bluuebunny/crossref_metadata_2025 huggingface dataset:
Optimization of codon usage is required for effective genetic immunization against Art v 1, the major allergen of mugwort pollen
Authors: Bauer R. , Himly M. , Dedic A. , Ferreira F. , Thalhamer J. , Hartl A.
Published: September 2003

Background:  As the major allergen of mugwort pollen, Art v 1 is an important target for specific immunotherapy. However, both recombinant protein as well as a gene vaccine for Art v 1 failed to be immunogenic in mice. In order to improve immunogenicity we focused on genetic immunization because interspecific differences of codon usage have been shown as an obstacle for effective induction of immune responses with gene vaccines encoding infectious pathogens.

Objective:  In order to find out, whether codon usage might also be used to improve genetic immunization with allergen genes, the response against a gene vaccine expressing the wild‐type gene of Art v 1 (pCMV‐wtArt) was compared with a synthetic codon‐optimized vector with human codon usage (pCMV‐humArt).

Methods:  Balb/c mice were injected intradermally with pCMV‐wtArt or pCMV‐humArt. In vitro expression levels of both constructs were compared in transfection experiments. Total immunoglobulin G (IgG), IgG1, IgG2a and IgE antibodies were analyzed by enzyme‐linked immunosorbent assay and the anaphylactic activity of the sera was determined by allergen‐specific degranulation of rat basophil leukemia‐2H3 cells.

Results:  No immune response was detectable with the gene vaccine expressing the wildtype Art v 1, but immunization with pCMV‐humArt revealed a strong and allergen‐specific induction of antibody responses. The antibodies recognized both the recombinant as well as the purified natural (glycosylated) Art v 1 molecule. The response type was Th1‐biased, as indicated by high levels of IgG2a antibodies. Expression analysis with B16 mouse melanoma cells transfected with pCMV‐humArt or pCMV‐wtArt revealed an impaired expression of the wild‐type vector but normal translation after recoding.

Conclusion: The results demonstrate that optimization of codon usage offers a simple way to improve immunogenicity and therefore should be routinely considered in the development of gene vaccines for the treatment of allergy.
Interleukin (IL)‐4 and to a lesser extent either IL‐13 or interferon‐gamma regulate the production of eotaxin‐2/CCL24 in nasal polyps
Authors: Lezcano‐Meza D. , Dávila‐Dávila B. , Vega‐Miranda A. , Negrete‐García M. C. , Teran L. M.
Published: September 2003

Background:  Eotaxin‐2/CCL24 is a potent eosinophil attractant that has been implicated in the recruitment of eosinophils in allergic disease. We have investigated whether the cytokines interleukin (IL)‐4, IL‐13, and interferon (IFN)‐gamma regulate eotaxin‐2/CCL24 in nasal polyps.

Methods:  Nasal polyps were cultured in the presence of the cytokines described above and the concentration of eotaxin‐2/CCL24 was measured in the culture supernatant.

Results:  IL‐4 was found to be the major stimulus for eotaxin‐2/CCL24 production from nasal polyps followed by IL‐13 and IFN‐gamma. IL‐4 induced eotaxin‐2/CCL24 in a dose‐dependent manner with concentrations as low as 0.1 ng/ml being able to induce eotaxin‐2/CCL24. By immunohistochemistry, eotaxin‐2/CCL24 immunoreactivity was localized to mononuclear cells in the IL‐4 stimulated nasal polyp tissue. Interestingly, nasal turbinates obtained from patients suffering from nonallergic rhinitis (vasomotor rhinitis) were also found to release eotaxin‐2/CCL24 both spontaneously and following cytokine stimulation with IL‐4 and IFN‐gamma being major inducers of this cytokine.

Conclusions:  All together these findings suggest that Th1 and Th2 cytokines may regulate eotaxin‐2/CCL24 production in nasal polyps and nonallergic rhinits.
Assessment of inhaled BDP‐dose dependency of exhaled nitric oxide and local and serum eosinophilic markers in steroids‐naive nonatopic asthmatics
Authors: Negro R. Dal , Micheletto C. , Tognella S. , Turco P. , Rossetti A. , Cantini L.
Published: September 2003

The aim of the present study was to assess the dose‐dependency from inhaled steroids of changes of airways inflammation [eosinophils count and eosinophil cationic protein (ECP)] measures in induced sputum and in serum, as well as that of exhaled nitric oxide. Twenty steroid‐naive patients with nonatopic asthma of mild to moderate degree [forced expiratory volume in 1 s (FEV1) = 70% of predicted] and with negative response to the standard tests for allergy were selected; after a 1‐week run‐in period they were randomized to receive a 12‐week treatment period of inhaled beclomethasone dipropionate dry powder given with the Pulvinal® inhaler (Clenil P®, Chiesi Farmaceutici S.p.A., Parma, Italy) in two different dose regimens, 400 μg bid (high dose) or 200 μg bid (low dose), over a double blind, parallel groups design. The following outcome measures were assessed in baseline and after 1, 6 and 12 weeks of treatment: FEV1 (l), eosinophils count in sputum (%), is ECP (μg/l), serum eosinophils count (%), serum ECP (μg/l) and exhaled NO (ppb). The results showed that all the considered parameters improved in both groups: the increase over baseline of FEV1 and the decrease of NO were significant at any time in the high‐dose group and only at week 12 in the low‐dose group (NS between groups), whereas the markers of eosinophilic activity showed more consistent reductions in the high‐dose than in the low‐dose group when measured in induced sputum (P < 0.05 between groups after 6 and 12 weeks for eosinophils count and after 12 weeks for ECP). Decreases over baseline of markers measured in serum were more rapid in the high‐dose group, without differences between groups. A marked trend towards a negative correlation was found between FEV1 and ECP, (r = −0.72, P < 0.05), between FEV1 and eosinophils in sputum (r = −0.31, NS) and between FEV1 and exhaled NO (r = −0.38, NS), all of them only in the high‐dose group. The results of the study demonstrate that changes of levels of eosinophilic activity in the airways are dependent from the daily dose of inhaled steroids when measured in induced sputum and that the local assessment can therefore represent a practical and noninvasive method to monitor the extent of airways inflammation.
Allergen‐dependent CD14 modulation and apoptosis in monocytes from allergic patients
Authors: Monteseirín J. , Bonilla I. , Chacón P. , Vega A. , Camacho M. J. , Guardia P. , Conde J. , Sobrino F.
Published: September 2003

