\s=b\Otitis media with effusion (OME) is a common middle ear inflammatory disease in the pediatric population. This article determines concentrations of three functionally and metabolically distinct inflammatory mediators in middle ear effusions (MEE) and corresponding plasma of children with OME. One hundred two patients (mean age, 4.9 years) with persistent OME were studied. Middle ear effusions were collected from all subjects and plasma from a subset at the time of tympanostomy tube insertion. Histamine was assayed radioisotopically, 13, 14-dihydro-15-keto-prostaglandin F2\g=a\(stable PGF2\g=a\ metabolite) by radioimmunoassay, and neutrophil chemotactic factor of anaphylaxis by modified Boyden chamber. Mean MEE levels of the mediators (39 \m=+-\ 13 ng/mL, 462 \m=+-\179 pg/mL, and 264% \ m=+-\ 57% positive control, respectively) were markedly higher than those of corresponding plasma (0.5 \m=+-\ 0.1 ng/mL, 285 \m=+-\127 pg/mL, and 47% \m=+-\5% positive control, respectively). The mean histamine content of mucoid effusions (43.2 \ m=+-\56.9 ng/mL) was significantly higher than that of purulent (22.5 \m=+-\10.5 ng/mL) and serous (17.9 \m=+-\ 16.8 ng/mL) effusions. Higher histamine levels were observed in effusions positive for Haemophilus influenzae when compared with those with other pathogenic isolates. The high concentrations of these mediators in MEE and their potential for inducing or sustaining the inflammatory process supports a role in the pathogenesis of OME. (Arch Otolaryngol Head Neck Surg 1988;114:1131-1133 Otitis media with effusion (OME) is a middle ear inflammatory dis¬ ease that is common in the pediatrie population. A variety of potent inflammatory mediators has been detected in middle ear effusions (MEE) of children with OME.1 More¬ over, a role for these mediators in the pathogenesis and maintenance of the disease condition was emphasized by the results of a number of recent experimental studies using animal models.2 In humans, IgE-dependent mediator release or allergy has also been incriminated as a risk factor in OME. This investigation determined MEE concentrations of two mast cellderived mediators, histamine and neutrophil chemotactic factor of anaphylaxis (NCF-A), and a stable metabolite of prostaglandin (PG)F2o. The latter two have not been previous¬ ly studied in OME. PATIENTS AND METHODSOne hundred two patients, aged 1 to 23 years (mean age, 4.9 years) with docu¬ mented persistent OME unresponsive to antimicrobial therapy were enrolled after obtaining informed consent. All patients had tympanostomy tubes inserted at which time MEEs were collected. Unilateral effu¬ sions were collected from 25 patients and bilateral effusions were collected from the remaining 17 patients. These samples were classified visually as serous, mucoid, or purulent; cultured immediately for aerobic bacteria; and a subset was submitted for complete cell count by Coulter Counter and differential. The remaining fluid (50 to 200
Pediatrics, Stony Brook, NY. Rapid-frozen liver biopsy tissue and freeze-substitution (FS) or cryo-thinsectioning offer new methods of morphology, biochemistry, and x-ray microanalysis. Three liver biopsies from patients with RS and three from children with other diseases were studied by a new technique: rapid freezing and freeze substitution with osmium tetroxide-acetone. As previously described in muscle, mitochondria1 (Mt) intracristal spaces are nonexistent and cristae appear as tip;ht seven-layered structures in both RS and non-RS liver. Pleomorphism of RS Mt is striking; Mt enlargement is not watery expansion, but a non-aqueous milieu with no disruption of cristae and no ice artefact. In severely injured liver cells in RS, two populations of Mt are present. In contrast to Mt, peroxlsomes contain large ice crystals, and their contents may be mainly aqueous. Large pools of smooth endoplasmic reticulum are discretely outlined, without ice artefact, as are Golgi lanellae. All hepatocyte organelles are easily recognizable by standard EM criteria, but membranes and spacial relationships differ from traditional !Ol processing. Sodium benzoate (SB) therapy in hypera&.onemic children is based on the elimination of excess nitrogen as hippurate, but its mechanism is speculatory. We gave 2 % SB in drinking water to normal (+/Y) and spf/Y mice. After acclimatization for 48 h, i.p. 3~-glycine and 14~-serine were given as sources of transported and biosynthesized glycine respectively. 