The Mycobacterium genus contains major human pathogens, like Mycobacterium tuberculosis and Mycobacterium leprae, which are the causative agents of Tuberculosis and Leprosy, respectively. They have evolved as successful human pathogens by adapting to the adverse conditions prevailing inside the host, which include host immune activation, nutrient depletion, hypoxia, and so on. During such adaptation for the survival and establishment of persistent infection inside the host, the pathogen, like M. tuberculosis, regulates its cell division. It is known that M. tuberculosis enters a state of non-replicating persistence (NRP) inside the host, to establish latent infection, which helps the survival of the pathogen under adverse host conditions such as hypoxia and nutrient depletion. The pathogen can reactivate itself, to come out of the NRP state, and establish active infection at a later stage, when conditions are suitable for its proliferation. The altered physiological state of the latent bacterium makes it tolerant to drugs, which are only effective against proliferating tubercle bacilli. In view of this unique behavioural physiology of tubercle bacilli, it is important to study the process of cell division and how it is regulated in the NRP and actively growing states. The work reported in the thesis is an attempt to understand these aspects of mycobacterial cell division. iii Chapter 1. Introduction: This chapter gives a detailed introduction to bacterial cell division and its regulation in various organisms, like Escherichia coli, Bacillus subtilis, Caulobacter crescentus, and others. In the background of this information, the major studies on mycobacterial cell division and its regulation are presented. Chapter 2. Materials and Methods: This chapter describes in detail all the materials and methods used in the experiments, which are presented in the four data chapters, 3-6. Chapter 3. Ultrastructural Study of the Formation of Septal Partition and Constriction in Mycobacteria and Delineation of its Unique Features: Mycobacteria have triple-layered complex cell wall, playing an important role in its survival under adverse conditions in the host. It is not known how these layers in the mother cell participate during cell division. Therefore, the ultrastructural changes in the different envelope layers of Mycobacterium tuberculosis, Mycobacterium smegmatis, and Mycobacterium xenopi, during the process of septation and septal constriction, were studied, using Transmission and Scanning Electron Microscopy. The unique aspects of mycobacterial septation and constriction were identified and were compared with those of E. coli and Bacillus subtilis septation. Further, based on all these observations, models were proposed for septation in M. tuberculosis and M. smegmatis. Chapter 4. Identification of Asymmetric Septation and Division in Mycobacteria and Its Role in Generating Cell Size Heterogeneity: Bacterial populations are known to harbour phenotypic heterogeneity that helps survival under stress conditions, as this heterogeneity comprises subpopulations that have differential susceptibility to stress conditions. The iv heterogeneity has been known to lead to the requirement for prolonged drug treatment for the elimination of the tolerant subpopulation. Hence, it is important to study the different mechanisms, which operate to generate population heterogeneity. Therefore, in this chapter, studies were carried out to find out whether asymmetric septation and division occur in mycobacteria to generate cell size heterogeneity. Subpopulations of mycobacterial mid-log phase cells of M. tuberculosis, M. smegmatis, and M. xenopi were found to undergo asymmetric division to generate cell size heterogeneity. The asymmetric division and the ultrastructure and growth features of the products of the division were studied. Chapter 5. Study of Mycobacterial Cell Division Using Growth-Synchronised Cells: In this chapter, different stages of cell septation and constriction were studied using growth-synchronised M. smegmatis cells. Phenethyl alcohol (PEA), which has been found to reversibly arrest mycobacterial cells, was used for growth synchronisation. The growth-synchronised mycobacterial cells, which were released from PEA block, were studied at different stages of septation and septal constriction, at the ultrastructural and molecular levels. Chapter 6. Identification of the Stage of Cell Division Arrest in NRP Mycobacteria: The exact stage at which the NRP tubercle bacilli are arrested in cell division is currently unknown. In Wayne’s in vitro model for hypoxia-responsive tubercle bacilli, gradual depletion of oxygen leads to hypoxic stress, inducing the bacilli to enter non-replicating persistence (NRP) state. Using this model, the stage of cell division arrest in M. tuberculosis was characterised at the ultrastructural and molecular levels. Hypoxia-stressed M. smegmatis was used as an experimental system for contrast. The thesis concludes with salient findings, a bibliography, and the list of publications.

