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BackgroundMyokines, released from the contracting muscle, enable communication between the working muscles and other tissue. Their release during physical exercise is assumed to depend on mode, duration, and intensity.ObjectiveThe aim is to examine the acute changes in circulating levels of the myokines IL-6, IL-10, and IL-1ra induced by a bout of resistance exercise and to consider potential moderators of the results.MethodsSystematic literature search was conducted for resistance exercise intervention studies measuring IL-6, IL-10, or IL-1ra before and immediately after resistance exercise in healthy individuals. Random effects meta-analysis was performed for each myokine.ResultsA small to moderate positive effect of resistance exercise for IL-6 and a moderate to large positive effect for IL-1ra were detected. This could not be shown for IL-10. No moderators of the results were detected.ConclusionThis systematic review and meta-analysis clearly showed the immediate positive effects of an acute resistance exercise session on IL-6 and IL-1ra levels.

Recovery from coronavirus disease 2019 (COVID-19) can be impaired by the persistence of symptoms or new-onset health complications, commonly referred to as Long COVID. In a subset of patients, Long COVID is associated with immune system perturbations of unknown etiology, which could be related to compromised immunoregulatory mechanisms.The objective of this scoping review was to summarize the existing literature regarding the frequency and functionality of Tregs in convalescent COVID-19 patients and to explore indications for their potential involvement in the development of Long COVID.A systematic search of studies investigating Tregs during COVID-19 convalescence was conducted on MEDLINE (The literature search yielded 17 relevant studies, of which three included a distinct cohort of patients with Long COVID. The reviewed studies suggest that the Treg population of COVID-19 patients can reconstitute quantitatively and functionally during recovery. However, the comparison between recovered and seronegative controls revealed that an infection-induced dysregulation of the Treg compartment can be sustained for at least several months. The small number of studies investigating Tregs in Long COVID allowed no firm conclusions to be drawn about their involvement in the syndrome's etiology. Yet, even almost one year post-infection Long COVID patients exhibit significantly altered proportions of Tregs within the CD4+ T cell population.Persistent alterations in cell frequency in Long COVID patients indicate that Treg dysregulation might be linked to immune system-associated sequelae. Future studies should aim to address the association of Treg adaptations with different symptom clusters and blood parameters beyond the sole quantification of cell frequencies while adhering to consensualized phenotyping strategies.

BackgroundRecovery from coronavirus disease 2019 (COVID-19) can be impaired by the persistence of symptoms or new-onset health complications, commonly referred to as Long COVID. In a subset of patients, Long COVID is associated with immune system perturbations of unknown etiology, which could be related to compromised immunoregulatory mechanisms.ObjectiveThe aim of this scoping review was to investigate if regulatory T cell (Treg) dysregulation is observable beyond the acute illness and if it might be involved in Long COVID immunopathology.DesignA systematic search of studies investigating Tregs during COVID-19 convalescence was conducted on MEDLINE (via Pubmed) and Web of Science.ResultsThe literature search yielded 17 relevant studies, of which three included a distinct cohort of patients with Long COVID. The reviewed studies suggest that the Treg population of COVID-19 patients can reconstitute quantitatively and functionally during recovery. However, the comparison between recovered and seronegative controls revealed that an infection-induced dysregulation of the Treg compartment can be sustained for at least several months. The small number of studies investigating Tregs in Long COVID allowed no firm conclusions to be drawn about their involvement in the syndrome’s etiology. Yet, even almost one year post-infection Long COVID patients exhibit significantly altered proportions of Tregs within the CD4+ T cell population.ConclusionsPersistent alterations in cell frequency in Long COVID patients indicate that Treg dysregulation might be linked to immune system-associated sequelae. Future studies should aim to address the association of Treg adaptations with different symptom clusters and blood parameters beyond the sole quantification of cell frequencies while adhering to consensualized phenotyping strategies.