24 h urine was analyzed for total hippurate, free glycine and orotate. Hippurate spots were separated by TLC and counted for radioactivity. Output of hippurate and free glycine increased significantly in SB treated spf/Y and +/Y as compared to untreated mice, while orotate excretion decreased (p < 0.013). Specific activity of 3~ and 1 4~ (DPM/LImol hippurate) showed significant increases in treated groups, indicating utilization of transported and biosynthesized glycine. However, 3~: 1 4~ ratio showed predominant use of 3~-~lycine. The ratio was higher in treated spf mice as codpared to untreated controls (3.1 f 0.7 Vs 1.5 i 0.04; p < 0.02); same increase was seen in +/Y (1.9 f 0.2 Vs 1.0 t 0.1; p < 0.01). The results indicate that transported glycine has a bigger contribution in the formation of hippurate in vivo than glycine synthesized from serine. Since glycine is highly amnoniagenic, we postulate that the conjugation of excess glycine transported from body pools would result in depletion of ammoniagenic potential. This would be supplemented by the effect of glycine biosynthesis through reactions requiring a metabolic input of NH3. IDIOSYNCRATIC HEPATOTOXIC RESPONSE TO SODIUM VAL-PROATE (SV) IN A SUB-POPULATION OF SPARSE-FUR (spf)245 MUTANT MICE. Iiaz Qureshi, Jaqques Letarte. Beatriz Tuchweber, Ibrahim Yousef and S.R. Qureshi',' Centre de Recherche pgdiatrique, HSpital Sainte-Justine, Montreal, Qu6. Canada.Groups of six normal +/Y and six spf/Y mice wlth X-linked ornithine transcarbamylase (OTC) deficiency were given SV in drinking wat...
(Spon. by I. Rezvani). Recent studies have shown that 1,25-D may be involved in the regulation of differentiation and function of the immune system. The syndrome of resistance to 1,25-D provides an opportunity for investigation of the role of 1,ZS-D in the immune system of human subjects. In this study we examined some aspects of the immune system in 5 patients with resistance to 1,ZS-D and alopecia who had defective receptors for 1,25-D in skin fibroblasts. All patients had normal numbers of blood T and B lymphocytes and normal helperjinducer (OKT4) to suppressor/cytotoxic (OKT8) ratio. Proliferative responses to phytohemagglutinin and concanavalin A were comparable to those of normal controls. Serum IgG, IgM and IgA were within the normal range for all patients. Staphylcoccal killing and monocyte chemotaxis were comparable to controls in the one patient examined. Although phagocytosis of Candida albicans by neutrophils was normal, candidacidal activity was significantly lower (32f 5% of injestedc C. albicans in 1 hr) than that observed in the controls (83t16f of injested C. albicans in 1 hr). The data indicate that 1,25-D participates in activation of intrcellular killing of some microvbes by human neutrophils and support the view that 1,25-D is involved in a wide range of cellular processes. HUMAN THYMOCYTE CHEMILUMINESCENCE-DEPENDENCE ON 974 IMMUNE BASIS FOR EUSTACHIAN TUBE (ET) OBSTRUCTION 975P h i l i p Fireman, Paul K. Stillwagon, David P. Skoner, Charles D. Bluestone, William J . Doyle (Spon. Thomas K. Oliver, J r . ) Children's Hospital, P i t t s b u r g h , Pennsylvania Recent s t u d i e s i n our l a b o r a t o r i e s have documented t h e development of a l l e r g i c r h i n i t i s (AR) and subsequent ET o b s t r u c t i o n a f t e r provocative i n t r a n a s a l pollen o r house dust mite antigen challenges i n hypersensitive s u b j e c t s . The induced ET obstruct i o n was antigen dose dependent and i n v e r s e l y r e l a t e d t o t h e pat i e n t ' s s p e c i f i c IgE antibody t i t e r . Since prolonged ET obstruct i o n u n d e r l i e s t h e pathogenesis of o t i t i s media with e f f u s i o n (OME), t h e s e d a t a suggest a r o l e f o r AR i n t h e e t i o l o g y of OME. To a s c e r t a i n i f ET o b s t r u c t i o b and/or O M E develop during n a t u r a l p o l l e n exposure, weekly b i l a t e r a l ET function was measured by t h e 9-step p r e s s u r e swallow tympanometric t e s t . Eight c h i l d r e n ages 5 t o 14 years with documented ragweed ARand r e c u r r e n t OME were s t u d i e d before, during and a f t e r t h e 1984 ragweed season which was estimated by d a i l y p o l l e n counts. Ragweed a l l e r g y was confirmed by p o s i t i v e s k i n t e s t with serum RAST ranging from 1 t o 10 percent. Symptom s c o r e and medication d i a r i e s were kept. Whereas9of 16 e a r s (56%) showed normal ET function before p o l l e n season and 10 of 16 (63%) a f t e r p o l l e n season, only 2 of 15 (13%) had normal ET function a t t h e p...
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