Luminal transport through intact endoplasmic reticulum limits the magnitude of localized Ca2+ signals.

The endoplasmic reticulum (ER) forms an interconnected network of tubules stretching throughout the cell. Understanding how ER functionality relies on its structural organization is crucial for elucidating cellular vulnerability to ER perturbations, which have been implicated in several neuronal pathologies. One of the key functions of the ER is enabling Ca[Formula: see text] signaling by storing large quantities of this ion and releasing it into the cytoplasm in a spatiotemporally controlled manner. Through a combination of physical modeling and live-cell imaging, we demonstrate that alterations in ER shape significantly impact its ability to support efficient local Ca[Formula: see text] releases, due to hindered transport of luminal content within the ER. Our model reveals that rapid Ca[Formula: see text] release necessitates mobile luminal buffer proteins with moderate binding strength, moving through a well-connected network of ER tubules. These findings provide insight into the functional advantages of normal ER architecture, emphasizing its importance as a kinetically efficient intracellular Ca[Formula: see text] delivery system.

Colonization, diversity, and seasonality of fishes at pelagic fish aggregating devices.

The pelagic zone of the ocean can be a challenging environment in which to conduct research and as a result we lack the robust baseline abundance and diversity data, compared to what is available in more accessible coastal habitats, to be able to track changes or stressors to the biota in this environment. Many large-scale fisheries target pelagic fish, and much of the information available on these species is based on fisheries-dependent data that may be biased towards hotspots and commercially valuable fishes. Here, a long-term video and visual fish survey was conducted on two subsurface moored fish aggregating devices (FADs) in the pelagic waters of the central Bahamas to determine the feasibility of using moored pelagic FADs as tools for collecting fish abundance and diversity data. A wide range of species was documented, including large migratory fish that are the focus of commercial and recreational fisheries, and smaller often overlooked species on which little abundance or seasonality information exists. We found that FADs colonize quickly and reach a peak stable (albeit seasonally cyclical) abundance and diversity within the first several months after deployment. Species richness was higher in video surveys, but abundance was higher in visual surveys, except for sharks. Our results highlight the need to tailor survey methods to fit the context and study objective, and provide further evidence for the importance of fisheries-independent data in monitoring pelagic species.

Inshore and offshore marine migration pathways of Atlantic salmon post-smolts from multiple rivers in Scotland, England, Northern Ireland, and Ireland.

The migratory behavior of Atlantic salmon (Salmo salar) post-smolts in coastal waters is poorly understood. In this collaborative study, 1914 smolts, from 25 rivers, in four countries were tagged with acoustic transmitters during a single seasonal migration. In total, 1105 post-smolts entered the marine study areas and 438 (39.6%) were detected on a network of 414 marine acoustic receivers and an autonomous underwater vehicle. Migration pathways (defined as the shortest distance between two detections) of up to 575 km and over 100 days at sea were described for all 25 populations. Post-smolts from different rivers, as well as individuals from the same river, used different pathways in coastal waters. Although difficult to generalize to all rivers, at least during the year of this study, no tagged post-smolts from rivers draining into the Irish Sea were detected entering the areas of sea between the Hebrides and mainland Scotland, which is associated with a high density of finfish aquaculture. An important outcome of this study is that a high proportion of post-smolts crossed through multiple legislative jurisdictions and boundaries during their migration. This study provides the basis for spatially explicit assessment of the impact risk of coastal pressures on salmon during their first migration to sea.

Prophylactic immunization to Helicobacter pylori infection using spore vectored vaccines.

BACKGROUND: Helicobacter pylori infection remains a major public health threat leading to gastrointestinal illness and increased risk of gastric cancer. Mostly affecting populations in developing countries no vaccines are yet available and the disease is controlled by antimicrobials which, in turn, are driving the emergence of AMR. MATERIALS AND METHODS: We have engineered spores of Bacillus subtilis to display putative H. pylori protective antigens, urease subunit A (UreA) and subunit B (UreB) on the spore surface. Following oral dosing of mice with these spores, we evaluated immunity and colonization in animals challenged with H. pylori. RESULTS: Oral immunization with spores expressing either UreA or UreB showed antigen-specific mucosal responses (fecal sIgA) including seroconversion and hyperimmunity. Following challenge, colonization by H. pylori was significantly reduced by up to 1-log. CONCLUSIONS: This study demonstrates the utility of bacterial spores for mucosal vaccination to H. pylori infection. The heat stability and robustness of Bacillus spores coupled with their existing use as probiotics make them an attractive solution for either protection against H. pylori infection or potentially for therapy and control of active infection.

