Fingerprinting of chlorinated paraffins and their transformation products in plastic consumer products.

Chlorinated paraffins (CPs) can be classified according to their length as short-chain (SC, C10-C13), medium-chain (MC, C14-C17) and long-chain (LC, C ≥ 18) CPs. Technical CP-mixtures can contain a wide range of carbon- (C-, nC = 10-30) and chlorine- (Cl-, nCl = 3-19) homologues. CPs are high-production volume chemicals (>106 t/y). They are used as flame-retardants, plasticizers and coolant fluids. Due to the persistence, bioaccumulation, long-range environmental transport potential and adverse effects, SCCPs are regulated as persistent organic pollutants (POPs) by the Stockholm Convention. Transformation of CPs can lead to the formation of unsaturated compounds such as chlorinated mono- (CO), di- (CdiO) and tri-olefins (CtriO). Such transformation reactions can occur at different stages of CP manipulation providing characteristic C-/Cl-homologue distributions. All this results in unique patterns that collectively create a fingerprint, which can be distinguished from CP-containing samples. Therefore, CP-fingerprinting can develop into a promising tool for future source apportionment studies and with it, the reduction of environmental burden of CPs and hazards to humans. Herein, CP-containing plastics were studied to establish fingerprints and develop this method. We analyzed four household items by reverse-phase liquid-chromatography coupled with a mass spectrometer with an atmospheric pressure chemical ionization source and an Orbitrap mass analyzer (RP-LC-APCI-Orbitrap-MS) operated at a resolution of 120000 (FWHM at m/z 200). MS-data of different CP-, CO-, CdiO- and CtriO-homologues were efficiently processed with an R-based automatic mass spectra evaluation routine (RASER). From the 16720 ions searched for, up to 4300 ions per sample were assigned to 340 C-/Cl-homologues of CPs and their transformation products. Specific fingerprints were deduced from the C-/Cl-homologues distributions, the carbon- (nC) and chlorine- (nCl) numbers and saturation degree. These fingerprints were compared with the ones obtained by a GC-ECNI-Orbitrap-MS method.

Simultaneous quantification of progestogens in plasma and serum by UHPLC-HRMS employing multiplexed targeted single ion monitoring.

Progesterone is the predominant gestagen in most mammals studied so far. It plays a substantial role in the regulation of the female reproductive cycle and in providing support for pregnancy maintenance. Despite its known functions, gaps in knowledge are present regarding its reduced metabolites that potentially exert biological activity. Therefore, a new UHPLC-HRMS method based on a Q Exactive™ mass spectrometer was developed to detect and quantify simultaneously progesterone, its hormone precursor pregnenolone and 10 reduced progestogens (20α-DHP, 20ß-DHP, 3α,5α-THP, 3α,5ß-THP, 3ß,5α-THP, 3ß,5ß-THP, 3α-DHP, 3ß-DHP, 5α-DHP and 5ß-DHP) in plasma and serum samples. Purification was achieved by an optimized solid phase extraction (SPE) and the analysis was conducted in positive electrospray ionization (ESI) mode with the application of multiplexed selected ion monitoring (msx-t-SIM). The method validation included the study of sensitivity, selectivity, curve fitting, carry-over, accuracy, precision, recovery and matrix effects. Despite the poor ionization properties of underivatized steroids, a high sensitivity in the range of pg/mL was achieved.

Progestogen profiling in plasma during the estrous cycle in cattle using an LC-MS based approach.

In many mammalian species, corpus luteum derived progesterone (P4) is the main functional gestagen during the estrous cycle and pregnancy. P4 can be metabolized into various metabolites, of which some are biologically active. While some metabolites target the classical nuclear progesterone receptor (PR), neurosteroids bind the receptors of type A γ-aminobutyric acid (GABAA-r) in the brain. According to the position of reduction within the molecule, metabolites of P4 can be characterized into C20-reduced progestogens (20α-dihydroprogesterone (20α-DHP) and 20ß-dihydroprogesterone (20ß-DHP)), C3-reduced progestogens (3α-dihydroprogesterone (3α-DHP) and 3ß-dihydroprogesterone (3ß-DHP)), 5α-reduced progestogens (5α-dihydroprogesterone (5α-DHP), allopregnanolone and isopregnanolone) and 5ß-reduced progestogens (5ß-dihydroprogesterone (5ß-DHP), pregnanolone and epipregnanolone). We questioned whether the reduced progestogens are present in bovine plasma during the estrous cycle and whether their profiles differed from the profile of the common precursor P4 around the time of luteolysis. The analytes were monitored in plasma samples using liquid chromatography mass spectrometry (LC-MS). While progestogens lagged behind the drop of P4 at luteolysis, they followed the profile of P4 during the estrous cycle. The abundance of P4 was predominant followed by allopregnanolone, pregnanolone, epipregnanolone and 20ß-DHP. Further studies will need to focus particularly on the period around luteolysis.

