Enumeration, variability, and transport of Escherichia coli in the foreshore reservoir and surface water at freshwater beaches

Beaches are of immense recreational, societal and economic value. This value, however, is considerably diminished by poor water quality. Fecal indicator bacteria (FIB) are measured at recreational beaches worldwide to assess the water quality. A beach closure or advisory is issued if FIB concentrations in surface water exceed recreational water quality standards. Due to the lengthy time required to enumerate FIB (24 – 96 hours), statistical and mechanistic models have been developed to predict water quality exceedances a priori and to better understand why and under what conditions water quality exceedances occur. These models as well as beach water quality management strategies are often based on limited mechanistic understanding of the fate and transport of FIB in the beach environment. For instance, FIB are known to accumulate at very high concentrations in foreshore sand and porewater at beaches (herein referred to as the foreshore reservoir). The dynamics of FIB accumulation in the foreshore reservoir and its subsequent release, including the impact on surface water quality exceedances, is unknown. It is also unclear how to best quantify the abundance of FIB in the reservoir including its partitioning between the sand and pore water. An increased understanding of the behavior of FIB at beaches is needed to improve the accuracy of predictive water quality models, develop effective measures to reduce water quality exceedances, improve water quality monitoring strategies, and ultimately to better protect human health at recreational beaches. This thesis focuses on addressing key knowledge gaps regarding the behavior and quantification of FIB in the foreshore reservoir. In the first study, seasonal and daily variabilities in FIB concentrations in the foreshore reservoir and surface water are evaluated including determining the influence of environmental factors, such as temperature, waves, and rainfall. In this study, seasonal variability in FIB concentrations in the surface water and foreshore reservoir were found to depend on environmental factors, with some beaches showing a gradual increasing trend through the summer, then decreasing towards the beginning of fall. However, daily variation showed that FIB variability is much more complex and FIB may not simply accumulate over the summer months as previously thought. Further, this study showed for the first time that FIB may be able to replicate in unseeded natural foreshore beach sand not subjected to external stimuli. The second study uses experimental and field data to evaluate the behavior of FIB in the beach environment during intensified wave conditions including the transfer of FIB from the foreshore reservoir to the surface water. This study showed that as wave height increased foreshore sand erosion resulted in elevated E. coli concentrations in surface water, as well as depletion of E. coli from the foreshore sand and pore water. E. coli initially attached to foreshore sand rather than initially residing in the pore water was found to be the main contributor to elevated surface water concentrations. Surface water E. coli concentrations were a function of not only wave height (and associated sand erosion) but also the time elapsed since a preceding period of high wave intensity. This finding is important for statistical regression models used to predict beach advisories. While calculations suggested that foreshore sand erosion may be the dominant mechanism for releasing E. coli to surface water during intensified wave conditions at a fine sand beach, comparative characterization of the E. coli distribution at a coarse sand-cobble beach suggested that interstitial pore water flow and discharge may be more important for coarser sand beaches. The third study compared the partitioning of FIB in the foreshore reservoir between the sand and pore water and evaluated different sampling methods for quantifying FIB in the foreshore reservoir at beaches with varying grain sizes. This study showed that the collection of the top 1 cm of unsaturated sand resulted in higher and more variable concentrations than the top 5 cm of sand. There were no statistical differences in E. coli concentrations when using different methods to sample the saturated sand. Overall, the unsaturated sand had the highest amount of E. coli when compared to saturated sand and pore water (considered on a bulk volumetric basis). Pore water sampled with a shovel resulted in the highest observed E. coli concentrations (only statistically significant at fine sand beaches) and lowest variability compared to other sampling methods. These findings presented will help determine the appropriate sampling strategy for characterizing FIB abundance in the foreshore reservoir as a means of predicting its potential impact on nearshore surface water quality and public health risk. Overall, this thesis presents valuable information to health departments, beach managers, and scientists interested in improving water quality and water quality predictions at recreational beaches. Findings from this thesis increase understanding of FIB behavior, especially in the foreshore reservoir, and can be used to improve predictive water quality models, develop strategies to reduce FIB levels at beaches, and identify where and when a foreshore reservoir may be an important source of FIB to the surface water at a beach

A Place for the Church within Professional Psychology

There are various reasons why Christian psychologists may resist fully participating in a church community. Among these are historic tensions between science and religion, the complexity of multiple role relationships, cynicism related to clients\u27 negative experiences in religious communities, and differing perspectives on attributions and human nature. Despite these obstacles, there are compelling reasons for psychologists to remain involved in church. Eight of these reasons are described--clustered into professional, relational, and transformational domains--and implications discussed

Structure–function studies of chemokine-derived carboxy-terminal antimicrobial peptides