2023 International Olympic Committee's (IOC) consensus statement on Relative Energy Deficiency in Sport (REDs).

Relative Energy Deficiency in Sport (REDs) was first introduced in 2014 by the International Olympic Committee's expert writing panel, identifying a syndrome of deleterious health and performance outcomes experienced by female and male athletes exposed to low energy availability (LEA; inadequate energy intake in relation to exercise energy expenditure). Since the 2018 REDs consensus, there have been >170 original research publications advancing the field of REDs science, including emerging data demonstrating the growing role of low carbohydrate availability, further evidence of the interplay between mental health and REDs and more data elucidating the impact of LEA in males. Our knowledge of REDs signs and symptoms has resulted in updated Health and Performance Conceptual Models and the development of a novel Physiological Model. This Physiological Model is designed to demonstrate the complexity of either problematic or adaptable LEA exposure, coupled with individual moderating factors, leading to changes in health and performance outcomes. Guidelines for safe and effective body composition assessment to help prevent REDs are also outlined. A new REDs Clinical Assessment Tool-Version 2 is introduced to facilitate the detection and clinical diagnosis of REDs based on accumulated severity and risk stratification, with associated training and competition recommendations. Prevention and treatment principles of REDs are presented to encourage best practices for sports organisations and clinicians. Finally, methodological best practices for REDs research are outlined to stimulate future high-quality research to address important knowledge gaps.

Investigating the behaviour of Atlantic salmon (Salmo salar L.) post-smolts during their early marine migration through the Clyde Marine Region.

It is thought that survival during migration is particularly poor for Atlantic salmon post-smolts immediately after entry into sea and particularly in the estuarine environment. Nonetheless, there is currently a lack of information on Atlantic salmon post-smolt movement behaviour in estuaries in the UK. This study used acoustic tagging to estimate loss rates and compare the behaviour of Atlantic salmon post-smolts migrating from two distinctly different rivers draining into the Clyde Estuary, the River Endrick (n = 145) and the Gryffe (n = 102). Contrary to most literature, post-smolts undertook rapid migrations through the estuary, potentially decreasing their exposure to predators/anthropogenic stressors and reducing their estimated loss rates (river: 1%-3% km-1 ; estuary: 0.20%-0.60% km-1 ). The low loss rates in the estuary occurred despite post-smolts engaging in passive reversal movements with the tide upon entering the estuary, possibly allowing them more time to adapt to the increased salinity. Atlantic salmon post-smolts from both the rivers used similar migration pathways exiting into the coastal marine zone during ebbing tide. This study provides novel information on the timing and migratory routes of Atlantic salmon post-smolts in the Clyde Estuary that can ultimately be used to inform management decisions on how to assess and reduce the potential impacts of current natural and anthropogenic stressors. Temporal repeatability of this study over multiple years is required to determine if there is variation in the factors driving the migratory patterns and loss rates of smolts in this system.

Notes on age determination, size and age structure, longevity and growth of co-occurring macrourid fishes.

Nineteen species of the deep-water fish family Macrouridae inhabit the Mid-Atlantic Ridge (MAR) of the North Atlantic. Size and age structure, longevity and length at age were explored for seven of the more abundant species: Coryphaenoides armatus, Coryphaenoides brevibarbis, Coryphaenoides carapinus, Coryphaenoides guentheri, Coryphaenoides mediterraneus, Coryphaenoides rupestris and Macrourus berglax. The otoliths from these species had growth increments resembling those accepted as annuli in other deep-water species for which validation experiments have been conducted. Based on the counts of these growth increments, age estimates were derived as the basis for studies of age structure, growth and longevity. All the species appeared to have relatively low growth rates, but the interspecific variation in longevity illustrates that there is a considerable variation within the family and among species inhabiting the same deep-sea environment. Most of the species likely complete full life cycles on the MAR, but it is probable that both the youngest and the largest-oldest individuals of some of the species were not sampled adequately.

Evidence for seasonal cycles in deep-sea fish abundances: A great migration in the deep SE Atlantic?

