Close association between oropharyngeal and rhinopharyngeal colonization with Staphylococcus aureus - clues to new insight of MRSA colonization of the oropharynx.

This study provides data on prevalence of Staphylococcus aureus in oropharynx, rhinopharynx and vestibulum nasi. Specimens were taken from these three pharyngeal sites in 346 patients and analysed for S. aureus. Abnormal pharyngeal findings and patient histories were recorded. S. aureus was found in 8.1%, 7.2% and 20.2% of all specimens from oropharynx, rhinopharynx and vestibulum nasi, respectively. A strong association between colonization of oropharynx and rhinopharynx was found, especially when vestibulum nasi was not colonized. These findings can be used in development of more effective meticillin-resistant Staphylococcus aureus decolonization regimes.

DAF-16-dependent and independent expression targets of DAF-2 insulin receptor-like pathway in Caenorhabditis elegans include FKBPs.

The daf-2 insulin-like receptor pathway regulates development and life-span in Caenorhabditis elegans. Reduced DAF-2 signaling leads to changes in downstream targets via the daf-16 gene, a fork-head transcription factor which is regulated by DAF-2, and results in extended life-span. Here, we describe the first identification of genes whose expression is controlled by the DAF-2 signaling cascade. dao-1, dao-2, dao-3, dao-4, dao-8 and dao-9 are down-regulated in daf-2 mutant adults compared to wild-type adults, whereas dao-5, dao-6 and dao-7 are up-regulated. The latter genes are negatively regulated by DAF-2 signaling and positively regulated by DAF-16. Positive regulation by DAF-2 on dao-1, dao-4 and dao-8 was mediated by DAF-16, whereas daf-16 mediates only part of DAF-2 signaling for dao-2 and dao-9. Regulation by DAF-2 is most likely DAF-16 independent for dao-3 and hsp-90. RNA levels of dao-5 and dao-6 showed elevated expression in daf-2 adults, as well as being strongly expressed in dauer larvae. In contrast, hsp-90 transcript levels are low in daf-2 mutant adults though they are enriched in dauer larvae, indicating overlapping but not identical mechanisms of efficient life maintenance in stress-resistant dauer larvae and long-lived daf-2 mutant adults. dao-1, dao-8 and dao-9 are homologs of the FK506 binding proteins that interact with the mammalian insulin pathway. dao-3 encodes a putative methylenetetrahydrofolate dehydrogenase. DAO-5 shows 33 % identity with human nucleolar phosphoprotein P130. dao-7 is similar to the mammalian ZFP36 protein. Distinct regulatory patterns of dao genes implicate their diverse positions within the signaling network of DAF-2 pathway, and suggest they have unique contributions to development, metabolism and longevity.

A dietary source of coenzyme Q is essential for growth of long-lived Caenorhabditis elegans clk-1 mutants.

Mutations in the clk-1 gene of the nematode Caenorhabditis elegans result in slowed development, sluggish adult behaviors, and an increased lifespan. CLK-1 is a mitochondrial polypeptide with sequence and functional conservation from human to yeast. Coq7p, the Saccharomyces cerevisiae homologue, is essential for ubiquinone (coenzyme Q or Q) synthesis and therefore respiration. However, based on assays of respiratory function, it has been reported that the primary defect in the C. elegans clk-1 mutants is not in Q biosynthesis. How do the clk-1 mutant worms have essentially normal rates of respiration, when biochemical studies in yeast suggest a Q deficiency? Nematodes are routinely fed Escherichia coli strains containing a rich supply of Q. To study the Q synthesized by C. elegans, we cultured worms on an E. coli mutant that lacks Q and found that clk-1 mutants display early developmental arrest from eggs, or sterility emerging from dauer stage. Provision of Q-replete E. coli rescues these defects. Lipid analysis showed that clk-1 worms lack the nematode Q(9) isoform and instead contain a large amount of a metabolite that is slightly more polar than Q(9). The clk-1 mutants also have increased levels of Q(8), the E. coli isoform, and rhodoquinone-9. These results show that the clk-1 mutations result in Q auxotrophy evident only when Q is removed from the diet, and that the aging and developmental phenotypes previously described are consistent with altered Q levels and distribution.

Structure and expression of daf-12: a nuclear hormone receptor with three isoforms that are involved in development and aging in Caenorhabditis elegans.