IntroductionAcute kidney injury (AKI) is associated with significant morbidity and mortality. The diagnosis is currently based on urine output and serum creatinine and there is a lack of biomarkers that directly reflect tubular damage. Here, we establish flow cytometric quantification of renal epithelial cells as a potential biomarker for quantifying the severity of tubular kidney damage and for predicting AKI outcome.MethodsA total of 84 patients with AKI were included in this study, divided into an exploratory cohort (n=21) and confirmatory cohort (n=63), as well as 25 controls. Urine of patients was collected and processed within 72 hours after AKI onset. Different urinary tubular epithelial cell (TEC) populations were identified and quantified by flow cytometry (FACS). Urinary cell counts were analyzed regarding AKI severity defined by KDIGO stage as well as renal recovery, length of hospital stay and occurrence of MAKE-30 events.ResultsUrinary TEC counts correlated with stages of AKI based on KDIGO classification and were significantly enriched in patients with AKI compared to healthy donors and inpatient controls in both cohorts. Furthermore, both proximal and distal TEC (pTEC, dTEC) counts performed well in identification of patients with AKI regardless of stage. Urinary amounts of pTEC and dTEC showed a strong correlation, with predominance of dTEC. Higher numbers of TEC were associated with extended length of hospital stay, while elevated pTEC counts were associated with the occurrence of MAKE-30 events. Follow-up measurements showed decreasing amounts of urinary TEC after AKI recovery over several days.ConclusionThe amount of urinary TEC directly reflects severity of tissue damage in human AKI. Our protocol furthermore provides a basis for a deeper phenotypic analysis of urinary TEC populations.

Recently, mass cytometry has enabled quantification of up to 50 parameters for millions of cells per sample. It remains a challenge to analyze such high-dimensional data to exploit the richness of the inherent information, even though many valuable new analysis tools have already been developed. We propose a novel algorithm “pattern recognition of immune cells (PRI)” to tackle these high-dimensional protein combinations in the data. PRI is a tool for the analysis and visualization of cytometry data based on a three or more-parametric binning approach, feature engineering of bin properties of multivariate cell data, and a pseudo-multiparametric visualization. Using a publicly available mass cytometry dataset, we proved that reproducible feature engineering and intuitive understanding of the generated bin plots are helpful hallmarks for re-analysis with PRI. In the CD4+T cell population analyzed, PRI revealed two bin-plot patterns (CD90/CD44/CD86 and CD90/CD44/CD27) and 20 bin plot features for threshold-independent classification of mice concerning ineffective and effective tumor treatment. In addition, PRI mapped cell subsets regarding co-expression of the proliferation marker Ki67 with two major transcription factors and further delineated a specific Th1 cell subset. All these results demonstrate the added insights that can be obtained using the non-cluster-based tool PRI for re-analyses of high-dimensional cytometric data.

Background Having destinations within walking distance can encourage older people to walk. Yet, not all destinations may be equally important. Little is known about the types of destinations fostering older adults’ walking for transport in small and medium-sized towns and rural communities. The aim of this study was to explore the associations between the availability of different destinations and walking for transport among older adults living in communities with less than 100,000 inhabitants. Methods Between May and September 2019, self-reported data from 2242 older adults (≥ 65 years) living in the Metropolitan Region Northwest (Germany) were collected within the project AFOOT – Securing urban mobility of an aging population. Data from 2137 study participants were eligible for this analysis. Logistic regression models were used to investigate the relationship between the perceived destination availability of 19 different destinations within a 20-min or 10-min walk from home, respectively, and the engagement in walking for transport. Crude and adjusted models were run separately for each destination and distance category. Exploratory subgroup analyses examined the associations between the availability of destinations within a 20-min walk from home and walking for transport stratified by gender, use of a walking aid, and car availability. Results The availability of each of the investigated destinations within a 20-min walk and of nearly all of these destinations within a 10-min walk from home was significantly positively associated with walking for transport in crude models. Most associations remained significant after adjustment for covariates. The strongest associations were found for the availability of small stores, pharmacy, and bakery. The availability of a bus stop showed the weakest associations and was not significantly associated with walking for transport after adjustment for covariates. Conclusions The provision of local amenities within walking distance may be a promising approach to foster older adults’ walking for transport in smaller communities with less than 100,000 inhabitants and to enable active and healthy aging in place. Further quantitative and qualitative research is needed to validate these findings and to better understand older adults’ walking behavior.

Der Beitrag von Sabine Baumgart und Andrea Rudiger reflektiert die Abhangigkeit der Stadtgrose in Bezug auf strategische Entwicklungsansatze im Rahmen des Stadtumbaus in Klein- und Mittelstadten in Nordrhein-Westfalen. Denn es zeigt sich, dass die Problemlagen, die strategischen Losungsansatze und auch die Operationalisierung sich in manchen Bereichen von denen groserer Stadte unterscheiden.

