Origin and compensation of imaging artefacts in localization-based super-resolution microscopy.

Interpretation of high resolution images provided by localization-based microscopy techniques is a challenge due to imaging artefacts that can be categorized by their origin. They can be introduced by the optical system, by the studied sample or by the applied algorithms. Some artefacts can be eliminated via precise calibration procedures, others can be reduced only below a certain value. Images studied both theoretically and experimentally are qualified either by pattern specific metrics or by a more general metric based on fluorescence correlation spectroscopy.

A note on the special fillet fatty acid composition of the dwarf carp (Cyprinus carpio carpio) living in thermal Lake Hévíz, Hungary.

Fatty acid (FA) composition of the fillet and the intestinal content of dwarf common carp (Cyprinus carpio carpio) living in Lake Hévíz was determined in wintertime collected samples and results were compared to widespread literature data on carp. Fillet FA profile of the thermally adapted (28 °C) Hévíz dwarf carps differed from profiles originated from divergent culture and feeding conditions in the overall level of saturation. Fillet myristic acid proportions largely exceeded all literature data in spite of poor dietary supply. Fillet fatty acid results indicate the effects of thermal adaptation (high saturation level) and the correlative effects of feed components rich in omega-3 fatty acids, with special respect to docosahexaenoic acid. With the application of discriminant factor analysis the Hévíz sample was accurately differentiated from the literature data on carp fillet fatty acid profile, mostly based on C14:0, C18:1 n9, C18:2 n6, C20:1 n9 and C20:4 n6 FAs. In summary, fillet FA profile suggested thermal adaptation, location specificity and the ingestion of algal and bacterial material.

Synthesis and biological effects of some kynurenic acid analogs.

The overactivation of excitatory amino acid receptors plays a key role in the pathomechanism of several neurodegenerative disorders and in ischemic and post-ischemic events. Kynurenic acid (KYNA) is an endogenous product of the tryptophan metabolism and, as a broad-spectrum antagonist of excitatory amino acid receptors, may serve as a protective agent in neurological disorders. The use of KYNA is excluded, however, because it hardly crosses the blood-brain barrier. Accordingly, new KYNA analogs which can readily cross this barrier and exert their complex anti-excitatory activity are generally needed. During the past 6 years, we have developed several KYNA derivatives, among others KYNA amides. These new analogs included one, N-(2-N,N-dimethylaminoethyl)-4-oxo-1H-quinoline-2-carboxamide hydrochloride (KYNA-1), that has proved to be neuroprotective in several models. This paper reports on the synthesis of 10 new KYNA amides (KYNA-1-KYNA-10) and on the effectiveness of these molecules as inhibitors of excitatory synaptic transmission in the CA1 region of the hippocampus. The molecular structure and functional effects of KYNA-1 are compared with those of other KYNA amides. Behavioral studies with these KYNA amides demonstrated that they do not exert significant nonspecific general side-effects. KYNA-1 may therefore be considered a promising candidate for clinical studies.

Value of ultrasound in preoperative local staging in early breast cancer.

PURPOSE: The purpose of this study was to determine the precision of breast ultrasound for the measurement of breast lesions compared to the histological measurement. A number of other dependent variables were also analyzed. MATERIAL AND METHODS: 460 patients with 445 malignant lesions were examined using breast ultrasound and the lesions were measured and compared to the histologically measured size. The data was further analyzed according to histology, tumor stage, age, grading and therapy. RESULTS: Metric comparison showed good correlation between sonography and the pathologically measured size of breast lesions, especially in tumor stage T 1 and T 2 and within ductal invasive carcinomas. Higher tumor stages lead to imprecise measurements and the histological type of lobular invasive carcinoma also results in imprecise measurements. Age and grading do not influence measurement precision. CONCLUSION: Breast ultrasound allows precise measurement of breast lesions especially at lower tumor stages. The higher the tumor stage, the more imprecise the measurement becomes. Multivariate analysis shows no cross impact between tumor stage and histological type with respect to the quality of measurement.

Glioblastoma multiforme with an unusual location and clinical course.

