Outcomes of Acute Pancreatitis in Hospitalized Patients With Generalized Anxiety Disorder.

Introduction Acute pancreatitis (AP) is a common cause of hospitalization in the United States. There is evidence that chronic stress increases the risk for more severe AP episodes. One common form of chronic stress is generalized anxiety disorder (GAD). The purpose of this research was to investigate the impact of GAD on the outcomes of adult patients admitted to the hospital with AP. Methods Utilizing the 2014 National Inpatient Sample database and International Classification of Diseases, Ninth Edition Revision (ICD) codes, AP patients were selected. Common inpatient outcomes of AP patients with and without GAD were examined. The outcomes studied were acute renal failure, acute respiratory failure, sepsis, acute deep vein thrombosis, myocardial infarction, intestinal perforation, and inpatient mortality. A multivariate logistic regression analysis was conducted to assess if GAD was an independent predictor for these outcomes. Results Among 82,156 adult patients hospitalized for AP during the 2014 year, 10,611 of them had coexisting GAD. AP patients with comorbid GAD were found to have an increased likelihood of acute renal failure (aOR = 1.19, 95% confidence interval (CI) = 1.11-1.28, p < 0.001), sepsis (aOR = 1.09, 95% CI = 1.01 -1.19, p = 0.037), acute deep vein thrombosis (aOR = 1.63, 95% CI = 1.06-2.50, p = 0.025), and inpatient mortality (aOR = 1.62, 95% C = I 1.27-2.08, p < 0.001). There was no statistically significant difference found between the two cohorts for the outcomes of myocardial infarction and intestinal perforation. Conclusion In patients hospitalized with AP, those with coexisting GAD were found to have an increased risk of developing acute renal failure, sepsis, acute deep vein thrombosis, and inpatient mortality. There may be benefits to identifying AP patients with comorbid GAD at the time of admission and monitoring them more carefully during their hospitalization to help identify early signs of complications or prevent the negative outcomes seen in this study.

Analysis of the economic burden of docusate sodium at a United States tertiary care center.

OBJECTIVES: The primary objective was to determine the financial resources allocated to docusate at a representative U.S. tertiary care center. Secondary objectives included comparing docusate utilization between two tertiary care centers, and exploring alternative uses for the funds spent on docusate. METHODS: The study population included all patients 18 years and older admitted to University Hospital in Newark, New Jersey. Every scheduled docusate prescription for the study population between January 1st, 2015 and December 31st, 2019 was collected. The annual total cost associated with docusate use per year was calculated. The 2015 data from this study and a 2015 McGill University Health Centre study were compared. Also, alternative uses for the money utilized on docusate were assessed. RESULTS: Over the study period, 37,034 docusate prescriptions and 265,123 docusate doses were recorded. The average cost of prescribing docusate was $25,624.14 per year and $49.37 per hospital bed per year. A comparison between the 2015 data of University Hospital and McGill showed that McGill prescribed 107 doses and spent $10.09 more per hospital bed than University Hospital. Finally, alternative uses for the average yearly spending on docusate equated to 0.35 the salary of a nurse, 0.51 the salary of a secretary, 20.66 colonoscopies, 27.00 upper endoscopies, 186.71 mammograms, 1,399.37 doses of polyethylene glycol 3350, 3,826.57 doses of lactulose, or 4,583.80 doses of psyllium. CONCLUSION: A single average size tertiary care hospital spent about $25,000 yearly on docusate despite its lack of clinical effectiveness. While this amount is small compared to an overall hospital budget, when considering likely comparable docusate use at the U.S's 6,090 hospitals, the economic burden of docusate becomes significant. The funds currently being used on docusate could be redirected to alternative, more cost-effective purposes.

Outcomes of Gastroparesis in Hospitalized Patients With Generalized Anxiety Disorder.