Doctorat en Sciences biomédicales et pharmaceutiques / info:eu-repo/semantics/nonPublished

La quantité de données disponibles n'a jamais été aussi grande. Se poser les bonnes questions, c'est-à-dire des questions qui soient à la fois pertinentes et dont la réponse est accessible est difficile. L'analyse topologique de données tente de contourner le problème en ne posant pas une question trop précise mais en recherchant une structure sous-jacente aux données. Une telle structure est intéressante en soi mais elle peut également guider le questionnement de l'analyste et le diriger vers des questions pertinentes. Un des outils les plus utilisés dans ce domaine est l'homologie persistante. Analysant les données à toutes les échelles simultanément, la persistance permet d'éviter le choix d'une échelle particulière. De plus, ses propriétés de stabilité fournissent une manière naturelle pour passer de données discrètes à des objets continus. Cependant, l'homologie persistante se heurte à deux obstacles. Sa construction se heurte généralement à une trop large taille des structures de données pour le travail en grandes dimensions et sa robustesse ne s'étend pas au bruit aberrant, c'est-à-dire à la présence de points non corrélés avec la structure sous-jacente.Dans cette thèse, je pars de ces deux constatations et m'applique tout d'abord à rendre le calcul de l'homologie persistante robuste au bruit aberrant par l'utilisation de la distance à la mesure. Utilisant une approximation du calcul de l'homologie persistante pour la distance à la mesure, je fournis un algorithme complet permettant d'utiliser l'homologie persistante pour l'analyse topologique de données de petite dimension intrinsèque mais pouvant être plongées dans des espaces de grande dimension. Précédemment, l'homologie persistante a également été utilisée pour analyser des champs scalaires. Ici encore, le problème du bruit aberrant limitait son utilisation et je propose une méthode dérivée de l'utilisation de la distance à la mesure afin d'obtenir une robustesse au bruit aberrant. Cela passe par l'introduction de nouvelles conditions de bruit et l'utilisation d'un nouvel opérateur de régression. Ces deux objets font l'objet d'une étude spécifique. Le travail réalisé au cours de cette thèse permet maintenant d'utiliser l'homologie persistante dans des cas d'applications réelles en grandes dimensions, que ce soit pour l'inférence topologique ou l'analyse de champs scalaires. / Massive amounts of data are now available for study. Asking questions that are both relevant and possible to answer is a difficult task. One can look for something different than the answer to a precise question. Topological data analysis looks for structure in point cloud data, which can be informative by itself but can also provide directions for further questioning. A common challenge faced in this area is the choice of the right scale at which to process the data.One widely used tool in this domain is persistent homology. By processing the data at all scales, it does not rely on a particular choice of scale. Moreover, its stability properties provide a natural way to go from discrete data to an underlying continuous structure. Finally, it can be combined with other tools, like the distance to a measure, which allows to handle noise that are unbounded. The main caveat of this approach is its high complexity.In this thesis, we will introduce topological data analysis and persistent homology, then show how to use approximation to reduce the computational complexity. We provide an approximation scheme to the distance to a measure and a sparsifying method of weighted Vietoris-Rips complexes in order to approximate persistence diagrams with practical complexity. We detail the specific properties of these constructions.Persistent homology was previously shown to be of use for scalar field analysis. We provide a way to combine it with the distance to a measure in order to handle a wider class of noise, especially data with unbounded errors. Finally, we discuss interesting opportunities opened by these results to study data where parts are missing or erroneous.