We present a unique case of a brain tumor patient with atypical location and progression. He was initially presented with mood and anxiety type symptoms together with aphasia and left-sided paraesthesias. Magnetic resonance imaging and CSF were negative and the patient was diagnosed with PTSD as he recently experienced a small motor vehicle accident. Two months after the first presentation, MRI revealed multifocal juxtacortical, leptomeningeal hyperdensities in the bilateral frontal lobes. MRI-guided frameless stereotactic biopsy defined a diagnosis of GBM 1 week prior to death which occurred within 4 months. Postmortally, formalin-fixed brain demonstrated that the main tumor mass was located in the fornix, infiltrating the ventricular system and disseminating over the cortex, cerebellum and spinal cord. The authors recommend closer scrutiny of psychiatric patients presenting CNS symptomatology, negative MRI, CT and CSF.

Nitrogen losses and chemical parameters during co-composting of solid wastes and liquid pig manure.

The aim of this research was to study nitrogen losses during the treatment of the liquid fraction (LF) of pig manure by co-composting and to establish the best conditions for compost production with higher nitrogen and low heavy metal contents. Windrows were constituted with the solid fraction (SF) of pig manure, different organic waste (SF of pig manure, sawdust and grape bagasse) as co-substrate and Populus spp. wood chips as bulking material and watered intensely with the LF. Results show that nitrogen losses ranged from 30% to 66% of initial nitrogen and were mainly governed by substrate to bulking mass ratio and liquid fraction to substrate (LF/S) ratio, and only secondarily by operational parameters. Nitrogen losses decreased from 55-65% at low LF/S ratios (1.7-1.9 m3/t total solids (TS)) to 30-39% at high LF/S ratios (4.4-4.7 m3/t TS). Therefore, integrating the LF in the composting process at high LF/S ratios favoured nitrogen recovery and conservation. Nitrogen in the fine fraction (ranging from 27% to 48% of initial nitrogen) was governed by operational parameters, namely pH and temperature. Final compost showed low content in most heavy metals, but Zn was higher than the limits for compost use in agriculture. Zn content in the obtained compost varied from 1863 to 3269 mg/kg dm, depending on several factors. The options for obtaining better quality composts from the LF of pig manure are selecting co-substrates with low heavy metal content and using them instead of the SF of pig manure.

Neutron reflectometry with the Multi-Blade 10B-based detector.

The Multi-Blade is a boron-10-based gaseous detector developed for neutron reflectometry instruments at the European Spallation Source in Sweden. The main challenges for neutron reflectometry detectors are the instantaneous counting rate and spatial resolution. The Multi-Blade has been tested on the CRISP reflectometer at the ISIS Neutron and Muon Source in the UK. A campaign of scientific measurements has been performed to study the Multi-Blade response in real instrumental conditions. The results of these tests are discussed in this paper.

Hydrolytic anaerobic reactor and aerated constructed wetland systems for municipal wastewater treatment - HIGHWET project.

The HIGHWET project combines the hydrolytic up-flow sludge bed (HUSB) anaerobic digester and constructed wetlands (CWs) with forced aeration for decreasing the footprint and improving effluent quality. The HIGHWET plant in A Coruña (NW of Spain) treating municipal wastewater consists of a HUSB and four parallel subsurface horizontal flow (HF) CWs. HF1, HF2 and HF3 units are fitted with forced aeration, while the control HF4 is not aerated. All the HF units are provided with effluent recirculation, but different heights of gravel bed (0.8 m in HF1 and HF2, and 0.5 m in HF3 and HF4) are implemented. Besides, a tobermorite-enriched material was added in the HF2 unit in order to improve phosphorus removal. The HUSB 76-89% of total suspended solids (TSS) and about 40% of chemical oxygen demand (COD) and biological oxygen demand (BOD). Aerated HF units reached above 96% of TSS, COD and BOD at a surface loading rate of 29-47 g BOD5/m2·d. An aeration regime ranging from 5 h on/3 h off to 3 h on/5 h off was found to be adequate to optimize nitrogen removal, which ranged from 53% to 81%. Average removal rates of 3.4 ± 0.4 g total nitrogen (TN)/m2·d and 12.8 ± 3.7 g TN/m3·d were found in the aerated units, being 5.5 and 4.1 times higher than those of the non-aerated system. The tobermorite-enriched HF2 unit showed a distinct higher phosphate (60-67%) and total phosphorus (54%) removal.

Murine monoclonal antibodies against carcinoembryonic antigen: immunological, pharmacokinetic, and targeting properties in humans.