During Caenorhabditis elegans early larval development environmental conditions promote a cascade of signaling molecules to direct growth to the reproductive adult or to arrest development as a dauer larva. Two parallel chemosensory signal transduction pathways, one of which is transforming growth factor (TGF)-beta-like, converge on the daf-12 gene to regulate dauer formation. A third insulin-like signaling pathway interacts with the daf-12 pathway to regulate both dauer formation and adult longevity. To further understand the role of daf-12 in these processes, we have molecularly characterized this gene. We establish rescue of the mutant dauer defective phenotype with a genomic clone. We show that three transcripts of different lengths, due to differential splicing, are made from the daf-12 gene. The deduced protein isoforms are similar to both DNA- and ligand-binding domains of nuclear hormone receptors. The three daf-12 transcripts are produced throughout development and expression increases during the preparation for and execution of dauer formation. Analysis of various daf mutant strains suggests that the isoform ratios of daf-12 steady-state mRNA are not changed by reduction of function in the TGF-beta and insulin signaling components of the dauer pathway. The daf-12 promoter directs expression of GFP in the pharynx. daf-12 is a C. elegans nuclear hormone receptor with multiple isoforms, is expressed throughout development in distinct cells, and functions under a variety of environmental conditions.

Fluid absorption related to ion transport in human airway epithelial spheroids.

Airway epithelium explants from cystic fibrosis (CF) patients and non-CF subjects formed monolayered spheres, with the apical ciliated cell membrane facing the bath and the basolateral cell membrane pointing toward a fluid-filled lumen. With the use of two microelectrodes, transepithelial potential difference and changes in potential difference in response to passage of current pulses were recorded, and epithelial resistance and the equivalent short-circuit current were calculated. Non-CF control potential difference and short-circuit current values were significantly lower than the CF values, and amiloride inhibited both values. Fluid transport rates were calculated from repeated measurements of spheroid diameters. The results showed that 1) non-CF and CF spheroids absorbed fluid at identical rates (4.4 microl x cm(-2) x h(-1)), 2) amiloride inhibited fluid absorption to a lower residual level in non-CF than in CF spheroids, 3) Cl(-)-channel inhibitors increased fluid absorption in amiloride-treated non-CF spheroids to a level equal to that of amiloride-treated CF spheroids, 4) hydrochlorothiazide reduced the amiloride-insensitive fluid absorption in both non-CF and CF spheroids, and 5) osmotic water permeabilities were equal in non-CF and CF spheroids ( approximately 27 x 10(-7) cm x s(-1) x atm(-1)).

Topical budesonide treatment reduces endothelial expression of intercellular adhesion molecules (vascular cell adhesion molecule-1 and P-selectin) and eosinophil infiltration in nasal polyps.

Infiltration of eosinophil granulocytes and endothelial expression of vascular cell adhesion molecule-1 (VCAM-1) and P-selectin was investigated in biopsies of polyps and inferior turbinates from 16 patients with nasal polyps, by the use of immunohistochemical staining and stereological quantification before, during and after topical treatment with budesonide (Rhinocort Turbuhaler). Before glucocorticoid treatment a higher density of eosinophil profiles (p < 0.005), making up a larger proportion of the total cellular infiltrate (p < 0.0005), was found in the polyps compared with the inferior turbinates. Endothelial VCAM-1 expression was higher in polyps than in inferior turbinates (p < 0.005), in contrast with the expression of P-selectin, which was more frequently expressed in the inferior turbinates (p < 0.05). Topical glucocorticoid treatment reduced the density of eosinophil profiles (p < 0.05) and the endothelial expression of VCAM-1 (p < 0.007) and P-selectin (p < 0.02) in polyps. Eosinophil counts and VCAM-1 expression returned to pre-treatment levels 8 weeks after discontinuation of budesonide treatment. The observed reduction in endothelial expression of cellular adhesion molecules may interfere with cellular recruitment in nasal polyps and thus contribute to the known anti-inflammatory effect of glucocorticoid in nasal polyposis.

Ion transport in epithelial spheroids derived from human airway cells.