Background   Gastroparesis is a common gastrointestinal pathology that has been increasing in prevalence and represents a significant cost to the United States healthcare system. Gastroparesis is associated with psychological dysfunction, including generalized anxiety disorder (GAD). GAD is known to be a prevalent and chronic manifestation of anxiety, which has been increasing in prevalence since the year 2020. Despite the association between gastroparesis and GAD, there has been limited research on the possible impact GAD may have on the morbidity and mortality of patients hospitalized for gastroparesis, which is further evaluated in this study.   Methods   Using the Nationwide Inpatient Sample from the year 2014, a retrospective study was conducted to assess the outcomes of hospitalized gastroparesis patients with and without a history of GAD. In this study, the analyzed outcomes included acute kidney injury (AKI), acute respiratory failure, sepsis, acute deep vein thrombosis, myocardial infarction, intestinal obstruction, and inpatient mortality. To assess whether GAD is an independent risk factor for the outcomes, a multivariate logistic regression analysis was used.   Results   There were 22,150 patients with gastroparesis assessed in this study; GAD was found to be a comorbid diagnosis in 4,196 of those patients. In the GAD cohort, there was an elevated risk for AKI (adjusted odds ratio 1.24, p < 0.001). The adjusted odds ratios for acute respiratory failure, sepsis, acute deep vein thrombosis, myocardial infarction, intestinal obstruction, and inpatient mortality did not meet the threshold for statistical significance.   Conclusion   In hospitalized gastroparesis patients, GAD is a risk factor for AKI. This finding may be attributed to prerenal azotemia due to an increased risk of nausea and vomiting associated with GAD, as well as the medications used to treat GAD such as escitalopram and duloxetine. In addition, the dual inflammatory states caused by the co-existence of both GAD and gastroparesis may also have a role in increasing the risk for AKI. The results of this study may become increasingly relevant given the increasing prevalence of GAD. .

Outcomes of Patients Hospitalized for Acute Diverticulitis With Comorbid Generalized Anxiety Disorder.

Introduction Diverticular disease and anxiety disorders are common in the general population. Prior research on diverticular disease showed that these patients have an increased frequency of anxiety and depression. The objective of this study was to explore the impact of generalized anxiety disorder (GAD) on the outcomes of adult patients admitted with acute diverticulitis. Methods Using the National Inpatient Sample database from the year 2014 and International Classification of Diseases, Ninth Edition Revision, Clinical Modification (ICD-9 CM) codes, acute diverticulitis patients were selected. The outcomes of diverticulitis patients with and without GAD were explored. The outcomes of interest included inpatient mortality, hypotension/shock, acute respiratory failure, acute hepatic failure, sepsis, intestinal abscess, intestinal obstruction, myocardial infarction, acute renal failure, and colectomy. A multivariate logistic regression analysis was performed to determine if GAD is an independent predictor for the outcomes. Results Among 77,520 diverticulitis patients in the study, 8,484 had comorbid GAD. GAD was identified as a risk factor for intestinal obstruction (adjusted odds ratio (aOR) 1.22, 95% CI: 1.05-1.43, p<0.05), and intestinal abscess (aOR 1.19, 95% CI: 1.10-1.29, p<0.05). GAD was found to be a protective factor for hypotension/shock (aOR 0.83, 95% CI: 0.76-0.91, p<0.05) and acute respiratory failure (aOR 0.76, 95% CI: 0.62-0.93, p<0.05). The aORs of sepsis, inpatient mortality, myocardial infarction, acute renal failure, and colectomy were not statistically significant. Conclusions Patients with acute diverticulitis who are also diagnosed with GAD are at increased risk for intestinal obstruction and intestinal abscess, which may be due to the influence GAD has on the gut microbiota as well as the impact of GAD pharmacotherapy on gut motility. There was also a decreased risk for acute respiratory failure and hypotension/shock appreciated in the GAD cohort which may be attributable to the elevated healthcare resource utilization seen generally in GAD patients, which may allow for presentation to the emergency department, hospitalization, and treatment earlier in the diverticulitis disease course.

Pre-existing Opioid Use Worsens Outcomes in Patients With Diverticulitis.

