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Cyclooxygenase inhibitory and antioxidant compounds from crabapple fruits.

Crabapple trees belong to the Malus genus (Rosaceae) and bear fruits that are sparingly consumed and used in the preparation of fruit beverages. Cyclooxygenase (COX) enzyme inhibitory and antioxidant bioassay-guided fractionation of the aqueous and methanol extracts of Malus x kornicensis and Malus x Indian Summer yielded (+)-catechin (1), (-)-epicatechin (2), cyanidin-3-O-beta-galactopyranoside (3), and amygdalin (4). Pure compounds 1-4 were obtained by HPLC, identified by LC-ES/MS, CD, and NMR spectroscopic methods and evaluated for their COX enzyme inhibitory and antioxidant activities. In COX-1 and -2 enzyme inhibitory assays, compounds 1-3 (all at 80 microM) showed activities of 20.4, 46.3%; 57.6, 47.9%; and 8.2, 13.7%, respectively, compared to naproxen (54.3, 41.3%; 10 microM), ibuprofen (47.5, 39.8%; 10 microM), Celebrex (46.2, 66.3%; 1.67 ppm), and Vioxx (23.8, 88.1%, 1.67 ppm). In the antioxidant assay, the catechins (1-2) and anthocyanin (3) (all at 40 microM) showed activities of 61.3, 62.5, and 60.1%, respectively. The synthetic antioxidants, tert-butylhydroquinone (TBHQ), butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), and vitamin E (all tested at 10 microM), gave 75.2, 80.1, 70.0, and 10.2% activities, respectively. The cyanogenic glycoside, amygdalin (4), and its hydrolysis products, mandelonitrile (5) and benzaldehyde (6), were not active in the antioxidant or COX enzyme inhibitory assays at 80 microM concentrations.

Amygdalin↗

Diterpenoid, steroid, and triterpenoid agonists of liver X receptors from diversified terrestrial plants and marine sources.

It has been demonstrated that liver X receptors (LXR) play a significant role in cholesterol homeostasis. Agonists of LXR are expected to increase cellular cholesterol efflux, lower LDL, and raise HDL levels. Screening of a natural product library of plant extracts using a LXR-SPA binding assay and bioassay-guided fractionation of a number of plant and marine gorgonian extracts led to the isolation of a number of active compounds. These included acanthoic acid (1) and alcohol (2), viperidone (3), polycarpol (4), rosacea acid (5), a cycloartane derivative (6), a new cycloartane analogue (7), betulinic acid (8), and gorgostane derivatives (9, 10, and 11). Of these compounds, 1, 4, and 11 exhibited potent binding affinity for alpha-receptor with IC(50) values of 0.25, 0.12, and 0.07 microM, respectively. Functionally they also showed strong coactivator association stimulation for LXRalpha receptor with EC50 values of 0.18, 0.03, and 0.05 microM, respectively. They also exhibited 15-, 8-, and 13-fold induction of the alpha-receptor in a transactivation assay in HEK-293 cells, respectively. In general these compounds were selective for the LXR alpha-receptor over the beta-receptor in all assays and were much better stimulators of the alpha-receptor than the endogenous steroid ligands.

Animals↗

Isolation and characterization of an antiviral flavonoid from Waldsteinia fragarioides.

The antiviral agent in a fraction from Waldsteinia fragarioides (Rosaceae) was purified using bioassay-guided fractionation of activity against herpes simplex type 1 virus. Structural eluciation by instrumental methods identified the active component to be the known flavonoid glycoside, isoquercitrin (3,3',4',5,7-pentahydroxyflavone-3 beta-O-glucoside), which had not previously been shown to possess antiviral activity.

Antiviral Agents↗

Primary structural features of rosaceous S-RNases associated with gametophytic self-incompatibility.

We isolated cDNA clones encoding five S-RNases (S1-, S3-, S5-, S6-, S7-RNases) from pistils of Pyrus pyrifolia (Japanese pear), a member of the Rosaceae. Their amino acid sequences were aligned with those of other rosaceous S-RNases sequenced so far. A total of 76 conserved amino acid residues were stretched throughout the sequence, but were absent from the 51-66 region which was designated the hypervariable (HV) region. The phylogenetic tree of rosaceous S-RNases showed that S-RNase polymorphism predated the divergence of Pyrus and Malus. Pairwise comparison of these S-RNases detected two highly homologous pairs, P. pyrifolia S1- and S4-RNases (90.0%) and P. pyrifolia S3- and S5-RNases (95.5%). The positions of amino acid substitutions between S1- and S4-RNases were spread over the entire region, but in the pair of S3- and S5-RNases, amino acid substitutions were found in the 21-90 region including the HV region. The substitutions in this restricted region appear to be sufficient to discriminate between S3 and S5 pollen and to trigger the self-incompatible reaction.

Alleles↗

An F-box gene linked to the self-incompatibility (S) locus of Antirrhinum is expressed specifically in pollen and tapetum.