The presence of soluble c-erbB-2 in saliva and serum among women with breast carcinoma: a preliminary study.

The protein c-erbB-2, also known as Her2/neu, is a prognostic breast cancer marker assayed in tissue biopsies from women diagnosed with malignant tumors. Present studies suggest that soluble fragments of the c-erbB-2 oncogene may be released from the cell surface and become detectable in patients with carcinoma of the breast. Consequently, the purpose of this study was to assay the c-erbB-2 protein in the saliva and serum of women with and without carcinoma of the breast and to determine whether the protein possesses any diagnostic value. To determine the diagnostic utility of this oncogene, the soluble form of the c-erbB-2 protein was assayed in the saliva and serum using ELISA in three different groups of women. The three groups consisted of 57 healthy women, 41 women with benign breast lesions, and 30 women diagnosed with breast cancer. To compare the relative diagnostic utility of the c-erbB-2 protein, CA 15-3 was also measured. The CA 15-3 measurements served as a "gold standard" by which to compare the c-erbB-2 protein's diagnostic effectiveness. We found c-erbB-2 protein in the saliva and serum of all three groups of women. The salivary and serological levels of c-erbB-2 in the cancer patients, however, were significantly higher (P < 0.001) than the salivary and serum levels of healthy controls and benign tumor patients. Additionally, the c-erbB-2 protein was found to be equal to or to surpass the ability of CA 15-3 to detect patients with carcinoma. The results of the pilot study suggest that the c-erbB-2 protein may have potential use in the initial detection and/or follow-up screening for the recurrence of breast cancer in women.

Effect of corn root exudates on the degradation of atrazine and its chlorinated metabolites in soils.

DIMBOA (3,4-dihydro-2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3-one), a major benzoxazinone of Poaceae plants, was isolated and purified from corn seedlings. The effect of isolated and purified DIMBOA on the degradation of atrazine [2-chloro-4-(ethylamino)-6-(isopropylamino)-s-triazine], and its toxic breakdown products, desethylatrazine [2-chloro-4-amino-6-(isopropylamino)-s-triazine; DEA] and desisopropylatrazine [2-chloro-4-(ethylamino)-6-amino-s-triazine; DIA], was studied in the absence of plants using batch experiments, while the effect of corn root exudates on these compounds was determined in hydroponic experiments. Degradation experiments were performed in the presence and absence of 50 microM, 1 mM, or 5 mM DIMBOA resulting in ratios of DIMBOA to pesticide of 1:1, 20:1, and 100:1. We observed a 100% degradation of atrazine to hydroxyatrazine within 48 h at a ratio of DIMBOA to atrazine of 100:1. DIMBOA had the largest effect on atrazine, while it was about three times less effective on DEA and DIA. Corn (Zea mays L. cv. LG 2185) was exposed to 10 mg L(-1) of either atrazine, DEA, or DIA for 11 d in a growth chamber experiment. Up to 4.3 micromol L(-1) d(-1) of hydroxyatrazine were formed in the nutrient solutions by plants exposed to atrazine, while the formation of hydroxylated metabolites from plants exposed to DEA and DIA was smaller and also delayed. The formation of hydroxylated metabolites increased in the solution with plant age in all atrazine, DEA, and DIA treatments. HMBOA (3,4-dihydro-2-hydroxy-7-methoxy-2H-1,4-benzoxazin-3-one), the lactam precursor of DIMBOA, and a tentatively identified derivative of MBOA (2,3-dihydro-6-methoxy-benzoxazol-2-one) were detected in the corn root exudates. Mass balance calculations revealed that up to 30% of the disappearance of atrazine and DEA, and up to 10% of DIA removal from the solution medium in our study could be explained by the formation of hydroxylated metabolites in the solution itself. Our results show that higher plants such as corn have the potential to promote the hydrolysis of triazine residues in soils by exudation of benzoxazinones.