Background: CD14 is a most important monocyte surface molecule. Recently, it has been reported that there is an important relationship between CD14 and immunoglobulin E, and that regulation of CD14 expression is an effector mechanism mediating apoptosis of monocytes.

Objective: The present study was designed to determine whether specific allergens were able to modulate CD14 expression and apoptosis by monocytes from allergic patients or whether specific immunotherapy (IT) might affect these processes.

Methods: One group of adult allergic asthmatic patients had received IT for the previous 3 years. Another similar group was not treated with IT. We challenged peripheral blood monocytes from both groups of asthmatic patients in vitro with the specific allergen that produced clinical symptoms in asthmatic patients. The cells were also challenged with allergen to which the patients were not sensitive. Monocytes from normal subjects were also challenged with allergens. Expression of CD14 on the monocyte surface was analyzed by flow cytometry, and soluble CD14 (sCD14) in culture supernatant by enzyme‐linked immunosorbent assay. The three groups of subjects were challenged with allergens, and apoptosis was analyzed by flow cytometry.

Results: When monocytes from non‐IT‐treated asthmatic patients were cultivated with the allergens to which the patients were sensitive, a significant up‐regulation on the monocyte surface was observed compared with results from the healthy group (P < 0.003) and from the IT asthmatic group (P < 0.003). A significantly higher sCD14 level was observed in the culture supernatant of the monocytes from the IT asthmatic group were observed compared with those from the healthy group (P < 0.001) and those from the non‐IT asthmatic group (P < 0.001). A significantly higher apoptosis level was observed in monocytes from the IT asthmatic group compared with those from the healthy group (P < 0.001) and those from the non‐IT asthmatic group (<0.001).

Conclusions: We present evidence that the expression of CD14 on the surface of monocytes and the apoptosis of the same cells can be modulated by an allergen‐dependent mechanism. These processes can be affected by IT.
Human serum IgE‐mediated mast cell degranulation shows poor correlation to allergen‐specific IgE content
Authors: Marchand F. , Mecheri S. , Guilloux L. , Iannascoli B. , Weyer A. , Blank U.
Published: September 2003

Background:  Although allergen‐specific IgE content in serum can be determined immunochemically, little is known about the relationship between this parameter and the strength of the degranulation response upon allergen triggering.

Objectives:  Analyse the degranulation capacity of immunochemically defined purified and serum IgE after challenge with anti‐IgE or allergen using a rat mast cell line (RBL) transfected with the α‐chain of the human high‐affinity IgE receptor (FcɛRI).

Methods:  Purified IgE specific for 4‐hydroxy‐3nitrophenylacetyl, purified IgE of unknown specificity, and sera from allergic patients sensitive to Dermatophagoides pteronyssinus and Dactylis glomerata were assessed. Degranulation was measured by a β‐hexosaminidase release assay after anti‐IgE or allergen‐specific challenge.

Results:  For purified monoclonal IgE a significant correlation (r = 0.97) was found between the proportion of bound allergen‐specific IgE and the strength of the degranulation response. In contrast, no correlation (r = 0.27) was detected after sensitization with serum IgE.

Conclusion:  Our studies demonstrate that mast cell activation mediated through IgE from allergic patients is a result of complex relationships that are not only dependent on allergen‐specific IgE content but also relate to the capacity to efficiently sensitize and trigger the signalling responses that lead to degranulation.
Prevalence of asthma with aspirin hypersensitivity in the adult population of Poland
Authors: Kasper L. , Sladek K. , Duplaga M. , Bochenek G. , Liebhart J. , Gladysz U. , Malolepszy J. , Szczeklik A.
Published: September 2003

Background: Acetylsalicylic acid (ASA) and other nonsteroid anti‐inflammatory drugs (NSAIDs) are reported to account for 21–25% of all adverse drug reactions. Some asthmatics may react to ASA and other NSAIDs with acute bronchoconstriction, profuse rhinorrhea and skin flushing. This is a distinct clinical syndrome called aspirin‐induced asthma (AIA). The prevalence of AIA among asthmatic patients in Poland has not been previously assessed.

Methods: A questionnaire survey of 12 970 adults of both sexes, randomly selected from the population of Poland.

Results: The prevalence of AIA in the general population of Poland was estimated as 0.6%. Thirty patients (4.3%; 95% CI: 2.8–5.8) of 703 asthmatics (5.4% of general population) reported symptoms attesting to hypersensitivity to aspirin. In 27% of them the reactions were precipitated by aspirin, whereas in the remaining subjects by other NSAIDs.

Conclusions: The prevalence of AIA in Poland is 4.3%, being somewhat lower than in Finland and Australia, where it was recently reported to account for 8.8 and 10.9% of the adult asthmatics, respectively. These figures indicate that aspirin hypersensitivity might be a significant community problem.
Serum inhibitors for human mast cell growth: possible role of retinol
Authors: Ishida S. , Kinoshita T. , Sugawara N. , Yamashita T. , Koike K.
Published: September 2003

Background: In vitro culture systems have been used to study the physiological and pathological characteristics of human mast cells. However, there are some differences in proliferation and maturation of mast cells between fetal bovine serum (FBS)‐containing and serum‐deprived cultures. Accordingly, we attempted to identify circulating factor(s) affecting the development of human mast cells.

