Altered Irinotecan Pharmacokinetics in Diet-Induced Obesity

Title: Altered Irinotecan Pharmacokinetics in Diet-Induced Obesity Authors: Pranav Shah*, Adarsh Gandhi and Romi Ghose Affiliation: University of Houston, Department of Pharmacological and Pharmaceutical Sciences, Houston, TX - 77030 Purpose: Irinotecan (CPT-11) is a topoisomerase I inhibitor that has been shown to be highly effective in treatment of colon, stomach, pancreas, and non-small cell lung cancers. It has recently been shown that CPT-11 administration is associated with liver toxicity and this effect is compounded by baseline obesity. It was found that patients with a BMI index of \u3e25 were twice as much susceptible to developing liver toxicity than patients with BMI index of Methods: For drug metabolism studies, liver S9 fractions were prepared from diet-induced obese (DIO, 60% Kcal diet fed mice) and lean mice (10% Kcal diet fed mice). UGT1A-mediated metabolism of SN-38 was determined in liver S9 fractions (2 mg/ml protein) incubated with SN-38 (15 µmol) for 60 min. For pharmacokinetic studies, mice were injected with a single oral dose of 10 mg/kg CPT-11 and blood samples were collected from 0-480 minutes. Plasma and feces samples were analyzed for CPT-11 and SN-38 concentrations using LC-MS/MS. Liver tissues were harvested for real-time PCR studies. The mRNA and serum TNF-α levels were measured in liver and plasma samples, respectively. Results: We found that the rate of formation of SN-38G was ~2 fold lower in the DIO mice compared to the lean controls. This corresponded with reduced expression of UGT1A1 in DIO mice livers. We did not observe significant changes in the area under the curve (AUC) or clearance of CPT-11 between the DIO and lean mice. However, plasma and fecal exposure (AUC) of SN-38 was increased by ~2 folds in the DIO mice compared to the lean controls. We also observed significantly higher mRNA and serum levels of TNF-α in the DIO mice as compared to the lean mice. Higher TNF-α levels are known to be associated with liver toxicity. Conclusion: CPT-11 dosage should be closely monitored for effective and safe chemotherapy in obese patients who are at a higher risk of developing liver toxicity

A Data Scientific Approach Towards Predictive Maintenance Application in Manufacturing Industry

Most industries have recently started to harness the power of data to assess their performance and improve their production systems for future competitiveness and sustainability. Therefore, utilization of data for obtaining insights through data-driven approaches is invading every domain of industrial applications. Predictive maintenance (PdM) is one of the highest impacted industrial use cases in data-driven applications due to its ability to predict machine failures by implementing machine learning algorithms. This study aims to propose a systematic data scientific approach to provide valuable insights by analysing industrial alarm and event log data, which might further be used for investigation in root cause understanding and planning of necessary maintenance activities. To do that, a Cross-Industry Standard Process for Data Mining (CRISP-DM) is followed as a reference model in this study. The results are presented by first understanding the relationship between alarms and product types being processed in the selected machines by using exploratory data analysis (EDA). Along with this, the behavior of problematic alarms is identified. Afterward, a predictive analysis formulated as a multi-class classification problem is performed using various Machine Learning (ML) models to predict the category of alarm and generate rules to be used for further investigation in maintenance planning. The performance of the developed models is evaluated based on the different metrics and the decision tree model is selected with the higher accuracy score among them. As a theoretical contribution, this study presents an implementation of predictive modeling in a structured way, which uses a systematic data scientific approach based on industrial alarm and event log data. On the other hand, as a practical contribution, this study provides a set of decision rules that can act as decision support for further exploration of possible in-depth root causes through the other contextual data, and hence it gives an initial foundation towards PdM application in the case company

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Role of Inflammation in Metabolism and Hepatotoxicity of Prototypical Drugs