The effects of projects, plans or programmes on human health are currently often dealt with unsatisfactorily in terms of the indicators and methods used in current German environmental assessments ...

Antibody conjugates applicable in both conventional flow and mass cytometry would offer interesting options for cross-platform comparison, as well as the enrichment of rare target cells by conventional flow cytometry (FC) sorting prior to deep phenotyping by mass cytometry (MC). Here, we introduce a simple method to generate dual fluorochrome/metal-labelled antibodies by consecutive orthogonal labelling. First, we compared different fluorochrome-conjugated antibodies specific for CD4, such as FITC, Vio667, VioGreen or VioBlue for their compatibility with the conventional secondary MAXPAR® labelling protocol. After labelling with 141 Pr, the fluorescence emission spectra of all fluorochromes investigated retained their characteristics, and CD4 dual conjugates (DCs) provided consistent results in immune phenotyping assays performed by FC and MC. The phenotypical composition of CD4+ T-cells was maintained after enrichment by FC sorting using different CD4 DCs. Finally, magnetic cell depletion was combined with FC sorting using CD19-VioBlue-142 Nd, CD20-VioGreen-147 Sm, CD27-Cy5-167 Er and CD38-Alexa488-143 Nd DC to enrich rare human plasmablasts to purities >80%, which allowed a subsequent deep phenotyping by MC. In conclusion, DCs have been successfully established for direct assay comparison between FC and MC, and help to minimise MC data acquisition time for deep phenotyping of rare cell subsets.

Maintenance of physical activity and mobility is a crucial factor for healthy ageing and ageing in place. The built environment can either promote or hinder active transportation, and also affect social participation and independence. Although public health researchers have been examining this relationship for years, in Germany it is neither a commonly addressed topic of public health authorities nor explicitly in the focus of urban planning departments. The AFOOT project pursues an inter- and transdisciplinary approach to identify entry points for health and equity assessment in urban planning procedures, with a particular focus on small- and medium-sized towns. The final goal is to develop a guide for intersectoral policy action. The project methodology includes document analysis, interviewing of experts, workshops and the simulation of implementation by role-playing games with local stakeholders. Against the background of the ‘Health in All Policies’ strategy, encouragement of a transdisciplina...

This chapter aims to overcome binary descriptions of transport systems in Nairobi and Dar es Salaam by framing the relations of existing local transport provision as “heterarchies”. Drawing on the case of Jakarta, Doreen Lee describes how “traffic provides the temporal infrastructure that governs the flow of living and leisure, and the patterning of individual desires and struggles by creating a sensation of ‘being traffic’”. The diversity of actors and stakeholders in the transport provision of informal and decentral modes of transport in combination with the variety of formal regulatory institutions, leads to a variety of aims, business models, passenger bases, fare systems, employment, and standards in both locations. The aims regarding improving the reach, accessibility, and flexibility of the transport system in Nairobi and Dar es Salaam serve to justify a reorganization of the current heterarchic organization of the transport sector and to introduce mass rapid transit systems” systems.

In the last decades, significant efforts have been made to investigate possible cytotoxic effects of metallic nanoparticles (NPs). Methodologies enabling precise information regarding uptake and intracellular distribution of NPs at the single cell level remain to be established. Mass cytometry (MC) has been developed for high-dimensional single cell analyses and is a promising tool to quantify NP-cell interactions. Here, we aim to establish a new MC-based quantification procedure to receive absolute numbers of NPs per single cell by using a calibration that considers the specific transmission efficiency (TE) of suspended NPs. The current MC-quantification strategy accept TE values of complementary metal solutions. In this study, we demonstrate the different transmission behavior of 50 nm silver NPs (AgNP) and silver nitrate solution. We have used identical AgNPs for calibration as for in vitro-differentiated macrophages (THP-1 cell line) in a time- and dose-dependent manner. Our quantification relies on silver intensities measuring AgNPs in the same detection mode as the cells. Results were comparable with the TE quantification strategy using AgNPs but differed when using ionic silver. Furthermore, intact and digested cell aliquots were measured to investigate the impact of MC sample processing on the amount of AgNPs/cell. Taken together, we have provided a MC-specific calibration procedure to precisely calculate absolute numbers of NPs per single cell. Combined with its unique feature of multiplexing up to 50 parameters, MC provides much more information on the single cell level than single cell-inductively coupled plasma mass spectrometry (SC-ICPMS) and, therefore, offers new opportunities in nanotoxicology.