In the present study, we describe a novel three-dimensional airway epithelial explant preparation and demonstrate its use for ion transport studies by electrophysiological technique. Suspension cultures of sheets of epithelial cells released by protease treatment from cystic fibrosis (CF) and non-CF nasal polyps developed free-floating, monolayered epithelial spheres, with the apical, ciliated cell membrane facing the bath and the basolateral cell membrane pointing toward a fluid-filled lumen. Microelectrode impalement of both non-CF and CF spheroids revealed lumen-positive transepithelial electrical potential differences (PDs) that were inhibited by amiloride, indicating that the spheroids were inflated due to amiloride-sensitive Na+ absorption followed by water. Transformation to a Cl- secretory state was achieved by addition of ATP to the bath, leading to the development of a diphenylamine-2-carboxylate-sensitive PD. A cAMP-induced increase in PD was seen in non-CF spheroids only. In response to hydrocortisone treatment, Na+ transport reflected by amiloride-sensitive PD increased and more so in CF than in non-CF spheres. We concluded that this preparation is a useful model for the airway surface epithelium and is suitable for studies of transport mechanisms and regulation.

Dendritic mast cells in the human nasal mucosa.

Human mast cells can be divided into two subtypes: MCTC cells, which contain tryptase and chymase, and MCT cells, which contain tryptase only. Herein we have used a combination of histamine, tryptase and chymase immunohistochemistry as a novel approach to the study of mast cells. Using this technique, we have discovered a new type of MCTC mast cell in biopsies of the nasal mucosa from healthy subjects and allergic patients. These mast cells have histamine-positive, dendrite-like cellular processes. Some cells have only one slender process, whereas other cells have several long processes extending from different parts of the cell body. Some of the cellular processes divide into two or three terminal branches, and histamine is sometimes found in small swellings along the course of the processes. Our findings contribute new aspects to the concept of mast cell heterogeneity. Thus, human mast cells may vary not only with respect to mediator content, but also with respect to gross morphologic features such as the presence of dendrite-like cellular processes. The recognition of this extreme heterogeneity may be an important step toward a better understanding of mast cell biology.

Two pleiotropic classes of daf-2 mutation affect larval arrest, adult behavior, reproduction and longevity in Caenorhabditis elegans.

The nematode Caenorhabditis elegans responds to overcrowding and scarcity of food by arresting development as a dauer larva, a nonfeeding, long-lived, stress-resistant, alternative third-larval stage. Previous work has shown that mutations in the genes daf-2 (encoding a member of the insulin receptor family) and age-1 (encoding a PI 3-kinase) result in constitutive formation of dauer larvae (Daf-c), increased adult longevity (Age), and increased intrinsic thermotolerance (Itt). Some daf-2 mutants have additional developmental, behavioral, and reproductive defects. We have characterized in detail 15 temperature-sensitive and 1 nonconditional daf-2 allele to investigate the extent of daf-2 mutant defects and to examine whether specific mutant traits correlate with each other. The greatest longevity seen in daf-2 mutant adults was approximately three times that of wild type. The temperature-sensitive daf-2 mutants fell into two overlapping classes, including eight class 1 mutants, which are Daf-c, Age, and Itt, and exhibit low levels of L1 arrest at 25.5 degrees. Seven class 2 mutants also exhibit the class 1 defects as well as some or all of the following: reduced adult motility, abnormal adult body and gonad morphology, high levels of embryonic and L1 arrest, production of progeny late in life, and reduced brood size. The strengths of the Daf-c, Age, and Itt phenotypes largely correlated with each other but not with the strength of class 2-specific defects. This suggests that the DAF-2 receptor is bifunctional. Examination of the null phenotype revealed a maternally rescued egg, L1 lethal component, and a nonconditional Daf-c component. With respect to the Daf-c phenotype, the dauer-defective (Daf-d) mutation daf-12(m20) was epistatic to daf-2 class 1 alleles but not the severe class 2 alleles tested. All daf-2 mutant defects were suppressed by the daf-d mutation daf-16(m26). Our findings suggest a new model for daf-2, age-1, daf-12, and daf-16 interactions.

Efficacy of an aqueous and a powder formulation of nasal budesonide compared in patients with nasal polyps.

