of the immunized animals indicated that the combination of the peptide antigen with the PLA microspheres induced a strong Th1 biased immune response to the antigen. The time of peptide incubation with the microspheres prior to administration did not affect the immune response, which further simplifies the preparation of this type of vaccine. The results justify further investigation of the possibility of inducing effective cellular immune responses against cancer cells overexpressing HER-2/neu molecules by simply mixing appropriate HER-2/neu peptide antigens with PLA microspheres.

Cellular FLIP (long isoform) overexpression in T cells drives Th2 effector responses and promotes immunoregulation in experimental autoimmune encephalomyelitis.

Cellular FLIP (c-FLIP) is an endogenous inhibitor of death receptor-induced apoptosis through the caspase 8 pathway. It is an NF-kappaB-inducible protein thought to promote the survival of T cells upon activation, and its down-regulation has been implicated in activation-induced cell death. We have generated transgenic mice overexpressing human c-FLIP long form (c-FLIP(L)) specifically in T cells using the CD2 promoter (TgFLIP(L)). TgFLIP(L) mice exhibit increased IgG1 production upon stimulation by a T cell-dependent Ag and a markedly enhanced contact hypersensitivity response to allergen. In addition to showing augmented Th2-type responses, TgFLIP(L) mice are resistant to the development of myelin oligodendrocyte glycoprotein 35-55 peptide-induced experimental autoimmune encephalomyelitis, a Th1-driven autoimmune disease. In vitro analyses revealed that T cells of TgFLIP(L) mice proliferate normally, but produce higher levels of IL-2 and show preferential maturation of Th2 cytokine-producing cells in response to antigenic stimulation. After adoptive transfer, these (Th2) cells protected wild-type recipient mice from experimental autoimmune encephalomyelitis induction. Our results show that the constitutive overexpression of c-FLIP(L) in T cells is sufficient to drive Th2 polarization of effector T cell responses and indicate that it might function as a key regulator of Th cell differentiation.

Synthesis and angiogenetic activity in the chick chorioallantoic membrane model of thymosin beta-15.

Thymosin beta-15 (Tbeta15), a 44 amino acid peptide (MW = 5173) localized in human prostate and breast cancer tissues was successfully synthesized in multigram quantities using Fmoc solid-phase peptide synthesis. The synthesized product was shown to have the right structure by ESI and MALDI mass spectral techniques and amino acid analysis. Relatively high yield was achieved, which might be due to enhanced acid stability of the p-cyanotrityl resin used. The effect of the synthesized Tbeta15 on the angiogenesis process was investigated using the chick chorioallantoic membrane (CAM) in vivo model. At concentrations above 1 microg/10 microl per disc, Tbeta15 exhibited a positive effect on angiogenesis, comparable to the effect of the intense angiogenetic factor phorbol 12-myristate 13-acetate at a standard concentration of 0.1 microg/10 microl per disc. The results of this study contribute to the further elucidation of the biological regulatory role of thymosin peptides and provide helpful information in the investigation of their possible therapeutic potential.