AbstractRecent reports which show that several chemokines can act as direct microbicidal agents have drawn renewed attention to these chemotactic signalling proteins. Here we present a structure–function analysis of peptides derived from the human chemokines macrophage inflammatory protein-3α (MIP-3α/CCL20), interleukin-8 (IL-8), neutrophil activating protein-2 (NAP-2) and thrombocidin-1 (TC-1). These peptides encompass the C-terminal α-helices of these chemokines, which have been suggested to be important for the direct antimicrobial activities. Far-UV CD spectroscopy showed that the peptides are unstructured in aqueous solution and that a membrane mimetic solvent is required to induce a helical secondary structure. A co-solvent mixture was used to determine solution structures of the peptides by two-dimensional 1H-NMR spectroscopy. The highly cationic peptide, MIP-3α51–70, had the most pronounced antimicrobial activity and displayed an amphipathic structure. A shorter version of this peptide, MIP-3α59–70, remained antimicrobial but its structure and mechanism of action were unlike that of the former peptide. The NAP-2 and TC-1 proteins differ in their sequences only by the deletion of two C-terminal residues in TC-1, but intact TC-1 is a very potent antimicrobial while NAP-2 is inactive. The corresponding C-terminal peptides, NAP-250–70 and TC-150–68, had very limited and no bactericidal activity, respectively. This suggests that other regions of TC-1 contribute to its bactericidal activity. Altogether, this work provides a rational structural basis for the biological activities of these peptides and proteins and highlights the importance of experimental characterization of peptide fragments as distinct entities because their activities and structural properties may differ substantially from their parent proteins

Loss of the p53/p63 Regulated Desmosomal Protein Perp Promotes Tumorigenesis

Dysregulated cell–cell adhesion plays a critical role in epithelial cancer development. Studies of human and mouse cancers have indicated that loss of adhesion complexes known as adherens junctions contributes to tumor progression and metastasis. In contrast, little is known regarding the role of the related cell–cell adhesion junction, the desmosome, during cancer development. Studies analyzing expression of desmosome components during human cancer progression have yielded conflicting results, and therefore genetic studies using knockout mice to examine the functional consequence of desmosome inactivation for tumorigenesis are essential for elucidating the role of desmosomes in cancer development. Here, we investigate the consequences of desmosome loss for carcinogenesis by analyzing conditional knockout mice lacking Perp, a p53/p63 regulated gene that encodes an important component of desmosomes. Analysis of Perp-deficient mice in a UVB-induced squamous cell skin carcinoma model reveals that Perp ablation promotes both tumor initiation and progression. Tumor development is associated with inactivation of both of Perp's known functions, in apoptosis and cell–cell adhesion. Interestingly, Perp-deficient tumors exhibit widespread downregulation of desmosomal constituents while adherens junctions remain intact, suggesting that desmosome loss is a specific event important for tumorigenesis rather than a reflection of a general change in differentiation status. Similarly, human squamous cell carcinomas display loss of PERP expression with retention of adherens junctions components, indicating that this is a relevant stage of human cancer development. Using gene expression profiling, we show further that Perp loss induces a set of inflammation-related genes that could stimulate tumorigenesis. Together, these studies suggest that Perp-deficiency promotes cancer by enhancing cell survival, desmosome loss, and inflammation, and they highlight a fundamental role for Perp and desmosomes in tumor suppression. An understanding of the factors affecting cancer progression is important for ultimately improving the diagnosis, prognostication, and treatment of cancer

Aligning and synchronization of MIS5 proxy records from Lake Ohrid (FYROM) with independently dated Mediterranean archives: implications for DEEP core chronology

The DEEP site sediment sequence obtained during the ICDP SCOPSCO project at Lake Ohrid was dated using tephrostratigraphic information, cyclostratigraphy, and orbital tuning through the marine isotope stages (MIS) 15-1. Although this approach is suitable for the generation of a general chronological framework of the long succession, it is insufficient to resolve more detailed palaeoclimatological questions, such as leads and lags of climate events between marine and terrestrial records or between different regions. Here, we demonstrate how the use of different tie points can affect cyclostratigraphy and orbital tuning for the period between ca. 140 and 70 ka and how the results can be correlated with directly/indirectly radiometrically dated Mediterranean marine and continental proxy records. The alternative age model presented here shows consistent differences with that initially proposed by Francke et al. (2015) for the same interval, in particular at the level of the MIS6-5e transition. According to this new age model, different proxies from the DEEP site sediment record support an increase of temperatures between glacial to interglacial conditions, which is almost synchronous with a rapid increase in sea surface temperature observed in the western Mediterranean. The results show how a detailed study of independent chronological tie points is important to align different records and to highlight asynchronisms of climate events. Moreover, Francke et al. (2016) have incorporated the new chronology proposed for tephra OH-DP-0499 in the final DEEP age model. This has reduced substantially the chronological discrepancies between the DEEP site age model and the model proposed here for the last glacial-interglacial transition

Sedimentological processes and environmental variability at Lake Ohrid (Macedonia, Albania) between 637 ka and the present

