The role of type III collagen in spontaneous cervical arterial dissections

OBJECTIVE: To determine the efficacy of lymphadenectomy after nephroureterectomy in patients with transitional cell carcinoma (TCC) of the upper urinary tract. PATIENTS AND METHODS: Between January 1986 and December 1995, 72 patients (mean age 67 years, range 45-82) underwent nephroureterectomy for primary TCC of the upper urinary tract. In 35 patients, a lymphadenectomy was also performed. The clinicopathological data were analysed retrospectively, focusing on the significance of lymphadenectomy. RESULTS: Lymph vessel invasion was found in 28 patients and its incidence was closely correlated with both tumour grade and pathological stage. Of the 35 patients who underwent lymphadenectomy, lymph node metastases were found in 13 patients, all of whom had lymph vessel invasion. There was no significant difference in the survival rate between patients with and without lymphadenectomy; however, among the 44 patients with no lymph vessel invasion, the survival rate of those with lymphadenectomy was significantly higher than in those without (P<0.05). CONCLUSION: Lymphadenectomy may provide a therapeutic advantage in patients with upper urinary tract TCC and no lymph vessel invasion. However, patients with lymph vessel invasion seem to have systemic disease; therefore, aggressive systemic adjuvant therapies rather than regional lymphadenectomy should be applied in these patients.     

Adaptation and validation of antibody-ELISA using dried blood spots on filter paper for epidemiological surveys of tsetse-transmitted trypanosomosis in cattle

The indirect enzyme-linked immunosorbent assay (ELISA) for the detection of anti-trypanosomal antibodies in bovine serum was adapted for use with dried blood spots on filter paper. Absorbance (450 nm) results for samples were expressed as percent positivity, i.e. percentage of the median absorbance result of four replicates of the strong positive control serum. The antibody-ELISA was evaluated in Zambia for use in epidemiological surveys of the prevalence of tsetse-transmitted bovine trypanosomosis. Known negative samples (sera, n = 209; blood spots, n = 466) were obtained from cattle from closed herds in tsetse-free areas close to Lusaka. Known positive samples (sera, n = 367; blood spots, n = 278) were obtained from cattle in Zambia's Central, Lusaka and Eastern Provinces, diagnosed as being infected with Trypanosoma brucei, T. congolense, or T. vivax using the phase-contrast buffy-coat technique or Giemsa-stained thick and thin blood smears. For sera (at a cut-off value of 23.0% positivity) sensitivity and specificity were 86.1 and 95.2%, respectively. For bloodspots (at a cut-off value of 18.8% positivity) sensitivity and specificity were 96.8 and 95.7%, respectively. The implications of persistence of antibodies following treatment or self-cure are discussed.     

Expression and cellular distribution of baculovirus-expressed bovine herpesvirus 1 (BHV-1) glycoprotein D (gD) sequences

Glycoprotein D (gD) of bovine herpesvirus 1 (BHV-1), a homolog of herpes simplex virus gD, represents a major component of the viral envelope and is a dominant immunogen. To study the antigenic properties of the different regions of gD, we have expressed the full-length gD encoding gene and overlapping fragments spanning various regions of the gD open reading frame in a baculovirus (Autographa californica nuclear polyhedrosis virus)--insect cell (Spodoptera frugiperda, SF-9) system. Maximum levels of expression for all proteins were obtained 48 to 72 h post infection of SF-9 cells by recombinant viruses. Full-length and truncated recombinant gD proteins reacted specifically with anti-gD monospecific serum as determined by immunoprecipitation and immunoblotting, indicating that the proteins retained their antigenicity. However, based on the reactivity with a panel of gD-specific monoclonal antibodies (Mabs), the full-length recombinant gD lacked proper expression for two highly neutralizing linear epitopes identified by Mabs R54 and 9D6. The rest of the epitopes appeared to be preserved and antigenically unaltered. Immunofluorescence studies of recombinant baculovirus infected SF-9 cells using gD monospecific serum, revealed no direct correlation between cellular localization of the expressed proteins and their amino acid sequences.     