Nasal polyps are commonly treated surgically. Intranasal administration of topical corticosteroids has gained increased acceptance as a treatment alternative. The aim of our study was to compare the efficacy of treatment of two formulations of budesonide with placebo on nasal polyps. At four Danish clinics 138 patients suffering from moderate or severe nasal polyps were randomized to a twice daily treatment with Rhinocort Aqua 128 micrograms, Rhinocort Turbuhaler 140 micrograms or placebo (Astra Draco, Sweden) for 6 weeks. Polyp size (primary efficacy variable), nasal symptoms, sense of smell, and patients' overall evaluation of treatment of efficacy were assessed by scores. Polyp size was reduced significantly in both budesonide treated groups compared with placebo, but there was no statistical difference between the two actively treated groups. Patients' nasal symptom scores was significantly more reduced in the Aqua compared to the Turbuhaler treated group, and both reduced symptom scores were significantly better compared to placebo. Sense of smell was significantly improved in the actively treated groups compared to placebo. The proportion of patients rating substantial or total control over symptoms after 6 weeks treatment was 60.9% and 48.2% in the Aqua and Turbuhaler-treated groups, respectively, which was significantly better compared with 29.8% in the placebo-treated group. Rhinocort Aqua and Rhinocort Turbuhaler were equally well tolerated.

Expression of intercellular adhesion molecule-1 on the vascular endothelium in nasal polyps before, during and after topical glucocorticoid treatment.

Nasal polyposis is a result of a chronic inflammatory disorder in the upper airways. In vitro studies have revealed that extravasation of leucocytes requires interactions between several sets of adhesion molecules expressed on the circulating leucocytes and the vascular endothelium. Therefore, the endothelial expression of intercellular adhesion molecule-1 (ICAM-1) in biopsies from polyps and inferior turbinates was investigated by the use of immunohistochemical staining. Biopsies were obtained from 11 patients suffering from nasal polyposis before, during and after treatment with 100 microg budesonide (Rhinocort Turbuhaler) in each nostril twice daily. Before, during and after treatment, ICAM-1 was expressed in the majority of vessels in polyps and mucosa of inferior turbinates. The intensity of endothelial ICAM-1 expression in polyps was significantly reduced during topical glucocorticoid treatment compared with the pretreatment and posttreatment levels (p < 0.005). In biopsies from the inferior turbinates, the intensity of the endothelial ICAM-1 expression was lower during treatment than after discontinuation of medical treatment (p < 0.005). In conclusion, topical budesonide treatment seems to downregulate ICAM-1 expression on the vascular endothelium in nasal polyps. Such an effect may interfere with leucocyte extravasation and partially account for the anti-inflammatory effect of local glucocorticoid treatment in human nasal polyposis.

Nasal polyps and their relation to polyps/hypertrophic polypoid mucosa in the paranasal sinuses: a macro-, endo-, and microscopic study of autopsy materials.

The present study comprised macro- and endoscopic screening of the nasosinusal complex in 56 autopsies, 24 with nasal polyps and 32 without. Seven had nasal polyposis (bilaterally more than two). Hypertrophic polypoid mucosa in the paranasal sinuses was not found in the cases with nasal polyposis and in only 2 (4%) of the total number (24) of cases with polyps. Microscopic examination of nine small polyps, the mucosa from which they originated, and control specimens showed accumulation of eosinophils in the mucosa from which the polyps originated, indicating localized inflammation.

[Chronic unilateral maxillary sinusitis caused by foreign bodies in the maxillary sinus]. / Kronisk unilateral sinuitis maxillaris forårsaget af fremmedlegemer i kaebehulen.

Chronic maxillary sinusitis is a common condition. The disease is most often caused by blockage of the sinus ostia or presence of an oroantral fistula. We report two cases of unilateral chronic maxillary sinusitis associated with foreign bodies in the maxillary sinus. In the first case the foreign body consisted of dental amalgam, in the second case of two silastic sponges left behind from a former operation because of "blowout" fracture. In the case of benign chronic maxillary sinusitis and foreign bodies, functional endoscopic sinus surgery (FESS) preceded by a CT-scan is an excellent approach, by which diagnosis as well as treatment can be achieved with minimal surgical trauma.

Site of origin of nasal polyps. Transcranially removed naso-ethmoidal blocks as a screening method for nasal polyps in autopsy material.

In autopsy material naso-ethmoidal blocks were removed in 19 patients. None of the patients had a preceding rhinoscopy. Before removal, the maxillary and sphenoidal sinuses were opened and screened for polyps. After removal the total naso-ethmoidal complex was carefully examined for nasal polyps. The site of origin of the individual polyp was registered and documented photographically. Nine polyps were found in five patients, of which four had no history of asthma, allergy or sinusitis. Two patients had unilateral solitary polyps, three had polyps bilaterally. All polyps were found in the meatus. Eighty-nine per cent of the polyps were related to the clefts from the ethmoid. Polyps were not observed inside the ethmoidal and other paranasal sinuses.