Animal migrations are of global ecological significance, providing mechanisms for the transport of nutrients and energy between distant locations. In much of the deep sea (>200 m water depth), the export of nutrients from the surface ocean provides a crucial but seasonally variable energy source to seafloor ecosystems. Seasonal faunal migrations have been hypothesized to occur on the deep seafloor as a result, but have not been documented. Here, we analyse a 7.5-year record of photographic data from the Deep-ocean Environmental Long-term Observatory Systems seafloor observatories to determine whether there was evidence of seasonal (intra-annual) migratory behaviours in a deep-sea fish assemblage on the West African margin and, if so, identify potential cues for the behaviour. Our findings demonstrate a correlation between intra-annual changes in demersal fish abundance at 1,400 m depth and satellite-derived estimates of primary production off the coast of Angola. Highest fish abundances were observed in late November with a smaller peak in June, occurring approximately 4 months after corresponding peaks in primary production. Observed changes in fish abundance occurred too rapidly to be explained by recruitment or mortality, and must therefore have a behavioural driver. Given the recurrent patterns observed, and the established importance of bottom-up trophic structuring in deep-sea ecosystems, we hypothesize that a large fraction of the fish assemblage may conduct seasonal migrations in this region, and propose seasonal variability in surface ocean primary production as a plausible cause. Such trophic control could lead to changes in the abundance of fishes across the seafloor by affecting secondary production of prey species and/or carrion availability for example. In summary, we present the first evidence for seasonally recurring patterns in deep-sea demersal fish abundances over a 7-year period, and demonstrate a previously unobserved level of dynamism in the deep sea, potentially mirroring the great migrations so well characterized in terrestrial systems.

As migrações dos animais são importantes para a ecologia global pois fornecem mecanismos para o transporte de nutrientes e energia entre diferentes locais. Em grande parte do oceano profundo (>200 m de profundidade), a exportação de nutrientes da superfície para os ecossistemas do fundo marinho é uma fonte de energia crucial, mas que varia entre estações. Consequentemente, calcula-se que ocorram migrações sazonais de animais no fundo marinho, mas tal nunca foi reportado. Neste estudo, nós analisamos dados fotográficos do observatório do fundo marinho DELOS colhidos ao longo de 7.5 anos, para determinar se existem indícios de comportamentos migratórios sazonais (intra-anuais) na comunidade de peixes de profundidade na costa oeste africana ao largo de Angola e, se se confirmar, tentar identificar o que desencadeia este comportamento. Os resultados obtidos mostram que há uma correlação entre as alterações intra-anuais da densidade de peixes demersais a 1,400 m de profundidade e as estimativas de produção primária obtidas por satélite. A densidade de peixes atinge o seu máximo no final de Novembro, com um pico menos acentuado em Junho, aproximadamente quatro meses após os respectivos picos de produção primária na superfície. As alterações na densidade de peixe ocorreram de uma forma tão rápida que não podem ser explicadas por recrutamento ou mortalidade, e como tal devem só podem ser geradas por uma alteração do comportamento. Dado a recorrência do padrão observado, e importância da estrutura trófica fundo-topo em ecossistemas do oceano profundo, nós colocamos a hipótese de que uma fracção grande da comunidade de peixes faz migrações sazonais nesta região, e propomos que a variação sazonal da produção primária na superfície esteja na sua origem. Este controlo trófico poderá levar a alterações na densidade de peixes no fundo marinho via, por exemplo, a produção secundária de presas e/ou disponibilidade de corpos em processo de decomposição. Resumindo, nós apresentamos aqui a primeira evidência de padrões sazonais recorrentes na densidade de peixes demersais de profundidade ao longo de um período de sete anos, e provamos existir um nível de dinamismo nunca dantes observado no oceano profundo, que poderá espelhar as grandes migrações comummente observadas em sistemas terrestres.

Computer-aided photo-identification of a rare stingray, Megatrygon microps.

A protocol for photo-identification of individual Megatrygon microps has been defined. One hundred and four identification photographs were taken between 2005 and 2019. Spot patterns on the dorsal surface were used to identify individuals. Unique scarring on eight M. microps re-observed provided an independent confirmation of pattern stability of up to 761 days. Previous studies lacked statistical testing used to validate this photo-identification approach. I3 S photo-matching software was used to successfully match images, identifying 69 individuals. A photo-matching software facilitates an open-source platform for identifying individual M. microps, allowing for better population assessments.

A new model study species: high accuracy of discrimination between individual freckled hawkfish (Paracirrhites forsteri) using natural markings.