Inflammation leads to altered drug metabolism and augments drug-induced hepatotoxicity. However, the mechanisms are not fully understood. Inflammation is known to alter the gene expression and activity of several key drug metabolizing enzymes (DMEs) further compromising the safety and efficacy of drugs. Inflammatory responses in the liver are primarily mediated by the Toll-like receptors (TLRs), found on the cell surface of hepatocytes and immune cells, such as Kupffer cells. TLR2 and TLR4, upon activation recruit the first adaptor molecule, the Toll-interleukin 1 receptor adaptor protein (TIRAP). TIRAP was shown to play a differential role in regulating gene expression of Cyp3a11 in lipopolysaccharide- (LPS, TLR4 agonist) or lipoteichoic acid- (LTA, TLR2 agonist) induced inflammation in mice. In this project, we studied the role of the TLR signaling pathway in regulating inflammation-mediated altered drug metabolism and hepatotoxicity of prototypical drugs in mice. Aim 1: We observed significant down-regulation of Cyp3a11 protein expression and activity (measured by midazolam (MDZ) metabolism) in LPS- or LTA-induced inflammation. This further correlated with reduced clearance and increased pharmacological action of MDZ in both, LPS- or LTA-treated mice. In contrast to LPS, TIRAP was involved only in LTA-mediated down regulation of Cyp3a11 and metabolism of MDZ. Aim 2: We studied the effects of inflammation and the TLR signaling pathway in regulating hepatotoxicity of chlorpromazine (CPZ). We selected CPZ due to its idiosyncratic hepatotoxicity reported in humans and animals. Sustained activation of the kinase, c-Jun-N-terminal kinase (JNK) is associated with cell death. However, the roles of JNK and TIRAP in regulating CPZ-induced hepatotoxicity in presence of inflammation are unknown. In our in vivo study, LPS or LTA treatment led to significant induction of CPZ hepatotoxicity with increase in serum tumor necrosis factor (TNF)-α and sustained activation of JNK upto 4 h. Histopathological analysis revealed marked depletion of hepatic glycogen in LPS/CPZ or LTA/CPZ groups. Also, CPZ-induced hepatotoxicity, increase in serum TNF-α and activation of JNK in presence of LPS or LTA were dependent on TIRAP. In our in vitro study, we observed significant induction of alanine aminotransferase (ALT) in CPZ-treated primary mouse hepatocytes in presence of LPS or LTA. We observed prolonged activation of JNK in presence TNF-α and TNF-α-mediated increase in ALT in presence of CPZ was attenuated by the JNK inhibitor. Overall, by focusing on the TLR signaling pathway, this dissertation provides novel insights in the underlying mechanisms involved in regulating drug metabolism and hepatotoxicity of prototypical drugs in inflammation.Pharmacological and Pharmaceutical Sciences, Department o

Differential Role of Toll-Interleukin 1 Receptor Domain-Containing Adaptor Protein in Toll-Like Receptor 2-Mediated Regulation of Gene Expression of Hepatic Cytokines and Drug-Metabolizing EnzymesS⃞

Pharmacological activities of drugs are impaired during inflammation because of reduced expression of hepatic drug-metabolizing enzyme genes (DMEs) and their regulatory nuclear receptors (NRs): pregnane X receptor (PXR), constitutive androstane receptor (CAR), and retinoid X receptor (RXRα). We have shown that a component of Gram-positive bacteria, lipoteichoic acid (LTA) induces proinflammatory cytokines and reduces gene expression of hepatic DMEs and NRs. LTA is a Toll-like receptor 2 (TLR2) ligand, which initiates signaling by recruitment of Toll-interleukin 1 receptor domain-containing adaptor protein (TIRAP) to the cytoplasmic TIR domain of TLR2. To determine the role of TIRAP in TLR2-mediated regulation of DME genes, TLR2(+/+), TLR2(−/−), TIRAP(+/+), and TIRAP(−/−) mice were given LTA injections. RNA levels of the DMEs (Cyp3a11, Cyp2b10, and sulfoaminotransferase), xenobiotic NRs (PXR and CAR), and nuclear protein levels of the central NR RXRα were reduced ∼50 to 60% in LTA-treated TLR2(+/+) but not in TLR2(−/−) mice. Induction of hepatic cytokines (interleukin-1β, tumor necrosis factor-α, and interleukin-6), c-Jun NH2-terminal kinase, and nuclear factor-κΒ was blocked in TLR2(−/−) mice. As expected, expression of hepatic DMEs and NRs was reduced by LTA in TIRAP(+/+) but not in TIRAP(−/−) mice. Of interest, cytokine RNA levels were induced in the livers of both the TIRAP(+/+) and TIRAP(−/−) mice, whereas LTA-mediated induction of serum cytokines was attenuated in TIRAP(−/−) mice. LTA-mediated down-regulation of DME genes was attenuated in hepatocytes from TLR2(−/−) or TIRAP(−/−) mice and in small interfering RNA-treated hepatocytes. Thus, the effect of TLR2 on DME genes in hepatocytes was mediated by TIRAP, whereas TIRAP was not involved in mediating the effects of TLR2 on cytokine expression in the liver