Background and objective Diverticulitis occurs in 10-25% of patients with diverticulosis. Although opioids can decrease bowel motility, there is scarce data on the effect of chronic opioid use on the outcomes of diverticulitis. In this study, we aimed to explore the outcomes of diverticulitis in patients with pre-existing opioid use. Methods Data between 2008 and 2014 from the National Inpatient Sample (NIS) database was extracted using the International Classification of Diseases, 9th Revision (ICD-9) codes. Univariate and multivariate analyses were used to generate odds ratios (OR). Elixhauser Comorbidity Index (ECI) scores predicting mortality and readmission were calculated based on weighted scores from 29 different comorbidities. Scores were compared between the two groups using univariate analysis. Inclusion criteria included patients with a primary diagnosis of diverticulitis. Exclusion criteria included patients less than 18 years of age, and a diagnosis of opioid use disorder in remission. Studied outcomes included inpatient mortality, complications (including perforation, bleeding, sepsis event, ileus, abscess, obstruction, and fistula), length of hospital stay, and total costs.  Results A total of 151,708 patients with diverticulitis and no active opioid use and 2,980 patients with diverticulitis and active opioid use were hospitalized in the United States from 2008 to 2014. Opioid users had a higher OR for bleeding, sepsis, obstruction, and fistula formation. Opioid users had a lower risk of developing abscesses. They had longer lengths of stay, higher total hospital charges, and higher Elixhauser readmission scores. Conclusion Hospitalized diverticulitis patients with comorbid opioid use are at an elevated risk of in-hospital mortality and sepsis. This could be attributed to complications from injection drug use predisposing opioid users to these risk factors. Outpatient providers caring for patients with diverticulosis should consider screening their patients for opioid use and try offering them medication-assisted treatment to reduce their risk of poor outcomes.

Outcomes of Patients Who Developed Clostridioides difficile Infection During Hospitalization and Had a History of Comorbid Post-Traumatic Stress Disorder.

INTRODUCTION: Clostridioides difficile (C. difficile), is a common cause of nosocomial diarrhea. Antibiotic use is a risk factor for developing C. difficile infection (CDI). Clinical presentations of CDI range from mild diarrhea to fulminant colitis. A history of anxiety increases the risk of developing irritable bowel syndrome following CDI. Post-traumatic stress disorder (PTSD) is a common form of anxiety and is associated with several medical comorbidities. This study explores the impact PTSD has on the outcomes of adult patients who develop CDI while hospitalized. METHODS: Hospitalized adults who had developed CDI were selected from the 2014 National Inpatient Sample database using the International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9) codes. The outcomes of CDI patients with and without comorbid PTSD were explored. The outcomes assessed in this study were inpatient mortality, sepsis, hypotension/shock, acute renal failure, acute respiratory failure, megacolon, colonic perforation, and intestinal abscess. Independent t-tests and chi-squared tests were used to compare means and proportions, respectively. A multivariate logistic regression analysis was utilized to determine whether PTSD is an independent predictor of the outcomes. RESULTS: Among 72,383 hospitalized adults who developed CDI in the year 2014, 465 also had a diagnosis of PTSD. PTSD was found to be an independent risk factor for inpatient mortality (adjusted odds ratio {aOR} 2.93, 95% confidence interval (CI): 1.39-6.21, p = 0.005), and sepsis (aOR 1.61, 95% CI: 1.24-2.07, p = 0.001). However, PTSD was not a risk factor for hypotension/shock (aOR 1.26, 95% CI: 0.97-1.63, p = 0.080), acute renal failure (aOR 1.02, 95% CI: 0.81-1.28, p = 0.895), or acute respiratory failure (aOR 1.15, 95% CI: 0.83-1.58, p = 0.412) in patients with CDI. Due to small sample sizes of patients who developed megacolon, colonic perforation, and intestinal abscess, further analysis of these outcomes was not performed. CONCLUSION: Inpatients who develop CDI with comorbid PTSD are at increased risk for sepsis and inpatient mortality. These findings may be due to the impact of PTSD's dysregulation of the hypothalamic-pituitary axis leading to low cortisol production, increased serum cytokine concentrations, and/or increased intestinal inflammation. Awareness of these increased risks when triaging CDI patients with PTSD and possibly increased psychiatric interventions to treat PTSD may be necessary to help reduce the risk of sepsis and inpatient mortality in this subgroup of patients.