Methods: We measured the serum levels of retinol and several cytokines. To elucidate the antiproliferative effects of the serum, a retinoic acid receptor (RARα) antagonist and neutralizing antibodies against cytokines were used.

Results: Similar to FBS, human serum dose‐dependently suppressed the growth of tryptase+ cells from CD34+ cord blood cells or 20‐week cultured mast cells under stimulation with stem cell factor (SCF). The serum‐mediated inhibition might be based on a decline in proliferation rate. Among inhibitors for mast cell growth, retinol and transforming growth factor (TGF)‐β1 were present at high levels in human serum. In contrast with anti‐TGF‐β1 antibody, an RARα antagonist counteracted the serum‐induced suppression of human mast cell proliferation.

Conclusions: Our results suggest that retinol and its derivatives act as a circulating regulator for human mast cell growth. The RARα antagonist may be a useful tool to obtain higher numbers of mast cells in FBS‐containing cultures.
Two CD14 promoter polymorphisms and atopic phenotypes in Czech patients with IgE‐mediated allergy
Authors: Bučková D. , Hollá L. I. , Schüller M. , Znojil V. , Vácha J.
Published: September 2003

Background: Immunoglobulin E (IgE)‐mediated allergy belongs to common chronic disorders resulting from an interaction between both genetic and environmental factors. The gene encoding CD14 is a positional candidate gene for allergic diseases as it is localized on chromosome 5q31.1, a region that is linked to asthma and bronchial hyperresponsiveness. Recently, several polymorphisms in the promoter region of this gene have been associated with atopic phenotypes in various populations.

Methods: We investigated relationship among atopic phenotypes and two polymorphisms [C(‐159)T and G(‐1359)T] in the promoter of the CD14 gene in the Czech population. Polymerase chain reaction with restriction fragment length polymorphism analyses was used to determine the CD14 genotypes in subjects with IgE‐mediated allergic diseases (n = 562) and random controls (n = 320).

Results: The CD14 allele or genotype distributions were similar in patients and control group. However, the frequency of the C allele of the C(‐159)T polymorphism was higher in patients with positive skin prick tests for moulds than in patients without reactivity to this antigen (P < 0.002, Pcorr<0.01). In addition, we found that patients with homozygous genotype (GG) of the G(‐1359)T polymorphism had marginally lower percentage of positive skin prick tests compared with the other genotypes (P < 0.029, Pcorr > 0.05).

Conclusions: Our study supports the idea that CD14 gene variants may act as disease modifiers of IgE‐mediated allergic diseases.
Childhood asthma and continuous exposure to cats since the first year of life with cats allowed in the child's bedroom
Authors: Oberle D. , von Mutius E. , von Kries R.
Published: September 2003

Background: There are controversial data as to interdependencies of exposure to furred pets in infancy and the prevalence of asthma and hay fever in children. Does the timing, intensity and type of pet exposure matter?

Methods: Cross‐sectional questionnaire data on 8216 German schoolchildren aged 5–7 years not living on a farm in ten rural districts in Bavaria in 1997 were analysed. The diagnosis of asthma and hay fever was ascertained with the International Study of Asthma and Allergies in Childhood (ISAAC) core questions. Wheeze and asthma were classified as ‘atopic’ in children who also had hay fever or atopic dermatitis. Prevalence and intensity of exposure to pets in the first year of life and at present were assessed via questionnaire.

Results: Although the study was of considerable size we found no convincing association between atopic disease and pet exposure in general. Exposure to cats from the first year of life to school entry, however, was associated with a reduced prevalence of atopic asthma, if cats were allowed to be in the child's bedroom: no case of atopic asthma in 296 children exposed and an aOR 0.11 (95% CI:0.01–0.52) for atopic wheeze in the last 12 months.

Conclusions: Allowing cats to be in the child's bedroom from the first year of life onwards may be an indicator of intensive exposure to cats and appears to prevent the development of childhood asthma.
Maternal smoking in pregnancy alters neonatal cytokine responses
Authors: Noakes P. S. , Holt P. G. , Prescott S. L.
Published: September 2003

Background: Maternal cigarette smoking in pregnancy is an important, common and avoidable exposure that has been linked with elevated cord blood (CB) immunoglobulin E levels and subsequent asthma and allergic disease in childhood. Despite this, there is still very little information about the immunological effects of maternal smoking on the fetus.

Methods: This aim of this study was to compare cord blood mononuclear cell (CBMC) cytokine responses to allergens [ovalbumin (OVA) or house dust mite (HDM)] and mitogens [concanavalin A (ConA) or phytohemaglutinen (PHA)] in neonates whose mothers smoked throughout pregnancy (n = 17) with responses of neonates whose mothers never smoked (n = 40). Cell cultures were stimulated for 24 h and supernatants collected for cytokine detection by enzyme‐linked immunosorbent assay [interleukin (IL)‐13, IL‐6, interferon (IFN)γ and IL‐10]. Cell pellets were also collected for cytokine mRNA detection (IL‐5, IL‐9, IFNγ).

Results: Maternal smoking in pregnancy was associated with significantly higher neonatal T helper type 2 (IL‐13 protein) responses to both HDM (P = 0.01) and OVA (P = 0.035). These effects remained statistically significant after allowing for confounding factors, including the effects of maternal atopy. Similar trends were also seen for IL‐9mRNA, IL‐5mRNA and IL‐6 responses, although these were not statistically significant. Although IFNγ mRNA responses to PHA (P = 0.015) and ConA (P = 0.025) were lower if mothers smoked in pregnancy, there were no differences in neonatal (Th1) IFNγ protein responses to allergens or mitogens.

