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PubMed2002

cDNA cloning and heterologous expression of the major allergens from peach and apple belonging to the lipid-transfer protein family.

BACKGROUND: Lipid-transfer proteins (LTPs) have been identified as major allergens of Rosaceae fruits in populations living in areas virtually free of Fagales trees, such as several Mediterranean communities. Pru p 3 and Mal d 3, the allergens from peach and apple, respectively, have a main clinical relevance in these areas. OBJECTIVE: To isolate and characterize cDNAs for Pru p 3 and Mal d 3, and to produce recombinant Pru p 3 in the yeast Pichiapastoris. METHODS: cDNAs for both allergens were isolated by polymerase chain reaction using nondegenerated primers. Expression of Pru p 3 was performed in P. pastoris using the pPIC 9 vector. The recombinant product was purified by gel-filtration chromatography followed by RP-HPLC. Immunodetection and immunoblot inhibition assays were carried out with sera from peach-allergic patients. RESULTS: The cDNAs for both Pru p 3 and Mal d 3 showed a 273 open reading frame coding for the 91 amino acid mature polypeptides. The deduced amino acid sequences exhibited N-terminal regions fully identical to those previously determined for the natural peach and apple allergens. Pru p 3 was expressed in P. pastoris at 20 mg/L of culture medium. The recombinant allergen showed the same N-terminal sequence (plus a glutamic acid added for proper extracellular expression) and apparent molecular size as natural Pru p 3. Both the recombinant and natural forms of Pru p 3 displayed similar immunoglobulin (Ig)E-binding capacity in immunodetection and immunoblot inhibition assays. CONCLUSIONS: Comparison of the complete primary structures of mature Pru p 3 and Mal d 3 deduced from their corresponding cDNA clones supports the close relationship between both allergens. Recombinant Pru p 3 binds IgE in vitro like its natural counterpart. Therefore, it can be a useful tool for specific diagnosis and structural studies.

Allergens↗
PubMed2001

Microbial ecology of human skin in health and disease.

Cultivation of human skin reveals numerous bacteria and at least one fungus to be normal inhabitants of this ecosystem; however, most of our knowledge about the microbiology of human skin was acquired decades ago. Modern techniques employing nucleic acid-based microbial identification methods demonstrate the limitations of cultivation for appreciating microbial diversity in many ecosystems. The application of modern molecular methods to the study of skin may offer new perspectives on the resident microfora, and new insights into the causes of antibiotic responsive dermatologic conditions, such as acne and rosacea.

Acne Vulgaris↗
PubMed2004

Mail surveys can achieve high response rates in a dermatology patient population.

In order to understand the burden of skin disease on patient populations, researchers need to be able to measure exposures and outcomes of interest in a population-based study. One method of obtaining such information is mailed patient surveys. This method of obtaining information, however, raises concerns regarding possible low response rates, and to the best of our knowledge, this method has not been attempted in large dermatology patient populations with a high response rate. We sought to determine whether using a slightly modified version of the Dillman Total Design Method as a mailed survey protocol would result in a high response rate in a dermatology population. A mail survey was sent using a slightly modified version of Dillman's Total Design Method to 4894 patients seen in the Dermatology Department of the University of Pennsylvania, who were diagnosed with nonmelanoma skin cancer, dermatophytosis, acne rosacea, seborrheic keratosis, or warts; 74.1% of the subjects responded to the mailings; 69.8% (n=3203) of patients returned a completed survey. Response rates (both as overall response and as survey completers only) were high across a wide spectrum of dermatologic illnesses and did not vary significantly by entry diagnosis. Dillman's Total Design Method appears to be an effective tool for researchers studying the burden of skin disease in a large sample of dermatology patients.

Aged↗
PubMed1999

The red face-an overview and delineation of the MARSH syndrome.

The problem of the red face in females is reviewed. After excluding common causes such as contact dermatitis, seborrhoeic eczema and photodermatitis the diseases affecting the remaining patients fall into three groups: marked erythema with no feeling of heat or sensitivity, usually erythromelanosis faciei; marked flushing and burning with intense sensitivity for which the term facial erythrodysaesthesia is proposed; the so-called MARSH syndrome in which an overlap of androgen-dominant symptoms occurs - melasma, acne, rosacea, seborrhoeic eczema, and hirsutism. The latter group may respond best to low dose oral isotretinoin.