Outcomes of Inflammatory Bowel Disease in Hospitalized Patients With Generalized Anxiety Disorder.

Background The development of inflammatory bowel disease (IBD), which encompasses ulcerative colitis and Crohn's disease, is multifactorial. Stress from anxiety is a risk factor for IBD. Generalized anxiety disorder (GAD) is twice as likely in IBD patients. This study explores the outcomes of patients hospitalized for IBD with comorbid GAD. Methods A retrospective analysis utilizing the 2014 USA National Inpatient Sample database was performed to assess the outcomes of hospitalized IBD patients with and without GAD. The outcomes analyzed were sepsis, acute hepatic failure, hypotension/shock, acute respiratory failure, acute deep vein thrombosis, acute renal failure, intestinal obstruction, myocardial infarction, ileus, inpatient mortality, colectomy, intestinal abscess, intestinal perforation, and megacolon. A multivariate logistic regression analysis was employed to explore whether GAD is a risk factor for these outcomes. Results Among 28,173 IBD hospitalized patients in the study, GAD was a comorbid diagnosis in 3,400 of those patients. IBD patients with coexisting GAD were found to be at increased risk for acute hepatic failure (adjusted odds ratio (aOR) 1.80, p = 0.006), sepsis (aOR 1.33, p < 0.001), acute respiratory failure (aOR 1.24, p = 0.018), inpatient mortality (aOR 1.87, p < 0.001), intestinal abscess (aOR 2.35, p = 0.013), and intestinal perforation (aOR 1.44, p = 0.019). The aORs for the remaining outcomes were not statistically significant. Conclusions In hospitalized IBD patients, GAD is a risk factor for sepsis, acute hepatic failure, acute respiratory failure, intestinal abscess, intestinal perforation, and inpatient mortality. IBD and GAD are becoming increasingly common, which will likely lead to a larger number of complications among inpatients with these comorbidities.

Rare Jejunal Large B-cell Lymphoma Mimicking a Crohn's Disease Without Terminal Ileum Involvement.

This case describes a 49-year-old man who presented with a several-month history of melena, and unintentional weight loss. Prior esophagogastroduodenoscopy and colonoscopy were unrevealing. Further evaluation with capsule endoscopy showed patchy erythematous mucosa in the jejunum creating suspicion for Crohn's Disease. Subsequent push enteroscopy found nodular and congested patchy mucosa of jejunum, and stigmata of bleeding in the proximal and mid-jejunum. Repeat colonoscopy showed a diffuse area of erythematous mucosa in the recto-sigmoid colon, and moderately congested mucosa in the ascending colon, but a normal terminal ileum. A small bowel biopsy eventually revealed large B-cell lymphoma. This is one of the first seven reported cases of small bowel lymphoma mimicking Crohn's Disease and the first to not have any ileal involvement.

A Rare Case of an Incidental Near-Complete Gastric Band Erosion Without Perforation.

Laparoscopic gastric banding has been favored for the treatment of morbid obesity because it is minimally invasive, effective, and reversible. One of the complications is gastric band erosion which can cause abdominal pain, hematemesis, and hematochezia. Erosions can be partial and can lead to intra-abdominal free air, peritonitis, and sepsis. Endoscopic removal of the gastric band can be done safely and effectively using a wire and a mechanical lithotripter. We describe a patient with a rare case of an incidental near-complete gastric band erosion without perforation and subsequent endoscopic removal.

Outcomes of Hospitalized Acute Alcoholic Hepatitis (AH) in Patients With Bipolar 1 Disorder (B1D).