Conclusions: These findings indicate that maternal cigarette smoking can modify aspects of fetal immune function and highlight the need for further studies in this area.
Small vulvar squamous cell carcinomas and adjacent tissues. A morphologic study
Authors: POULSEN HEMMING , JUNGE JETTE , VYBERG MOGENS , HORN THOMAS , LUNDVALL FINN
Published: September 2003

Vulvar squamous cell carcinomas are of different subtypes and degrees of differentiation, and may be associated with adjacent lichen sclerosus and/or varying degrees of dysplasia. The aim of this investigation was to study small carcinomas with a diameter of less than 2 cm in order to find a possible relation between subtypes of carcinomas and adjacent epithelial changes.

Fourteen cases of small vulvar squamous cell carcinomas were totally embedded in paraffin. Serial sectioning made a detailed mapping of all different lesions possible, and a two‐ and three‐dimensional imaging was obtained in each case.

Seven patients with keratinizing squamous cell carcinomas (median age 65) had adjacent lichen sclerosus. All carcinomas were completely surrounded by areas of VIN1. VIN2 and VIN3 were not found. Seven patients without lichen sclerosus (median age 58) showed squamous cell carcinomas of the keratinizing type (n=2) or the basaloid type (n=5). Five of these cases were incompletely surrounded by varying degrees of dysplasia, mainly VIN2 and VIN3.

Two different pathogenetic pathways for the development of vulvar squamous cell carcinoma are likely.
Strong associations between gene function and codon usage
Authors: FUGLSANG ANDERS
Published: September 2003

The association between codon usage and gene function was analyzed in the complete genomes of Eschericia coli, Bacillus subtilis, Lactococcus lactis and Campylobacter jejuni, using the functional annotation provided by NCBI. Two distinctly different ways of quantifying codon usage were used in the analysis. By using contingency tables it was found that for most amino acids a highly significant association with gene function exists for all species, indicating that codon usage at the level of individual amino acids is generally closely coordinated with gene function. By computing the effective number of codons in the annotated genes and comparing the median values in groups of different gene functions it was shown for all species that codon bias gene by gene also differs.
Usefulness of different techniques in the study of the epidemiology of salmonellosis
Authors: RUIZ M. , RODRÍGUEZ J. C. , SIRVENT E. , ESCRIBANO I. , CEBRIÁN L. , ROYO G.
Published: September 2003

Our objective was to evaluate the usefulness of serotyping, phagotyping, antibiotyping and RAPD (random amplified polymorphic DNA) in the study of the epidemiology of salmonellosis in Elche (Spain). We examined 1232 clinical isolates of non‐typhoid Salmonella serotypes. Serotyping adequately differentiates 12.3% of the isolates and phagotyping raises this percentage to 37%. Antibiotyping is a suitable complement in certain circumstances, such as in the case of specific outbreaks. RAPD is a fast and simple technique which is within the reach of most laboratories, complementing the results obtained by antibiotyping and serotyping, in order to be able to control outbreaks of salmonellosis in everyday practice.
Patterns of mutations in target genes in septicemia isolates of Escherichia coli and Klebsiella pneumoniae with resistance or reduced susceptibility to ciprofloxacin
Authors: FENDUKLY F. , KARLSSON I. , HANSON H. S. , KRONVALL G. , DORNBUSCH K.
Published: September 2003

Thirty‐two Escherichia coli and 21 Klebsiella pneumoniae septicemia isolates with varying degrees of resistance to ciprofloxacin were analyzed for the presence of point mutations within the quinolone‐resistance target genes. The number of mutations observed in the resistant isolates agreed with the level of ciprofloxacin resistance in both species. Such isolates were also resistant to nalidixic acid. Isolates with borderline susceptibility to ciprofloxacin, on the other hand, behaved differently in the two species. In E. coli all the isolates harbored at least one mutation and these isolates were also resistant to nalidixic acid, while no mutations were detected in the K. pneumoniae isolates, and susceptibility to nalidixic acid was unpredictable. Therefore, nalidixic acid cannot be used as a class representative. Time‐kill curve studies on an isolate with borderline susceptibility from each species showed higher degrees of resistance to ciprofloxacin in comparison to that of the wild‐type E. coli. A previously unreported parC mutation, S57→T, was detected in a resistant E. coli isolate and might expand the QRDR of this gene. Normalized resistance interpretations of histograms confirmed the setting of microbiological zone breakpoints for ciprofloxacin testing.
Selective lack of the C16:0 fatty acid isoform of sulfatide in pancreas of type II diabetic animal models
Authors: BLOMQVIST MARIA , OSTERBYE THOMAS , MÅNSSON JAN‐ERIC , HORN THOMAS , BUSCHARD KARSTEN , FREDMAN PAM
Published: September 2003

Sulfatide (3′‐sulfogalactosyl‐ceramide) is a glycosphingolipid mainly located in the nervous system, but has also been found in the islets of Langerhans. Previous studies have suggested that sulfatide is involved in insulin processing and secretion. In this study, sulfatide expression and metabolism in pancreas and isolated islets of the type II diabetes models, ob/ob‐ and db/db mouse, was investigated using TLC‐ELISA, metabolic labelling and electron microscopy. As in non‐diabetic Lewis rat and human pancreas, sulfatide was located in secretory granules of the beta cells. However, the type II diabetic animal models and their background strains had an altered sulfatide expression, involving the lack of the C16:0 sulfatide fatty acid isoform, compared to non‐diabetic Lewis rat, BALB/c mouse and human pancreatic tissue, in which the two dominating pancreatic sulfatide isoforms C16:0 and C24:0 are expressed. Correspondingly, in isolated ob/ob islets, sulfatide synthesis excluded the production of C16:0 sulfatide. Insulin administration to ob/ob mouse, which lowers beta cell activity, resulted in significantly increased sulfatide expression in pancreas (p=0.0003), but still no expression of the C16:0 sulfatide isoform. In vitro, the C16:0 sulfatide was shown to be the isomer involved in the preservation of insulin crystals. Thus, it is hypothesized that the selection of sulfatide isomers in pancreas might be a genetic factor contributing to disease development in type II diabetic animal models.
Identification of Friend murine retrovirus‐infected immune cells and studies of the effects of sex and steroid hormones in the early phase of infection
Authors: BRULAND TORUNN , LAVIK LISS ANNE S. , DAI HONG YAN , DALEN ARE
Published: September 2003

