Mechanistic Insights Into Neutrophil Turnover During Gram-Negative Pneumonic Sepsis

Sepsis is one of the oldest and most elusive syndromes in medicine. It is characterized by a systemic inflammatory response leading to acute organ dysfunction. In the United States alone there are over 750,000 cases of sepsis each year, with a mortality rate between 20-50%. Sepsis is the second leading cause of death in the ICU and the tenth leading cause of death overall in the United States. Over half of all ICU resources are consumed in the treatment of sepsis. There are currently no effective therapies available against sepsis, all attempts to develop an effective therapy have failed. Respiratory infections are the leading cause of sepsis, leading to what is called pneumonic sepsis. The Gram-negative bacteria Klebsiella pneumoniae (KPn) is associated with upwards of 20% of all pneumonic sepsis cases. Utilizing a mouse model of pneumonic sepsis induced by intranasal infection of KPn, I studied the role that mammalian C-type lectin receptors (CLRs) play in the development and clearance of pneumonic sepsis. CLRs are primarily expressed on phagocytic immune cells of myeloid origin and are considered pattern recognition receptors that have the ability to shape the immune response by recognizing both pathogen associated molecular patterns and host damage associated molecular patterns in many different pathological conditions. From the pathogen perspective, I studied the effect KPn has on the immune cells and how that may impact the ability of KPn establish the infection. In my first project, I examined the role of two CLRs, Macrophage Galactose Type Lectin 1 (MGL1) and Clec4e otherwise known as Mincle, in the development and clearance of pneumonic sepsis caused by KPn. This project was built on previous observations made by my advisor Dr. Jyotika Sharma which showed these two CLRs upregulated in the lung during KPn induced pneumonic sepsis. In regards to MGL1, I am the first author on the paper that we recently published in the Journal of Immunology “Macrophage Galactose-Type Lectin-1 Deficiency Is Associated with Increased Neutrophilia and Hyperinflammation in Gram-Negative Pneumonia” (J Immunol. 2016 Apr 1;196(7):3088-96. doi: 10.4049/jimmunol.1501790.). In this paper we showed that MGL1-/- mice were more susceptible to KPn infection, a phenotype that did not correlate with the systemic and local bacterial burden; the ability of macrophages and neutrophils to phagocytose and kill bacteria or neutrophil NETs. We demonstrated that the mechanism underlying the increased mortality of MGL1-/- mice was increased ability of neutrophils to infiltrate the lungs causing their overwhelming accumulation and severe neutrophil-mediated pathology in the lungs in the absence of MGL1. Mechanistic insights into the potential negative regulatory function of MGL1 in neutrophil infiltration, their clearance by efferocytosis as well as the process of granulopoiesis are some of the future directions of this work that are being perused in the laboratory. In the paper I co-authored on Mincle, “Protective Role of Mincle in Bacterial Pneumonia by Regulation of Neutrophil Mediated Phagocytosis and Extracellular Trap Formation” (Infect Dis. (2014) 209 (11): 1837-1846.doi: 10.1093/infdis/jit820), we showed that Mincle mediates two important bacterial clearance mechanisms i.e. bacterial phagocytosis and extracellular trap (NET) formation by neutrophils. As a result, Mincle-/- mice are more susceptible to KPn infection as compared to the Mincle sufficient wild-type mice. This project is being led by Dr. Atul Sharma, a postdoctoral fellow in the lab. My second project examined the effect of KPn infection on the clearance of neutrophils by a process called efferocytosis. While performing experiments for the role of MGL1 in efferocytosis, I found that in comparison with the uninfected neutrophils, KPn infected cells were engulfed less efficiently by macrophages. As efferocytosis is a receptor mediated process, I discovered that while KPn infection increases the expression of repressive molecules called “Don’t Eat Me” signals on neutrophils, it is the modulation of distribution of a key “Eat Me” signal, phosphatidylserine (PtdSer) and the subsequent delay of apoptosis in neutrophils that is partially involved in KPn mediated inhibition of efferocytosis. Accordingly, KPn infected neutrophils also induce an alternative rout of programmed cell death called necroptosis. It is the combination of the “eat me” signal downregulation and the induction of necroptosis in KPn infected neutrophils that inhibit their efferocytic uptake by macrophages. A part of this data has been communicated for publication. Current work that I have led in the lab on this project involves determining what signaling mechanisms KPn is activating to decrease PtdSer as well as activation of necroptosis to inhibit the efferocytosis of infected PMNs. The overarching goal of this work was to increase the overall knowledge on the role CLRs play in pneumonic sepsis in terms of neutrophil turnover and how KPn subverts these processes to its own advantage. I believe that this knowledge can be used to identify novel targets for effective therapy of sepsis as well as other inflammatory conditions