Mass cytometry is increasingly employed in larger immune profiling studies involving data acquisitions across several days and multiple sites. For gaining a maximum of information from respective data by computational analyses, several techniques have been developed to minimize noise in mass cytometric data sets, such as sample banking, standardized instrument setup, sample barcoding, and signal normalization. However, the repeated preparation of cocktails composed of isotope-tagged antibodies remained a significant source of error. We here show that premixed antibody cocktails fail to deliver expected staining patterns when stored at 4°C for 4 weeks. As a solution, we developed and tested a cryopreservation method for highly multiplexed antibody cocktails for mass cytometry including lanthanide, palladium, and platinum conjugates that yielded stable staining patterns for at least 9 months when stored at temperatures below -80°C. Using frozen aliquots of antibody cocktails is an economic and flexible approach to significantly improve data consistency in large mass cytometry studies with repetitive staining/measurement cycles spanning several days or involving multiple data acquisition sites. © 2019 International Society for Advancement of Cytometry.

Polystyrene beads are broadly applied in flow cytometry. Implementing bead-based assays in mass cytometry is desired but hampered by the lack of an elemental label required for their detection. In this study, we introduce stable osmium tetroxide labeling as a universal approach for generating functionalized beads readily detectable by mass cytometry. We demonstrate the utility of osmium-labeled beads for signal spillover compensation in mass cytometry, and, strikingly, their application in quantitative Ab-binding capacity assays combined with high-dimensional profiling of human PBMC enabled the systematic assessment of receptor expression profiles across large numbers of cellular phenotypes. This analysis confirmed increased monocytic Siglec-1 expression in active systemic lupus erythematosus patients and, additionally, revealed interrelated reductions of CD4 expression by regulatory and memory CD4 T cells and HLA-DR expression by myeloid dendritic cells, pointing toward defective cross-talk at the immunological synapse that may limit immune responses in systemic lupus erythematosus. By converting conventional flow cytometry beads into beads suitable for mass cytometry, our approach paves the way toward the broad implementation of bead-based assays in high-dimensional cell profiling studies by mass cytometry in biomedical research.

Keywords: mass cytometry; panel design; mass-response characteristic; human blood leukocytes; immune phenotyping; immune monitoring; autoimmunity; rheumatism; systemic lupus erythematosus (SLE); biomarker and clinical trial

Mass cytometry has pioneered >40-parameter single-cell analyses that allow for the characterization of complex cellular networks at unprecedented depth. Up to 135 parameters can be simultaneously detected, but limited availability of metal tags suitable for labeling of specific probes prevents optimal exploitation of the analytical capacity of mass cytometers. To this end, we here establish the application of elemental silver nanoparticles (AgNP) of different size for reporting cell surface antigens on human leukocytes in mass cytometry assays. The mass channels at 107 Da and 109 Da are uniquely occupied by silver isotopes and do not interfere with other mass cytometry reagents. Streptavidin-coated AgNP (SA-AgNP) facilitated distinct and specific detection of various antigens, such as CD8, CD244 and CD294 on peripheral blood leukocytes pre-incubated with respective biotinylated primary antibodies. Signal intensities elicited by 40 nm-sized AgNP allowed specific detection of the low abundance antigen CD25 on both, peripheral blood regulatory T cells and CD25lo CD127+ CD4+ T cells, enabling their distinct clustering in viSNE plots. SA-AgNP were of high elemental purity, showed minor background binding to cells in immunoassays, and were compatible with previously established staining protocols for PBMC and leukocytes, facilitating their use in complex mass cytometry panels. Considering the synthesis of AgNP from isotopically purified silver, the usage of AgNP extends the analytical capacity of mass cytometry panels by one, prospectively two, additional parameters, suitable for the detection of cellular targets of low abundance. © 2016 International Society for Advancement of Cytometry.