We have examined three 131I-labeled murine monoclonal antibodies (MAbs) against carcinoembryonic antigen (CEA), NP-2, NP-3, and NP-4, after i.v. injection in patients with diverse cancers. Although the MAbs had a similar tumor-targeting ability, several important features were discovered that have led us to the selection of one of these MAbs for further clinical evaluation. We found that it is important to evaluate MAbs with a high immunoreactivity. For example, the MAb NP-2 was used initially in patients with an immunoreactivity between 35 and 50%. Although the tumor-imaging properties of this MAb compared favorably with the affinity-purified, goat anti-CEA antibody that we used previously, further purification of NP-2 to an immunoreactivity greater than 70% uncovered a previously unknown cross-reactivity with human granulocytes. It was also discovered that the MAbs differed in their ability to complex with CEA in the blood. Plasma samples were analyzed by gel filtration at 1 or 24 h after injection. The formation of complexes with circulating CEA was dependent on the CEA:MAb ratio in the blood. NP-3 complexed to a greater degree with CEA than NP-4, but NP-2 did not complex with CEA even at CEA:NP-2 ratios of 55 to 1. NP-3 commonly showed enhanced uptake in the colon by external scintigraphy, and examination of the radioactivity in the stool showed that most of the radioactivity was associated with whole IgG and large-sized fragments of NP-3. We also compared the rate of elimination of radioactivity from the blood for all of the MAbs and compared the clearance of NP-3 to NP-4 at three different ranges of MAb protein doses (less than 1.0 mg, 1 to 5 mg, and 5 to 20 mg). The blood clearance rate for NP-3 was fastest among the other MAbs at protein doses exceeding 1.0 mg. Patients given less than 1.0 mg of NP-4 had a significantly (P less than 0.005) shorter elimination half-life than patients given more than 1.0 mg of NP-4. By virtue of NP-4's good targeting properties in patients and its limited complexation with circulating CEA, it was selected as the MAb of choice for CEA tumor imaging.

Radiosensitivity detected by the micronucleus test is not generally increased in sporadic prostate cancer patients.

The micronucleus test (MNT) has shown increased micronuclei (MN) frequencies in BRCA associated and sporadic breast cancer patients, Ataxia telangiectasia and Nijmegen Breakage Syndrome patients, demonstrating a common cellular phenotype of increased radiosensitivity. Some genes, causative of these diseases, have also recently been associated with prostate cancer. In order to investigate if prostate cancer exhibits the cellular phenotype of increased radiosensitivity, we performed MNT analysis on 22 sporadic prostate cancer patients and 43 male controls. We determined the baseline MN frequency, in order to see in vivo chromosomal damage without radiation, and induced (after irradiation with 2 Gy) frequency of MN, both in binucleated cells (BNC) obtained from cultured peripheral blood lymphocytes. An automated image analysis system was used to score the MN employing two different classifiers (Classifier A and B) for detection of BNC. The mean baseline frequencies were 48/43 MN/1000 BNC (A/B) for the controls and 42/50 (A/B) for prostate cancer patients. The induced MN frequencies amounted to 107/111 MN/1000 BNC (A/B) for controls and 111/114 MN/1000 BNC (A/B) for prostate cancer patients. The obtained MN frequencies did not result in a statistically significant difference between unselected cases and controls. However, restricting the analysis to young patients (50-60 years, N = 7) and age-matched controls (N = 7) revealed marginally significant higher MN frequencies in patients. We conclude that increased radiosensitivity is not a property of prostate cancer patients in general.

Methane and carbon dioxide emissions from constructed wetlands receiving anaerobically pretreated sewage.

The aim of this research was to determine methane and carbon dioxide emissions from a hybrid constructed wetland (CW) treating anaerobically pre-treated sewage. The CW was constituted of two horizontal flow (free water surface followed by a subsurface) units. A long-term study was carried out as both CW units were monitored for three campaigns in Period 1 (0.9-1.5years after start-up), and four campaigns in Period 2 (4.5-5.8years after start-up). The closed chamber method with collecting surfaces of 1810cm(2) was used. For this system, variability due to position in the transverse section of CW, plant presence or absence and recommended sampling period was determined. Overall methane emissions ranged from 96 to 966mgCH4m(-2) d(-1), depending on several factors as the operation time, the season of the year and the position in the system. Methane emissions increased from 267±188mgCH4m(-2)d(-1) during the second year of operation to 543±161mgCH4m(-2)d(-1) in the sixth year of operation. Methane emissions were related to the age of the CW and the season of the year, being high in spring and becoming lower from spring to winter. Total CO2 emissions ranged mostly from 3500 to 5800mgCO2m(-2)d(-1) during the sixth year of operation, while nitrous oxide emissions were below the detection limit of the method.