In many flowering plants, self-fertilization is prevented by an intraspecific reproductive barrier known as self-incompatibility (SI), that, in most cases, is controlled by a single multiallelic S locus. So far, the only known S locus product in self-incompatible species from the Solanaceae, Scrophulariaceae and Rosaceae is a class of ribonucleases called S RNases. Molecular and transgenic analyses have shown that S RNases are responsible for pollen rejection by the pistil but have no role in pollen expression of SI, which appears to be mediated by a gene called the pollen self-incompatibility or Sp gene. To identify possible candidates for this gene, we investigated the genomic structure of the S locus in Antirrhinum, a member of the Scrophulariaceae. A novel F-box gene, AhSLF-S2, encoded by the S2 allele, with the expected features of the Sp gene was identified. AhSLF-S2 is located 9 kb downstream of S2 RNase gene and encodes a polypeptide of 376 amino acids with a conserved F-box domain in its amino-terminal part. Hypothetical genes homologous to AhSLF-S2 are apparent in the sequenced genomic DNA of Arabidopsis and rice. Together, they define a large gene family, named SLF (S locus F-box) family. AhSLF-S2 is highly polymorphic and is specifically expressed in tapetum, microspores and pollen grains in an allele-specific manner. The possibility that Sp encodes an F-box protein and the implications of this for the operation of self-incompatibility are discussed.

Alleles↗

Chromosome walking in the Petunia inflata self-incompatibility (S-) locus and gene identification in an 881-kb contig containing S2-RNase.

Self-incompatibility (SI) in the Solanaceae, Rosaceae and Scrophulariaceae is controlled by the polymorphic S locus, which contains two separate genes encoding pollen and pistil determinants in SI interactions. The S-RNase gene encodes the pistil determinant, whereas the pollen determinant gene, named the pollen S gene, has not yet been identified. Here, we set out to construct an integrated genetic and physical map of the S locus of Petunia inflata and identify any additional genes located at this locus. We first conducted chromosome walking at the S2 locus using BAC clones that contained either S2-RNase or one of the nine markers tightly linked to the S locus. Ten separate contigs were constructed, which collectively spanned 4.4 Mb. To identify additional genes located at the S2 locus, a 328-kb region (part of an 881-kb BAC contig) containing S2-RNase was completely sequenced. Approximately 76% of the region contained repetitive sequences, including transposon-like sequences. Other than S2-RNase, an F-box gene, named PiSLF2 (S2-allele of P. inflata S-locus F-box gene), was the only predicted gene whose deduced amino acid sequence was similar to the sequences of known proteins in the database. Two different cDNA selection methods were used to identify additional genes in the 881-kb contig; 11 groups of cDNA clones were identified in addition to those for S2-RNase and PiSLF2. RT-PCR analysis of expression profiles and PCR analysis of BAC clones and genomic DNA confirmed that seven of these 11 newly identified genes were located in the 881-kb contig.

Chromosome Walking↗

Prevalence of pollinosis in the Basque Country.

BACKGROUND: The prevalence of allergic diseases, mainly pollinosis, has increased within the last decades. Our study aimed to determine the prevalence of sensitization to Poaceae pollen in the Basque Country. This is a region of northern Spain, with an area of 7261 km2 and a population of 2109009 inhabitants. Two different climatic regions may be distinguished in the Basque Country (the Atlantic and the Oceanic). METHODS: A transversal study was carried out on 2216 subjects, aged 10-40 years. A personal interviewwas carried out in order to compile study data, by means of a questionnaire that had been previously validated with a clinical history and an allergy study. RESULTS AND CONCLUSIONS: Our results show the prevalence of pollinosis in the Basque Country to be 10.6% (C.I. 95% 9.35-11.96%) without significant differences between men and women. The prevalence in the Atlantic climate area (9.71%) was lower than in the Oceanic climate area (13.61%). There were no differences between persons living in a rural environment (10.87%) and those living in an urban setting (10.51%). Pollinosis was more frequent in individuals aged 10-20 (11.41%) and 20-30 (12.54%) than those aged 30-40 years (7.43%). Three features significantly distinguished pollinic from nonpollinic patients: 1) a more common complaint of symptoms after ingestion of Rosaceae fruits or nuts (10.2%) 2) a family history of atopy (8.8%) 3) a greater occurrence of bronchial asthmatic symptoms (23%).

Adolescent↗

An allergen from Olea europaea pollen (Ole e 7) is associated with plant-derived food anaphylaxis.