Erythema↗
PubMed2003

Five cases of cutaneous Rosai-Dorfman disease.

Rosai-Dorfman disease (RDD), previously known as sinus histiocytosis with massive lymphadenopathy, is a benign, idiopathic histiocytic proliferative disorder. It commonly affects lymph nodes, but any organ of the body may be involved. Histological findings include characteristic large, pale, histiocytic cells (Rosai-Dorfman cells) exhibiting cytophagocytosis. Immunohistochemically, these histiocytes are positive for S-100 protein and CD68, but stain negatively for CD1a. On electron microscopy, Birbeck granules are absent. RDD limited to the skin is rare, less than 30 cases having been reported to date. We present five further cases of purely cutaneous RDD. Three presented as solitary nodules and one as a large, well-circumscribed plaque. The fifth patient, who was HIV positive, had a rosacea-like facial eruption.

Adult↗
PubMed2003

Comparative analysis of the self-incompatibility (S-) locus region of Prunus mume: identification of a pollen-expressed F-box gene with allelic diversity.

BACKGROUND: Self-incompatibility (SI) in the Solanaceae, Rosaceae and Scrophulariaceae is gametophytically controlled by a single polymorphic locus, termed the S-locus. To date, the only known S-locus product is a polymorphic ribonuclease, termed S-RNase, which is secreted by stylar tissue and thought to act as a cytotoxin that degrades the RNA of incompatible pollen tubes. However, understanding how S-RNase causes S-haplotype specific inhibition of pollen tubes has been hampered by the lack of a cloned pollen S-determinant gene. RESULTS: To identify the pollen S-determinant gene, we investigated the genomic structure of the S-locus region of the S1- and S7-haplotypes of Prunus mume (Japanese apricot), and identified 13 genes around the S-RNase gene. Among them, only one F-box gene, termed SLF (S-locus F-box), fulfilled the conditions for a pollen S-determinant gene: (i) together with the S-RNase gene, it is located within the highly divergent genomic region of the S-locus, (ii) it exhibits S-haplotype specific diversity among three analysed S-haplotypes, and (iii) it is specifically expressed in pollen, but not in the styles or leaves. CONCLUSION: The results indicate that SLF is a prime candidate for the pollen S-determinant gene of SI.

Amino Acid Sequence↗
PubMed1999

Molecular phylogeography of Dryas integrifolia: glacial refugia and postglacial recolonization.

Chloroplast DNA variation in the Arctic plant species Dryas integrifolia (Rosaceae) was analysed in relation to both the present-day geographical distribution of populations and to Pleistocene fossil records of this species. The phylogeographical structure was weak but the analysis of haplotype diversity revealed several groups of haplotypes having present-day geographical ranges that overlap locations postulated from geographical and fossil evidence to have been glacial refugia. Based on this information we infer that two important refugial sources of Arctic recolonization by this species were Beringia and the High Arctic. Two other putative refugia, located southeast of the ice sheet and along coastal regions of the eastern Arctic may have served as sources for recolonization of smaller portions of the Arctic. The genetic substructure in the species is mostly due to variation among populations regardless of the ecogeographical region in which they are found. Spatial autocorrelation at the regional scale was also detected. High levels of diversity both within populations and ecogeographical regions are probably indicative of population establishment from several sources possibly combined with recent gene flow.

Journal Article↗
PubMed2004

Genetic effects of chronic habitat fragmentation on tree species: the case of Sorbus aucuparia in a deforested Scottish landscape.

Sustainable forest restoration and management practices require a thorough understanding of the influence that habitat fragmentation has on the processes shaping genetic variation and its distribution in tree populations. We quantified genetic variation at isozyme markers and chloroplast DNA (cpDNA), analysed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in severely fragmented populations of Sorbus aucuparia (Rosaceae) in a single catchment (Moffat) in southern Scotland. Remnants maintain surprisingly high levels of gene diversity (HE) for isozymes (HE = 0.195) and cpDNA markers (HE = 0.490). Estimates are very similar to those from non-fragmented populations in continental Europe, even though the latter were sampled over a much larger spatial scale. Overall, no genetic bottleneck or departures from random mating were detected in the Moffat fragments. However, genetic differentiation among remnants was detected for both types of marker (isozymes Theta n = 0.043, cpDNA Theta c = 0.131; G-test, P-value < 0.001). In this self-incompatible, insect-pollinated, bird-dispersed tree species, the estimated ratio of pollen flow to seed flow between fragments is close to 1 (r = 1.36). Reduced pollen-mediated gene flow is a likely consequence of habitat fragmentation, but effective seed dispersal by birds is probably helping to maintain high levels of genetic diversity within remnants and reduce genetic differentiation between them.

