Carbohydrate masking of an antigenic epitope of influenza virus haemagglutinin independent of oligosaccharide size

Comparison of the haemagglutinins (HA) of the pathogenic avian influenza viruses A/FPV/Dutch/27 (H7N7) and A/FPV/Rostock/34 (H7N1) revealed 94.7% nucleotide and 93.8% amino acid sequence homologies. Six of the seven N-glycosidic oligosaccharides of the Rostock HA are at the same positions as the six carbohydrates of the Dutch strain. The additional oligosaccharide side chain of the Rostock strain, which is of the complex type, is attached to asparagine149 in antigenic epitope B. The accessibility of this antigenic epitope has been analysed by using rabbit antisera raised against synthetic peptides comprising amino acids 143-162. The carbohydrates of the HA of the Rostock strain have been modified (i) to truncated cores by expression in insect cells using a baculovirus vector, (ii) to oligomannosidic side chains by growth in the presence of the trimming inhibitor methyldeoxynojirimycin and (iii) to a single N-acetylglucosamine residue by removal of the oligomannosidic sugar with endo-β-N-acetylglucosaminidase H. Neither the authentic nor the modified oligosaccharides allowed antibody binding, as indicated by enzyme-linked immunosorbent assay (ELISA) and Western blot analyses. Reactivity was observed, however, after complete removal of the carbohydrate from HA of the Rostock strain by digestion with peptide-N-glycosidase F. HA of the Dutch strain was reactive without prior peptide-N-glycosidase F treatment. These results demonstrate that a single N-acetylglucosamine at asparagine149 is sufficient to prevent recognition of the peptide epitop

Expression of the synthetic gene of an artificial DDT-binding polypeptide in Escherichia coli

This paper reports the expression of an artificial functional polypeptide in bacteria. The gene of a designed 24-residue DDT-binding polypeptide (DBP) was inserted between the BamHI and PstI cleavage sites of plasmid pUR291. The hybrid plasmid, pUR291-DBP, was cloned in Escherichia coli JM109. After induction by isopropyl-β-D-thiogalactopyranoside a fusion protein was expressed in which DBP was linked to the COOH-termiuus of β-galactosidase. DBP, which is stable to trypsin, was obtained by tryptic digestion of the fusion protein and subsequent fractionation of the tryptic peptides by reversed-phase h.p.l.c. Recombinant and chemically synthesized DBP showed identical chromatographic properties, amino acid composition, and chymotryptic digestion patterns. Both the β-galactosidase-DBP fusion and isolated recombinant DBP bound DDT. The fusion protein was 25 times as potent as the designed 24-residue DBP in activating a cytochrome P-450 model system using equimolar catalytic amounts of the two protein

Die Wiederbegrünung städtischen Ödlandes, dargestellt am Beispiel Leipzigs

Die Hercynia publiziert Originalbeiträge mit dem Schwerpunkt Ökologie (mit ihren vielseitigen Aspekten der Biodiversität), Botanik, Zoologie, Geologie und Geografie, den anwendungsorientierten Bereichen des Natur- und Umweltschutzes, sowie der Land- und Forstwirtschaft

Achieving Biocompatible SABRE : An in vitro Cytotoxicity Study

Production of a biocompatible hyperpolarized bolus for signal amplification by reversible exchange (SABRE) could open the door to simple clinical diagnosis via magnetic resonance imaging. Essential to successful progression to preclinical/clinical applications is the determination of the toxicology profile of the SABRE reaction mixture. Herein, we exemplify the cytotoxicity of the SABRE approach using invitro cell assays. We conclude that the main cause of the observed toxicity is due to the SABRE catalyst. We therefore illustrate two catalyst removal methods: one involving deactivation and ion-exchange chromatography, and the second using biphasic catalysis. These routes produce a bolus suitable for future invivo study

Die Vegetation des Lauchs bei Eilenburg: ein Beitrag zur Kenntnis nordwestsächsischer Pflanzengesellschaften, insbesondere des Hartholz- Auenwaldes der Mulde

WALTER, B.; GUTTE, P.: Vegetation of the forest „Lauch” near Eilenburg - contribution to the knowledge of plant communities in northwestern Saxony. - Hercynia 36 (2003): 47–73.Based on 86 phytosociological records 17 plant communities from the „Lauch” - a forest near Eilenburg (Germany, NW-Saxony) - are described, especially the associations Querco-Ulmetum and Galio- Carpinetum, which are the dominating forest-communities. The importance of the region for the protection of nature is stressed, and suggestions for measures of cultivation and development are proposed.Die Hercynia publiziert Originalbeiträge mit dem Schwerpunkt Ökologie (mit ihren vielseitigen Aspekten der Biodiversität), Botanik, Zoologie, Geologie und Geografie, den anwendungsorientierten Bereichen des Natur- und Umweltschutzes, sowie der Land- und Forstwirtschaft