Male mice are more susceptible than female mice to the murine retrovirus FIS‐2. We previously reported that sex‐related factors influence early virus replication via mechanisms involving a glucocorticoid response element (GRE) in the long terminal repeat (LTR) enhancer region. In the present study, we investigated further the influence of sex and steroid hormones on early murine retrovirus dissemination and immune functions. In male mice we found a correlation between an early expansion of the CD8+ cell subset and rapid infection of lymphocytes, including CD8+ cells. Virus load in blood declined faster in females than in males, and the postpeak declines coincided with more rapidly generation of antibodies against virus‐positive cells. Moreover, female‐derived T‐cells responded better to in vitro mitogen stimuli than male‐derived T‐cells. Physiological concentrations of progesterone and dexamethasone induced a dose‐dependent inhibition of T‐cell proliferation. Administration of progestin in vivo did not modify early FIS‐2 production in female mice. Male castrated mice, who were notably less involved in aggressive behaviour and fighting compared to male control mice, had a significant delay of virus dissemination. We suggest that testosterone‐dependent aggression, with successive enhanced stress hormone levels, may influence both FIS‐2 replication and immune responses during infection.
Cytokine and surface receptor diversity of NK cells in resistant C3H/HeN and susceptible BALB/c mice with chronic Pseudomonas aeruginosa lung infection
Authors: CALUM HENRIK , MOSER CLAUS , JENSEN PETER ØSTRUP , SHIRAI RYO , HØIBY NIELS
Published: September 2003

The purpose of the present study was to investigate whether NK cells from resistant C3H/HeN mice and susceptible BALB/c mice showed different release of cytokines and expression of surface molecules during chronic P. aeruginosa lung infection using alginate‐embedded P. aeruginosa mimicking the infection in cystic fibrosis. Lung cell suspensions were depleted of lymphocytes by magnetic cell sorting. The concentrations of IFN‐γ, IL‐1β and GM‐CSF were estimated by ELISA at day 1 and 2 after infection. Non‐infected mice were used as controls. Flow cytometry was used to estimate the surface expression of the LFA‐1 and Fc receptors on NK cells. At day 2, IFN‐γ levels increased in C3H/HeN mice but decreased in BALB/c mice. The GM‐CSF levels increased only in the C3H/HeN mice at day 1 and 2. Surface expression of LFA‐1 on the NK cells was higher in C3H/HeN mice at day 1 and 2. In contrast, the expression of Fc receptors was significantly lower on NK cells in C3H/HeN mice at day 1 and 2. In conclusion, the present results show phenotypic differences in NK cells in the two mice strains in chronic P. aeruginosa lung infection, indicating different modulating effects in the Th1/Th2 balance.
Apocrine change in Fine‐needle aspiration biopsy: nuclear morphometry and DNA image cytometry
Authors: ELZAGHEID ADEM , KUOPIO TEIJO , KORHONEN ANNA‐MAIJA , COLLAN YRJO
Published: September 2003

The aim of this study was to investigate the potential of computerized nuclear morphometry and DNA image cytometry in characterizing the apocrine change of mammary epithelium in fine‐needle aspiration biopsy (FNAB). The effect of two different sample processing techniques on the results was also studied. Mean nuclear areas in air‐dried smears ranged from 59.0 μm2 to 151.0 μm2 and in ethanol‐fixed samples from 32.3 μm2 to 63.4 μm2. The DNA histograms of apocrine cells usually showed a dominant peak in the diploid region. In some cases the mode of the peak was slightly shifted to the right or left in respect to the control peak. One case had a tetraploid cell population, suggesting atypical apocrine change. After histological investigation this case was diagnosed as infiltrating carcinoma. The patient had earlier been treated with x‐ray irradiation for a mediastinal lymphoma. Findings of nuclear morphometry and DNA cytometry in apocrine metaplasia are here described in a systematic study for the first time. The data suggest that these methods may help in distinguishing premalignant and malignant apocrine lesions from typical apocrine metaplasia of mammary epithelial cells.
Evaluation of different disk diffusion/media combinations for detection of methicillin resistance in Staphylococcus aureus and coagulase‐negative staphylococci
Authors: SKOV ROBERT , LARSEN ANDERS RHOD , FRIMODT‐MØLLER NIELS , ESPERSEN FRANK
Published: September 2003

In order to find a disk diffusion method with both high sensitivity and specificity for determination of methicillin resistance primarily for S. aureus but also for coagulase‐negative staphylococci we screened several methodological variants using a material of 66 S. aureus comprising of 11 methicillin‐susceptible, 18 borderline‐resistant, and 37 methicillin‐resistant strains. Only four of the combinations studied performed with both high sensitivity and specificity. Two of these, the Columbia agar +4.5% NaCl and Mueller Hinton agar +2% NaCl combined with a 5 μg oxacillin disk, confluent inoculum and 24 h incubation at 35 °C were further evaluated using 105 MRSA and 91 mecA‐negative S. aureus and 193 clinical isolates of coagulase‐negative staphylococci. The Columbia agar +4.5% NaCl performed excellently for both S. aureus and coagulase‐negative staphylococci. For Columbia agar +4.5% NaCl using a 5 μg oxacillin disk we suggest an interpretive zone diameter of R≤15 mm and S≥16 mm for S. aureus and R≤24 mm and S≥26 mm for coagulase‐negative staphylococci. The Mueller Hinton agar +2% NaCl performed well for coagulase‐negative staphylococci but for S. aureus at least three (3%) very major errors were found, making this method less attractive.
Analysis of protein tyrosine kinase expression in melanocytic lesions by tissue array
Authors: Shen Steven S. , Zhang Peter S. , Eton Omar , Prieto Victor G.
Published: September 2003