Variations between distinct natural markings of freckled hawkfish (Paracirrhites forsteri) could allow in situ identification of individuals from underwater photography. Receiver operating characteristic analysis was used to assess the ability of the Interactive Individual Identification System (I3 S) software to assist in discriminating between images of P. forsteri individuals. This study's results show the high discriminant ability of I3 S to differentiate between unlike individuals and identify images of the same individual. The ability to use automatic computer-aided assistance in the study of this species will enable future research to explore behaviour and movements of individuals in the wild.

Endoplasmic reticulum morphology regulation by RTN4 modulates neuronal regeneration by curbing luminal transport.

Cell functions rely on intracellular transport systems distributing bioactive molecules with high spatiotemporal accuracy. The endoplasmic reticulum (ER) tubular network constitutes a system for delivering luminal solutes, including Ca2+, across the cell periphery. How the ER structure enables this nanofluidic transport system is unclear. Here, we show that ER membrane-localized reticulon 4 (RTN4/Nogo) is sufficient to impose neurite outgrowth inhibition in human cortical neurons while acting as an ER morphoregulator. Improving ER transport visualization methodologies combined with optogenetic Ca2+ dynamics imaging and in silico modeling, we observed that ER luminal transport is modulated by ER tubule narrowing and dilation, proportional to the amount of RTN4. Excess RTN4 limited ER luminal transport and Ca2+ release, while RTN4 elimination reversed the effects. The described morphoregulatory effect of RTN4 defines the capacity of the ER for peripheral Ca2+ delivery for physiological releases and thus may constitute a mechanism for controlling the (re)generation of neurites.

Investigating the functional hierarchy of Bacillus megaterium PV361 spore germinant receptors.

Spores of Bacillus megaterium QM B1551 germinate rapidly when exposed to a number of single-trigger germinant compounds, including glucose, proline, leucine, and certain inorganic salts. However, spores of strain PV361, a plasmidless QM B1551 derivative that lacks the GerU germinant receptor (GR) responsible for mediating germination in response to single-trigger compounds, can germinate efficiently when incubated in nutritionally rich media, presumably via activation of additional germinant receptors. In this work, we have identified five chromosomally encoded GRs and attempted to characterize, by mutational analysis, germinant recognition profiles associated with the respective receptors in strain PV361. Of strains engineered with single GR insertion-deletions, only GerK-null spores displayed significant defective germination phenotypes when incubated in 5% (wt/vol) beef extract or plated on rich solid medium. Cumulative decreases in viability were observed in GerK-null spores that also lacked GerA or GerA2, indicating that these GRs, which exerted little effect on spore germination when disrupted individually, have a degree of functionality. Unexpectedly, an efficient germination response to combinations of germinants was restored in GerA(+) spores, which lack all other functional GRs, providing evidence for negative cooperativity between some GRs within the spore. Tetrazolium-based germinative assays conducted with purified spores indicated that these newly characterized B. megaterium GRs are cognate for a wide and chemically diverse range of germinant molecules, but unlike GerU, can only be induced to trigger germination when stimulated by at least two different germinants.

Activity-regulated growth of motoneurons at the neuromuscular junction is mediated by NADPH oxidases.

Neurons respond to changes in the levels of activity they experience in a variety of ways, including structural changes at pre- and postsynaptic terminals. An essential plasticity signal required for such activity-regulated structural adjustments are reactive oxygen species (ROS). To identify sources of activity-regulated ROS required for structural plasticity in vivo we used the Drosophila larval neuromuscular junction as a highly tractable experimental model system. For adjustments of presynaptic motor terminals, we found a requirement for both NADPH oxidases, Nox and dual oxidase (Duox), that are encoded in the Drosophila genome. This contrasts with the postsynaptic dendrites from which Nox is excluded. NADPH oxidases generate ROS to the extracellular space. Here, we show that two aquaporins, Bib and Drip, are necessary ROS conduits in the presynaptic motoneuron for activity regulated, NADPH oxidase dependent changes in presynaptic motoneuron terminal growth. Our data further suggest that different aspects of neuronal activity-regulated structural changes might be regulated by different ROS sources: changes in bouton number require both NADPH oxidases, while activity-regulated changes in the number of active zones might be modulated by other sources of ROS. Overall, our results show NADPH oxidases as important enzymes for mediating activity-regulated plasticity adjustments in neurons.

