Hydrolysis of dansyl-peptide substrates by leucine aminopeptidase: origin of dansyl fluorescence changes during hydrolysis

The origin of the fluorescence changes observed in stopped-flow experiments of the hydrolysis of three 5-(dimethylamino)naphthalene-1-sulfonyl-(dansyl) peptide substrates by porcine kidney cytosol leucine aminopeptidase has been investigated. The substrates used all have the potential to accept energy from aromatic residues of the enzyme via resonance energy transfer when they are bound as enzyme-substrate complexes, indicating that fluorescence changes due to the buildup and decay of such intermediates are possible. However, the fluorescence of these substrates differs from that of the products, and direct excitation of their dansyl groups during hydrolysis can also be responsible for the observed fluorescence changes due to changes in the concentrations of free substrate and product. The dansyl fluorescence changes observed with excitation wavelengths near 280 nm are not accompanied by quenching of the enzyme fluorescence, as would be expected if there were enzyme-to-substrate energy transfer. The magnitude of the maximal fluorescence change at a fixed concentration of substrate is also independent of the enzyme concentration. Furthermore, the excitation profile for the fluorescence changes shows that they arise from direct excitation of the dansyl group. Thus, there is no energy transfer in these reactions, and the fluorescence changes observed arise from direct excitation of the dansyl group and reflect the instantaneous concentration of substrate. This behavior contrasts sharply with that for the reaction of carboxypeptidase A with dansyl-Gly-Tyr, which has been studied as a positive control for an energy-transfer system.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of stream-crossing by a pipeline on the benthic macroinvertebrate communities of a small mountain stream

Aquatic environmental impact associated with stream-crossing by a pipeline was monitored at Archibald Creek, B.C. for two years. Water chemistry and benthic macroinvertebrates were used as monitoring tools. Results indicated that impacts arising from stream-crossing were short-term and non-residual.Funded by a contract from Canadian Arctic Gas Study Limited, calgary, to Aquatic Environments Limited, Calgary.Aquatic Environments Limited, Calgary.Aquatic Environments Limited, Calgary.     

Über das Invertierungsvermögen der Speicheldrüsen und des Mitteldarmes von Bienen verschiedenen Alters

Zusammenfassung Die gewonnenen Ergebnisse zeigen mit völliger Klarheit, daß die Schlunddrüse ein stark ausgesprochenes Invertierungsvermögen besitzt, das sich jedoch in hohem Grade, je nach dem Alter der Bienen ändert. Im Gegensatz zum Mitteldarm, der Invertase auszuscheiden beginnt von dem Augenblicke des Ausschlüpfens der jungen Biene, bedarf die Schlunddrüse eines gewissen Zeitraumes zu ihrer Entwicklung. Die von uns angestellten Versuche können indessen die Frage noch nicht endgültig entscheiden, in welchem Moment die Schlunddrüse zu funktionieren beginnt. Wir haben die Sekretionstätigkeit der Drüse an verhältnismäßig alten Bienen — von 10 Tagen an — beobachtet und die maximalen Ziffern der Zuckerinversion bei 30tägigen Bienen ermittelt. Hieraus lassen sich noch keine verallgemeinernden Schlüsse ziehen. Unsere Versuche begannen im August und dauerten bis Mitte September, fanden also am Ende der Saison statt. Das Versuchsvolk war schwach und die Zahl der jungen Bienen und der Brut gering (auf einem Rahmen). Als Beleg dazu genügt der Hinweis, daß die Bienen ihre Orientierungsausflüge im Alter von 18 Tagen unternahmen. Alle diese Faktoren könnten zweifellos das Tempo der Entwicklung des Organismus der Biene — und der Sehlunddrüse im besonderen — beeinflussen. Es ist sehr wahrscheinlich, daß eine Untersuchung über die Veränderung des Invertierungsvermögens der Schiunddrüsen von Bienen, die zu verschiedenen Zeiten der Frühlings- und Sommersaison ausgeschlüpft waren, auch eine verschiedene Schnelligkeit der Entwicklung der Drüse ergeben würde.Was die Brustdrüse anbetrifft, so geben unsere Versuche kein genügendes Material zur Beurteilung, welche Rolle sie im Kohlehydratstoffwechsel der Biene spielt. Nur eines steht fest, daß sie an der Invertierung des Zuckers in keinem Falle beteiligt ist.Der Mitteldarm scheint auch mit dem Alter der Biene sein Invertierungsvermögen zu steigern, wenn auch nicht in solchem Umfange als die Schlunddrüse.Die von uns gewonnenen Resultate erlauben, einige Schlüsse über mehrere Fragen der Bienenbiologie zu ziehen, wie z. B. die Frage über den Anteil der Schlunddrüse an der Futtersaftbereitung usw. Doch müssen solche spezielle Fragen an einer anderen Stelle ausführlich besprochen werden.Zum Schlusse sprechen wir Dr. Resnitschenko sowie Dr. Alpatov für die freundliche Hilfe bei der Ausführung der Arbeit unseren Dank aus.     