Lake Ohrid (Macedonia, Albania) is thought to be more than 1.2 million years old and host more than 300 endemic species. As a target of the International Continental scientific Drilling Program (ICDP), a successful deep drilling campaign was carried out within the scope of the Scientific Collaboration on Past Speciation Conditions in Lake Ohrid (SCOPSCO) project in 2013. Here, we present lithological, sedimentological, and (bio-)geochemical data from the upper 247.8 m composite depth of the overall 569 m long DEEP site sediment succession from the central part of the lake. According to an age model, which is based on 11 tephra layers (first-order tie points) and on tuning of bio-geochemical proxy data to orbital parameters (second-order tie points), the analyzed sediment sequence covers the last 637 kyr. The DEEP site sediment succession consists of hemipelagic sediments, which are interspersed by several tephra layers and infrequent, thin (< 5 cm) mass wasting deposits. The hemipelagic sediments can be classified into three different lithotypes. Lithotype 1 and 2 deposits comprise calcareous and slightly calcareous silty clay and are predominantly attributed to interglacial periods with high primary productivity in the lake during summer and reduced mixing during winter. The data suggest that high ion and nutrient concentrations in the lake water promoted calcite precipitation and diatom growth in the epilimnion during MIS15, 13, and 5. Following a strong primary productivity, highest interglacial temperatures can be reported for marine isotope stages (MIS) 11 and 5, whereas MIS15, 13, 9, and 7 were comparably cooler. Lithotype 3 deposits consist of clastic, silty clayey material and predominantly represent glacial periods with low primary productivity during summer and longer and intensified mixing during winter. The data imply that the most severe glacial conditions at Lake Ohrid persisted during MIS16, 12, 10, and 6, whereas somewhat warmer temperatures can be inferred for MIS14, 8, 4, and 2. Interglacial-like conditions occurred during parts of MIS14 and 8

Sedimentological processes and environmental variability at Lake Ohrid (Macedonia, Albania) between 637 ka and the present

Lake Ohrid (Macedonia, Albania) is thought to be more than 1.2 million years old and host more than 300 endemic species. As a target of the International Continental scientific Drilling Program (ICDP), a successful deep drilling campaign was carried out within the scope of the Scientific Collaboration on Past Speciation Conditions in Lake Ohrid (SCOPSCO) project in 2013. Here, we present lithological, sedimentological, and (bio-)geochemical data from the upper 247.8 m composite depth of the overall 569 m long DEEP site sediment succession from the central part of the lake. According to an age model, which is based on 11 tephra layers (first-order tie points) and on tuning of bio-geochemical proxy data to orbital parameters (second-order tie points), the analyzed sediment sequence covers the last 637 kyr. The DEEP site sediment succession consists of hemipelagic sediments, which are interspersed by several tephra layers and infrequent, thin (< 5 cm) mass wasting deposits. The hemipelagic sediments can be classified into three different lithotypes. Lithotype 1 and 2 deposits comprise calcareous and slightly calcareous silty clay and are predominantly attributed to interglacial periods with high primary productivity in the lake during summer and reduced mixing during winter. The data suggest that high ion and nutrient concentrations in the lake water promoted calcite precipitation and diatom growth in the epilimnion during MIS15, 13, and 5. Following a strong primary productivity, highest interglacial temperatures can be reported for marine isotope stages (MIS) 11 and 5, whereas MIS15, 13, 9, and 7 were comparably cooler. Lithotype 3 deposits consist of clastic, silty clayey material and predominantly represent glacial periods with low primary productivity during summer and longer and intensified mixing during winter. The data imply that the most severe glacial conditions at Lake Ohrid persisted during MIS16, 12, 10, and 6, whereas somewhat warmer temperatures can be inferred for MIS14, 8, 4, and 2. Interglacial-like conditions occurred during parts of MIS14 and 8. © Author(s) 2016

A human STAT3 gain-of-function variant confers T cell dysregulation without predominant Treg dysfunction in mice

Primary immune regulatory disorders (PIRD) represent a group of disorders characterized by immune dysregulation, presenting with a wide range of clinical disease, including autoimmunity, autoinflammation, or lymphoproliferation. Autosomal dominant germline gain-of-function (GOF) variants in STAT3 result in a PIRD with a broad clinical spectrum. Studies in patients have documented a decreased frequency of FOXP3+ Tregs and an increased frequency of Th17 cells in some patients with active disease. However, the mechanisms of disease pathogenesis in STAT3 GOF syndrome remain largely unknown, and treatment is challenging. We developed a knock-in mouse model harboring a de novo pathogenic human STAT3 variant (p.G421R) and found these mice developed T cell dysregulation, lymphoproliferation, and CD4+ Th1 cell skewing. Surprisingly, Treg numbers, phenotype, and function remained largely intact; however, mice had a selective deficiency in the generation of iTregs. In parallel, we performed single-cell RNA-Seq on T cells from STAT3 GOF patients. We demonstrate only minor changes in the Treg transcriptional signature and an expanded, effector CD8+ T cell population. Together, these findings suggest that Tregs are not the primary driver of disease and highlight the importance of preclinical models in the study of disease mechanisms in rare PIRD