Nous proposons dans cette thèse de caractériser quantitativement la variabilité à différentes échelles au cours de l'embryogenèse. Pour ce faire, nous utilisons une combinaison de modèles mathématiques et de résultats expérimentaux. Dans la première partie, nous utilisons une petite cohorte d'oursins digitaux pour construire une représentation prototypique du lignage cellulaire, reliant les caractéristiques des cellules individuelles avec les dynamiques à l'échelle de l'embryon tout entier. Ce modèle probabiliste multi-niveau et empirique repose sur les symétries des embryons et sur les identités cellulaires; cela permet d'identifier un niveau de granularité générique pour observer les distributions de caractéristiques cellulaires individuelles. Le prototype est défini comme le barycentre de la cohorte dans la variété statistique correspondante. Parmi plusieurs résultats, nous montrons que la variabilité intra-individuelle est impliquée dans la reproductibilité du développement embryonnaire. Dans la seconde partie, nous considérons les mécanismes sources de variabilité au cours du développement et leurs relations à l'évolution. En nous appuyant sur des résultats expérimentaux montrant une pénétrance incomplète et une expressivité variable de phénotype dans une lignée mutante du poisson zèbre, nous proposons une clarification des différents niveaux de variabilité biologique reposant sur une analogie formelle avec le cadre mathématique de la mécanique quantique. Nous trouvons notamment une analogie formelle entre l'intrication quantique et le schéma Mendélien de transmission héréditaire. Dans la troisième partie, nous étudions l'organisation biologique et ses relations aux trajectoires développementales. En adaptant les outils de la topologie algébrique, nous caractérisons des invariants du réseaux de contacts cellulaires extrait d'images de microscopie confocale d'épithéliums de différentes espèces et de différents fonds génétiques. En particulier, nous montrons l'influence des histoires individuelles sur la distribution spatiales des cellules dans un tissu épithélial. / We propose in this thesis to characterize variability quantitatively at various scales during embryogenesis. We use a combination of mathematical models and experimental results. In the first part, we use a small cohort of digital sea urchin embryos to construct a prototypical representation of the cell lineage, which relates individual cell features with embryo-level dynamics. This multi-level data-driven probabilistic model relies on symmetries of the embryo and known cell types, which provide a generic coarse-grained level of observation for distributions of individual cell features. The prototype is defined as the centroid of the cohort in the corresponding statistical manifold. Among several results, we show that intra-individual variability is involved in the reproducibility of the developmental process. In the second part, we consider the mechanisms sources of variability during development and their relations to evolution. Building on experimental results showing variable phenotypic expression and incomplete penetrance in a zebrafish mutant line, we propose a clarification of the various levels of biological variability using a formal analogy with quantum mechanics mathematical framework. Surprisingly, we find a formal analogy between quantum entanglement and Mendel’s idealized scheme of inheritance. In the third part, we study biological organization and its relations to developmental paths. By adapting the tools of algebraic topology, we compute invariants of the network of cellular contacts extracted from confocal microscopy images of epithelia from different species and genetic backgrounds. In particular, we show the influence of individual histories on the spatial distribution of cells in epithelial tissues.

MENVSC / Department of Hydrology and Water Resources / Sewage sludge (biosolids) from wastewater treatment plants (WWTPs) has been widely used as a soil improver in Europe, United States of America and some developing countries including South Africa. It has its benefits for farmers as a good source of organic matter and minerals, however, sludge after treatment still contains pathogenic organisms, heavy metals and persistent organic pollutants (POPs). The POP and heavy metal contaminants that accumulate in sludge may transfer through the food chain and cause adverse effects on human beings. In this study, a field experiment was carried out on farmland fertilized with sewage sludge from a wastewater treatment plant (WWTP) that vasically receives domestic wastewater and storm water. Vegetable spinach (Spinacia oleracea) was used for this study and was planted on a farmland under controlled conditions. Ten ridges each of dimensions 20 m × 0.3 m was made and dry sludge weights of 5, 10, 15, 20, 25, 30, 35, 40, 45 and 50 kg were applied as manure on each of the ridges, respectively. Representative samples of sludge and soil were taken for analysis of heavy metals and POPs. At maturity, in twelve weeks, the root and leave samples of the vegetable were taken from all the ridges including the control. The soil, sludge, and vegetable samples were analyzed for total heavy metal content (Cd, Cr, Cu, Ni, Pb, Co, Zn, Al, Fe, Mn), speciated heavy metal content and POP (PAH, PCB). Soil and sludge samples were also analyzed for total organic content, pH, cation exchange capacity (CEC), conductivity and alkalinity. The analysis for total heavy metals and speciated heavy metal content was carried out using inductively coupled plasma optical emission spectrophotometer (ICP-OES), and CEC analysis was carried out using atomic absorption spectrophotometer (AAS). A two-dimensional gas chromatograph with time of flight mass spectrometry detector (GC X GC TOFMS) was used for POP measurements. pH measurement was made using a pH meter and conductivity measurement using a conductivity meter. Alkalinity and total organic content analysis was performed using titrimetric apparatus. The highest total heavy metal concentration of 378.9 mg / kg was recorded in Fe metal in soil and Leaf sample while the lowest total metal concentration of 0.0003 mg / kg was in Cu metal in root sample. The highest heavy metal concentration of 1002 mg / kg in speciated forms was in Mn metal in F1 fraction and the lowest of 0.0004 mg / kg was in Cd metal in F5 fraction. PAHs were only found in soil samples and their concentrations ranged from 2.53 mg / kg to 146.5 mg / kg. There were no PCB detected in all the samples analysed. The results indicated that the trace metals concentrations found in the exchangeable fraction were higher than those observed in any of the preceding extractions except in the case of Cd, Cr, Fe and Pb where Fe-Mn oxide and organic matter fractions predominated and were closely followed by exchangeable fraction.