Global Warming and Community Outreach (Semester Unknown) IPRO 331: Global Warming and Community Outreach IPRO331 Poster F09

a. Utilize previous presentations to inform and educate the public about the cause, impact, and responses to global warming. b. Divide the overall issue into major aspects including the ones set by previous IPRO’s while adding our own. c. Present material to larger and more diverse audiences in order to create widespread awareness about global warming. d. Focus on solid, scientific data from credible sources that define why and how global warming is occurring, rather than discussing the politics and economics that surround the issue. e. Add the most recent updates to the presentation such as climate engineering. f. Use previous research and presentations but taking a more systematic approach when creating presentations. g. Devise a creative ways to interact with the audience such as skits and demonstrations. h. Provide surveys at the end of each presentation to receive feedback. i. Update an existing IPRO 331 website and enhance it with updated research. Also to make is more accessible to the general public and make them aware of it. This will allow anyone anywhere to access information on global warming. j. Enhance team members’ public speaking abilities via interactions during class along with mock presentations throughout the semester. k. Each member is expected to organize and present at three different locations per month at least throughout the semester.Deliverable

Global Warming and Community Outreach (Semester Unknown) IPRO 331: Global Warming and Community Outreach IPRO331 MidTerm Presentation F09

a. Utilize previous presentations to inform and educate the public about the cause, impact, and responses to global warming. b. Divide the overall issue into major aspects including the ones set by previous IPRO’s while adding our own. c. Present material to larger and more diverse audiences in order to create widespread awareness about global warming. d. Focus on solid, scientific data from credible sources that define why and how global warming is occurring, rather than discussing the politics and economics that surround the issue. e. Add the most recent updates to the presentation such as climate engineering. f. Use previous research and presentations but taking a more systematic approach when creating presentations. g. Devise a creative ways to interact with the audience such as skits and demonstrations. h. Provide surveys at the end of each presentation to receive feedback. i. Update an existing IPRO 331 website and enhance it with updated research. Also to make is more accessible to the general public and make them aware of it. This will allow anyone anywhere to access information on global warming. j. Enhance team members’ public speaking abilities via interactions during class along with mock presentations throughout the semester. k. Each member is expected to organize and present at three different locations per month at least throughout the semester.Deliverable

Global Warming and Community Outreach (Semester Unknown) IPRO 331: Global Warming and Community Outreach IPRO331 Final Presentation F09

a. Utilize previous presentations to inform and educate the public about the cause, impact, and responses to global warming. b. Divide the overall issue into major aspects including the ones set by previous IPRO’s while adding our own. c. Present material to larger and more diverse audiences in order to create widespread awareness about global warming. d. Focus on solid, scientific data from credible sources that define why and how global warming is occurring, rather than discussing the politics and economics that surround the issue. e. Add the most recent updates to the presentation such as climate engineering. f. Use previous research and presentations but taking a more systematic approach when creating presentations. g. Devise a creative ways to interact with the audience such as skits and demonstrations. h. Provide surveys at the end of each presentation to receive feedback. i. Update an existing IPRO 331 website and enhance it with updated research. Also to make is more accessible to the general public and make them aware of it. This will allow anyone anywhere to access information on global warming. j. Enhance team members’ public speaking abilities via interactions during class along with mock presentations throughout the semester. k. Each member is expected to organize and present at three different locations per month at least throughout the semester.Deliverable

Global Warming and Community Outreach (Semester Unknown) IPRO 331: Global Warming and Community Outreach IPRO331 Final Report F09

a. Utilize previous presentations to inform and educate the public about the cause, impact, and responses to global warming. b. Divide the overall issue into major aspects including the ones set by previous IPRO’s while adding our own. c. Present material to larger and more diverse audiences in order to create widespread awareness about global warming. d. Focus on solid, scientific data from credible sources that define why and how global warming is occurring, rather than discussing the politics and economics that surround the issue. e. Add the most recent updates to the presentation such as climate engineering. f. Use previous research and presentations but taking a more systematic approach when creating presentations. g. Devise a creative ways to interact with the audience such as skits and demonstrations. h. Provide surveys at the end of each presentation to receive feedback. i. Update an existing IPRO 331 website and enhance it with updated research. Also to make is more accessible to the general public and make them aware of it. This will allow anyone anywhere to access information on global warming. j. Enhance team members’ public speaking abilities via interactions during class along with mock presentations throughout the semester. k. Each member is expected to organize and present at three different locations per month at least throughout the semester.Deliverable