DNA, Chloroplast↗
PubMed2002

Molecular analysis of the stylar-expressed Solanum chacoense small asparagine-rich protein family related to the HT modifier of gametophytic self-incompatibility in Nicotiana.

Gametophytic self-incompatibility (GSI) systems involving the expression of stylar ribonucleases have been described and extensively studied in many plant families including the Solanaceae, Rosaceae and Scrophulariaceae. Pollen recognition and rejection is governed in the style by specific ribonucleases called S-RNases, but in many self-incompatibility (SI) systems, modifier loci that can modulate the SI response have been described at the genetic level. Here, we present at the molecular level, the isolation and characterization of two Solanum chacoense homologues of the Nicotiana HT modifier that had been previously shown to be necessary for the SI reaction to occur in N. alata (McClure et al., 1999). HT homologues from other solanaceous species have also been isolated and a phylogenetic analysis reveals that the HT genes fall into two groups. In S. chacoense, these small proteins named ScHT-A and ScHT-B are expressed in the style and are developmentally regulated during anthesis identically to the S-RNases as well as following compatible and incompatible pollination. To elucidate the precise role of each HT isoform, antisense ScHT-A and RNAi ScHT-B lines were generated. Conversion from SI to self-compatibility (SC) was only observed in RNAi ScHT-B lines with reduced levels of ScHT-B mRNA. These results confirm the role of the HT modifier in solanaceous SI and indicate that only the HT-B isoform is directly involved in SI.

Amino Acid Sequence↗
PubMed2000

Demodex folliculorum in pregnancy.

BACKGROUND: Demodex folliculorum (DF), which is a resident in human pilosebaceous follicles, has been implicated in rosacea, blepharitis, pityriasis folliculorum, pustular folliculitis, and skin lesions of immunosuppressed patients on chemotherapy or with acquired immunodeficiency syndrome (AIDS). OBJECTIVE: We aimed to determine the influence of pregnancy on the carriage of DF. METHODS: Mite density in 40 pregnant women was compared with that of age- and sex-matched controls. Two standardized skin surface biopsies were taken from each subject. Results In pregnant women, DF was found at 0.55/cm2, compared with a mean value of 1.2/ cm2 in nonpregnant controls. The difference between the two groups was not statistically significant (Mann-Whitney test, P = 0.22). CONCLUSIONS: Our findings indicate that pregnancy may not have a direct effect on mite density.

Adolescent↗
PubMed1999

Presence of asparagine-linked N-acetylglucosamine and chitobiose in Pyrus pyrifolia S-RNases associated with gametophytic self-incompatibility.

S-RNases encoded by the S-locus of rosaceous and solanaceous plants discriminate between the S-alleles of pollen in gametophytic self-incompatibility reactions, but it is not clear how. We report the structures of N-glycans attached to each of the N-glycosylation sites of seven S-RNases in Pyrus pyrifolia of the Rosaceae. The structures were identified by chromatographic analysis of pyridylaminated sugar chains prepared from S4-RNase and by liquid chromatography/electrospray ionization-mass spectrometric analysis of the protease digests of reduced and S-carboxymethylated S-RNases. S4-RNase carries various types of sugar chains, including plant-specific ones with beta1-->2-linked xylose and alpha1-->3-linked fucose residues. More than 70% of the total N-glycans of S4-RNase are, however, an N-acetylglucosamine or a chitobiose (GlcNAcbeta1-->4GlcNAc), which has not been found naturally. The N-acetylglucosamine and chitobiose are mainly present at the N-glycosylation sites within the putative recognition sites of the S-RNase, suggesting that these sugar chains may interact with pollen S-product(s).