INTRODUCTION: Alcoholic hepatitis (AH) is a common cause of hospital admissions and is associated with a high mortality rate. AH occurs frequently in patients with heavy alcohol use. Alcohol use disorder (AUD) commonly presents with comorbid psychiatric disorders such as bipolar disorder. Bipolar disorder patients are also known to be at an increased risk for chronic liver diseases. Bipolar 1 disorder (B1D) is often considered the most severe presentation among different types of bipolar disorder. This study assesses the clinical outcomes of patients admitted for AH with concomitant B1D. METHODS: Adult patients with AH were identified within the 2014 National Inpatient Sample (NIS) database. International Classification of Diseases, Ninth Edition Revision, Clinical Modification (ICD-9 CM) codes were used to select for all of the diagnoses for this study. AH patients were subdivided into those with and without B1D. The outcomes of interest were sepsis, hepatic encephalopathy, acute respiratory failure, acute kidney injury, ischemic stroke, hepatic failure, coagulopathy, and inpatient mortality. A multivariate logistic regression analysis was performed to explore whether B1D is an independent predictor for the outcomes. RESULTS: Among 4,453 patients with AH identified, 166 patients also had B1D. AH patients with comorbid B1D were seen to be younger (42.9 years old vs. 46.2 years old, p < 0.05) and more commonly female (55.4% vs. 36.5%, p < 0.05). The B1D subgroup of AH patients were found to less likely develop acute hepatic failure (adjusted odds ratio (aOR) 0.13, 95% confidence interval (CI): 0.02-0.97, p < 0.05). The adjusted odds ratios for the remaining outcomes were not statistically significant. CONCLUSIONS: Our study indicates that B1D may be an independent protective factor against acute hepatic failure in patients hospitalized with AH. This finding can be explained by frequent laboratory monitoring and psychiatric assessments performed by psychiatrists treating B1D patients, as well as the impact B1D has on cortisol release induced by hyperactivity of the hypothalamic-pituitary-adrenal (HPA) axis.

Malnutrition Is Associated With Worse Outcomes of Inpatient Endoscopic Retrograde Cholangiopancreatography.

Objectives Endoscopic retrograde cholangiopancreatography (ERCP) is frequently used to manage pancreaticobiliary disorders in an inpatient setting. Malnutrition is prevalent among hospitalized patients, and it is generally associated with poor clinical outcomes. However, there is a lack of studies on how malnutrition affects the outcomes of inpatient ERCP. Thus, we investigated the outcomes of inpatient ERCP among patients with malnutrition. Methods Adult patients who underwent ERCP from the 2014 National Inpatient Sample database were selected to conduct retrospective analysis. Patient demographics and outcomes of ERCP were compared between the groups with and without malnutrition. The outcomes of interest were inpatient mortality, length of stay, total hospital charge, and ERCP complications, including pancreatitis, cholecystitis, cholangitis, sepsis, hemorrhage, and intestinal perforation. Results Patients with malnutrition had longer length of stay (15.5 days vs. 6.7 days, p < 0.05) and higher total hospital charge ($149,699 vs. $71,723, p < 0.05). Malnutrition was an independent risk factor for inpatient mortality (adjusted odds ratio (aOR) 2.54, 95% confidence interval (CI): 1.70-3.82, p < 0.05), sepsis (aOR 2.20, 95% CI: 1.82-2.65, p < 0.05), hemorrhage (aOR 1.64, 95% CI: 1.05-2.56, p < 0.05), and intestinal perforation (aOR 4.29, 95% Cl:1.61-11.46, p < 0.05). Conclusions Our study indicates that patients with malnutrition are more likely to have worse outcomes, such as increased inpatient mortality, sepsis, hemorrhage, and intestinal perforation. Understanding the nutrition status of patients undergoing ERCP can be a useful approach for risk stratification and determining if closer surveillance of the complications is warranted.

Outcomes of Upper Gastrointestinal Bleeding in Hospitalized Patients With Generalized Anxiety Disorder.

