VOC sampling locations table for Ground Zero area, Jan 2002

NYC 9/11 Public Portal Document

Elkhart, IN). Cell differentials were performed by one 2. Nasal histopathology. Dr. James Wagner of person (SHG) counting 500 cells per slide. After lavage, Michigan State University (MSU) instructed EPA the lungs were removed and stored at -80° C for future personnel in this procedure, utilized on mice from the assays (to be determined). nose-only inhalation exposure (experiment B). Assays for total protein, albumin, lactate Immediately after death, the head of each animal was dehydrogenase (LDH) andN-acetyl-P-D-glucosaminidase removed from the carcass and both nasal passages were (NAG) are routine measures of lung injury (Henderson et fixed by slowly flushing retrograde through the al., 1985) and were carried out on an aliquot of BAL nasopharynx with 1-2 ml 4% paraformaldehyde. The supernatant as previously described using a Cobas Fara II nasal cavity was then immersed in a large volume of the centrifugal spectrophotometer (Gavett et al., 1997). Four fixative for at least 24h until further processing. The fixed other BAL supernatant aliquots were prepared from each nasal cavities were placed in 0.1 M PBS (pH 7.2, 4 °C) sample; one of these was supplemented with 10% fetal and shipped overnight to Dr. Wagner at MSU. Nasal bovine serum to prevent loss of cytokines and other cavities were decalcified in a 13% solution of formic acid proteins in low protein concentration fluids, and the 4 for 5 days, and then rinsed in distilled water for Ih. After aliquots from each sample were stored at -80° C. These decalcification, three transverse tissue blocks of the nasal samples are available for analysis of cytokines and other cavity, cut perpendicular to the hard palate, were selected proteins (to be determined). for light microscopic analysis. The first tissue block was sectioned from the proximal aspect of the nasal cavity M. Histopathology immediately posterior to the upper incisor tooth (T1). The I. Lung histopathology. In experiments A and B, second transverse tissue block was taken at the level of the mice which were tested for Meh responsiveness were incisive papilla (T2) and the third and most distal tissue subsequently assessed for lung histopathology, while in block was taken at the level of the second palatial ridge experiment C, all mice were tested for Meh responsiveness (T3). The tissue blocks were embedded in paraffin, and 6 and were lavaged before assessment of lung pm-thick sections were cut from the anterior surface of histopathology. Mice were anesthetized with urethane and each block. Sections were histochemically stained with killed as described above for BAL. Lungs were removed hematoxylin and eosin for morphologic identification of and fixed by tracheal perfusion in a fume hood with ice nasal tissues. Nasal tissues (three sections/mouse) from a cold 4% paraformaldehyde at 25 cm pressure for 15 total of 48 mice tested for Meh responsiveness (8 minutes. The trachea was then tied off and placed in a vial mice/exposure group/time point) were microscopically of4% paraformaldehyde at 4 °C. After 24 hours, the lungs examined by Dr. Jack Harkema (MSU). Nasal lesions were drained and placed in phosphate buffered saline at 4 were graded on the following scale: 1 = minimal, 2 = °C. mild, 3 = moderate, and 4 = marked inflammation. The lungs were transferred to Experimental Pathology Laboratories (Research Triangle Park, NC), where fixed N. Statistical Analysis lungs were processed to paraffin blocks, sectioned at an All statistical analysis were done using SAS approximate thickness of 5 p, placed on glass slides and procedures (Cary, NC). There were generally three types stained with hematoxylin and eosin (H&E). Longitudinal of responses collected: 1) PenH responses recorded coronal sections were cut on a lateral plane to include repeatedly for each animal as area under the curve (AUC) mainstem bronchi for viewing a maximal amount of lung for various concentration exposures to Meh; 2) responses area. Two additional unstained lung sections were to DLCO were analyzed from a single response from 4 prepared for future use. Histopathologic observations for animals; and 3) individual measurements measured once individual animals in each experiment were tabulated, and for each animal as a univariate variable. Experimental the degree of severity of inflammatory changes and the designs varied with each experiment and each part of an presence of PM-related pigment were graded on a scale of experiment. Statistical designs used were replicated one to five (1 = minimal, 2 = slight/mild, 3 = moderate, 4 completely random designs for experiment A. Crossed- = moderately severe, 5 = severe/high). The pathologist designs were used for experiment B and C involving knew which animals comprised a group, which group was treatments (TRT) and days (DAY), Randomized block the saline or air-exposed control group, the day after designs were used for DLCO experiments. treatment, and the doses given to the experimental groups, When initial multivariate repeated measures analysis but did not know the identities of the individual PM of variance (MANOVA) test showed significant samples other than by a unique number or letter. interactions between dose of Meh and TRT or DAY in the

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NYC-WTC_000149155