Characterization of the human serum depletome by label-free shotgun proteomics.

While it is known that immunoaffinity depletion of abundant proteins in serum removes additional proteins beyond those targeted, there has been little characterization of the co-depleted proteins in the high abundant fraction, which we refer to here as the "depletome". We present evidence of co-depletion of non-targeted proteins in human serum using a top-20 immunodepletion column, as shown by label-free liquid chromatography mass spectrometry (LC-MS(E)) profiling. This led to identification of 147 proteins which were specific for this fraction and comprised proteins with functions predominantly in binding and transport of nucleotides, metal ions, carbohydrates and lipids. These results suggest that further studies on this commonly ignored serum fraction may provide new insights into clinical proteomics.

Oxidative stress, inflammation and recovery of muscle function after damaging exercise: effect of 6-week mixed antioxidant supplementation.

There is no consensus regarding the effects of mixed antioxidant vitamin C and/or vitamin E supplementation on oxidative stress responses to exercise and restoration of muscle function. Thirty-eight men were randomly assigned to receive either placebo group (n = 18) or mixed antioxidant (primarily vitamin C & E) supplements (n = 20) in a double-blind manner. After 6 weeks, participants performed 90 min of intermittent shuttle-running. Peak isometric torque of the knee flexors/extensors and range of motion at this joint were determined before and after exercise, with recovery of these variables tracked for up to 168 h post-exercise. Antioxidant supplementation elevated pre-exercise plasma vitamin C (93 ± 8 µmol l(-1)) and vitamin E (11 ± 3 µmol l(-1)) concentrations relative to baseline (P < 0.001) and the placebo group (P ≤ 0.02). Exercise reduced peak isometric torque (i.e. 9-19% relative to baseline; P ≤ 0.001), which persisted for the first 48 h of recovery with no difference between treatment groups. In contrast, changes in the urine concentration of F(2)-isoprostanes responded differently to each treatment (P = 0.04), with a tendency for higher concentrations after 48 h of recovery in the supplemented group (6.2 ± 6.1 vs. 3.7 ± 3.4 ng ml(-1)). Vitamin C & E supplementation also affected serum cortisol concentrations, with an attenuated increase from baseline to the peak values reached after 1 h of recovery compared with the placebo group (P = 0.02) and serum interleukin-6 concentrations were higher after 1 h of recovery in the antioxidant group (11.3 ± 3.4 pg ml(-1)) than the placebo group (6.2 ± 3.8 pg ml(-1); P = 0.05). Combined vitamin C & E supplementation neither reduced markers of oxidative stress or inflammation nor did it facilitate recovery of muscle function after exercise-induced muscle damage.

Reactive Oxygen Species Mediate Activity-Regulated Dendritic Plasticity Through NADPH Oxidase and Aquaporin Regulation.

Neurons utilize plasticity of dendritic arbors as part of a larger suite of adaptive plasticity mechanisms. This explicitly manifests with motoneurons in the Drosophila embryo and larva, where dendritic arbors are exclusively postsynaptic and are used as homeostatic devices, compensating for changes in synaptic input through adapting their growth and connectivity. We recently identified reactive oxygen species (ROS) as novel plasticity signals instrumental in this form of dendritic adjustment. ROS correlate with levels of neuronal activity and negatively regulate dendritic arbor size. Here, we investigated NADPH oxidases as potential sources of such activity-regulated ROS and implicate Dual Oxidase (but not Nox), which generates hydrogen peroxide extracellularly. We further show that the aquaporins Bib and Drip, but not Prip, are required for activity-regulated ROS-mediated adjustments of dendritic arbor size in motoneurons. These results suggest a model whereby neuronal activity leads to activation of the NADPH oxidase Dual Oxidase, which generates hydrogen peroxide at the extracellular face; aquaporins might then act as conduits that are necessary for these extracellular ROS to be channeled back into the cell where they negatively regulate dendritic arbor size.

Otolith chemoscape analysis in whiting links fishing grounds to nursery areas.

Understanding life stage connectivity is essential to define appropriate spatial scales for fisheries management and develop effective strategies to reduce undersized bycatch. Despite many studies of population structure and connectivity in marine fish, most management units do not reflect biological populations and protection is rarely given to juvenile sources of the fished stock. Direct, quantitative estimates that link specific fishing grounds to the nursery areas, which produced the caught fish are essential to meet these objectives. Here we develop a continuous-surface otolith microchemistry approach to geolocate whiting (Merlangius merlangus) and infer life stage connectivity across the west coast of the UK. We show substantial connectivity across existing stock boundaries and identify the importance of the Firth of Clyde nursery area. This approach offers fisheries managers the ability to account for the benefits of improved fishing yields derived from spatial protection while minimising revenue loss.