New developments in automated cytogenetic imaging: unattended scoring of dicentric chromosomes, micronuclei, single cell gel electrophoresis, and fluorescence signals.

The quantification of DNA damage, both in vivo and in vitro, can be very time consuming, since large amounts of samples need to be scored. Additional uncertainties may arise due to the lack of documentation or by scoring biases. Image analysis automation is a possible strategy to cope with these difficulties and to generate a new quality of reproducibility. In this communication we collected some recent results obtained with the automated scanning platform Metafer, covering applications that are being used in radiation research, biological dosimetry, DNA repair research and environmental mutagenesis studies. We can show that the automated scoring for dicentric chromosomes, for micronuclei, and for Comet assay cells produce reliable and reproducible results, which prove the usability of automated scanning in the above mentioned research fields.

Evaluation of a remote radioiodination system for radioimmunotherapy.

This report describes a remote radioiodination system which is inexpensive, easy to assemble, disposable, and capable of radioiodinating curie levels of activity safely. In addition to the safety afforded by this system, an immobilized oxidant and anion exchange resin are used to generate electrophilic iodine and remove free iodine, respectively. Reducing agents are not used and, therefore, when radioiodinating F(ab')2 fragments, degradation does not occur. In contrast, chloramine-T, sodium metabisulfite (CT/SMB) iodinations of F(ab')2 fragments resulted in products with up to 40% Fab' fragments. Radiolabeling yields (65.8% +/- 8.1%) and antibody immunoreactivity (68.8% +/- 8.0%) were not statistically different (p less than 0.001) from those obtained in remote CT/SMB iodinations. The system is currently being used to radioiodinate both IgG and F(ab')2 monoclonal antibodies with up to 450 mCi 131I for clinical radioimmunotherapy trials.

Sustained complete autonomic blockade.

1. A 3-hr period of complete autonomic blockade was induced in anaesthetized sheep and dogs using a continuous intravenous infusion of trimetaphan camphorsulphonate and atropine. Circulatory, respiratory and metabolic parameters were studied over 24 hr or until the animal died, whichever came first.2. After termination of the blockade all animals remained hypotensive and within a few hours all died in irreversible shock which was not accompanied by any of the otherwise characteristic respiratory or metabolic consequences of circulatory failure.3. Administration of trimetaphan alone for a similar period was innocuous.4. When during complete autonomic blockade a continuous infusion of dopamine or hypertensin was additionally administered mortality rate was halved but all survivors remained hypotensive.

Donor marrow progenitors (CFU-Mix, BFU-E and CFU-GM) and haemopoietic engraftment following HLA matched sibling bone marrow transplantation.

Bone marrow multipotent (CFU-Mix) and unipotent (CFU-GM and BFU-E) progenitor cells in the donor marrow inoculums were measured in 24 histocompatible sibling bone marrow transplants. The number of donor marrow nucleated cells, CFU-Mix, CFU-GM and BFU-E given per kilogram (kg) of recipient's body weight were 2.4 +/- 0.6 X 10(8), 3.6 +/- 4.2 X 10(3), 4.9 +/- 3.3 X 10(-4) and 4.3 +/- 4.1 X 10(4) respectively (mean +/- S.D.). Fast engraftment patients, as assessed by rise in peripheral blood neutrophils (greater than or equal to 0.5 and greater than or equal to 1.0 X 10(9)/l) and platelets (greater than 20 and greater than 50 X 10(9)/l), received a significantly greater amount of CFU-Mix/kg (greater than 3 X 10(3)/kg, p less than 0.025) and CFU-GM/kg (greater than 3 X 10(4)/kg, p less than 0.05 except for plat greater than or equal to 20 X 10(9)/l) than the slow recovery patients. Significant correlations were found between the donor CFU-Mix/kg infused and neutrophil recovery to 1 X 10(9)/l and platelet to 50 X 10(9)/l (Spearman's rank correlation coefficient r = 0.38, p = 0.04 and r = 0.58, p = 0.003, respectively). The amount of donor CFU-GM/kg given also correlated significantly to neutrophil (1 X 10(9)/l) and platelet (50 X 10(9)/l) recovery, (r = 0.33 and r = 0.37, respectively, p less than or equal to 0.05). There was no association between BFU-E, and marrow nucleated cells infused per kg and haemopoietic recovery. A number of clinical parameters were also examined to determine other factors that may influence the rate of engraftment. Acute graft vs host disease (greater than or equal to grade II) and methotrexate therapy post-transplant delayed the platelet regeneration. The results of the present report indicate that in vitro measurement of donor CFU-Mix and CFU-GM progenitors infused, correlate with the speed of granulocyte and platelet recovery in clinical allogeneic bone marrow transplants.