A Case Study of the Development and Implementation of NIACC’s John Pappajohn Entrepreneurial Center

American community colleges are vital centers of educational opportunity. They are the “neighborhood schools of higher education,” as noted by community college scholar, James Palmer; they are close to home and inclusive of their communities (RCCA, 2016, p. 1). Expected to improve their local, regional, and state economies, community colleges develop and implement educational opportunities that meet the immediate and projected needs of local businesses and industry (Dougherty & Bakia, 2000; Kasper, 2003). In 2014, the American Association of Community Colleges (AACC) reported that 45 percent of all undergraduates in the United States enrolled in a community college (AACC, 2016, p. 1). In Iowa alone, approximately one of every four Iowans—between the ages of 18 and 64—annually enrolls within a credit or non-credit community college course. During the 2014-2015 academic year (AY), Iowa’s community colleges served 138,642 credit students and 232,480 non-credit students (IADeptofEd, 2017a, p. 9), and awarded 795,235 Associate Degrees (A.A.) and 494,995 certificates (AACC, 2016, p. 2). Consequently, community colleges are viewed as major drivers of economic growth (Milliron & de los Santos, 2004), and “…fulfill a multiplicity of roles within their communities, offering a myriad of educational programs and services” (Bragg, 2001, p. 93). Community colleges provide specialized training, coursework, and educational opportunities that make them key contributors to local community and economic development efforts (Kasper, 2003). Within this study, there is particular interest in the decisions, plans, and execution of the John Pappajohn Entrepreneurial Center (JPEC) at North Iowa Area Community College (NIACC)—respectively, the implementation of the center. Through a case study method, seven people were interviewed with an eye to understanding the development, continued efforts, and best practices of NIACC’s JPEC. This qualitative approach captured the perspective of the key professionals who played an active role in the development of the JPEC at NIACC, allowing for richness of each participant’s experiences. A case study of a single site interviewing multiple individuals was utilized. The purpose was to speak with key participants involved in the NIACC JPEC development. Specific research questions (RQ) addressed: how and why the center was formed, identification of what the necessary components for implementation were, determination of what considerations are necessary for its continuation, what obstacles are associated with entrepreneurship center (EC) implementation within community colleges, and what are the unique aspects of community college ECs. I utilized an exploratory case study approach that allowed the researcher the opportunity to identify the development, implementation, and organizational workings of NIACC’s John Pappajohn Entrepreneurial Center—in its real-world context. Through theme analysis of the recorded and transcribed interview transcripts, eight primary themes were identified, including: JPEC Beginnings, Structure, Education, Leadership, Measuring Success, Community Involvement, Obstacles, and Uniqueness. This information may be helpful to Iowa’s additional 14 community colleges, as well as those beyond the state of Iowa, to provide guidance throughout the process of entrepreneurial center development