The MacKinnon Lists Technique: An efficient new method for rapidly assessing biodiversity and species abundance ranks in the marine environment.

Widespread and ever-increasing anthropogenic impacts in the marine environment are driving a need to develop more efficient survey methods for monitoring changes in marine biodiversity. There is a particular urgent need for survey methods that could more rapidly and effectively detect change in species richness, abundance and community composition. Here, test the suitability of the Mackinnon Lists Technique for use in the marine environment by testing its effectiveness for rapid assessment of fish communities. The MacKinnon Lists Technique is a time-efficient and cost-effective sampling method developed for studying avian tropical biodiversity, in which several list samples of species can be collected from a single survey. Using the well-established MaxN approach on data from deployments of a Baited Remote Underwater Video Systems for comparison, we tested the suitability of the MacKinnon Lists Technique for use in marine environments by analysing tropical reef fish communities. Using both methods for each data set, differences in community composition between depths and levels of protection were assessed. Both methods were comparable for diversity and evenness indices with similar ranks for species. Multivariate analysis showed that the MacKinnon Lists Technique and MaxN detected similar differences in community composition at different depths and protection status. However, the MacKinnon Lists Technique detected significant differences between factors when fewer videos (representing reduced survey effort) were used. We conclude that the MacKinnon Lists Technique is at least as effective as the widely used MaxN method for detecting differences between communities in the marine environment and suggest can do so with lower survey effort. The MacKinnon Lists Technique has the potential to be widely used as an effective new tool for rapid conservation monitoring in marine ecosystems.

Substrate-selective supramolecular tandem assays: monitoring enzyme inhibition of arginase and diamine oxidase by fluorescent dye displacement from calixarene and cucurbituril macrocycles.

A combination of moderately selective host-guest binding with the impressive specificity of enzymatic transformations allows the real-time monitoring of enzymatic reactions in a homogeneous solution. The resulting enzyme assays ("supramolecular tandem assays") exploit the dynamic binding of a fluorescent dye with a macrocyclic host in competition with the binding of the substrate and product. Two examples of enzymatic reactions were investigated: the hydrolysis of arginine to ornithine catalyzed by arginase and the oxidation of cadaverine to 5-aminopentanal by diamine oxidase, in which the substrates have a higher affinity to the macrocycle than the products ("substrate-selective assays"). The depletion of the substrate allows the fluorescent dye to enter the macrocycle in the course of the enzymatic reaction, which leads to the desired fluorescence response. For arginase, p-sulfonatocalix[4]arene was used as the macrocycle, which displayed binding constants of 6400 M(-1) with arginine, 550 M(-1) with ornithine, and 60,000 M(-1) with the selected fluorescent dye (1-aminomethyl-2,3-diazabicyclo[2.2.2]oct-2-ene); the dye shows a weaker fluorescence in its complexed state, which leads to a switch-off fluorescence response in the course of the enzymatic reaction. For diamine oxidase, cucurbit[7]uril (CB7) was used as the macrocycle, which showed binding constants of 4.5 x 10(6) M(-1) with cadaverine, 1.1 x 10(5) M(-1) with 1-aminopentane (as a model for the thermally unstable 1-aminopentanal), and 2.9 x 10(5) M(-1) with the selected fluorescent dye (acridine orange, AO); AO shows a stronger fluorescence in its complexed state, which leads to a switch-on fluorescence response upon enzymatic oxidation. It is demonstrated that tandem assays can be successfully used to probe the inhibition of enzymes. Inhibition constants were estimated for the addition of known inhibitors, i.e., S-(2-boronoethyl)-L-cysteine and 2(S)-amino-6-boronohexanoic acid for arginase and potassium cyanide for diamine oxidase. Through the sequential coupling of a "product-selective" with a "substrate-selective" assay it was furthermore possible to monitor a multistep biochemical pathway, namely the decarboxylation of lysine to cadaverine by lysine decarboxylase followed by the oxidation of cadaverine by diamine oxidase. This "domino tandem assay" was performed in the same solution with a single reporter pair (CB7/AO).