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Activated sludge (AS) is a biological treatment process that is employed in wastewater treatment plants. Filamentous bacteria in AS plays an important role in the settling ability of the sludge. Proper settling of the sludge is essential for normal functionality of the wastewater plants, where filamentous bulking is always a persistent problem preventing sludge from settling. The performance of AS plants is conventionally monitored by physico-chemical procedures. An alternative way of monitoring the AS in wastewater treatment process is to use image processing and analysis. Good performance of the image segmentation algorithms is important to quantify flocs and filaments in AS. In this article, an algorithm is proposed to perform segmentation of filaments in the phase contrast images using phase stretch transform. Different values of strength (S) and warp (W) are tested to obtain optimum segmentation results and decrease the halo and shade-off artefacts encountered in phase contrast microscopy. The performance of the algorithm is assessed using DICE coefficient, accuracy, false positive rate (FPR), false negative rate (FNR) and Rand index (RI). Sixty-one gold approximations of ground truth images were manually prepared to assess the segmentation results. Thirty-two of them were acquired at 10× magnification and 29 of them were acquired at 20× magnification. The proposed algorithm exhibits better segmentation performance with an average DICE coefficient equal to 52.25%, accuracy 99.74%, FNR 41.8% and FPR 0.14% and RI 99.49%, based on 61 images., (© The Author(s) 2018. Published by Oxford University Press on behalf of The Japanese Society of Microscopy. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Image processing and analysis is an effective tool for monitoring and fault diagnosis of activated sludge (AS) wastewater treatment plants. The AS image comprise of flocs (microbial aggregates) and filamentous bacteria. In this paper, nine different approaches are proposed for image segmentation of phase-contrast microscopic (PCM) images of AS samples. The proposed strategies are assessed for their effectiveness from the perspective of microscopic artifacts associated with PCM. The first approach uses an algorithm that is based on the idea that different color space representation of images other than red-green-blue may have better contrast. The second uses an edge detection approach. The third strategy, employs a clustering algorithm for the segmentation and the fourth applies local adaptive thresholding. The fifth technique is based on texture-based segmentation and the sixth uses watershed algorithm. The seventh adopts a split-and-merge approach. The eighth employs Kittler's thresholding. Finally, the ninth uses a top-hat and bottom-hat filtering-based technique. The approaches are assessed, and analyzed critically with reference to the artifacts of PCM. Gold approximations of ground truth images are prepared to assess the segmentations. Overall, the edge detection-based approach exhibits the best results in terms of accuracy, and the texture-based algorithm in terms of false negative ratio. The respective scenarios are explained for suitability of edge detection and texture-based algorithms.

The highly conserved G-M-N motif of the CorA-Mrs2-Alr1 family of Mg(2+) channels has been shown to be essential for Mg(2+) transport. We performed random mutagenesis of the G-M-N sequence of Saccharomyces cerevisiae Mrs2p in an unbiased genetic screen. A large number of mutants still capable of Mg(2+) influx, albeit below the wild-type level, were generated. Growth complementation assays, performed in media supplemented with Ca(2+) or Co(2+) or Mn(2+) or Zn(2+) at varying concentrations, lead to identification of mutants with reduced growth in the presence of Mn(2+) and Zn(2+). We hereby conclude that (1) at least two, but predominantly all three amino acids of the G-M-N motif must be replaced by certain combinations of other amino acids to remain functional, (2) replacement of any single amino acid within the G-M-N motif always impairs the function of Mrs2p, and (3) we show that the G-M-N motif determines ion selectivity, likely in concurrence with the negatively charged loop at the entrance of the channel thereby forming the Mrs2p selectivity filter.

Porphyromonas gingivalis, a periodontal pathogen, has the ability to lyse erythrocytes. The hemolytic activity of P. gingivalis A7436 was purified as a 45-kDa protein from the culture supernatant of a 3-days old culture using nickel-nitrilotriacetic acid chromatography. Erythrocytes treated with purified P. gingivalis hemolysin showed the presence of pores and extracellular debris by scanning electron microscopy. Active immunization of mice with 15 micrograms hemolysin induced neutralizing antibodies to hemolysin. Heating at 60 degrees C and treatment with trypsin and dithiothreitol abolished hemolytic activity, while incubation with the protease inhibitor Na-p-tosyl-L-lysine chloromethyl ketone caused no effect. We report here for the first time purification of a hemolysin from P. gingivalis A7436. The amino acid sequence of an internal peptide of hemolysin showed sequence similarity with fimbrillin from P. gingivalis HG564. However, the amino acid composition of purified hemolysin was different from that of P. gingivalis fimbrillin. Also, the ability to lyse but not agglutinate erythrocytes and to bind to nickel-nitrilotriacetic acid differentiates P. gingivalis hemolysin from fimbrillin.

A simple method designed to allow coagulation studies in heparinized blood was evaluated. Citrated plasma containing heparin was incubated with an anion-exchange resin commercially available in tablet form (Heparsorb). After centrifugation of the mixture, coagulation studies were carried out on the supernatant plasma. One heparin neutralizer (HN) tablet was capable of removing as much as 43 units of heparin from 1 ml plasma within 10 min of incubation. When normal plasma was exposed to the heparin neutralizer, little or no change in the activities of clotting factors XII, XI, VIII, VII, X, V, and II was observed. However, the resin caused a substantial (44%) loss of factor IX-activity from normal plasma. This loss of factor IX-activity was not observed with plasma from patients receiving coumadin therapy. The results of coagulation tests (aPTT, PT, TT) and determination of fibrinogen/fibrin degradation products performed with HN-treated plasmas from nine patients receiving heparin therapy for thromboembolic disease and seven patients undergoing cardiopulmonary bypass operations were virtually identical to those obtained for the same persons before heparin was administered. The method, which is suitable for the routine clinical laboratory, may be useful in the hemostatic evaluation of critically ill patients who experience bleeding complications while receiving heparin therapy, in the laboratory control of coumadin therapy during the heparin-coumadin overlap period, and in the rapid identification of heparin contamination of blood specimens as a cause of an unexplained aPTT prolongation.