Acetylglucosamine↗
PubMed2001

Delayed diffuse lamellar keratitis after laser in situ keratomileusis.

Two cases are reported of delayed diffuse lamellar keratitis after uneventful laser in situ keratomileusis. The first patient presented with an epithelial defect 6 weeks after laser in situ keratomileusis. Three days later the defect was healed but diffuse lamellar keratitis was noted. This was treated with topical dexamethasone and ketorolac with complete resolution of the diffuse lamellar keratitis over 3 weeks. The second patient presented with an epithelial defect and gross diffuse lamellar keratitis 10 weeks after laser in situ keratomileusis. Treatment was with topical dexamethasone and ciprofloxacin with gradual resolution of the diffuse lamellar keratitis. Common to both patients was a background of rosacea, implanted debris with no initial reaction, and epithelial defects leading to diffuse lamellar keratitis. It is suggested that these two cases represent epithelial defect associated corneal infiltration, which resembles classical diffuse lamellar keratitis with the spread of inflammatory cells through a path of least resistance.

Administration, Topical↗
PubMed1999

Visualization of harpin secretion in planta during infection of apple seedlings by Erwinia amylovora.

Erwinia amylovora is a Gram-negative pathogenic bacterium that infects pear and apple trees as well as other plants from the Rosaceae family. E. amylovora pathogenicity is dependent on a functional Hrp type III secretion system. Harpin, a protein playing a major role in virulence, has been shown to be exported in vitro via the type III secretion apparatus. The data presented here focus on harpin detection in planta after infection of apple seedlings with the wild-type strain CFPB 1430. Using a specific harpin antiserum, harpin was not detected inside the host plant cells, but was found associated with the bacteria and secreted. The extracellular localization of harpin is in agreement with the physiological effects induced by purified harpin when applied as an exogenous elicitor. Harpin was not found associated with the host plant cell wall, a result that weakens its postulated role in cell wall loosening. A differential labelling was observed at the bacterial level: for some bacteria, harpin was exclusively cytoplasmic, whereas in others, it appeared as small patches over the bacterial outer membrane or associated with extracellular linear structures. All the bacteria present within the same area were similarly labelled, suggesting co-ordination in the secretion process. All observations suggest that harpin is synthesized in the bacterial cytoplasm and that secretion occurs from this cytoplasmic pool upon sensing of a plant or bacterial signal.

Bacterial Outer Membrane Proteins↗
PubMed2001

Beneficial effect of Helicobacter pylori eradication in dermatologic diseases.

BACKGROUND: A relationship between skin diseases, particularly rosacea and chronic urticaria, and H. pylori infection has been suggested. METHODS: We preformed a prospective evaluation of the effect of H. pylori eradication in patients with a variety of chronic skin diseases. Patients were followed monthly for at least one year after cure of the infection. The effect of therapy was scored using a three point scale: complete remission (> 90% improvement), partial remission (50-90% improvement) or no improvement (< 50 improvement). The relationship between response and anti-H. pylori Ig G and E to specific H. pylori antigens was analyzed by Western blot analysis. RESULTS: Eighty-eight H. pylori-infected patients with skin disease were enrolled. Treatment was successful in 73% of patients with chronic urticaria as 23% (6 of 26) had complete and 50% had partial remission. Sixty-two percent (18 of 29) with pruritus cutaneus had partial remission as well as 30% with prurigo chronica multiformis had complete remission. Western blotting was done on 24 patients with skin disease and a 44K H. pylori antigen was detected by Ig E analysis in 100% (5 of 5) patients with complete remission compared to 23% (3 of 13) in those without skin disease. CONCLUSIONS: These results suggest it may be prudent to test patients with chronic urticaria, prurigo chronica multiformis, pruritus cutaneus, and eczema nummulare for H. pylori infection and to eradicate the infection in those whose test is positive.

Anti-Bacterial Agents↗
PubMed2002

Surgical management of rhinophyma: report of eight patients treated with electrosection.