Genes that regulate both development and longevity in Caenorhabditis elegans.

The nematode Caenorhabditis elegans responds to conditions of overcrowding and limited food by arresting development as a dauer larva. Genetic analysis of mutations that alter dauer larva formation (daf mutations) is presented along with an updated genetic pathway for dauer vs. nondauer development. Mutations in the daf-2 and daf-23 genes double adult life span, whereas mutations in four other dauer-constitutive genes positioned in a separate branch of this pathway (daf-1, daf-4, daf-7 and daf-8) do not. The increased life spans are suppressed completely by a daf-16 mutation and partially in a daf-2; daf-18 double mutant. A genetic pathway for determination of adult life span is presented based on the same strains and growth conditions used to characterize Daf phenotypes. Both dauer larva formation and adult life span are affected in daf-2; daf-12 double mutants in an allele-specific manner. Mutations in daf-12 do not extend adult life span, but certain combinations of daf-2 and daf-12 mutant alleles nearly quadruple it. This synergistic effect, which does not equivalently extend the fertile period, is the largest genetic extension of life span yet observed in a metazoan.

DNA content in adenoid cystic carcinomas.

BACKGROUND: A retrospective study was performed on 51 patients with adenoid cystic carcinomas to see whether DNA ploidy, tumor stage, and histopathologic grading correlated with prognosis. METHODS: Histopathologic grading was performed according to Szanto et al and DNA content was estimated from archived material using the technique by Hedley et al. RESULTS: Thirty-nine tumors were DNA diploid and 12 were DNA aneuploid. Histologic grade III was more often associated with DNA aneuploidy than the lower grades (p = 0.011). DNA ploidy also correlated with clinical stage (p = 0.011). Log-rank analysis and Cox regression analysis of treatment failures revealed significant findings for S-phase value and DNA ploidy. CONCLUSIONS: Our results indicate that DNA ploidy estimations, S-phase value, and histologic grading are prognostic factors in adenoid cystic carcinomas. These examinations should therefore be incorporated in the evaluation of patients with adenoid cystic carcinomas.

[Results of hearing preservation via middle fossa approach in surgery for acoustic neurinoma]. / Resultater af hørebevarende acusticusneurinomkirurgi via fossa media-adgang.

The results of hearing preservation surgery in a series of 10 patients with 10 unilateral acoustic neuromas are reported. Mortality was 0%, complications including rhinoliquorrhoea occurred in 10%, and the tumour was removed in 100%. Hearing preservation was obtained in 80%. Normal or nearly normal postoperative facial nerve function evaluated one year after surgery was achieved in all cases. According to House-Brackmann classification, 60% had HB-1 and 40% had HB-2. The fact that no audiovestibular or electrophysiological examinations are specific for acoustic neuroma can not be overemphasized. Early diagnosis of these small tumors in patients with good hearing has made this kind of surgery possible.

En bloc removal of the ethmoid and ostiomeatal complex in cadavers, with a practical application.

Two different approaches, i.e. the anterior approach and the combined approach, for en bloc removal of the ethmoidal and ostiomeatal complex are described. In both techniques the contours of the face are completely maintained. Preliminary studies of prevalence of nasal polyps found by using these methods are described.

Aging and resistance to oxidative damage in Caenorhabditis elegans.

The dauer larva state and the age-1 mutation, both of which extend life-span in Caenorhabditis elegans, were tested for hyperresistance to cellular damage that may be relevant to aging. The age-1 strain TJ401 displayed hyperresistance to oxidative stress relative to its parental strain. The activities of two enzymes that protect cells from oxidative damage, superoxide dismutase (SOD) and catalase, showed an age-dependent increase in mutant animals, which was not seen in the parental strain. These increases in activities paralleled the time course of the hyperresistance. The results are consistent with the age-1 gene product functioning as a negative regulator of SOD and catalase activities. In wild-type and age-1 dauer larvae, elevated levels of SOD activity, but not of catalase activity, were present when compared with young adults. The common increase in SOD activity prompted cloning the C. elegans Cu/Zn SOD gene. Its position on the physical map of the genome was in the region to which the age-1 gene has been genetically mapped, but it is unlikely that a mutation at the SOD locus confers the Age phenotype. Results support the free radical theory of aging by suggesting that the increased resistance to oxidative stress may be among the causes of increased longevity in both strain TJ401 and in the dauer larva.