The use of soluble, salivary c-erbB-2 for the detection and post-operative follow-up of breast cancer in women: the results of a five-year translational research study.

A surge of new technological developments, coupled with the limitations of existing disease-detection methodologies, is propelling the field of medical diagnostics forward at unprecedented rates. Advancements in proteomics and nanotechnology are paving the way for diagnostic tests that will be capable of rapid multi-analyte detection in both laboratory and non-laboratory settings. Technological advancements have also benefited biomarker research to the point where saliva is now recognized as an excellent diagnostic medium that can be collected simply and non-invasively. Salivary biomarkers have been identified that may provide diagnostic information about a variety of cancers and other diseases. In particular, proof-of-principle has been demonstrated for salivary c-erbB-2, whose elevation has been shown to correlate strongly with breast malignancy in women. The purpose of this manuscript is to review the past literature and present the current research focused on the use of saliva as a diagnostic medium for the detection of malignancies that are remote from the oral cavity.

Neighboring group participation in the electrospray ionization tandem mass spectra of polyamine toxins of spiders. Part 1: α, ω-diaminoalkane compounds.

The fragmentation pattern of N-(4-aminobutyl)-3-(4-hydroxyphenyl)prop-2-enamide (1) is characterized by an ion formed by neighboring group participation. The phenolic hydrogen is transferred to the primary amino group via a 15-membered ring. This has been proven by H/D exchange reactions and by analyzing homologues and derivatives.

Correlation of stereoselectivity and ion response in electrospray mass-spectrometry. electrospray ionization-mass spectrometry as a tool to predict chemical behavior?

Several alkali cation complexes of chiral alkoxymethyl-substituted acylsilanes, prepared in situ by the admixture of alkali iodides to acylsilane sample solutions, were investigated by electrospray ionization-mass spectrometry. Competition experiments suggest that the relative complex stabilities of such species in acetonitrile solution follow the order [Formula: see text], which is different from the order of the relative stabilities of the complexes in the gas phase ([Formula: see text]) as qualitatively determined by a tandem mass spectrometry-type experiment. Additionally, a rough correlation of relative ion responses in the mass spectrometry with relative stereoselectivities-derived from chelate-controlled reactions performed with the respective silanes-was found. The latter observation suggests that the electrospray ionization-mass spectrometry technique is a potentially useful method to predict chemical behavior, and it demands little experimental effort.

Influence of radiation sterilization on the stability of trifluorothymidine.

The influence of radiation sterilization on the stability of trifluorothymidine (TFT) was investigated. TFT was irradiated under ambient atmosphere with a 60Co-source and with an electron accelerator at 25, 50, and 100 kGy, respectively. The radiation-induced effects were determined by chromatographic and spectroscopic methods as well as potentiometrically with a fluoride selective electrode. TFT was moderately stable to ionizing radiation. The degradation induced by electron-beam irradiation was significantly (P=95%) smaller than by gamma-irradiation. The radiolysis products amounted to about 0.25% after electron-beam irradiation at 25 kGy, and to about 0.50% after gamma-irradiation, respectively. The main irradiation product was 5-trifluoromethyluracil (TFMU). In addition five further impurities were detected with HPLC. Identification of degradation products was performed using HPLC-ESI-MS. A degradation path of TFT after radiation sterilization was shown.

Transradial coronary angioplasty and stent implantation in acute myocardial infarction: initial experience.

Transradial coronary angioplasty and stent implantation in have been associated with reduced complications, length of stay and hospital costs when compared to the transfemoral approach. Fourteen high-risk patients with acute myocardial infarction underwent transradial coronary angioplasty and stent placement. All diagnostic and interventional procedures were successfully completed using 6 French guide catheters and ACS Tristar stents (Guidant Corporation, Santa Clara, California) up to 4 mm in diameter. Thirteen patients received glycoprotein IIb/IIIa inhibitors. There were no procedural or access site complications. The mean length of stay was 3.5 days and the mean time interval from initial radial cannulation compared favorably with 14 acute myocardial infarction patients undergoing transfemoral angioplasty and stent placement. Transradial angioplasty in acute myocardial infarction appears to be a safe and feasible option. The procedure time is not increased in experienced hands, and the combination of rare access site complications and early ambulation may lead to decreased morbidity and lower costs. Transradial angioplasty in acute myocardial infarction may be an attractive option in thrombolytic therapy patients (facilitated percutaneous coronary intervention) or those who require aggressive anticoagulation or antiplatelet therapy.