Background:  It has been proposed that melanoma progression involves a multistep process from benign nevi (BN), dysplastic nevi (DN), radial and vertical growth phase melanoma (MM) to metastatic melanoma (MMM). Protein tyrosine kinases (PTKs) may participate in this progression.

Methods:  Tissue microarray blocks of 89 melanocytic lesions were evaluated by immunohistochemistry for the expression of selected PTKs: c‐kit, c‐abl, abl‐related gene (ARG), platelet‐derived growth factor receptors α (PDGFR‐α) and β (PDGFR‐β).

Results:  Seventeen of 31 (55%) MMM lacked expression of c‐kit versus 100% expression (18/18) in DN and 96% expression (22/23) in MM; similarly, only 59% (10/17) of BN showed expression of c‐kit. PDGFR‐β expression levels were similar in BN, DN, and MM, but lower in MMM. There was a trend toward lower expression of abl and ARG from BN to MMM. There was a marked decrease in staining intensity of ARG from BN to DN, MM, and MMM.

Conclusion:  Our results support that BN is different from DN and MM and that these two are different from MMM. Metastasis appears to be associated with loss of c‐kit and PDGFR‐β expression. Since malignant melanoma expresses PTK, it may be a candidate for treatment with anti‐PTK, such as STI‐571 (Gleevec).
Scarring alopecia associated with mastocytosis
Authors: Xu Xiaowei , Solky Benjamin , Elenitsas Rosalie , Cotsarelis George
Published: September 2003

Background:  Mastocytosis is comprised of a group of heterogeneous diseases involving various organs. Urticaria pigmentosa is the most common manifestation of cutaneous mastocytosis; others include mastocytoma, diffuse mastocytosis, and telangiectasia macularis eruptiva perstans.

Methods:  We describe a case of indolent mastocytosis presenting as scarring alopecia. The scalp biopsy revealed a perifollicular and dermal inflammatory infiltrate composed predominantly of mast cells, which was confirmed by tryptase and Giemsa stains.

Results:  The preponderance of mast cells in the biopsy prompted testing for urine N‐methylhistamine levels, which were elevated and confirmed the diagnosis of mastocytosis. This is the first report of mastocytosis presenting as scarring alopecia.

Conclusions:  This case suggests that the diagnosis of mastocytosis should be entertained in patients presenting with scarring alopecia accompanied by an intense mast cell infiltrate on scalp biopsy and also supports the notion that mast cells may be involved in the pathogenesis of alopecia.
Myxoid Spitz nevus
Authors: Hoang Mai P.
Published: September 2003

Background:  The deposition of extracellular mucin has not been described in Spitz nevus and herein such a case is reported.

Methods:  A 6‐year‐old male presented with a growing 1.0‐cm pigmented lesion on his left anterior knee. The lesion was excised.

Results:  The histologic sections demonstrated a symmetrical and dome‐shaped proliferation of spindle and epithelioid melanocytes with sharp lateral demarcation at the dermal–epidermal junction and within the superficial dermis. Features of Spitz nevus such as retraction from the epidermis, eosinophilic bodies, and uniform cytologic atypia were seen. Of interest, there was marked mucin deposition within the epidermal clefts and between the neoplastic cells.

The mucin was highlighted by alcian blue and colloidal iron stains, and it was negative with mucicarmine and periodic acid–Schiff stains.

Conclusion:  Awareness that soitz nevi can rarely have extracellular mucin helps in avoiding diagnostic pitfalls.
Subungual pleomorphic fibroma
Authors: Hsieh Ya‐Ju , Lin Yang‐Chih , Wu Yu‐Hung , Su Hsin‐Yi , Billings Steven D. , Hood Antoinette F.
Published: September 2003

Background:  Pleomorphic fibroma is a benign fibroblastic tumor characterized by pleomorphic, hyperchromatic cells or giant multinucleated cells embedded in a collagenous stroma. These cytologic features may lead to an incorrect diagnosis of malignancy. Most cases reported in the literature are located on trunk or extremities; the presentation as a subungual mass is rare.

Methods:  We report an unusual case of a subungual pleomorphic fibroma in 66‐year‐old woman. Clinical information was obtained. Histologic examination and immunohistochemical studies were performed.

Results:  A 66‐year‐old woman presented with a longstanding (40 years), subungual mass that deformed the nail of the left middle finger. Microscopic examination revealed a paucicellular tumor composed of hyperchromatic spindled, pleomorphic, floret‐like giant cells embedded in haphazardly arranged collagen bundles in the dermis. No mitotic figures were seen. The tumor cells were vimentin‐positive but did not stain with antibodies to S‐100, cytokeratin, smooth muscle actin, factor XIIIa or CD34 negative. The diagnosis of a pleomorphic fibroma was made. Follow‐up shows no evidence of tumor, 36 months after excision.