Background Upper gastrointestinal bleeding (UGIB) has a high morbidity and mortality. Social deprivation is a risk factor for UGIB and is associated with anxiety. The primary pharmaceutical therapeutic agents for anxiety are selective serotonin reuptake inhibitors. Anxiety is prevalent in the general population and generalized anxiety disorder (GAD) is a common form of anxiety. This study explores the impact of GAD on the outcomes of adult patients hospitalized with UGIB. Methods Adult UGIB patients were selected utilizing the National (Nationwide) Inpatient Sample database from year 2014 and International Classification of Diseases, Ninth Revision, Clinical Modification (ICD-9-CM) codes. The outcomes of UGIB patients with and without GAD were investigated. The outcomes explored include inpatient mortality, hypotension/shock, acute renal failure, acute hepatic failure, acute respiratory failure and acute myocardial infarction. A multivariate logistic regression analysis was used to determine if GAD is an independent predictor of the outcomes. Results Among 19,850 UGIB patients studied, 2357 had comorbid GAD. GAD was identified as a risk factor for acute renal failure (adjusted odds ratio [aOR] 1.37, 95% confidence interval [CI] 1.30-1.57, p < 0.05) and inpatient mortality (aOR 1.50, 95% CI 1.01-2.06, p < 0.05). The aORs of hypotension/shock, acute hepatic failure, acute respiratory failure and acute myocardial infarction were not statistically significant. Conclusion UGIB patients with comorbid GAD are at elevated risk of inpatient mortality and acute renal failure. These results may gain increasing relevance as GAD prevalence has increased since the start of the coronavirus disease 2019 (COVID-19) pandemic.

Cannabis Use Is Associated With an Increased Risk of Intestinal Obstruction in Patients Hospitalized With Diverticulitis.

Objectives Diverticulitis is a common cause of hospitalization. The use of substances such as tobacco and alcohol can predispose patients to diverticulitis, and smoking is also associated with an increased risk of diverticulitis complications. Cannabis availability is growing in the United States, but there is a lack of data on the effects of cannabis use on the outcomes of diverticulitis. Thus, we investigated the effects of cannabis use on diverticulitis outcomes. Methods A retrospective analysis was conducted using 2014 data from the National Inpatient Sample. Patient demographics and outcomes of diverticulitis were compared between the groups with and without a history of cannabis use. The outcomes of interest were inpatient mortality, length of stay, total hospital charge, intestinal obstruction, shock/hypotension, colectomy, intestinal abscess, intestinal fistula, and intestinal perforation. Results Among 48,214 patients with diverticulitis, 447 patients had a history of cannabis use. Patients with a history of cannabis use were younger, more likely to be male, less likely to be White, had a lower Charlson Comorbidity Index, and had shorter hospital stays. There were no significant differences in inpatient mortality and total hospital charge. After adjusting for age, sex, race, and the Charlson Comorbidity Index, cannabis use was an independent risk factor for intestinal obstruction in patients hospitalized with diverticulitis. There were no statistically significant differences in other outcomes. Conclusions This study indicates that patients hospitalized with diverticulitis with a history of cannabis use are more likely to have an intestinal obstruction. Inhibition of gastrointestinal motility by cannabis in the setting of diverticular inflammation may explain this finding.

Inpatient Outcomes of Acute Pancreatitis Among Patients With Systemic Lupus Erythematosus: A Nationwide Analysis.

Objectives This study explores the characteristics and outcomes, including inpatient mortality, length of stay, and pancreatitis complications in patients hospitalized with acute pancreatitis (AP) with coexisting systemic lupus erythematosus (SLE). Methods Patients hospitalized with AP from the National Inpatient Sample from 2014 were selected. Patient characteristics and outcomes of AP were compared between the groups with and without SLE. Age, sex, race, Elixhauser Comorbidity Index (ECI), and etiologies of pancreatitis were measured. The outcomes of interest were inpatient mortality, length of stay, and complications, including respiratory failure, acute renal failure, myocardial infarction, hypotensive shock, sepsis, stroke, and ileus. Chi-squared tests and independent t-tests were used to compare proportions and means, respectively. Multivariate logistic regression analysis was performed to determine if SLE is an independent predictor for the outcomes, adjusting for age, sex, race, ECI, and etiologies of pancreatitis. Results Among 434,280 AP patients identified in the study, 3,015 patients had SLE. Among patients hospitalized with AP, those with SLE were younger, more likely to be female, more likely to be non-White, had higher ECI, and stayed longer in the hospital. Patients without SLE were more likely to have a history of cholelithiasis, alcohol abuse, and hypertriglyceridemia. AP patients presenting with SLE were at higher risk for respiratory failure, acute renal failure, hypotensive shock, stroke, and sepsis. Higher inpatient mortality was also associated with coexisting SLE. Conclusions Patients admitted for AP with SLE have worse outcomes compared to those without SLE. Understanding the potential effects of SLE on AP and optimizing patient care in this population accordingly may improve the quality of care and outcomes.