N-nitrosylation potential of mono-N-desethylamiodarone at physiological pH.

Amiodarone (AMI) is frequently used for the treatment of supraventricular arrhythmias. The parent drug is rapidly dealkylated to mono-N-desethylamiodarone (MDEA) and the plasma concentrations of AMI and MDEA are comparable. MDEA is a secondary amine and may thus undergo formation to the corresponding N-nitrosamine in combination with coadministered nitrovasodilators. Previous studies have shown that nitrovasodilators release the vasoactive NO? which may nitrosylate thiol or secondary amine groups in aqueous solutions. Therefore, the nitrosylation potential of MDEA at physiological pH was investigated. N-Nitroso-monodesethylamiodarone (NO-MDEA) was synthesized, characterized and used as a reference product for the detection of the corresponding N-nitrosamine. HPLC and NMR results have shown that the NO-MDEA product is an equilibrium of two configurational isomers (syn and anti). NO-release was generated by sodium nitroprusside (SNP) which was exposed to light. The formation to NO-MDEA was assayed by HPLC-UV. It has been found that MDEA is nitrosylated in the higher nanomolar range and that varying oxygenation of the reaction mixture did not significantly affect the reaction yields. The addition of thiols such as serum albumin (0.6mM), l-cysteine (2.5mM) or N-acetylcysteine (2.5mM) inhibited the NO-MDEA formation indicating that they may prevent N-nitrosamine formation in vivo. However, as S-nitrosothiols may also release NO?, in long term exposure to elevated levels of nitric oxide the nitrosylation of secondary amines may be taken into account.

Oral contraceptive usage and the expression of CA 15-3 and c-erbB-2 in the saliva of healthy women.

OBJECTIVE: This study was conducted to determine if oral contraceptive use affected the salivary levels of 2 cancer biomarkers, cancer antigen 15-3 (CA 15-3), and c-erbB-2 (erb) among women. STUDY DESIGN: A total of 40 women were recruited for this study, 20 controls and 20 using oral contraceptives. The average ages were 27.4 years (control) and 27.1 years (using estrogen supplements). Stimulated whole saliva specimens were collected from each woman. CA 15-3 levels were determined using enzyme immunoassay (EIA) while c-erbB-2 levels were determined by standard ELISA methods. RESULTS: Findings from this study indicate that there were no significant differences between the control and study groups. CONCLUSIONS: Therefore, saliva may be a useful tool for monitoring women with high estrogen levels due to either tumor load or any treatment regimen, i.e., the levels of the cancer biomarkers CA 15-3 and erb are not affected by increased levels of estrogen in the body.

Reliability assessment of soluble c-erbB-2 concentrations in the saliva of healthy women and men.

BACKGROUND: The protein c-erb B-2, also known as Her2/neu, is a prognostic breast cancer marker assayed in tissue biopsy specimens from women diagnosed with malignant tumors. Current studies suggest that soluble fragments of the c-erb B-2 oncogene may be released from the cell surface and become detectable in patients with a carcinoma of the breast. Consequently, the purpose of this study is to assay soluble c-erb B-2 protein in the saliva of healthy men and women to determine the reliability of the assay. METHODS: To determine the diagnostic utility of this oncogene, we assayed the soluble form of the c-erb B-2 protein in the saliva with an enzyme-linked immunosorbent assay. The study population consisted of 10 healthy women and 9 healthy men who were serially sampled for saliva 3 times a day for a 5-day period. Saliva was collected from each subject at 9 AM, 4 PM, and 9 PM during the 5-day period. RESULTS: We found the presence of c-erb B-2 protein in the saliva of both groups of subjects. The salivary levels of c-erb B-2 were not significantly different when compared for gender differences. Likewise, the results suggest that sampling during various times of the day for salivary c-erb B-2 levels has no effect on marker concentration. Reliability analyses showed that supervised salivary collections were more reliable than unsupervised collections. CONCLUSIONS: The results of this pilot study suggest that the assay for salivary c-erb B-2 protein is reliable and might have potential use in the initial detection and follow-up screening for the recurrence of breast cancer in both men and women.