Conclusion:  Pleomorphic fibroma of the subungual region is an unusual cutaneous tumor with histologic features that may cause confusion with true sarcomas. This is only the second case reported of a subungual pleomorphic fibroma. Pleomorphic fibroma should be considered in the differential diagnosis of pleomorphic subungual tumors.
Chronic vulvar purpura: persistent pigmented purpuric dermatitis (lichen aureus) of the vulva or plasma cell (Zoon's) vulvitis?
Authors: Li Qing , Leopold Kathy , Carlson J. Andrew
Published: September 2003

Background:  Lichen aureus is localized variant of persistent pigmented purpuric dermatitis that typically affects the legs and can be associated with delayed hypersensitivity reactions or vascular abnormalities. Plasma cell vulvitis (Zoon's vulvitis) is a rare condition that frequently contains hemosiderin deposits and is suspected to be a mucosal reaction pattern due to variety of insults, most often local irritation or trauma.

Case Report:  A 50‐year‐old female with longstanding complaints of spotting, vulvar dryness, irritation, and dyspareunia presented with circumscribed, purpuric, erythematous vulvar patches. Past estrogen cream treatment evoked symptoms of discomfort. On biopsy, siderophages and extravasated red blood cells were found in conjunction with a lichenoid, lymphocyte and plasma cell infiltrate, and dilated dermal and intraepithelial vessels.

Conclusions:  Reported herein is an unusual vulvar dermatosis that is best classified as a localized variant of persistent pigmented dermatosis (lichen aureus) but overlaps clinically and histologically with Zoon's vulvitis. This constellation of findings may represent a site‐specific mucosal reaction to an erosive process that could either be inflammatory (hypersensitivity reaction) and/or traumatic in nature.
Cutaneous sclerosing perineurioma of the digits: an uncommon soft‐tissue neoplasm. Report of two cases with immunohistochemical analysis
Authors: Canales‐Ibarra Claudia , Magariños Gabriel , Olsoff‐Pagovich Paul , Ortiz‐Hidalgo Carlos
Published: September 2003

Background:  Cutaneous sclerosing perineurioma is a recently characterized, uncommon tumor composed of perineurial cells, which exhibits immunoreactivity for epithelial membrane antigen (EMA). These lesions occur preferentially in children and young adults and usually develop as dermal or subcutaneous nodules in the hands or palms.

Methods:  We report two cases of cutaneous sclerosing perineurioma in young patients without stigmata of neurofibromatosis. Histologically, these lesions were well‐circumscribed masses and were characterized by a variable number of epithelioid and spindle cells with wavy nuclei end elongated cytoplasmic processes embedded in a dense collagen stroma.

Results:  These cells showed focal whorling formation, demonstrated robust immunoreactivity for EMA and CD99, and were uniformly negative for S‐100 protein, actin (HHF‐35), CD34, cytokeratin AE1‐3, and CD57.

Conclusion:  We comment on the differential diagnosis of fibrous cutaneous lesions based on immunohistochemistry.
Widespread cutaneous and perioral metastases of mesothelioma
Authors: Cassarino David S. , Xue Wayne , Shannon Kevin J.
Published: September 2003

Background:  Cutaneous involvement by mesothelioma represents a rare occurrence from an uncommon neoplasm. Most cases have resulted from local extension of an underlying body cavity mesothelioma or from surgical site contamination. We could only find six previously reported cases of distant cutaneous metastases of mesothelioma.

Methods:  We describe a case of metastatic mesothelioma involving multiple skin sites and the lip in a 64‐year‐old man with an underlying primary pleural mesothelioma.

Results:  A 64‐year‐old man presented with a lip lesion clinically diagnosed as keratoacanthoma vs. squamous cell carcinoma. Evaluation of the lip biopsy revealed tubulo‐glandular and acinar‐like arrangements of plump epithelioid cells with a hobnailed appearance, vesicular nuclei, and prominent nucleoli. The initial impression of metastatic carcinoma was revised to probable mesothelioma upon discovery of a previous history of pleural mesothelioma. Positive immunohistochemical stains for anti‐cytokeratin, anti‐calretinin, and HBME‐1 and negative stains for anti‐CEA, Leu‐M1, B72.3, and Ber‐Ep4 confirmed the diagnosis.

Conclusion:  We report a rare example of multiple cutaneous metastases of mesothelioma. We also demonstrate the usefulness of relatively new and specific immunomarkers for mesothelioma vs. adenocarcinoma. Metastatic mesothelioma to the skin or lip is a very rare occurrence but should be considered in the differential diagnosis of malignant epithelioid neoplasms.
A novel epidermal nevus syndrome with congenital cylindromatous turban tumor
Authors: Regalado Jacinto J.
Published: September 2003

Background:  Epidermal nevi (in the broad sense of epithelial nevi) may give rise to benign or malignant skin tumors. They may also be associated with anomalies of other organ systems in an epidermal nevus syndrome.

Results:  This article describes a preterm infant with nevus sebaceus of the scalp and face, a large turban tumor with features of malignant cylindroma and multiple non‐cutaneous defects. These included skeletal, hematopoietic, hepatobiliary, and urinary anomalies. Severe secondary lesions were present (pulmonary hypoplasia due to oligohydramnios; cerebral infarcts probably related to the turban tumor). Karyotype was normal, and family history was negative.

Conclusions:  This unique case is unlike any reported epidermal nevus syndrome. Similarly, there is no prior report of a congenital cylindroma, certainly not as a turban tumor, which implies very rapid growth. The presence of both overgrowth and undergrowth phenomena (e.g. hypoplastic urinary tract and biliary atresia) may reflect dysregulation of paracrine growth factors, presumably due to genetic mutation.
Intertriginous epidermal dysmaturation from pegylated liposomal doxorubicin
Authors: English Joseph C. , Toney Rachel , Patterson James W.
Published: September 2003

Background:  A new formulation of doxorubicin (pegylated liposomal doxorubicin) has been developed to attenuate the systemic side effects produced by this agent. Although pegylated liposomal doxorubicin has a much lower risk for cardiotoxicity, it has been associated with cutaneous side effects that differ from, and may also be more frequent than, those produced by the free form of the drug.

