Study of aggregation in surface sludge deposits from 14 full-scale French constructed wetlands using particle size distribution and dynamic vapor sorption analyses.

French vertical flow constructed wetlands (French VFCWs) are widely used for the treatment of wastewaters from small communities. In the system, unsettled wastewater is percolated through two successive stages of filter-cells planted with reeds. This causes the formation of a surface sludge layer. This layer plays positive roles in the treatment performance, but also leads to clogging. The objective of this study was to contribute to the description of the sludge deposits characteristics and their dynamics of evolution, which may control the development of clogging. Representative samples of sludge deposits were taken from 14 French VFCWs full-scale plants and analyzed for particle size, dynamic vapor sorption and other parameters of composition to compare their structure and evaluate the factors of influence. Results showed that ageing of the surface deposits layer over the years of operation in each plant induced the formation and integration of microaggregates within the initial macrostructure of fresh organic matter (OM). The humification process of the OM was found to play a key role in the aggregation process. The injection of FeCl3 operated to precipitate phosphates before filtration was found to accelerate the aggregation process in the early phase (<1 year) of operation of the sludge.

Kinetic processes in supercooled monosaccharides upon melting: Application of dielectric spectroscopy in the mutarotation studies of D-ribose.

Dielectric spectroscopy has been recently used to monitor mutarotation in undercooled D-fructose. This method can be viewed as a universal method to study mutarotation phenomenon in the whole family of monosaccharides. In this paper, we studied kinetics of mutaration of anhydrous D-ribose at ambient pressure as well as pressure effect on the rate constant of this process. Ribose mutarotation behavior is compared to the one obtained for D-fructose. In addition, we attempted to determine the "direction" of mutarotation in undercooled monosaccharides after quenching the melted sample. To this end, analysis of dipole moments of different tautomers of D-fructose and D-ribose have been performed. Conformational analysis of studied carbohydrates was done with use of density functional theory. Geometry optimizations as well as calculations of dipole moments were done on the 6-311++G(d,p)/B3LYP level. Finally, it turned out that data obtained from the mutarotation experiment might be helpful in understanding the origin of gamma-process occurring in the whole family of carbohydrates.

Comparative characterization of surface sludge deposits from fourteen French Vertical Flow Constructed Wetlands sewage treatment plants using biological, chemical and thermal indices.

Due to their design and mode of operation, French Vertical Flow Constructed Wetlands (VFCWs) accumulate suspended solids from the inflow wastewater in the form of a sludge layer at the surface of the first filter. In order to maintain the treatment performance over the long term, the characteristics of the sludge deposits and their evolution have to be well described. In this objective, a panel of sludge deposit samples taken from 14 French VFCW sewage treatment plants was investigated. Elemental composition and organic matter content, nature and reactivity were analyzed. Results clearly revealed two categories of sludge deposits, namely the "young-age plants" type (1 year of operation and less) and the "mature plants" type (3 years of operation and more). Sludge deposits from the "mature plants" exhibited same biological, physical and chemical properties. Their organic matter was globally less abundant, more humified and less biodegradable than in the young-age plants type. Their overall contents in trace metals were also higher, although in a limited manner. The effect of additional treatments, particularly FeCl3 injection for phosphorus precipitation, was observable in the "young-age plants" group. Finally, the sludge deposits sampled from one particular plant with specific operating conditions were found to exhibit very different characteristics from those of either groups identified. This observation underlined the influence of local conditions on the typology of the sludge deposits.

Association of CAST gene polymorphism with carcass value and meat quality in two synthetic lines of sheep.

The aim of study was to estimate genotype frequencies of the polymorphism in the intron 12 of CAST gene of growing lambs, and to evaluate associations between the CAST genotype and carcass and meat traits. A total number of 317 rams representing the synthetic BCP (163) and SCP (154) meaty lines were genotyped by the RFLP/MspI/NcoI/Hin6I. Three genotypes including "aa", "ae" and "ac" with most frequencies of 0.48, 0.27 and 0.19, were observed in whole population. Lambs with the "aa" CAST genotype showed the highest muscle percentage and the lowest fat percentage in the hind legs. On the other hand, the loins of rams with "ac" genotype had the highest intramuscular fat content, indicative of beneficial health-related properties and technological usefulness. However, differences between CAST genotype stated for the sensory properties, texture and thawing and cooking losses of the meat were insignificant. Breeding efforts aimed at the improvement of lamb meat quality should prioritize the selection of the "ac" genotype.

Changes in growth hormone secretion and leptin receptor mRNA expression under the influence of leptin and adrenocorticotropin in pituitary cells of early weaned ewe lambs.

