Jay A. Werkhaven

Human Nasopharyngeal-Associated Lymphoreticular Tissues: Functional Analysis of Subepithelial and Intraepithelial B and T Cells from Adenoids and Tonsils

Subepithelial and intraepithelial lymphocytes of human adenoids and tonsils were characterized and directly compared to determine the potential contribution of these tissues to mucosal and systemic immune responses. The distribution of T and B cell subsets, cytokine patterns, and antibody (Ab) isotype profiles were similar for adenoids and tonsils. Both tissues contained predominantly B cells ( approximately 65%), approximately 5% macrophages, and 30% CD3(+) T cells. The T cells were primarily of the CD4(+) subset ( approximately 80%). Tonsillar intraepithelial lymphocytes were also enriched in B cells. The analysis of dispersed cells revealed a higher frequency of cells secreting IgG than IgA and the predominant Ig subclass profiles were IgG1 > IgG3 and IgA1 > IgA2, respectively. In situ analysis also revealed higher numbers of IgG- than IgA-positive cells. These IgG-positive cells were present in the epithelium and in the subepithelial zones of both tonsils and adenoids. Mitogen-triggered T cells from tonsils and adenoids produced both Th1- and Th2-type cytokines, clearly exhibiting their pluripotentiality for support of cell-mediated and Ab responses. Interestingly, antigen-specific T cells produced interferon-gamma and lower levels of interleukin-5. These results suggest that adenoids and tonsils of the nasopharyngeal-associated lymphoreticular tissues represent a distinct component of the mucosal-associated lymphoreticular tissues with features of both systemic and mucosal compartments.

Growth of Respiratory Syncytial Virus in Primary Epithelial Cells from the Human Respiratory Tract

ABSTRACT Respiratory syncytial virus (RSV) is the most important cause of lower respiratory tract disease in infants and children. To study RSV replication, we have developed an in vitro model of human nasopharyngeal mucosa, human airway epithelium (HAE). RSV grows to moderate titers in HAE, though they are significantly lower than those in a continuous epithelial cell line, HEp-2. In HAE, RSV spreads over time to form focal collections of infected cells causing minimal cytopathic effect. Unlike HEp-2 cells, in which wild-type and live-attenuated vaccine candidate viruses grow equally well, the vaccine candidates exhibit growth in HAE that parallels their level of attenuation in children.

Prevention of Anterior Glottic Restenosis in a Canine Model with Topical Mitomycin-C

To develop a model for recurrent anterior glottic stenosis and to test the efficacy of topical mitomycin-C in preventing restenosis, we induced anterior glottic stenosis with a CO2 laser in 5 dogs. In 3 dogs, recurrence was established after surgical lysis. Subsequently, the 3 dogs received a single topical 3-minute treatment with a 1% solution of mitomycin-C after a second surgical lysis. In a parallel experiment, the other 2 dogs received a single topical 3-minute treatment with a 1% solution of mitomycin-C after the initial surgical lysis. An anterior glottic web was induced in all 5 dogs with the CO2 laser. The 3 dogs experienced restenosis at the anterior glottis after surgical lysis alone. Mitomycin-C prevented anterior glottic restenosis in 2 of the 3 dogs treated twice and in both of the dogs treated once (p = .02). We conclude that a recurrent stenosis of the anterior glottis may be induced reproducibly in the canine model with the CO2 laser. Application of topical mitomycin-C after lysis of an anterior glottic stenosis produces a statistically significant reduction in the rate of restenosis as compared to surgical lysis alone.

Analgesia after bilateral myringotomy and placement of pressure equalization tubes in children: acetaminophen versus acetaminophen with codeine.

Despite the brief nature of the procedure with limited tissue trauma, some form of analgesia is required in most children after bilateral myringotomy and placement of pressure equalization (PE) tubes.Previous studies have demonstrated the relative inefficacy of acetaminophen and nonsteroidal antiinflammatory drugs (NSAIDs), with 30%-55% of patients requiring supplemental postoperative analgesia. We undertook a prospective study evaluating the efficacy of the preoperative administration of oral acetaminophen (15 mg/kg) versus acetaminophen (10 mg/kg) and codeine (1 mg/kg). Fifty ASA grade I or II patients were randomized to receive oral midazolam premedication (0.7 mg/kg) mixed in either acetaminophen or acetaminophen with codeine elixir. Anesthesia was induced and maintained with halothane in nitrous oxide and oxygen. Postoperative pain was assessed at four times during the postoperative course using an objective pain scale. The two groups were similar with respect to age, weight, gender, duration of anesthesia, and duration of the surgical procedure. The patients who received acetaminophen with codeine had lower pain scores at all four points when compared with patients who received acetaminophen. None of the 25 patients who received acetaminophen with codeine required supplemental analgesics compared with 12 of 25 who received acetaminophen. No adverse effects were noted in either group. We conclude that the preoperative administration of acetaminophen with codeine provides superior analgesia after bilateral myringotomy and placement of PE tubes. (Anesth Analg 1995;81:496-500)