Application of nested PCR and mass spectrometry for DNA-based virus detection: HBV-DNA detected in the majority of isolated anti-HBc positive sera

DNA preparations from three different groups of serum samples were examined for HBV-DNA via a nested polymerase chain reaction assay (lower detection limit: 10 viral genomes in 100 microliters serum): Group I consisted of 11 uninfected control sera, group II consisted of sera obtained from 11 HBV infected patients and group III consisted of 21 isolated anti-HBc positive samples. The 21 samples from group III were HBV-DNA negative according to a conventional non-nested PCR assay and hybridization with a 32P-labelled probe. Using nested PCR and mass spectrometry, HBV-DNA was detected in none of group I and in all of group II samples. In 11 out of 21 (52%) of the isolated anti-HBc positive sera from group III, HBV-DNA was detected. No correlation was observed between HBV-DNA positivity and anti-HBc titers. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry provided a fast, sensitive and non-radioactive assay for the detection of PCR products without the need for gel electrophoresis or hybridization with labelled probes.     

A new class of tobacco chitinases homologous to bacterial exo-chitinases displays antifungal activity

A novel chitinase gene of tobacco was isolated and characterized by DNA sequence analysis of a genomic clone and a cDNA clone. Comparative sequence analysis of both clones showed an identity of 94%. The proteins encoded by these sequences do not correspond to any of the previously characterized plant chitinases of classes I-IV and are designated as class V chitinases. Comparison of the chitinase class V peptide sequence with sequences in the Swiss Protein databank revealed significant sequence similarity with bacterial exo-chitinases from Bacillus circulans, Serratia marcescens and Streptomyces plicatus. It was demonstrated that class V chitinase gene expression is induced after treatment of tobacco with different forms of stress, like TMV-infection, ethylene treatment, wounding or ultraviolet irradiation. Two related chitinase class V proteins of 41 and 43 kDa were purified from Samsun NN tobacco leaves inoculated with tobacco mosaic virus. The proteins were purified by Chelating Superose chromatography and gel filtration. In vitro assays demonstrated that class V chitinases have endo-chitinase activity and exhibit antifungal activity toward Trichoderma viride and Alternaria radicina. In addition, it was shown that class V chitinase acts synergistically with tobacco class I beta-1,3-glucanase against Fusarium solani germlings.     

Burst Detection by Water Demand Nowcasting Based on Exogenous Sensors

Bursts of drinking water pipes not only cause loss of drinking water, but also damage below and above ground infrastructure. Short-term water demand forecasting is a valuable tool in burst detection, as deviations between the forecast and actual water demand may indicate a new burst. Many of burst detection methods struggle with false positives due to non-seasonal water consumption as a result of e.g. environmental, economic or demographic exogenous influences, such as weather, holidays, festivities or pandemics. Finding a robust alternative that reduces the false positive rate of burst detection and does not rely on data from exogenous processes is essential. We present such a burst detection method, based on Bayesian ridge regression and Random Sample Consensus. Our exogenous nowcasting method relies on signals of all nearby flow and pressure sensors in the distribution net with the aim to reduce the false positive rate. The method requires neither data of exogenous processes, nor extensive historical data, but only requires one week of historical data per flow/pressure sensor. The exogenous nowcasting method is compared with a common water demand forecasting method for burst detection and shows sufficiently higher Nash-Sutcliffe model efficiencies of 82.7% - 90.6% compared to 57.9% - 77.7%, respectively. These efficiency ranges indicate a more accurate water demand prediction, resulting in more precise burst detection.

     

Using column generation to solve parallel machine scheduling problems with minmax objective functions

In this paper we consider the parallel machine scheduling problem of minimizing an objective function of the minmax type, like maximum lateness, subject to release dates, deadlines, and/or generalized precedence constraints. We use a destructive strategy to compute a lower bound. Here we test the feasibility of a decision problem by applying column generation to compute a bound on the number of machines that we need to feasibly accommodate all jobs. After having derived the lower bound, we try to find a matching upper bound by identifying a feasible schedule with objective function value equal to this lower bound. Our computational results show that our lower bound is so strong that this is almost always possible. We are able to solve problems with up to 160 jobs and 10 machines in 10 minutes on average.     