Hypervirulent Klebsiella pneumoniae infection in a woman with a CDH1 gene mutation.

The CDH1 gene, which encodes E-cadherin, may be associated with cancer when mutated, but the significance of mutations in the context of infection is unknown. In this report, we describe a case of disseminated hypervirulent Klebsiella pneumoniae infection in a 49 year old Caucasian woman with a documented CDH1 mutation.

Context-dependent medicinal effects of anabasine and infection-dependent toxicity in bumble bees.

BACKGROUND: Floral phytochemicals are ubiquitous in nature, and can function both as antimicrobials and as insecticides. Although many phytochemicals act as toxins and deterrents to consumers, the same chemicals may counteract disease and be preferred by infected individuals. The roles of nectar and pollen phytochemicals in pollinator ecology and conservation are complex, with evidence for both toxicity and medicinal effects against parasites. However, it remains unclear how consistent the effects of phytochemicals are across different parasite lineages and environmental conditions, and whether pollinators actively self-medicate with these compounds when infected. APPROACH: Here, we test effects of the nectar alkaloid anabasine, found in Nicotiana, on infection intensity, dietary preference, and survival and performance of bumble bees (Bombus impatiens). We examined variation in the effects of anabasine on infection with different lineages of the intestinal parasite Crithidia under pollen-fed and pollen-starved conditions. RESULTS: We found that anabasine did not reduce infection intensity in individual bees infected with any of four Crithidia lineages that were tested in parallel, nor did anabasine reduce infection intensity in microcolonies of queenless workers. In addition, neither anabasine nor its isomer, nicotine, was preferred by infected bees in choice experiments, and infected bees consumed less anabasine than did uninfected bees under no-choice conditions. Furthermore, anabasine exacerbated the negative effects of infection on bee survival and microcolony performance. Anabasine reduced infection in only one experiment, in which bees were deprived of pollen and post-pupal contact with nestmates. In this experiment, anabasine had antiparasitic effects in bees from only two of four colonies, and infected bees exhibited reduced-rather than increased-phytochemical consumption relative to uninfected bees. CONCLUSIONS: Variation in the effect of anabasine on infection suggests potential modulation of tritrophic interactions by both host genotype and environmental variables. Overall, our results demonstrate that Bombus impatiens prefer diets without nicotine and anabasine, and suggest that the medicinal effects and toxicity of anabasine may be context dependent. Future research should identify the specific environmental and genotypic factors that determine whether nectar phytochemicals have medicinal or deleterious effects on pollinators.

Tissue-specific methylation of human insulin gene and PCR assay for monitoring beta cell death.

The onset of metabolic dysregulation in type 1 diabetes (T1D) occurs after autoimmune destruction of the majority of pancreatic insulin-producing beta cells. We previously demonstrated that the DNA encoding the insulin gene is uniquely unmethylated in these cells and then developed a methylation-specific PCR (MSP) assay to identify circulating beta cell DNA in streptozotocin-treated mice prior to the rise in blood glucose. The current study extends to autoimmune non-obese diabetic (NOD) mice and humans, showing in NOD mice that beta cell death occurs six weeks before the rise in blood sugar and coincides with the onset of islet infiltration by immune cells, demonstrating the utility of MSP for monitoring T1D. We previously reported unique patterns of methylation of the human insulin gene, and now extend this to other human tissues. The methylation patterns of the human insulin promoter, intron 1, exon 2, and intron 2 were determined in several normal human tissues. Similar to our previous report, the human insulin promoter was unmethylated in beta cells, but methylated in all other tissues tested. In contrast, intron 1, exon 2 and intron 2 did not exhibit any tissue-specific DNA methylation pattern. Subsequently, a human MSP assay was developed based on the methylation pattern of the insulin promoter and human islet DNA was successfully detected in circulation of T1D patients after islet transplantation therapy. Signal levels of normal controls and pre-transplant samples were shown to be similar, but increased dramatically after islet transplantation. In plasma the signal declines with time but in whole blood remains elevated for at least two weeks, indicating that association of beta cell DNA with blood cells prolongs the signal. This assay provides an effective method to monitor beta cell destruction in early T1D and in islet transplantation therapy.