Single cell analysis of human tissues and solid tumors with mass cytometry

Mass cytometry measures 36 or more markers per cell and is an appealing platform for comprehensive phenotyping of cells in human tissue and tumor biopsies. While tissue disaggregation and fluorescence cytometry protocols were pioneered decades ago, it is not known whether established protocols will be effective for mass cytometry and maintain cancer and stromal cell diversity.

Advanced Microspot Microslad for the CO2 Laser

New advances in instrumentation have facilitated the development of a second generation carbon dioxide (CO2) laser microspot micromanipulator. The 710 Acuspot has unique advantages over the previous generation of microspots. The compact design is easier to handle and has attachment points for sterile draping. The unit produces a spot size of 250 microns at a 400-mm focal length and 160 microns at 250 mm; the maximum de-focus is 3.2 mm at all focal lengths. An innovative dichrolc mirror allows use of the laser units own HeNe laser as the aiming beam, eliminating possible aiming error introduced with a virtual image-aiming system. The dichrolc mirror also allows better light transmission, resulting in a brighter field of view. As with first generation microspots, the laser beam path is coincident with the microscope optical path, eliminating parallax; this feature has been especially advantageous in pediatric and otologic cases. Having used this unit for 6 months on more than 50 patients, we now consider the use of a microspot to be our delivery system of choice for most microlaryngeal laser surgical applications.

Use of AlloDerm in type I tympanoplasty: a comparison with native tissue grafts.

Objectives: AlloDerm, an acellular human dermis allograft, has been shown to be an effective option as a tympanic membrane (TM) graft in animals and humans and has several potential advantages, including eliminating donor site morbidity, reducing operative time, and preserving native tissues for later use. We compared AlloDerm and native tissue grafts in type I tympanoplasty with regard to operative time, graft success rate, and audiologic outcome.

Prevention of pressure ulcers after pediatric tracheotomy using a Mepilex Ag dressing

Skin irritation and ulceration beneath the tracheostomy tube or ties secondary to pressure and shearing forces on the skin frequently complicate pediatric tracheotomy in the immediate postoperative period. The aim of this study is to determine the effectiveness of Mepilex Ag dressings in reducing posttracheotomy wound complications.

Detection of Viruses in Human Adenoid Tissues by Use of Multiplex PCR

By PCR, we detected a high frequency of viruses in adenoids obtained from children without acute respiratory symptoms. Our results suggest that persistent/latent viral infection in the respiratory tract confounds interpretation of the association of pathogen detection by PCR with acute respiratory infection in these sources.

Tracheotomy in the First Year of Life: Outcomes in Term Infants, the Vanderbilt Experience

OBJECTIVE: In an era emphasizing critical care of preterm infants, we characterize the indications and outcomes of tracheotomies performed in the first year of life in term infants compared to preterm infants. METHODS: Retrospective study of 127 tracheotomies performed in the first year of life at a tertiary-care childrens hospital between 1988-2004. RESULTS: Mean gestational ages of the term and preterm groups were 38.97 and 29.71 weeks, respectively (P < 0.001). Indications for tracheotomy were upper airway abnormalities in 53% for the term group. The number of subsequent airway procedures required was 1.39 in the term group, achieving decannulation in 36.3%, with a 20.5% mortality rate. CONCLUSION: Compared to preterm infants, the term decannulation rate was favorable, as chronic lung disease was uncommon. However, non-tracheotomy-related mortalities remained high. SIGNIFICANCE: Tracheotomies are often performed for relief of upper airway obstruction, and congenital and acquired comorbidities not related to tracheotomy are associated with adverse outcomes in term infants. EBM rating: C-4 ©2006 American Academy of Otolaryngology–Head and Neck Surgery Foundation, Inc. All rights reserved.