Einzelhandel findet heute nicht nur in den Stadt- und Stadtteilzentren bzw. in den Einkaufszentren am Rande der Stadte statt, sondern zunehmend auch im Internet. Der Beitrag untersucht, welche Determinanten ausschlaggebend sind, im stationaren Einzelhandel bzw. im Online-Handel einzukaufen. Die Ergebnisse basieren auf einer Befragung von rund 2.900 Personen in den sechs nordrhein-westfalischen Stadtregionen Aachen, Bochum, Bonn, Dortmund, Munster und Koln. In Abhangigkeit der Entfernung zur jeweiligen Innenstadt wurde in insgesamt 26 Untersuchungsgebieten befragt. In allen sechs Stadtregionen zeigt sich, dass nicht die raumlichen, sondern ausgewahlte demographische und soziookonomische Faktoren das Einkaufsverhalten bestimmen. Das Geschlecht, das Alter und die Lebensstile spielen beim Online-Einkauf eine wesentliche Rolle, das Einkommen ist keine beeinflussende Grose.

The impact of nanoparticles, NPs, at the single cell level has become a major field of toxicological research and different analytical methodologies are being investigated to obtain biological and toxicological information to better understand the mechanisms of cell–NP interactions. Here, inductively coupled plasma mass spectrometry in the single cell measurement mode (SC-ICP-MS) is proposed to study the uptake of silver NPs, AgNPs, with a diameter of 50 nm by human THP-1 monocytes in a proof-of-principle experiment. The main operating parameters of SC-ICP-MS have been optimized and applied for subsequent quantitative analysis of AgNPs to determine the number of particles in individual cells using AgNP suspensions for calibration. THP-1 cells were incubated with AgNP suspensions with concentrations of 0.1 and 1 μg mL−1 for 4 and 24 hours. The results reveal that the AgNP uptake by THP-1 monocytes is minimal at the lower dose of 0.1 μg mL−1 (roughly 1 AgNP per cell was determined), whereas a large cell-to-cell variance dependent on the exposure time is observed for a 10 times higher concentration (roughly 7 AgNPs per cell). The method was further applied to monitor the AgNP uptake by THP-1 cells differentiated macrophages incubated at the same AgNP concentration levels and exposure times demonstrating a much higher AgNP uptake (roughly from 9 to 45 AgNPs per cell) that was dependent on exposure concentration and remained constant over time. The results have been compared and validated by sample digestion followed by ICP-MS analysis as well as with other alternative promising techniques providing single cell analysis.

This paper presents the process of the translation and instrumentalisation of colonial urban planning principles into contemporary urban planning laws and instruments in Dar es Salaam. Based on a historical reading of the urban planning institutions and empirical references about current urban planning practices in this city it develops three main claims. First, there is a continuation of colonial planning institutions in the post-colonial Dar es Salaam. The shift of power from the colonial authority to the national state of Tanzania did not greatly impact on the planning institutions and practices in this city. Second, colonial urban planning legacies still dominate the planning institutions and practices in the post-colonial Dar es Salaam, however in different forms. They are now shaped by different sets of actors, follow economic logics, benefit only small groups of the economically privileged at the cost of the majority, and support the accumulation of power of the nation-state authorities. Third, the urban consequences and the urban planning institutions and practices of Dar es Salaam are interrelated; each is a result of the process of translation and instrumentalisation of colonial urban planning principles in the post-colonial setting accompanied by poor management and governance processes as well as the contradictions in the land tenure system that characterise this city. Acknowledging the urban consequences as being conditioned by the intense interplay between planning institutions and practice, and urban management and governance, this paper shows how the continued presence of colonial planning principles in the shaping of post-colonial planning practices may contribute to present-day urban consequences in Dar es Salaam.