In the United Nations Convention on the Law of the Sea, the environmental protection of the marine environment was first addressed in a comprehensive manner on an international level. However, the Convention distinguishes between four different sorts of pollution depending on which source the pollution originates from. Still, one of these sources play a more crucial role in the protection of the marine environment than the other since that source is estimated to stand for 80 percent of all the marine pollution; namely marine pollution from land-based sources. As the throw-away culture has led to products being disposed of at a faster rate than ever before, in particular plastic products, the amount of land-based debris has also substantially increased over the last decades. This increased disposal rate of products in combination with poor waste treatment has consequently led to many kinds of wastes ending up in the ocean and causing severe harm, not only to the marine environment and its living species, but also to humans that eat the fish and use the many other ecosystem services of the Sea. In this thesis, some prominent international conventions on marine pollution from land-based sources are examined; namely the United Nations Convention on the Law of the Sea, the Basel Convention on the Control of Transboundary Movements of Hazardous Wastes and their Disposal, as well as the Stockholm Convention on Persistent Organic Pollutants. To achieve United Nations Sustainable Development Goal number 14.1 to significantly reduce marine pollution from land-based sources by 2025, this thesis claims that international laws addressing this sort of pollution need to be implemented. Moreover, this thesis rests on the belief that regional implementation is a crucial component in making states align with international law. However, while regional implementation has been ambitious in the European Union Law, many regions still lack enforceable frameworks that aim to reduce and prevent marine pollution from land-based sources.

Indiana University-Purdue University Indianapolis (IUPUI) / Chronic inflammatory disorders are characterized by an increase in excitability of small diameter sensory neurons located in dorsal root ganglia (DRGs). This sensitization of neurons is a mechanism for chronic inflammatory pain and available therapies have poor efficacy and severe adverse effects when used chronically. Prostaglandin E₂ (PGE₂) is an inflammatory mediator that plays an important role in sensitization by activating G-protein coupled receptors (GPCRs) known as E-series prostaglandin receptors (EPs) coupled to the protein kinase A (PKA) pathway. EPs are known to downregulate upon prolonged exposure to PGE₂ or in chronic inflammation, however, sensitization persists and the mechanism for this is unknown. I hypothesized that persistence of PGE₂-induced hypersensitivity is associated with a switch in signaling caused by prolonged exposure to PGE₂ or the neurotrophin nerve growth factor (NGF), also a crucial inflammatory mediator. DRG cultures grown in the presence or absence of either PGE₂ or NGF were used to study whether re-exposure to the eicosanoid is able to cause sensitization and activate PKA. When cultures were grown in the presence of NGF, PGE₂-induced sensitization was not attenuated by inhibitors of PKA. Activation of PKA by PGE₂ was similar in DRG cultures grown in the presence or absence of NGF when phosphatase inhibitors were added to the lysis and assay buffers, but significantly less in cultures grown in the presence of NGF when phosphatase inhibitors were not added. In DRG cultures exposed to PGE₂ for 12 hours-5 days, sensitization after re-exposure to PGE₂ is maintained and resistant to PKA inhibition. Prolonged exposure to the eicosanoid caused complete loss of PKA activation after PGE₂ re-exposure. This desensitization was homologous, time dependent, reversible, and insurmountable by a higher concentration of PGE₂. Desensitization was attenuated by reduction of expression of G-protein receptor kinase 2 and was not mediated by PKA or protein kinase C. The presented work provides evidence for persistence of sensitization by PGE₂ as well as switch from the signaling pathway mediating this sensitization after long-term exposure to NFG or PGE₂.