Methods:  We report a case of interface dermatitis with keratinocyte dysmaturation, limited to the intertriginous areas, in a patient who received pegylated liposomal doxorubicin, and we review the literature.

Results:  It is possible that the enhanced delivery of doxorubicin in liposomal form promotes cutaneous side effects of the drug. We consider the possible mechanisms for keratinocyte dysmaturation produced by this form of doxorubicin.

Conclusions:  Our findings further suggest that inflammatory changes seen on biopsy may not always allow a reliable histopathologic distinction between this chemotherapy effect and graft‐versus‐host disease.
Differential expression of α1 (IV) and α5 (IV) collagen chains in basal‐cell carcinoma
Authors: Quatresooz Pascale , Martalo Oriane , Piérard Gérald E.
Published: September 2003

Background:  Basement membrane alterations are common in malignancies, and they may indicate tumoral aggressiveness. Distinct patterns of tumoral coverage by collagen IV were reported in nodular and aggressive basal‐cell carcinomas (BCCs). Differential expressions of α (IV) collagen chains were also shown on frozen sections. The aim of our work was to document the immunohistochemical expression of α1, α3, and α5 (IV) collagen chains in BCC after routine fixation and processing.

Methods:  The patterns of distribution of α1 (IV), α3 (IV), and α5 (IV) collagen chains were studied in 20 formalin‐fixed and paraffin‐embedded BCCs showing different infiltrative patterns. One trichoblastoma was used as control.

Results:  In nodular BCCs, the expression of α5 (IV) collagen chain was downregulated and uneven. By contrast, α1 (IV) collagen chain expression was preserved around these tumors similar to the surrounding skin. However, the α1 (IV) collagen chain expression was discontinuous or absent in BCC areas showing an infiltrative pattern of extension. The α3 chain was absent both underneath all BCCs and non‐neoplastic skin.

Conclusions:  The basement membrane alterations around nodular BCCs involved more precisely the α5 (IV) collagen chains. Defects in α1 (IV) collagen chain expression seemed to be associated with a tumoral invasive and infiltrative pattern. The biological significance of these findings is unclear.
Characteristic immunohistochemical and ultrastructural findings indicate that Kindler's syndrome is an apoptotic skin disorder
Authors: Lanschuetzer C. M. , Muss W. H. , Emberger M. , Pohla‐Gubo G. , Klausegger A. , Bauer J. W. , Hintner H.
Published: September 2003

Background:  Kindler's syndrome is a rare genodermatosis mainly characterized by the onset of skin blistering in early childhood, web formation of fingers and toes, photosensitivity, and progressive poikiloderma. There is still debate whether this disease represents a distinctive entity in the spectrum of congenital bullous poikilodermas or a variant of dystrophic epidermolysis bullosa.

Objective:  To evaluate the recently proposed and debated characteristic immunohistochemical and ultrastructural features of Kindler's syndrome.

Patient/methods:  Immunofluorescence (IF) antigen mapping and transmission electron microscopy (TEM) were performed on a skin specimen from non‐sun‐exposed inner aspect of the upper arm of a 49‐year‐old patient with characteristic clinical features of Kindler's syndrome.

Results:  IF studies revealed focally an extensively broadened, partly reticular staining pattern in the dermoepidermal basement membrane zone (BMZ) with antibodies against laminin‐5 and type IV as well as type VII collagen. Anti‐α6 and β4 integrin staining revealed small gaps in the linear reactivity in the BMZ. Abundant keratin bodies, as detected by anti‐immunoglobulin M (IgM) staining, were focally present in the dermis, indicating prominent epidermal apoptosis. This was verified by a histochemical apoptosis stain [terminal deoxynucleotidyl transferase‐mediated dUTP nick‐end labeling (TUNEL) reaction]. Transmission electron microscopic examination showed manifold reduplications of the lamina densa (with attached anchoring fibrils) as well as a keratin body surrounded by a fibroblast in the upper dermis.

Conclusion:  We present characteristic immunohistochemical and ultrastructural features of Kindler's syndrome identical to those described by Shimizu et al. (Arch Dermatol 1997; 133: 1111) and provide evidence that Kindler's syndrome might primarily be an apoptotic disorder of basal keratinocytes.
A single ring of charged amino acids at one end of the pore can control ion selectivity in the 5‐HT3 receptor
Authors: Thompson Andrew J , Lummis Sarah C R
Published: September 2003

To determine the mechanisms by which cation‐ or anion‐specific channels select between these ions, we have examined the role of electrostatic factors in a typical ligand‐gated ion channel, the 5‐hydroxytryptamine3 (5‐HT3) receptor, by removal and/or insertion of rings of conserved charged residues at either end of the pore.

Neutralization of the negatively charged ring at the intracellular end of the channel (E−1′A) results in a nonselective channel (PNa/PCl=0.89).

Insertion of positively charged residues at the extracellular end of the pore either results in a nonfunctional receptor (A24′K) or one that remains cation‐selective (PNa/PCl=110; S19′R).

Combining the removal of a negatively charged ring (E−1′A) with the insertion of a positively charged ring (S19′R), however, results in a channel that is predominantly anion‐selective (PNa/PCl=0.37).

The data suggest that for the cation‐selective 5‐HT3 receptor, the control of selectivity exerted by charged rings at either end of the pore is dominated by the ring of negatively charged residues at the intracellular side of the channel. As changing the charge at this position has also been shown to change ionic selectivity in anion‐selective receptors, these data suggest that electrostatic factors can control selectivity in the whole Cys‐loop family.

British Journal of Pharmacology (2003) 140, 359–365. doi:10.1038/sj.bjp.0705424