Early weaning of ewe lambs strongly stimulates the hypothalamic-pituitary-adrenal axis and is associated with suppressed growth rate despite the increased food intake. At the same time, plasma leptin concentration increases only slightly or undetectably. To better understand this atypical interdependence among somatic stress, leptin, and lamb growth rate, we analyzed impact of leptin and/or adrenocorticotropic hormone (ACTH) on growth hormone (GH) secretion as well as the effect of ACTH on mRNA expression of two splice variants of leptin receptor (LEPRa, LEPRb) in pituitary cells isolated from early weaned ewe lambs. The GH secretion under the influence of leptin and/or ACTH depended on the timing of exposure and hormone concentration. After 6 - 30 h, GH secretion increased under 10-11 - 10-8 M leptin (P ≤ 0.05). However, after 24 - 30 h, GH secretion significantly increased only in cells exposed to both leptin and ACTH compared to culture with leptin only. Simultaneously, there was a significant (P ≤ 0.05) decrease in leptin receptor mRNA expression under the influence of ACTH at 10-8 - 10-6 M after 12 - 30 and 24 - 30 h for LEPRa and LEPRb, respectively. ACTH-related downregulation of LEPR mRNA was associated with a significant (P ≤ 0.05) reduction in leptin-stimulated GH secretion, also after 24 - 30 hours. Thus, the timing of ACTH exposure, followed by decreased leptin receptor mRNA, converged with the timing of decreased GH secretion under the influence of leptin with ACTH. The ACTH-induced downregulation of LEPR mRNA therefore may underlie the decrease in GH. These results show a direct role for leptin, ACTH, and leptin receptor expression in modulation of pituitary GH secretion in early weaned ewe lambs. During the early weaning-induced stress response, the ACTH-mediated decrease in sensitivity of pituitary cells to leptin may abolish a stimulatory effect of leptin on GH secretion and explain in part, the reduction in lamb growth rate.

Analytical indicators to characterize Particulate Organic Matter (POM) and its evolution in French Vertical Flow Constructed Wetlands (VFCWs).

The design of French VFCWs leads to the formation of a sludge layer at the surface of the first filters due to the retention of suspended solids from the percolation of unsettled wastewater. This layer plays a major role in the system but still little is known on its characteristics and evolutions. In this study, suspended solids and sludge deposits sampled from two French VFCW plants were analyzed by different methods in the objective to assess the evolution of particulate organic matter (POM) along the treatment chain and within the sludge layer, and identify relevant analytical indicators of these phenomena. The treatment chain included an aerobic trickling filter followed by FeCl3 injection and two successive stages of filters. Thermal analyses showed that OM contents of suspended solids decreased along the treatment chain. POM in inflow suspended solids was predominantly composed of reactive, biodegradable compounds which were partly hydrolyzed and mineralized notably at the trickling filter stage. 3D fluorescence spectra collected from aqueous POM extracts confirmed the evolution of organic matter from low-molecular reactive compounds to more complex and stable structures such as humic-like substances. FTIR confirmed the mineralization of POM's reactive constituents along the treatment chain by the decrease in the intensities of the characteristics bands of aliphatic compounds or proteins, and its humification in the sludge deposits through the relative increase of the bands at 1634cm-1 (vC=O) and 1238cm-1 (δC=O and/or δOH). Isotopic ratios δ2H/1H and δ15N/14N were found to be good indicators of POM evolutions. The higher values of δ2H/1H and δ15N/14N ratios measured in sludge deposits as compared to inflow suspended solids were related to POM humification and to microbial processes of POM hydrolysis and mineralization, respectively.

Identification of acyl phosphatidylglycerol as a minor phospholipid of Pseudomonas BAL-31.

Compound X, a minor phospholipid of Pseudomonas BAL-31 and bacteriophage PM2, has been identified as X-3-phosphatidyl-1'-(3'-acyl)-glycerol, or acyl phosphatidylglycerol. The water-soluble product obtained by mild alkaline hydrolysis showed the same RF value as that of glycerophosphoryl-glycerol. The chemical analysis gave the ratio 1 : 3 : 2 for phosphate-acyl ester-glycerol. The position of the third acyl group was determined by nuclear magnetic resonance techniques.

High-affinity receptor for interferon-gamma (IFN-gamma), a ubiquitous protein occurring in different molecular forms on human cells: blood monocytes and eleven different cell lines have the same IFN-gamma receptor protein.