An oral vaccine for type 1 diabetes based on live attenuated Salmonella.

Type 1 diabetes (T1D) is a metabolic disease that is initiated by the autoimmune destruction of pancreatic insulin-producing beta cells that is accompanied by the development of antigen-specific antibodies and cytotoxic T lymphocytes (CTLs). Several studies have shown that vaccination with diabetic autoantigens provides some protection against this process. In this report we describe a new oral vaccine that utilizes live attenuated Salmonella for simultaneous delivery of autoantigens in conjunction with immunomodulatory cytokine genes to immune cells in the gut mucosa. Recent data showed that live attenuated Salmonella is a safe, simple and effective vector for expression of antigens and cytokines by antigen-presenting cells (APCs) of gut-associated lymphatic tissue (GALT). This novel strategy was tested by fusion of the diabetic autoantigen preproinsulin with Salmonella secretory effector protein (SseF) of pathogenicity island-2 (SPI2). In this way the autoantigen is only expressed inside the host immune cells and translocated to the host cell cytosol. In addition Salmonella was used to deliver the gene for the immunomodulatory cytokine transforming growth factor beta (TGFß) for host cell expression. Oral co-vaccination of 8 week-old non-obese diabetic (NOD) mice with three weekly doses of both the autoantigen and cytokine significantly reduced the development of diabetes, improved the response to glucose challenge, preserved beta cell mass, and reduced the severity of insulitis compared with controls and autoantigen alone. Combination therapy also resulted in increased circulating levels of IL10 four weeks post-vaccination and IL2 for 12 weeks post-vaccination, but without effect on proinflammatory cytokines IL6, IL12(p70), IL17 and IFNγ. However, in non-responders there was a significant rise in IL12 compared with responders. Future studies will examine the mechanism of this vaccination strategy in more detail. In conclusion, Salmonella-based oral vaccines expressing autoantigens combined with imunomodulatory cytokines appears to be a promising therapy for prevention of T1D.

Development of a quantitative methylation-specific polymerase chain reaction method for monitoring beta cell death in type 1 diabetes.

DNA methylation is a mechanism by which cells control gene expression, and cell-specific genes often exhibit unique patterns of DNA methylation. We previously reported that the mouse insulin-2 gene (Ins2) promoter has three potential methylation (CpG) sites, all of which are unmethylated in insulin-producing cells but methylated in other tissues. In this study we examined Ins2 exon 2 and found a similar tissue-specific methylation pattern. These methylation patterns can differentiate between DNA from insulin-producing beta cells and other tissues. We hypothesized that damaged beta cells release their DNA into circulation at the onset of type 1 diabetes mellitus (T1DM) and sought to develop a quantitative methylation-specific polymerase chain reaction (qMSP) assay for circulating beta cell DNA to monitor the loss of beta cells. Methylation-specific primers were designed to interrogate two or more CpG in the same assay. The cloned mouse Ins2 gene was methylated in vitro and used for development of the qMSP assay. We found the qMSP method to be sensitive and specific to differentiate between insulin-producing cells and other tissues with a detection limit of 10 copies in the presence of non-specific genomic DNA background. We also compared different methods for data analysis and found that the Relative Expression Ratio method is the most robust method since it incorporates both a reference value to normalize day-to-day variability as well as PCR reaction efficiencies to normalize between the methylation-specific and bisulfite-specific components of the calculations. The assay was applied in the streptozotocin-treated diabetic mouse model and detected a significant increase in circulating beta cell DNA before the rise in blood glucose level. These results demonstrate that this qMSP assay can be used for monitoring circulating DNA from insulin-producing cells, which will provide the basis for development of assays to detect beta cell destruction in early T1DM.