BACKGROUND: Rhinophyma is the end stage of rosacea, affecting almost exclusively male patients, characterized by a progressive deformity in nasal shape. Depending on the extent of the deformity, rhinophyma may distort the appearance of those affected and patients may become psychosocially disabled. OBJECTIVE: To present the results of electrosurgical technique of sequential delamination with a slightly damped current in rhinophyma. METHODS: Eight healthy male patients affected by moderate to severe rhinophyma were recruited. A radiofrequency electrosurgical unit was used in the cut/coagulate mode at 40 W (3.8 MHz) to remove thin layers of tissue until the nose shape was re-created. RESULTS: All patients achieved acceptable cosmetic results. Cutaneous reepithelialization was achieved at day 14 after surgery. Erythema subsided within 4-8 weeks and healing was complete with minimal scar formation by 8-10 weeks. None of the patients developed hypertrophic scarring and hypopigmentation was minimal. CONCLUSION: Electrosection is as effective as laser treatments and less expensive. Although living in a laser era, electrosurgical remodeling of the nose is a good alternative for the treatment of rhinophyma.

Aged↗
PubMed2003

Biofeedback, cognitive-behavioral methods, and hypnosis in dermatology: is it all in your mind?

Biofeedback can improve cutaneous problems that have an autonomic nervous system component. Examples include biofeedback of galvanic skin resistance (GSR) for hyperhidrosis and biofeedback of skin temperature for Raynaud's disease. Hypnosis may enhance the effects obtained by biofeedback. Cognitive-behavioral methods may resolve dysfunctional thought patterns (cognitive) or actions (behavioral) that damage the skin or interfere with dermatologic therapy. Responsive diseases include acne excoriée, atopic dermatitis, factitious cheilitis, hyperhidrosis, lichen simplex chronicus, needle phobia, neurodermatitis, onychotillomania, prurigo nodularis, trichotillomania, and urticaria. Hypnosis can facilitate aversive therapy and enhance desensitization and other cognitive-behavioral methods. Hypnosis may improve or resolve numerous dermatoses. Examples include acne excoriée, alopecia areata, atopic dermatitis, congenital ichthyosiform erythroderma, dyshidrotic dermatitis, erythromelalgia, furuncles, glossodynia, herpes simplex, hyperhidrosis, ichthyosis vulgaris, lichen planus, neurodermatitis, nummular dermatitis, postherpetic neuralgia, pruritus, psoriasis, rosacea, trichotillomania, urticaria, verruca vulgaris, and vitiligo. Hypnosis can also reduce the anxiety and pain associated with dermatologic procedures.

Biofeedback, Psychology↗
PubMed2002

Allergic contact dermatitis to latanoprost.

An 85-year-old male with glaucoma presented with a 1-1/2 year history of tearing; red eyes; and pruritic, edematous, eczematous eyelids. Treatment for presumed ocular rosacea and seborrhea was unhelpful. Patch testing to our standard 64 antigens yielded a positive reaction to Balsam of Peru. Notably, benzalkonium chloride, thimerosal, and other preservatives elicited negative reactions. Repeat open application testing elicited positive results to Xalatan (latanoprost) 0.005% ophthalmic solution (Pharmacia & Upjohn, Kalamazoo, MI). A second session of patch tests to 10 personal products, in addition to Xalatan 0.005% solution and the Xalatan vehicle (provided by the manufacturer), elicited a strong positive reaction only to the full preparation of Xalatan 0.005% solution. This report describes the first known case of ACD to latanoprost, a new prostaglandin analog that is widely prescribed for treatment of glaucoma.

Aged↗
PubMed2001

Tacrolimus clinical studies for atopic dermatitis and other conditions.

The first topical immunomodulator approved for human use, tacrolimus ointment (Protopic, Fujisawa, Healthcare, Inc, Deerfield, IL), has been shown to be effective and safe in the treatment of children (aged 2 years and older) and adults with atopic dermatitis (AD). Clinical trials conducted worldwide have involved 12,000 patients, with safety and efficacy data available for up to 3 years of treatment. In addition to its beneficial effects in the management of AD, topical tacrolimus has also been reported to be of benefit in other immunologically mediated skin diseases including: hand dermatitis, contact dermatitis, eyelid dermatitis, erosive lichen planus, steroid-induced rosacea, pyoderma gangrenosum, and graft-versus-host disease. This article reviews the clinical experience of topical tacrolimus in the treatment of AD and other skin conditions.

Administration, Cutaneous↗