High-affinity receptors for human IFN-gamma were analyzed using 13 different cells, including blood monocytes. Scatchard analysis showed one high-affinity binding site for each cell. One cross-linked complex between IFN-gamma and the receptor was detected, although their apparent molecular masses were variable in different cells, as also confirmed in immunoblots of membrane extracts. Variations in molecular masses were abolished if N-linked glycosylation was absent. Stable tryptic fragments contained the intact binding site for IFN-gamma and antibody epitopes characteristic of the extracellular domain of the IFN-gamma receptor of Raji cells and were of different sizes only if glycosylated. In addition, Northern analysis showed the same mRNA encoding the high-affinity IFN-gamma receptor in each cell analyzed. Thus, all cells including blood monocytes express the same high-affinity IFN-gamma receptor protein. N-linked sugars may give structural stability to the IFN-gamma receptor and are unlikely to be directly involved in IFN-gamma binding.

Improved robust T-wave alternans detectors.

New statistical and spectral detectors, the modified matched pairs t test, the extended spectral method and the modified spectral method, were proposed for T-wave alternans (TWA) detection gaining robustness according to trend and single-frequency interferences. They were compared to classic detectors such as matched pairs t test, unpaired t test, spectral method, generalized likelihood ratio test and estimated TWA amplitude within a simulation framework and applied to real data. The optimal detection threshold was selected by using a full Monte-Carlo simulation where signals, with and without alternans episodes, were corrupted by Gaussian noise with different power and single-frequency interferences with different tones. All the combinations of noise and frequency were selected and repeated 500 times in order to compute probability of detection ([Formula: see text]) and the false alarm probability ([Formula: see text]), providing ROC curves. The study group consisted of 50 patients with implantable cardioverter-defibrillator (age: [Formula: see text]; LVEF: [Formula: see text]), who were paced (ventricular pacing) at 100 bpm. Two-minute recordings were analyzed. The XYZ orthogonal lead system was used. The best performance was reached by using the modified matched pairs t test (in comparison with the spectral method and other reference methods).

Import of human and Thermoplasma 20S proteasomes into nuclei of HeLa cells requires functional NLS sequences.

Proteasomes are present both in the nucleus and cytoplasm of eukaryotic cells. Their localization is regulated and changes during the cell cycle. Nuclear localization signal (NLS) type sequences were identified in proteasomes from various organisms. In addition, acidic complementary sequences were identified (cNLS) which could interact with the positively charged NLS, masking or unmasking them and thereby modulating nuclear import. In this paper we show that fluorescently labeled human erythrocyte 20S proteasomes accumulate in the nucleus of digitonin-permeabilized cells. This translocation is ATP-dependent and occurs through the nuclear pore complex as is shown by blocking of the nuclear pores with wheat germ agglutinin. In addition, we used 20S proteasomes from Thermoplasma acidophilum as a model system. Recombinant 20S proteasomes from the archaebacterium Thermoplasma acidophilum are imported into nuclei of HeLa and 3T3 cells similar to their eukaryotic counterpart. We constructed mutants in the putative NLS and cNLS region to study their effect on import. The NLS mutant was not imported into nuclei and showed cytoplasmic staining only. This indicates that this sequence is indeed responsible for nuclear targeting. Mutational studies of the cNLS do not support the involvement of this sequence in regulation of nuclear transport.

Construction of yeast artificial chromosome (YAC) clone banks covering three genome equivalents and isolation of YACs containing the human gene encoding tumor necrosis factor receptor beta.

Yeast artificial chromosome (YAC) banks covering in total about three haploid genome equivalents were constructed using a human Epstein-Barr-virus-transformed B lymphocytic cell line. Two clone banks were made: 20,000 clones with average inserts of 350 kb in the pYAC4 vector and 9850 clones with average inserts of 180 kb using vectors pJS89 and pJS91. Direct comparison of pYAC4 with pJS89 and pJS91 showed pYAC4 to be the most suitable cloning vector. Two partial banks with average insert sizes of 220 kb for human endothelial cell DNA and epithelial HEp2 cell DNA were also constructed, each covering 10% of the haploid genome. A rapid, three-step PCR screening procedure for isolation of individual YAC clones was developed and used to identify two clones encoding TNF-R beta. These clones cover about 200 kb and have 170 kb in common. TNF-R beta is 9.3 kb long and contains two introns within the protein-coding sequence.

Design, synthesis, and evaluation of 2 beta-alkenyl penam sulfone acids as inhibitors of beta-lactamases.