Multiparametric fluorescence and mass cytometry offers new perspectives to disclose and to monitor the high diversity of cell populations in the peripheral blood for biomarker research. While high-end cytometric devices are currently available to detect theoretically up to 120 individual parameters at the single cell level, software tools are needed to analyze these complex datasets automatically in acceptable time and without operator bias or knowledge. We developed an automated analysis pipeline, immunoClust, for uncompensated fluorescence and mass cytometry data, which consists of two parts. First, cell events of each sample are grouped into individual clusters. Subsequently, a classification algorithm assorts these cell event clusters into populations comparable between different samples. The clustering of cell events is designed for datasets with large event counts in high dimensions as a global unsupervised method, sensitive to identify rare cell types even when next to large populations. Both parts use model-based clustering with an iterative expectation maximization algorithm and the integrated classification likelihood to obtain the clusters. A detailed description of both algorithms is presented. Testing and validation was performed using 1) blood cell samples of defined composition that were depleted of particular cell subsets by magnetic cell sorting, 2) datasets of the FlowCAP III challenges to identify populations of rare cell types and 3) high-dimensional fluorescence and mass-cytometry datasets for comparison with conventional manual gating procedures. In conclusion, the immunoClust-algorithm is a promising tool to standardize and automate the analysis of high-dimensional cytometric datasets. As a prerequisite for interpretation of such data, it will support our efforts in developing immunological biomarkers for chronic inflammatory disorders and therapy recommendations in personalized medicine. immunoClust is implemented as an R-package and is provided as source code from www.bioconductor.org.

Raumplanung hat die Aufgabe der Entwicklung, Ordnung und Sicherung des Raumes. Dies basiert auf dem Grundsatz einer umfassenden nachhaltigen Entwicklung, die „die sozialen, wirtschaftlichen und umweltschutzenden Anforderungen auch in Verantwortung gegenuber kunftigen Generationen miteinander in Einklang bringt, und eine dem Wohl der Allgemeinheit dienende sozialgerechte Bodennutzung“ [1] gewahrleistet [2].

From city to megacity: Dhaka in Bangladesh is an example of how people living in peripheral settlements strive to secure their survival. But this “informal urbanisation” presents urban planners with enormous challenges.

Vom System Stadt zum System Megacity: Das Beispiel Dhakas in Bangladesch zeigt, wie Menschen mit Siedlungen an der Stadtperipherie ihr Uberleben sichern wollen. Doch die „informelle Urbanisierung” stellt Stadt- und Raumplaner vor gewaltige Herausforderungen.

Public health, as part of community well-being, is linked to topics concerning urban visions, strategies and actors. As instruments and procedures for the development of current approaches to health promotion deliver a contribution to community well-being the interface between public health bodies and urban planning institutions within their statutory and non-statutory frameworks is of particular interest. Reflections on historical traces of collaborative urban visions as opportunities and threats as well as experiences of administrative collaboration between urban planning and public health institutions establish an argumentative contribution to joint strategies of urban planning and public health. There are the legal framework, coherent joint visions, inter- and trans-disciplinary checklists for evaluation to establish and departmental concepts for health and inter-ministerial coordination should be enforced.

Gesundheit bzw. Gesundheitsvorsorge ist ein traditionelles Thema in der Raumplanung, da sie sowohl in der Raumordnung als auch der Bauleitplanung zu den explizit zu berucksichtigenden Grundsatzen der Raumordnung bzw. stadtebaulichen Belangen gehort. Zur Einflussnahme auf die Verhaltnisse, in denen Menschen wohnen und arbeiten, stehen der raumlichen Planung auf den unterschiedlichen Planungs- und Entscheidungsebenen eine Vielzahl von Instrumenten zur Verfugung. Um uber den Gesundheitsschutz auch Gesundheitsvorsorge und vor allem die Forderung von Gesundheit pro-aktiv zu gestalten, bedarf es einer verbesserten Verknupfung von Methoden, Instrumenten und Verfahren sowie der Kooperation der raumlichen Planung mit dem Offentlichen Gesundheitsdienst.

Mass cytometry has pioneered40-parameter single-cell analyses that allow for the characterization of complex cellular networks at unprecedented depth. Up to 135 parameters can be simultaneously detected, but limited availability of metal tags suitable for labeling of specific probes prevents optimal exploitation of the analytical capacity of mass cytometers. To this end, we here establish the application of elemental silver nanoparticles (AgNP) of different size for reporting cell surface antigens on human leukocytes in mass cytometry assays. The mass channels at 107 Da and 109 Da are uniquely occupied by silver isotopes and do not interfere with other mass cytometry reagents. Streptavidin-coated AgNP (SA-AgNP) facilitated distinct and specific detection of various antigens, such as CD8, CD244 and CD294 on peripheral blood leukocytes pre-incubated with respective biotinylated primary antibodies. Signal intensities elicited by 40 nm-sized AgNP allowed specific detection of the low abundance antigen CD25 on both, peripheral blood regulatory T cells and CD25

(2011). Informal Urbanization—Historical and Geographical Perspectives. disP - The Planning Review: Vol. 47, No. 187, pp. 12-23.