<p dir="ltr">The growing movement toward sustainable use of ocean resources is driven by the pressing need to alleviate environmental and human stressors on the planet and its oceans. From monitoring the food web to supporting sustainable fisheries and observing environmental shifts to protect against the effects of climate change, ocean observations significantly impact the Blue Economy. Acknowledging the critical role of Autonomous Underwater Vehicles (AUVs) in achieving persistent ocean exploration, this research addresses challenges focusing on the limited energy and storage capacity of AUVs, introducing a comprehensive underwater docking solution with a specific emphasis on enhancing the terminal homing phase through innovative vision algorithms leveraging neural networks.</p><p dir="ltr">The primary goal of this work is to establish a docking procedure that is failure-tolerant, scalable, and systematically validated across diverse environmental conditions. To fulfill this objective, a robust dock detection mechanism has been developed that ensures the resilience of the docking procedure through \comment{an} improved detection in different challenging environmental conditions. Additionally, the study addresses the prevalent issue of data sparsity in the marine domain by artificially generating data using CycleGAN and Artistic Style Transfer. These approaches effectively provide sufficient data for the docking detection algorithm, improving the localization of the docking station.</p><p dir="ltr">Furthermore, this work introduces methods to compress the learned docking detection model without compromising performance, enhancing the efficiency of the overall system. Alongside these advancements, a station-keeping algorithm is presented, enabling the mobile docking station to maintain position and heading while awaiting the arrival of the AUV. To leverage the sensors onboard and to take advantage of the computational resources to their fullest extent, this research has demonstrated the feasibility of simultaneously learning docking detection and marine wildlife classification through multi-task and transfer learning. This multifaceted approach not only tackles the limitations of AUVs' energy and storage capacity but also contributes to the robustness, scalability, and systematic validation of underwater docking procedures, aligning with the broader goals of sustainable ocean exploration and the blue economy.</p>

Depuis l'émergence de la pandémie de SARS-CoV-2 en décembre 2019, plus de 675 millions de cas confirmés ont été signalés dans le monde, dont 4,6 millions de cas au Canada uniquement. Bien que la plupart des individus récupèrent sans séquelles, 10 à 20 % des survivants signalent des symptômes persistants au-delà de quatre semaines après une infection par le SARS-CoV-2, tels que la fatigue, les altérations cognitives, la toux, l'anxiété, la dépression, la douleur thoracique et autres, connus sous le nom de COVID longue ou de condition post-SARS-CoV-2 (PCC). Par conséquent, la physiopathologie, le diagnostic et la prise en charge de la PCC sont devenus un axe de recherche majeur. Pour contribuer à la compréhension de la PCC, nous avons mené le projet IPCO (Institut de Recherches cliniques de Montréal (IRCM) Post-COVID-19 Research Clinic), en posant comme hypothèses 1 que les personnes infectés par le SARS-CoV-2 au Québec présenteraient des signes et symptômes fréquents et variés post-phase aiguë, affectant différents systèmes d'organes, et 2 Les niveaux élevés de D-dimères dans PCC ne sont pas pertinents pour les événements thromboemboliques 3 que Chez les individus atteints de la PCC, la vaccination contre la COVID-19 réduirait les symptômes de la PCC en diminuant l'inflammation. Pour évaluer ces hypothèses, nous avons recruté des participants âgés de plus de 18 ans, un à 18 mois après l'infection aiguë, présentant au moins un symptôme persistant, et programmé des visites de base et de suivi à 3-6 mois, 1 an et 2 ans post-infection aiguë. Chaque visite comprenait des évaluations cliniques, des prélèvements, des évaluations en laboratoire, des questionnaires sur l'alimentation et le bien-être, ainsi que des évaluations de la physiologie pulmonaire et cardiaque. Sur la base d'une étude allemande qui a catégorisé les symptômes du PCC et les individuals en trois groupes de sévérité, nous avons classé nos participants en trois niveaux de sévérité : non/légère (score du PCC <10,75), modérée (10,75 < score du PCC < 26,25) et sévère (score du PCC > 26,25). Cette thèse présente les résultats