A general method for synthesis of 2 beta-alkenyl penam sulfones has been developed. The new compounds inhibited most of the common types of beta-lactamase. The level of activity depended very strongly on the nature of the substituent in the 2 beta-alkenyl group. The inhibited species formed with the beta-lactamase from Citrobacter freundii 1205 was sufficiently stable for X-ray crystallographic studies. These, together with UV absorption spectroscopy and studies of chemical degradation, suggested a novel reaction mechanism for the new inhibitors that might account for their broad spectrum of action. The (Z)-2 beta-acrylonitrile penam sulfone Ro 48-1220 was the most active inhibitor from this class of compound. The inhibitor enhanced the action of, for example, ceftriaxone against a broad selection of organisms producing beta-lactamases. The organisms included strains of Enterobacteriaceae that produce cephalosporinases, which is an exceptional activity for penam sulfones.

Structure-based design of beta-lactamase inhibitors. 1. Synthesis and evaluation of bridged monobactams.

Bridged monobactams are novel, potent, mechanism-based inhibitors of class C beta-lactamases, designed using X-ray crystal structures of the enzymes. They stabilize the acyl-enzyme intermediate by blocking access of water to the enzyme-inhibitor ester bond. Bridged monobactams are selective class C beta-lactamase inhibitors, with half-inhibition constants as low as 10 nM, and are less effective against class A and class B enzymes (half-inhibition constants > 100 microM) because of the different hydrolysis mechanisms in these classes of beta-lactamases. The stability of the acyl-enzyme complexes formed with class C beta-lactamases (half-lives up to 2 days were observed) enabled determination of their crystal structures. The conformation of the inhibitor moiety was close to that predicted by molecular modeling, confirming a simple reaction mechanism, unlike those of known beta-lactamase inhibitors such as clavulanic acid and penam sulfones, which involve secondary rearrangements. Synergy between the bridged monobactams and beta-lactamase-labile antibiotics could be observed when such combinations were tested against strains of Enterobacteriaceae that produce large amounts of class C beta-lactamases. The minimal inhibitory concentration of the antibiotic of more than 64 mg/L could be decreased to 0.25 mg/L in a 1:4 combination with the inhibitor.

TLD postal dose intercomparison for megavoltage units in Poland.

The aim of the TLD pilot study was to investigate and to reduce the uncertainties involved in the measurements of absorbed dose and to improve the consistency in dose determination in the regional radiotherapy centres in Poland. The intercomparison was organized by the SSDL. It covered absorbed dose measurements under reference conditions for Co-60, high energy X-rays and electron beams. LiF powder type MT-N was used for the irradiations and read with the Harshaw TLD reader model 2000B/2000C. The TLD system was set up and an analysis of the factors influencing the accuracy of absorbed dose measurements with TL-detectors was performed to evaluate and minimize the measurement uncertainty. A fading not exceeding 2% in 12 weeks was found. The relative energy correction factor did not exceed 3% for X-rays in the range 4-15 MV, and 4% for electron beams between 6 and 20 MeV. A total of 34 beams was checked. Deviation of +/- 3.5% stated and evaluated dose was considered acceptable for photons and +/- 5% for electron beams. The results for Co-60, high energy X-rays and electron beams showed that there were two, three and no centres, respectively, beyond acceptance levels. The sources of errors for all deviations out of this range were thoroughly investigated, discussed and corrected, however two deviations remained unexplained. The pilot study resulted in an improvement of the accuracy and consistency of dosimetry in Poland.

Surface plasmon resonance sensor for domoic acid based on grafted imprinted polymer.

A molecularly imprinted polymer (MIP) film for domoic acid (DA) was synthesised by direct photo-grafting onto a gold chip suitable for a surface plasmon resonance (SPR) based bioanalytical instrument system, the BIAcore 3000. The gold surface was first functionalised with a self-assembled monolayer of 2-mercaptoethylamine and subsequent carbodiimide chemistry was performed for covalent attachment of the photoinitiator, 4,4'-azobis(cyanovaleric acid). This ensured that the formation of the MIP thin film, comprising 2-(diethylamino) ethyl methacrylate as functional monomer and ethylene glycol dimethacrylate as cross-linker, occurred only at the surface level. Optimisation and control over the grafting procedure were achieved using contact angle measurements and atomic force microscope (AFM) imaging. The surface grafting resulted in the formation of thin and homogeneous MIP film with thickness of 40 nm. A competitive binding assay was performed with free DA and its conjugate with horseradish peroxidase, which was used as a refractive label. The sensor was evaluated for its sensitivity, cross-reactivity, and robustness by using a BIAcore 3000. Likewise, monoclonal antibodies acting as natural receptors for the toxin were studied with the same BIAcore system. Results of a comparison between the artificial and natural receptors are reported. In contrast to monoclonal antibodies, the regeneration of MIP chip did not affect its recognition properties and continuous measurement was possible over a period of at least 2 months.