Multiparametric fluorescence and mass cytometry offers new perspectives to disclose and to monitor the high diversity of cell populations in the peripheral blood for biomarker research. While high-end cytometric devices are currently available to detect theoretically up to 120 individual parameters at the single cell level, software tools are needed to analyze these complex datasets automatically in acceptable time and without operator bias or knowledge. We developed an automated analysis pipeline, immunoClust, for uncompensated fluorescence and mass cytometry data, which consists of two parts. First, cell events of each sample are grouped into individual clusters. Subsequently, a classification algorithm assorts these cell event clusters into populations comparable between different samples. The clustering of cell events is designed for datasets with large event counts in high dimensions as a global unsupervised method, sensitive to identify rare cell types even when next to large populations. Both parts use model-based clustering with an iterative expectation maximization algorithm and the integrated classification likelihood to obtain the clusters. A detailed description of both algorithms is presented. Testing and validation was performed using 1) blood cell samples of defined composition that were depleted of particular cell subsets by magnetic cell sorting, 2) datasets of the FlowCAP III challenges to identify populations of rare cell types and 3) high-dimensional fluorescence and mass-cytometry datasets for comparison with conventional manual gating procedures. In conclusion, the immunoClust-algorithm is a promising tool to standardize and automate the analysis of high-dimensional cytometric datasets. As a prerequisite for interpretation of such data, it will support our efforts in developing immunological biomarkers for chronic inflammatory disorders and therapy recommendations in personalized medicine. immunoClust is implemented as an R-package and is provided as source code from www.bioconductor.org.

The history of urban development and the advance of planning as a tool for guiding urban growth have always been closely linked with public health issues. Recognising the links between a city’s urban layout and its public health status has in the past often been the beginning of the establishment of planning principles, whether in Mesopotamia to improve air circulation, in medieval times to protect cities from fire hazards, or in the industrial age to fight epidemics such as cholera by adhering to sanitary guidelines. The interdependencies between socio-spatial development and the public health status of urban settlements are especially important nowadays in the fast growing cities of developing and transforming regions, where city authorities often have problems accommodating rapidly growing populations and an expanding economy in such a way that public health risks are minimised. In light of the severity of public health risks, especially in growing cities, the WHO (http://www.who.int) in 2010 launched the campaign ‘Urban planning as a critical link to building a healthy 21st century’. The campaign emphasises promotion of urban planning, improvement of urban living conditions, and participatory governance so as to enhance the resilience of cities to disaster and provide a better urban living environment (ibid.). The megacity Dhaka, the subject of our research, is a striking case showing the deterioration of the urban environment under uncontrolled and rapid urbanisation with detrimental effects on public health.

SUMMARYUsing a proteomics approach, we examined the post-translational changes in cytosolic proteins when isolated Malpighian tubules of Aedes aegyptiwere stimulated for 1 min with the diuretic peptide aedeskinin-III (AK-III,10–7 mol l–1). The cytosols of control (C)and aedeskinin-treated (T) tubules were extracted from several thousand Malpighian tubules, subjected to 2-D electrophoresis and stained for total proteins and phosphoproteins. The comparison of C and T gels was performed by gel image analysis for the change of normalized spot volumes. Spots with volumes equal to or exceeding C/T ratios of ±1.5 were robotically picked for in-gel digestion with trypsin and submitted for protein identification by nanoLC/MS/MS analysis. Identified proteins covered a wide range of biological activity. As kinin peptides are known to rapidly stimulate transepithelial secretion of electrolytes and water by Malpighian tubules, we focused on those proteins that might mediate the increase in transepithelial secretion. We found that AK-III reduces the cytosolic presence of subunits A and B of the V-type H+ ATPase, endoplasmin, calreticulin, annexin,type II regulatory subunit of protein kinase A (PKA) and rab GDP dissociation inhibitor and increases the cytosolic presence of adducin, actin,Ca2+-binding protein regucalcin/SMP30 and actin-depolymerizing factor. Supporting the putative role of PKA in the AK-III-induced activation of the V-type H+ ATPase is the effect of H89, an inhibitor of PKA,on fluid secretion. H89 reverses the stimulatory effect of AK-III on transepithelial fluid secretion in isolated Malpighian tubules. However,AK-III does not raise intracellular levels of cAMP, the usual activator of PKA, suggesting a cAMP-independent activation of PKA that removes subunits A and B from the cytoplasm in the assembly and activation of the V-type H+ ATPase. Alternatively, protein kinase C could also mediate the activation of the proton pump. Ca2+ remains the primary intracellular messenger of the aedeskinins that signals the remodeling of the paracellular complex apparently through protein kinase C, thereby increasing transepithelial anion secretion. The effects of AK-III on active transcellular and passive paracellular transport are additive, if not synergistic, to bring about the rapid diuresis.

