Pulmonary and Cardiorenal Cyclooxygenase-1 (COX-1), -2 (COX-2), and Microsomal Prostaglandin E Synthase-1 (mPGES-1) and -2 (mPGES-2) Expression in a Hypertension Model

Hypertensive mice that express the human renin and angiotensinogen genes are used as a model for human hypertension because they develop hypertension secondary to increased renin-angiotensin system activity. Our study investigated the cellular localization and distribution of COX-1, COX-2, mPGES-1, and mPGES-2 in organ tissues from a mouse model of human hypertension. Male (n = 15) and female (n = 15) double transgenic mice (h-Ang 204/1 h-Ren 9) were used in the study. Lung, kidney, and heart tissues were obtained from mice at necropsy and fixed in 10% neutral buffered formalin followed by embedding in paraffin wax. Cut sections were stained immunohistochemically with antibodies to COX-1, COX-2, mPGES-1, and mPGES-2 and analyzed by light microscopy. Renal expression of COX-1 was the highest in the distal convoluted tubules, cortical collecting ducts, and medullary collecting ducts; while proximal convoluted tubules lacked COX-1 expression. Bronchial and bronchiolar epithelial cells, alveolar macrophages, and cardiac vascular endothelial cells also had strong COX-1 expression, with other renal, pulmonary, or cardiac microanatomic locations having mild-to-moderate expression. mPGES-2 expression was strong in the bronchial and bronchiolar epithelial cells, mild to moderate in various renal microanatomic locations, and absent in cardiac tissues. COX-2 expression was strong in the proximal and distal convoluted tubules, alveolar macrophages, and bronchial and bronchiolar epithelial cells. Marked mPGES-1 was present only in bronchial and bronchiolar epithelial cells; while mild-to-moderate expression was present in other pulmonary, renal, or cardiac microanatomic locations. Expression of these molecules was similar between males and females. Our work suggests that in hypertensive mice, there are (a) significant microanatomic variations in the pulmonary, renal, and cardiac distribution and cellular localization of COX-1, COX-2, mPGES-1, and mPGES-2, and (b) no differences in expression between genders

The effects of neutrophil-mediated damage in acute Mannheimia (Pasteurella) haemolytica pneumonia

ICAM-1 expression in cattle with Bovine Leukocyte Adhesion Deficiency (BLAD) vs. cattle without BLAD in Mannheimia haemolytica (MH) pneumonia at 2 or 4 hours PI was assessed using in situ hybridization. There was a significant increase in ICAM-1 in bronchiolar and alveolar epithelium and endothelium of arteries and veins in both BLAD and non-BLAD MH-inoculated cattle compared to controls. Highest intensity of ICAM-1 was present in bronchioles while bronchi had the lowest. Most alveolar macrophages and neutrophils in infected lungs expressed ICAM-1. ICAM-1 expression was generally increased in infected BLAD cattle at 2 hours PI compared to non-BLAD but not at 4 hours PI. The increased expression of ICAM-1 during acute MH pneumonia in cattle suggests that ICAM-1 is upregulated and may play a role in leukocyte infiltration;Next we studied the effects of a selectin inhibitor, TBC1269, on conjugated dienes (CD), inducible nitric oxide synthase (iNOS), and programmed cell death (PCD) in MH pneumonia in neonatal calves at 6 hours PI. Levels of CD increased (P < 0.05) after MH-inoculation in treated and untreated calves. A significant increase in PCD of leukocytes was observed in treated compared with the untreated calves. iNOS was expressed by epithelial cells and leukocytes. Expression of iNOS was decreased in areas where airway epithelium was associated with abundant inflammation. The shift from epithelial to leukocyte iNOS production in inflamed areas suggests that there is a balance between free radical generation by leukocytes and epithelial cell NO production. This shift is unaffected by selectin inhibition;Finally, neutrophil infiltration and ICAM-1 expression after TBC1269 treatment was assessed. There were higher (P < 0.05) neutrophil counts in lungs of both treated and untreated calves compared with the control group at 2 or 6 hours PI as determined by image analysis. Bronchioles and alveoli had the highest counts compared to other microscopic locations. ICAM-1 was significantly greater in bronchiolar and alveolar epithelium in both treated and untreated calves. TBC1269-treatment did not affect ICAM-1 expression. This work suggests that selectin inhibition does not inhibit neutrophil infiltration. ICAM-1 and CD18-mediated adherence may override the role of selectins during MH pneumonia.</p

Medical and Clinical Pathology Pre-Screening Visit and Enrolment Seasonal Variability in Healthy Volunteers

In the pharmaceutical industry, selection of healthy volunteers is one of the foundations in phase I clinical trials and is a difficult and costly process. The objectives of this study were to evaluate the seasonal variability in recruiting healthy volunteers and examine the value of utilizing a pre-screen visit for healthy volunteers to generate a database pool that will be used in the routine screening process of phase I clinical research studies. We retrospectively studied a total of 1115 male and female volunteers who were scheduled for a medical and clinical pathology pre-screen visit over a one year period. Written consents were obtained from all individuals who participated in the study. Medical pre-screen visit included a full medical history and examination and electrocardiogram and clinical pathology (clinical chemistry, hematology and urinalysis). There was apparent seasonal variability in the participation of individuals in the pre-screen visit. Increased values of clinical chemistry values such as alanine aminotransferase and aspartate aminotransferase accounted for the majority of the clinically relevant increased values. Increased values of WBC’s and both platelets and mean corpuscular volume accounted for the lowest values. In urinalysis, the most prevalent abnormal values were increased WBC’s and red blood cells. No apparent differences were seen between sexes.Conducting medical and clinical pathology pre-screening visit is important as a source for healthy volunteer database pool to participate in phase I trials