Evaluation of circulating immune complexes in pregnant women.

The level of blood immune complexes was determined in 100 healthy women with a normal course of pregnancy. The results of our studies indicate that using the test of EA rosette forming inhibition it is possible to detect circulating immune complexes in pregnant women. We have not demonstrated a quantitative interdependence of immune complexes level upon a number of past pregnancies.

Circulating immune complexes in patients with melanoma.

Sera from 51 melanoma patients and from 55 healthy donors were examined for the presence of immune complexes (ICs) by the EA-rosette forming cell inhibition assay. Using a cut-off point of 5% rosette inhibition, ICs were found in 144 out of 213 sera from patients with melanoma (67.6%) and in 11 out of 55 control subjects (20%, p less than 0.001). The percentage of positive results was significantly lower in patients with no evidence of disease (60%) than in patients with clinical symptoms of their tumor (localized disease -- 80%, p less than 0.02, metastatic disease -- 78%, p less than 0.05). The mean inhibitory rate in patients with metastatic disease (21%) was higher than in patients with localized tumor (15.1%, the difference not significant) and than in patients with no evidence of disease (10%, p less than 0.01). The high levels of ICs or a tendency for their increasing in most cases accompanied the dissemination of the neoplasm. The stabilization of ICs on the low levels was found to be a prognostically favorable sign.

Risk factors associated with incisor injury in elementary school children.

This study examined risk factors associated with incisor injury in 3396 third and fourth grade school children in Alachua County, Florida. One of six orthodontists completed a standardized examination form for each child to assess severity of incisor injury, gender, age, race, skeletal relationships, morphologic malocclusion, incisor exposure, interlabial gap, TMJ sounds, chin trauma, and history of lower facial trauma. One in five (19.2%) exhibited some degree of incisor injury. This was limited to a single tooth in 73.1% of those with injury, while enamel injury predominated (89.4%). The majority of the injuries (75.4%) were localized in the maxillary arch, with central incisors the most frequently traumatized. Chi-square tests of association indicated that gender, race, school, orthodontist, history of lower facial trauma, chin trauma, profile, and maxillary and mandibular horizontal positions were associated with incisor injury (P < 0.05). Wilcoxon rank sum tests identified differences in age, overjet, time of screening, and interlabial gap between those with and without injury (P < 0.05). Results of logistic regression analyses indicated risk of incisor injury was greater for children who had a prognathic maxilla, a history of trauma, were older, were male, and had greater overjet and mandibular anterior spacing.

[Circulating immune complexes in the blood serum of patients with untreated cervical cancer--their occurrence and quantitative analysis]. / Krazace kompleksy immunologiczne w surowicy nie leczonych chorych na raka szyjki macicy--wystepowanie i ocena ilosciowa.

Circulating immunological complexes (KI) are one of the parameters of humoral immunity. The occurrence of KI was found in the course of neoplastic diseases both in animals and humans. They are formed as a result of interaction of neoplastic antigens with specific tumor antibodies. Immunological complexes may be favourable or unfavourable to the modulation of effector mechanisms directed against neoplastic cells. In spite of extensive research, the participation of KI in immunological neoplastic response has not been clearly defined. The quantitative analysis of KI in the serum of patients with neoplasms revealed that the levels of KI may correlate with the degree of the progression of the disease, which has been shown for example in malignant melanoma, embryonal neuroma and carcinoma of the breast. The investigation of KI in the serum of patients with cervical carcinoma is rarely reported. The present paper shows 71 patients with squamous epithelial cervical carcinoma in various stages of progression, patients not receiving therapy. The evaluation of KI was made by means of two methods: test of inhibition of forming rosettes EA and inhibition of forming rosettes EAC. The authors of the few reports on the subject agree (which was also confirmed in our research) that the serum of patients with cervical carcinoma reveals the presence of KI. The frequency of occurrence and levels of KI may correlate with the degree of progression of a neoplastic process.

[Psychic factors in case histories of patients with alopecia areata--preliminary report]. / Czynniki psychiczne w wywiadzie pacjentów z lysieniem plackowatym--doniesienie wstepne.

We tested the significance of psychic factors in the etiopathogenesis of alopecia areata. We analysed the patient on the basis of a detailed examination based on the case history, including important events in his/her life, personality traits, serious events and the loss of emotional attachment. General important events happened to 80% of the patients, personality traits able to cause the disease were present in 73%, serious events in 62% and the loss of emotional attachment was also found in 62% of the patients. We tested 60 patients (31 women and 29 men). The evidence obtained from the detailed examination based on case histories indicated significantly frequent occurrence of the psychic factors preceding the occurrence of alopecia areata.