Pneumoperitoneum: diagnosis from chest radiographs.

Children under intensive care for various illnesses are usually followed closely with radiographic examinations of the chest. The recognition of a pneumoperitoneum as a complicating feature of these illneses depends on subtle radiographic signs that are frequently visible on the chest film studies. We have assembled and illustrated these findings in the hope that pediatricians will be better able to recognize this complication when it occurs.

Gastric stapling: a significant risk factor in the treatment of tricyclic antidepressant overdose.

Attention is called to prior gastric stapling as a risk factor for emesis and aspiration in those patients who are treated with single or repeated oral activated charcoal for tricyclic antidepressant overdose. When administered activated charcoal as part of the standard protocol for drug poisoning, these individuals have the potential for developing severe and repeated emesis with subsequent aspiration of this compound into the respiratory tract with development of the Acute Lung Injury syndrome.

Health Promotion Schools of Excellence: a model program for Kentucky and the nation.

This article reviews the development and progress of an innovative, comprehensive school health project in the Jefferson County school district known as the Health Promotion Schools of Excellence (HPSE). This project features unique working relationships between public and private entities seeking a common goal: "A Healthier Community." The goals and development of HPSE have been formulated to coincide with the emerging directives of the Kentucky Education Reform Act (KERA) as well as the growing pressures to control health care costs through effective preventive measures. The results of testing following the first and second years of the project show an encouraging trend of improved physical fitness levels for all students (grades K-12) and improving levels of health knowledge and attitudes in elementary level children (grades 4-8) as well as school faculty and staff. These initial results, although encouraging, will only be meaningful if they translate over time into a healthier, more responsible cohort of adolescents and young adults when compared to their peers not involved in the project. The project, while only in its third year of development, has already been recognized at regional and national levels as a successful model of a comprehensive school health program. As the project continues and grows, the authors anticipate developing one of the nation's largest and most comprehensive longitudinal data bases of childhood and adolescent health information.

Heavy metal removal in an UASB-CW system treating municipal wastewater.

The objective of the present study was to investigate for the first time the long-term removal of heavy metals (HMs) in a combined UASB-CW system treating municipal wastewater. The research was carried out in a field pilot plant constituted for an up-flow anaerobic sludge bed (UASB) digester as a pretreatment, followed by a surface flow constructed wetland (CW) and finally by a subsurface flow CW. While the UASB showed (pseudo) steady state operational conditions and generated a periodical purge of sludge, CWs were characterised by the progressive accumulation and mineralisation of retained solids. This paper analyses the evolution of HM removal from the water stream over time (over a period of 4.7 year of operation) and the accumulation of HMs in UASB sludge and CW sediments at two horizons of 2.7 and 4.0 year of operation. High removal efficiencies were found for some metals in the following order: Sn > Cr > Cu > Pb > Zn > Fe (63-94%). Medium removal efficiencies were registered for Ni (49%), Hg (42%), and Ag (40%), and finally Mn and As showed negative percentage removals. Removal efficiencies of total HMs were higher in UASB and SF units and lower in the last SSF unit.

Post-ischemic treatment with L-kynurenine sulfate exacerbates neuronal damage after transient middle cerebral artery occlusion.

Since brain ischemia is one of the leading causes of adult disability and death, neuroprotection of the ischemic brain is of particular importance. Acute neuroprotective strategies usually have the aim of suppressing glutamate excitotoxicity and an excessive N-methyl-d-aspartate (NMDA) receptor function. Clinically tolerated antagonists should antagonize an excessive NMDA receptor function without compromising the normal synaptic function. Kynurenic acid (KYNA) an endogenous metabolite of the tryptophan metabolism, may be an attractive neuroprotectant in this regard. The manipulation of brain KYNA levels was earlier found to effectively enhance the histopathological outcome of experimental ischemic/hypoxic states. The present investigation of the neuroprotective capacity of L-kynurenine sulfate (L-KYNs) administered systemically after reperfusion in a novel distal middle cerebral artery occlusion (dMCAO) model of focal ischemia/reperfusion revealed that in contrast with earlier results, treatment with L-KYNs worsened the histopathological outcome of dMCAO. This contradictory result indicates that post-ischemic treatment with L-KYNs may be harmful.