Assessing the Ethical Company

What makes a given organization ethical? Who decides? Can you measure ethics? What criteria should be used for assessment and how? This paper examines these questions by looking at a set of principles for business, a model of ethical organizational culture growing out of those principles and the development and use of an assessment tool built on that model. In the early 1990s, against the horizon of what were then the early stages of globalization, the Center for Ethical Business Cultures (CEBC) at the University of St. Thomas fostered the development of The Minnesota Principles, principles “developed by…business leaders to foster the fairness and integrity of business relationships in the emerging global marketplace.” These principles were designed to serve as a foundation for dialogue by business leaders around the world. Since that time, the principles have had a global impact. The Minnesota Principles provided the substantial basis for the Caux Round Table Principles for Business, now available in sixteen languages and used around the world

Microdissection and Measurement of Polytene Chromosomes Using the Atomic Force Microscope

A method to isolate specific regions of the Drosophila polytene chromosome using an atomic force microscope (AFM) was explored. The AFM was used for the microdissection of the locus of interest with much greater precision than standard microdissection techniques. The amplification of DNA isolated in this fashion by the polymerase chain reaction (PCR) is discussed. A study of the effect of hydration level on gross chromosome structure was carried out. It was shown that chromosome swelling is dependent upon humidity or the buffered medium. The significance of this swelling with respect to studies of chromosome structure under physiological conditions is considered

Modern risk management through the lens of the ethical organizational culture

Most recent efforts to create guidance for modern risk management practices emphasize the importance of connecting risk management policy and practice with an organization’s culture and values. However, identifying or establishing that connection is not widely discussed or understood. What does it mean to state that risk management is an expression of an organization’s values? This article discusses the basis for identifying the connection between organizational values through the lens of the Ethical Organizational Culture and attempts to draw out linkages with current risk management thinking on the subject. The stablishment of a basis of identifying organizational values and their link to risk management policy and practice is illustrated through a case analysis of the Veritas Institute’s Self-Assessment and Improvement methodology

Modern Risk Management: Managing risk through the ethical business culture model

What does it mean to state that risk management is an expression of an organisation’s values? This article discusses the basis for identifying the connection between organisational values through the lens of ethical business culture and attempts to draw out linkages with current risk management thinking. The approach described allows an analytic approach to risk management to be inserted into an ethical assessment method

Building Ethical Business Cultures: BRIC by BRIC

As the economies of Brazil, Russia, India, and China (BRICs) continue to grow both in size and clout, and their resident multinational corporations become major players in global markets, questions pertaining to trust and integrity, and of universally shared standards for ethical business behavior become important concerns for numerous stakeholders. Whether or not managers and employees behave ethically depends on how one defines ethical behavior and applies it to an organization’s culture. We start this article by discussing attributes of ethical business behavior and cultures in each of the four BRICs countries, and then present results of our recent large scale survey-based studies, comparing managers’ and employees’ perceptions of ethical cultures in BRICs and in economically developed Western economies

Visualization of circular DNA molecules labeled with colloidal gold spheres using atomic force microscopy

We have imaged gold‐labeled DNA molecules with the atomic force microscope(AFM). Circular plasmid DNA was labeled at internal positions by nick‐translation using biotinylated dUTP. For visualization, the biotinylated DNA was linked to streptavidin‐coated colloidal gold spheres (nominally 5 nm diam) prior to AFM imaging. Reproducible images of the labeled DNA were obtained both in dry air and under propanol. Height measurements of the DNA and colloidal gold made under both conditions are presented. The stability of the DNA‐streptavidin colloidal gold complexes observed even under propanol suggests that this labeling procedure could be exploited to map regions of interest in chromosomal DNA

Analyzing Chromosomes, Ion Channels and Novel Nucleic Acid Structures by AFM

The atomic force microscope (AFM) is proving to be a powerful tool for analysis of biological samples. We provide three examples of the application of AFM to the study of biological questions. First, polytene chromosomes from Drosophila are imaged and manipulated by the AFM. Second, the localization of calcium channels on the release face of a nerve terminal is described. Finally, analyses of a new form of DNA, the G-wire, is presented. These examples illustrate the wide variety of biological questions to which AFM can contribute