Finding a robust assignment of flights to gates at Amsterdam Airport Schiphol

In this paper we investigate the gate assignment problem as it appears at Amsterdam Airport Schiphol (AAS). Currently, the gate planners spend many hours on adjusting the automatically generated planning during the day of operation to make it proof against small deviations from the schedule. To alleviate this problem, we aim at finding a robust solution, given the planned arrivals and departures for the next day. We present a completely new integer linear programming formulation that is based on so-called gate plans. Each gate plan consists of a subset of the flights that can be assigned to a single gate of the corresponding type; gates with identical characteristics are aggregated in gate types. The gate assignment problem then boils down to selecting the best subset of gate plans such that each flight belongs to one selected gate plan, and such that the number of selected gate plans for a certain type of gate is equal to the number of gates of this type. In the first phase, we solve the LP-relaxation through column generation, and we describe specific features to find a very good solution to the ILP quickly. This solution is then handed to the planners at AAS in order to assign gate plans to physical gates. This consists of a number of relatively small problems that can be solved by hand and in which additional operational constraints can be incorporated. We also present the possibility of directly assigning flights to physical gates using the column generation formulation, where we then take into account other criteria as well. Computational results with real-life data provided by AAS are promising and indicate that the algorithm is able to solve real-life instances within rather small running times.     

Quantitative aspects of two methods to dissolve collagen-free muscle proteins from acetone dry powders of Guelders ring sausage

Summary The purpose of this study was to find experimental conditions for the complete solubility of collagen-free muscle proteins (CFMP) using acetone powder of Guelders ring sausage. Preliminary experiments were carried out to choose the best procedure for preparing the acetone dry powder. Two different methods of acetone extraction of minced sausage were compared. The acetone dry mass (ADM) method using continuous extraction in a Soxhlet [2] apparatus gave better results than the acetone powder (ACP) method, which used a blender [1]. The ADM method was used for further investigations. ADM was extracted with two types of sodium dodecyl sulphate (SDS), containing solvents A and B. Solvent A contains a Tris-boric acid buffer (pH 8.2) with 1.5% (m/v) SDS and 0.05% (m/v) dithioerythreitol [3]. Solvent B is a borate-chloric acid buffer (pH 9.0) with 2.0% (m/v) SDS and 1.0% (m/v) mercapto-ethanol [2]. Both solvents showed a linear relationship between the quantities of CFMP in ADM and the dissolved CFMP. The linear relationships were found between quantities of 10.0 and 30.0 mg (solution A) and of 5.0 and 30.0 mg ADM (solution B) per ml solvent. The solubility of CFMP was better in solvent B than in solution A. Completely dissolved CFMP from ADM was only obtained in the case of 5.0 mg ADM in 1.0 ml solution B. These conditions will be used in liquid chromatography experiments, the results of which will be reported later.

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Quantitative Aspekte zweier Verfahren für das Auflösen kollagenfreier Muskelproteine aus acetontrockenen Pulvern der Gelderschen Rauchwurst

Zusammenfassung Der Zweck dieser Untersuchungen ist, die experimentellen Bedingungen für die vollstän-dige Löslichkeit des kollagenfreien Muskelproteins (CFMP) aus dem Acetonpulver der Gelderschen Rauchwurst zu finden. Durch Vorversuche wurde die beste Arbeitsweise für das Zubereiten des Acetonpulvers gewählt. Zwei verschiedene Extraktionsverfahren mit zerkleinertem Wurstmaterial wurden miteinander verglichen. Die Methode mit der Acetontrockenmasse (ADM) mittels kontinuierlicher Extraktion [2] führte zu besseren Ergebnissen als die Methode mit Acetonpulver (ACP), wozu ein Mischgerät [1] verwendet wurde. Die ADM-Methode wurde für weitere Untersuchungen angewendet. ADM wurde mit zwei verschiedenen Extraktionslösungen von Natrium-Dodecylsulfat (SDS) (A und B) extrahiert. Lösung A enthalt einen Tris-Borsäure Puffer (pH 8,2) mit SDS (1,5%) und Dithioerithritol (0,05%) [3]. Die Lösung B enthält einen Borat-Salzsäure Puffer (pH 9,0) mit SDS (2,0%) und Mercapto-Ethanol (1,0%) [2]. Beide Extraktionslösungen zeigen ein lineares Verhalten zwischen den Mengen vom CFMP in ADM und in aufgelöstem CFMP. Diese Linearität wurde von 10,030,0 mg ADM (Lösungsmittel A) und von 5,030,0 mg ADM (Lösungsmittel B) gefunden. Die Lös-lichkeit in Lösung B ist gegeniiber Lösung A besser. Ein vollständig gelöstes CFMP aus ADM wurde nur bei der Extraktion von 5,0 mg ADM in 1,0 ml der Losung B erhalten. Diese Bedingung soll in unseren künftigen flüssigchromatographischen Experimenten verwendet werden.

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Supported by a grant from the Hoofdinspectie Levensmiddelen of the Ministry of Welzijn, Volksgezondheid en Cultuur