Effect of Phosphate on the Corrosion of Carbon Steel and on the Composition of Corrosion Products in Two-Stage Continuous Cultures of Desulfovibrio desulfuricans

A field isolate of Desulfovibrio desulfuricans was grown in defined medium in a two-stage continuous culture apparatus with different concentrations of phosphate in the feed medium. The first state (V1) was operated as a conventional chemostat (D = 0.045 h⁻¹) that was limited in energy source (lactate) or phosphate. The second stage (V2) received effluent from V1 but no additional nutrients, and contained a healthy population of transiently starved or resting cells. An increase in the concentration of phosphate in the medium fed to V1 resulted in increased corrosion rates of carbon steel in both V1 and V2. Despite the more rapid corrosion observed in growing cultures relative to that in resting cultures, corrosion products that were isolated under strictly anaerobic conditions from the two culture modes had similar bulk compositions which varied with the phosphate content of the medium. Crystalline mackinawite (Fe₉S₈), vivianite [Fe₃(PO₄)₂ · 8H₂O], and goethite [FeO(OH)] were detected in amounts which varied with the culture conditions. Chemical analyses indicated that the S in the corrosion product was almost exclusively in the form of sulfides, while the P was present both as phosphate and as unidentified components, possibly reduced P species. Some differential localization of S and P was observed in intact corrosion products. Cells from lactate-limited, but not from phosphate-limited, cultures contained intracellular granules that were enriched in P and Fe. The results are discussed in terms of several proposed mechanisms of microbiologically influenced corrosion.     

Staff Nurses’ Perceptions and Experiences about Structural Empowerment: A Qualitative Phenomenological Study

The aim of the study reported in this article was to investigate staff nurses’ perceptions and experiences about structural empowerment and perceptions regarding the extent to which structural empowerment supports safe quality patient care. To address the complex needs of patients, staff nurse involvement in clinical and organizational decision-making processes within interdisciplinary care settings is crucial. A qualitative study was conducted using individual semi-structured interviews of 11 staff nurses assigned to medical or surgical units in a 600-bed university hospital in Belgium. During the study period, the hospital was going through an organizational transformation process to move from a classic hierarchical and departmental organizational structure to one that was flat and interdisciplinary. Staff nurses reported experiencing structural empowerment and they were willing to be involved in decision-making processes primarily about patient care within the context of their practice unit. However, participants were not always fully aware of the challenges and the effect of empowerment on their daily practice, the quality of care and patient safety. Ongoing hospital change initiatives supported staff nurses’ involvement in decision-making processes for certain matters but for some decisions, a classic hierarchical and departmental process still remained. Nurses perceived relatively high work demands and at times viewed empowerment as presenting additional. Staff nurses recognized the opportunities structural empowerment provided within their daily practice. Nurse managers and unit climate were seen as crucial for success while lack of time and perceived work demands were viewed as barriers to empowerment.