A new pathway for cysteine biosynthesis has been elucidated in Mycobacterium tuberculosis. This pathway involves a protein-bound thiocarboxylate (CysO−SH) as the sulfide donor, similar to thiamin biosynthesis. Cysteine synthase M (CysM) catalyzes the addition of cysteine to the carboxy terminus of the protein-bound thiocarboxylate to generate a CysO−cysteine adduct. A protease, Mec+, hydrolyzes the CysO−cysteine adduct to release cysteine and regenerate CysO. Mec+ contains a JAMM motif, and this work provides the first functional characterization of the JAMM motif in prokaryotes. MoeZ, a paralogue of ThiF, has been shown to transfer sulfur onto CysO.

26 S proteasomes fulfill final steps in the ubiquitin-dependent degradation pathway by recognizing and hydrolyzing ubiquitylated proteins. As the 26 S proteasome mainly localizes to the nucleus in yeast, we addressed the question how this 2-MDa multisubunit complex is imported into the nucleus. 26 S proteasomes consist of a 20 S proteolytically active core and 19 S regulatory particles, the latter composed of two subcomplexes, namely the base and lid complexes. We have shown that 20 S core particles are translocated into the nucleus as inactive precursor complexes via the classic karyopherin alphabeta import pathway. Here, we provide evidence that nuclear import of base and lid complexes also depends on karyopherin alphabeta. Potential classic nuclear localization sequences (NLSs) of base subunits were analyzed. Rpn2 and Rpt2, a non-ATPase subunit and an ATPase subunit of the base complex, harbor functional NLSs. The Rpt2 NLS deletion yielded wild type localization. However, the deletion of the Rpn2 NLS resulted in improper nuclear proteasome localization and impaired proteasome function. Our data support the model by which nuclear 26 S proteasomes are assembled from subcomplexes imported by karyopherin alphabeta.

Gender Mainstreaming stellt eine aktuelle Herausforderung an die Implementation in Verwaltungshandeln dar. Die Integration der Geschlechterperspektive in alle Politikbereiche hat nun auch den Bereich der Stadtebaupolitik erreicht und kann hier an Erkenntnisse „frauenbezogener“ Planungsthemen von gut dreisig Jahren anknupfen. Lagen die inhaltlichen Schwerpunkte zunachst bei der Wohnung und beim Wohnumfeld, kamen dann die raumliche Beziehung zwischen Wohnung und Arbeitsplatz und damit die Schnittstellen von Erwerbsarbeit und informeller Arbeit ebenso hinzu wie die Themenfelder soziale Infrastruktur und Mobilitat. Mit der Verknupfung der raumlichen Planung von Arbeitsstatten und Gewerbe mit gleichstellungspolitischen Aspekten liegen bisher wenige Erfahrungen vor. Eine inhaltliche und prozessuale Verzahnung mit laufenden Verwaltungsverfahren formeller oder informeller raumlicher Planungen bietet sich hier an. Erste Erfahrungen werden derzeit in der Stadt Norderstedt in Schleswig-Holstein mit einem Leitprojekt „Gleichstellungspolitisch orientiertes Gewerbeflachenmanagement“ in Verknupfung mit einer Rahmenplanung und im Kontext des Regionalen Entwicklungskonzepts fur die Metropolregion Hamburg gesammelt. In diesem Zusammenhang wird im Jahr 2004 ein entsprechender Leitfaden vorliegen. Die folgenden Ausfuhrungen wurden teilweise in diesem Zusammenhang erarbeitet.