de trois sous-études IPCO. Dans l'étude descriptive, nous avons observé que la fatigue, les problèmes de mémoire et les maux de tête étaient les symptômes de PCC les plus courants, la majorité de nos participants étant des femmes et ayant été traités en ambulatoire pendant la phase aiguë. Dans l'étude transversale, nous avons constaté des différences significatives dans les mesures de santé et de bien-être à tous les moments, mais aucune différence significative dans les résultats des tests physiologiques entre les groupes PCC non/léger, modéré et sévère. Dans l'étude longitudinale, les marqueurs de l'inflammation se sont améliorés au fil du temps, mais le taux métabolique basal et la masse grasse ont augmenté. Dans la deuxième étude, nous avons observé une forte prévalence de participants ayant des niveaux de D-dimères, qui n'étaient pas associés à des événements thromboemboliques, et aucune corrélation entre le niveau de D-dimères et les niveaux de cytokines et de chimiokines. Dans la troisième étude, nous avons observé que les participants vaccinés présentaient significativement moins de symptômes de PCC. Notre étude fournit une meilleure compréhension de la physiopathologie du PCC et de l'effet de la vaccination sur le profil clinique et inflammatoire du PCC, ce qui pourrait aider à la conception d'outils de gestion clinique et de recherche futurs. / Since the emergence of the SARS-CoV-2 pandemic in December 2019, over 675 million confirmed cases have been reported globally, with 4.6 million cases in Canada alone. Although most individuals recover without residual disease, 10-20% of survivors report symptoms persisting beyond four weeks after SARS-CoV-2 infection, such as fatigue, cognitive impairments, cough, anxiety, depression, chest pain, and others known as long-COVID or post SARS-CoV-2 condition (PCC). Consequently, the pathophysiology, diagnosis, and management of PCC have become a significant focus of research. To contribute to the understanding of PCC, we conducted the IPCO (Institut de Recherches cliniques de Montréal (IRCM) Post-COVID-19 Research Clinic) project, hypothesizing that 1 SARS-CoV-2 infected individuals in Quebec would present frequent and varied signs and symptoms post-acute phase, affecting different organ systems, and that 2 high D-dimer level in PCC is irrelevant to thromboembolic events , and 3 in individuals with PCC, COVID-19 vaccination would decrease PCC symptoms by reducing inflammation. To evaluate these hypotheses, we enrolled participants aged >18 years, one to 18 months post-acute infection, with at least one persistent symptom, and scheduled baseline and follow-up visits at 3-6 months, 1 year, and 2 years post-acute infection. Each visit involved clinical evaluations, sampling, laboratory evaluations, diet and well-being questionnaires, and pulmonary and cardiac physiology evaluations. Based on a German study that categorized PCC symptoms and individuals into three severity groups, we classified our participants into three severity levels: non/mild (PCC score < 10.75), moderate (10.75 < PCC score < 26.25), and severe (PCC score > 26.25). This thesis reports the results of three IPCO studies. In the descriptive study, we observed that fatigue, memory problems, and headaches were the most common PCC symptoms, with the majority of our participants being female and managed as outpatients during the acute phase. In the cross-sectional study, we noted significant differences in health and well-being measurements at all time points, but no significant difference in physiological tests' results between different severity groups. In the longitudinal study, markers of inflammation improved over time, but the basal metabolic rate and body fat increased. In the second study, we observed a high prevalence of participants having D-dimer levels in blood, which were not associated with thromboembolic events, and no correlation between D-dimer levels and blood cytokine/ chemokine levels. In the third study, we observed that vaccinated participants had significantly fewer PCC symptoms, fewer organ systems affected, higher well-being scores, and lower blood cytokine/chemokine levels than the non-vaccinated group. We also observed correlations between certain cytokines/chemokines, as well as between clinical parameters and certain cytokines/chemokines. Our study provides a better understanding of the pathophysiology of PCC and effect of vaccination on the clinical and inflammatory profile of PCC, which could assist future research and clinical management tool design.