Salivary sIgA concentrations and stimulated whole saliva flow rates among women undergoing chemotherapy for breast cancer: an exploratory study.

A study was conducted to evaluate the secretion of stimulated whole saliva (SWS) and secretory IgA (sIgA) among three groups of women. One group was a healthy control group, the second group consisted of women diagnosed with metastatic breast cancer before being placed on chemotherapy, while the third group consisted of women on chemotherapy taking CMF (cyclophosphamide, methotrexate, and fluorouracil) for at least one month. There were seven patients in each group. SWS was collected. Salivary sIgA concentrations were determined by enzyme-linked immunoabsorbant assay. The results of the study showed that the mean SWS flow rate for the chemotherapy group (x = 0.96 mL/min) was significantly lower (p > 0.03) than that of the control group (x = 2.33 mL/min) and lower than that of the group with cancer (x = 1.81 mL/min). Additionally, the results showed that the mean sIgA concentrations for the chemotherapy group (x = 10.9 ng/mg of protein) were slightly lower than those of the control group (x = 13.7 ng/mg of protein) and lower than those of the group with cancer (x = 12.6 ng/mg of protein). The results of this study suggest that women placed on CMF for treatment of carcinoma of the breast may have reduced stimulated salivary production.

Assessment of the ochratoxin A production ability of Aspergillus tubingensis.

Aspergillus tubingensis is a black Aspergillus frequently isolated from different agricultural products, including grapes. Conflicting results have been published in recent years about its ability to produce ochratoxin A (OTA), a potent nephrotoxic and carcinogenic mycotoxin. This study re-examined six A. tubingensis strains deposited in international culture collections for OTA production. OTA could not be detected in any A. tubingensis extract using HPLC coupled with a fluorescence detector (FLD), whereas it was easily detected in ochratoxigenic A. niger extracts used as positive control. The same outcome was obtained using LC-MS. The presence of other metabolites with retention times similar to the OTA signal in the A. tubingensis extracts or background noise of the growth media may be reasons for the misinterpretation of the chromatograms obtained by HPLC-FLD.

[The evaluation of animal welfare of housing systems for small and large groups of fattening rabbits]. / Die Beurteilung der Tierartgerechtheit von Aufstallungssystemen für kleine und grosse Mastkaninchen-Gruppen.

In the present study three new group housing systems for fattening rabbits kept in smaller and larger units were evaluated with respect to the animal welfare as stated in the Swiss Regulations about animal protection. Judgement of animal welfare was based on the results of two practical examinations in which abnormal behaviour and injuries were elevated. To determine proper keeping evaluation criteria and their limits were set. In the examined housing systems no problems concerning locomotory and occupation behaviour, resting and hiding places were found. But in a few larger groups social problems occurred by the end of the fattening period. Hence heavy injuries were found before day 80 of life. These problems concerning aggressive behaviour and injuries may be explained by the accelerated sexual development (determined by strain, by housing conditions or by the presence of adult females) or the difficulty for the young rabbits to establish steady dominance hierarchies in this social environment and under the given conditions. These findings raise the question which factors are responsible for the difficulties.

[Raising pairs of young non-reproducing female rabbits in cages]. / Paarhaltung nichtreproduzierender Hauskaninchen-Zibben im Käfig.

From the age of three months 16 young females of the Hybrid strain ZIKA were kept in pairs in a cage which measured 60 x 68 x 56 cm and which had an elevated tier (30 x 60 cm). Over a period of seven months these pairs were observed (2 h) and examined morphologically once a month. A video recording during 24 h was conducted. These observations showed that the frequency of aggressive and sexual behaviours was not very high and was not the same with all pairs. The frequency of aggressive, sexual and subdominant behaviour varied during the investigation, sometimes even within a month. There were a lot of amicable behaviour patterns (allogrooming, friendly nose-to-nose, nose-to-body contacts) and the females spent a lot of time in body-contact. Except with one pair there were hardly found any injuries despite the observed aggressive encounters. The dominance hierarchy in the different pairs was clear from the beginning and remained so during the experiment. The dominant females showed aggressive and sexual behaviour more frequently, all subdominant animals went away more frequently. The results suggest that keeping females in pairs before breeding starts or keeping them in pairs as laboratory rabbits is basically possible though the relation has to be checked all the time. Sufficient possibilities for occupation and a good structure of room are important. Because the degree of sexual influence of the females is not clear yet, further clarifications are needed.