SPECT/CT and pulmonary embolism

Acute pulmonary embolism (PE) is diagnosed either by ventilation/perfusion (V/P) scintigraphy or pulmonary CT angiography (CTPA). In recent years both techniques have improved. Many nuclear medicine centres have adopted the single photon emission CT (SPECT) technique as opposed to the planar technique for diagnosing PE. SPECT has been shown to have fewer indeterminate results and a higher diagnostic value. The latest improvement is the combination of a low-dose CT scan with a V/P SPECT scan in a hybrid tomograph. In a study comparing CTPA, planar scintigraphy and SPECT alone, SPECT/CT had the best diagnostic accuracy for PE. In addition, recent developments in the CTPA technique have made it possible to image the pulmonary arteries of the lungs in one breath-hold. This development is based on the change from a single-detector to multidetector CT technology with an increase in volume coverage per rotation and faster rotation. Furthermore, the dual energy CT technique is a promising modality that can provide functional imaging in combination with anatomical information. Newer high-end CT scanners and SPECT systems are able to visualize smaller subsegmental emboli. However, consensus is lacking regarding the clinical impact and treatment. In the present review, SPECT and SPECT in combination with low-dose CT, CTPA and dual energy CT are discussed in the context of diagnosing PE

Simultaneous hyperpolarized 13C-pyruvate MRI and 18F-FDG-PET in cancer (hyperPET):feasibility of a new imaging concept using a clinical PET/MRI scanner

In this paper we demonstrate, for the first time, the feasibility of a new imaging concept - combined hyperpolarized (13)C-pyruvate magnetic resonance spectroscopic imaging (MRSI) and (18)F-FDG-PET imaging. This procedure was performed in a clinical PET/MRI scanner with a canine cancer patient. We have named this concept hyper PET. Intravenous injection of the hyperpolarized (13)C-pyruvate results in an increase of (13)C-lactate, (13)C-alanine and (13)C-CO(2) ((13)C-HCO(3)) resonance peaks relative to the tissue, disease and the metabolic state probed. Accordingly, with dynamic nuclear polarization (DNP) and use of (13)C-pyruvate it is now possible to directly study the Warburg Effect through the rate of conversion of (13)C-pyruvate to (13)C-lactate. In this study, we combined it with (18)F-FDG-PET that studies uptake of glucose in the cells. A canine cancer patient with a histology verified local recurrence of a liposarcoma on the right forepaw was imaged using a combined PET/MR clinical scanner. PET was performed as a single-bed, 10 min acquisition, 107 min post injection of 310 MBq (18)F-FDG. (13)C-chemical shift imaging (CSI) was performed just after FDG-PET and 30 s post injection of 23 mL hyperpolarized (13)C-pyruvate. Peak heights of (13)C-pyruvate and (13)C-lactate were quantified using a general linear model. Anatomic (1)H-MRI included axial and coronal T1 vibe, coronal T2-tse and axial T1-tse with fat saturation following gadolinium injection. In the tumor we found clearly increased (13)C-lactate production, which also corresponded to high (18)F-FDG uptake on PET. This is in agreement with the fact that glycolysis and production of lactate are increased in tumor cells compared to normal cells. Yet, most interestingly, also in the muscle of the forepaw of the dog high (18)F-FDG uptake was observed. This was due to activity in these muscles prior to anesthesia, which was not accompanied by a similarly high (13)C-lactate production. Accordingly, this clearly demonstrates how the Warburg Effect directly can be demonstrated by hyperpolarized (13)C-pyruvate MRSI. This was not possible with (18)F-FDG-PET imaging due to inability to discriminate between causes of increased glucose uptake. We propose that this new concept of simultaneous hyperpolarized (13)C-pyruvate MRSI and PET may be highly valuable for image-based non-invasive phenotyping of tumors. This methods may be useful for treatment planning and therapy monitoring

Senecio cannabifolius Less.: eine für Deutschland neue Art

Gutte, P.; Jäger, E. J.: Senecio cannabifolius Less. – new for Germany. – Hercynia N. F. 37 (2004): 169 – 173.Senecio cannabifolius Less., a high forb from East Asia, was found to be established in a flood plain forest in Leipzig. The possibility of spread of this neophyte is discussed by comparison of its native distribution with the native and synanthropic range of Fallopia sachalinensis.Die Hercynia publiziert Originalbeiträge mit dem Schwerpunkt Ökologie (mit ihren vielseitigen Aspekten der Biodiversität), Botanik, Zoologie, Geologie und Geografie, den anwendungsorientierten Bereichen des Natur- und Umweltschutzes, sowie der Land- und Forstwirtschaft

Enhance Crawler For Efficiently Harvesting Deep Web Interfaces

Scenario in web is varying quickly and size of web resources is rising, efficiency has become a challenging problem for crawling such data. The hidden web content is the data that cannot be indexed by search engines as they always stay behind searchable web interfaces. The proposed system purposes to develop a framework for focused crawler for efficient gathering hidden web interfaces. Firstly Crawler performs site-based searching for getting center pages with the help of web search tools to avoid from visiting additional number of pages. To get more specific results for a focused crawler, projected crawler ranks websites by giving high priority to more related ones for a given search. Crawler accomplishes fast in-site searching via watching for more relevant links with an adaptive link ranking. Here we have incorporated spell checker for giving correct input and apply reverse searching with incremental site prioritizing for wide-ranging coverage of hidden web sites