Gestillte Kinder sind über die Muttermilch mit persistenten organischen Schadstoffen (POPs) belastet. Tierexperimentelle Studien deuten auf eine besondere Empfindlichkeit des sich entwickelnden Immunsystems für POPs hin. Mögliche Effekte dieser Verbindungen auf den kindlichen Organismus sind bisher kaum untersucht. Es wurden potenzielle Einflüsse verschiedener POPs (PCDDs, PCDFs, PCBs, beta-HCH, HCB und pp-DDE) auf natürliche Killerzellen (NK-Zellen) und NK-Aktivität bei gestillten im Vergleich zu nicht gestillten Kindern untersucht. NK-Zellen sind eine Lymphozytensubpopulation (CD3-CD56/16+), die über ihre zytotoxische Aktivität allogene, virusinfizierte und maligne Zellen ohne vorherige Sensibilisierung töten kann. Es wurden 66 gesunde Kinder im Alter von 11-12 Monaten untersucht, davon waren 50 Kinder mindestens 4 Monate lang voll gestillt und 16 Kinder nicht gestillt. Aus einer Region mit bekannter erhöhter PCDD/PCDF-Belastung stammten 13 gestillte Kinder. Die NK-Zellzahlen wurden mittels Immunphänotypisierung am Durchflusszytometer bestimmt. Die Aktivität der NK-Zellen wurde mit einem nicht-radioaktiven, durchflusszytometrischen Zytotoxizitäts-Assay gemessen. Die POP-Konzentrationen im Blutfett der Probanden wurden kommerziell bestimmt. Weder bei den NK-Zellzahlen, noch bei der NK-Aktivität konnten zwischen den gestillten und nicht-gestillten Kindern signifikante Unterschiede im t-Test nachgewiesen werden. In Korrelationsanalysen zeigten sich keine signifikanten Einflüsse der POP-Konzentrationen auf NK-Zellzahlen und NK-Aktivität. Im Laufe der Untersuchung zeigte sich, dass der eingesetzte Zytotoxizitäts-Assay nur semiquantitative Daten lieferte. Die vorliegenden Ergebnisse und weitere Befunde bezüglich des Immunsystems der Probanden weisen darauf hin, dass die relativ hohe Belastung lange gestillter Säuglinge mit POPs nicht zu einer biologischen Wirkung im kindlichen Organismus führt. Angesichts der nachgewiesenen positiven Effekte des Stillens, kann diesbezüglich die bestehende Stillempfehlung bekräftigt werden. / Breast-fed infants are exposed to persistent organic pollutants (POPs) via breast milk. Animal studies indicate a special sensitivity of the maturing immune system to POPs. Possible effects of these compounds on the infantile organism are so far barely examined. Potential influences of several POPs (PCDDs, PCDFs, PCBs, beta-HCH, HCB and pp-DDE) on natural killer (NK) cells and NK activity of breast-fed infants in comparison to formula-fed infants were investigated. NK cells are a subset of lymphocytes (CD3-CD56/16+) that can kill allogeneic, virus-infected and malignant cells via their cytotoxic activity without prior sensitization. The study group consisted of 66 healthy infants examined at age 11 to 12 months, of which 50 infants were breast-fed and 16 infants were formula-fed. 13 breast-fed infants came from a region with known increased PCDD/PCDF-burden. Numbers of NK cells were measured by flow cytometric immunophenotyping. NK activity was analysed by a non-radioactive flow cytometric cytotoxicity assay. POP concentrations in the blood fat of the probands were calculated commercially. There were no significant differences between breast-fed and formula-fed infants concerning number and activity of NK cells in the t-test. Analysis of correlation showed no significant influences of POP concentrations on the number and activity of NK cells. In the course of the study the data obtained by the employed cytotoxicity assay proved to be only semiquantitative. The presented findings and further results concerning the immune system of the study subjects suggest that the relatively high burden of long-term breast-fed infants with POPs does not lead to a biological effect in the infantile organism. Regarding the proven positive effects of breast-feeding the existing recommendation to breastfeed can be encouraged.