BACKGROUND: Several cross-reacting proteins have been identified as responsible of the co-occurrence of pollinosis and plant-derived food allergy. This association has been mainly described in the birch-apple syndrome but other pollens such as Olea europaea and other fruits may also contain homologous proteins. OBJECTIVE: To evaluate the associations between sensitization to allergens of Olea europaea pollen and confirmed plant-derived food allergy, in addition to investigate if any pattern of clinical hypersensitivity of food allergy reaction (oral allergy syndrome (OAS) or anaphylaxis) and/or any fresh fruit or nut allergy, are associated to one or several Olea pollen allergen(s). METHODS: One-hundred and thirty-four consecutive patients diagnosed with pollinosis by Olea were studied. Of these patients only 40, reported adverse reaction to plant-derived food. Twenty-one (group A) were classified as OAS and 19 (group B) as anaphylaxis. Skin-tests with six Olea pollen allergens and several groups of fruits, were performed. Double-blind placebo-controlled food challenge (DBPCFC), confirmed the diagnostics of food allergy with the exception of patients who suffered previous anaphylactic reaction. RESULTS: All patients, showed a positive skin prick test (SPT), against one or more of Olea europaea allergens. Sensitization to Ole e 7, was more frequent (P = 0.02) in patients from group B. A total of 84 DBPCFC were performed with 44% positive results. Challenge confirmed at least the 50% of positive SPT in any case (peach: 68.42%; pear: 50%; melon: 71.42% and kiwi: 53.84%). In patients from group B, significant association with O. europaea pollen allergens were found between positive SPT to Rosaceae fruits and Ole e 3 (P = 0.045) and Ole e 7 (P = 0.03); Cucurbitaceae and Ole e 7 (P = 0.03) and Actinidiaceae with Ole e 3 (P = 0.04). CONCLUSIONS: The results of this study, establish a new spectrum of associations between pollens and plant-derived foods: sensitization to olive profilin (Ole e 2) is not more frequent in OAS patients. Patients with anaphylactic reaction after eating fruit are also sensitized to Ole e 7, a LTP present in Olea pollen, and suffer pollinic symptoms. Finally a polcalcin (Ole e 3) could be also associated to Olea pollen respiratory and food allergy.

Adult↗

Reactivity to potential cross-reactive foods in fruit-allergic patients: implications for prescribing food avoidance.

BACKGROUND: Prescribing therapeutic elimination diets in patients with fruit allergy should include recommendations on which other foods of the same family or group may be safely consumed. Evidence-based data on the management of fruit allergy are lacking; therefore, advice may vary from just avoiding the offending fruit, to overly restrictive diets of the entire botanical family. The aims of this investigation were to assess clinical reactivity to potential cross-reactive foods in fruit-allergic patients, and the implications for prescribing specific therapeutic elimination diets. METHODS: Sixty-five adults diagnosed with clinical allergy to one or more fruits were evaluated for IgE-mediated allergy to other related foods, which might share cross-reactive antigens. Those with actual allergy to some Rosaceae fruit (including peach, apple, apricot, plum, and almond) underwent skin prick testing (SPT), food-specific IgE assessment by the Pharmacia CAP-FEIA system, and oral challenges with the entire group. For those with allergy to other fruits (chestnut, melon, banana, kiwi, or avocado) immunologic and clinical reactivity to all five were evaluated. Since a number of people in the study also had proven clinically allergy to some nut or legume (peanut, sunflower seed, walnut, pistachio, or hazelnut), the reactivity to the entire set was investigated. RESULTS: Thirty-four of those tested (52%) were found to be clinically allergic to more than one fruit, so 125 allergic reactions occurred in the 65 patients. Peach, melon, kiwi, apple, and banana accounted for 72% of allergic reactions. Forty-two (65%) had sensitization to pollens, causing seasonal allergic rhinitis and/or asthma, and 18 (28%) were sensitized to latex. The 65 with IgE-mediated fruit allergy underwent 351 additional SPT and food-specific IgE determinations with potential cross-reactive foods considered in this study; 223 (64%) of these results were positive. The routine challenges with potential cross-reactive foods uncovered 18 further reactions in 14 (22%) out of 65 (to avocado in seven; apricot in three; plum in one; almond and peanut in one; banana and hazelnut in one; avocado, banana and kiwi in one). Only 8% (18/223) of positive results for SPT, or food-specific IgE levels of greater than 0.35 kU(A)/l to potential cross-reactive foods investigated in this study, were clinically relevant. CONCLUSION: Elimination diets that rely on total group avoidance, or only on the results of allergy testing, might have resulted in unnecessary restriction of 205 foods in the 65 people studied. The lack of routine oral challenges with related fruits--before prescribing dietary restriction--may have allowed 18 food reactions in one-fifth (14/65) of patients to be overlooked.

Adolescent↗

Occupational dermatitis in dental personnel: contact dermatitis with special reference to (meth)acrylates in 174 patients.

Between 1995 and 1998, 174 dental personnel were referred as patients to the Department of Occupational and Environmental Dermatology, Stockholm. After clinical examination, 131 were patch tested with the Swedish standard series and 109 with a dental screening series. Furthermore, 137 were tested for IgE-mediated allergy to natural rubber latex (NRL). Hand eczema was diagnosed in 109/174 (63%), 73 (67%) being classified as irritant contact dermatitis and 36 (33%) as allergic. Further diagnoses included other eczemas, urticaria, rosacea, psoriasis, tinea pedis, bullous pemphigoid or no skin disease. 77/131 (59%) had positive reactions to substances in the standard series and 44/109 (40%) to substances exclusive to the dental series. 24/109 (22%) patients had positive reactions to (meth)acrylates, the majority with reactions to several test preparations. Reactions to HEMA (2-hydroxyethyl methacrylate), EGDMA (ethyleneglycol dimethacrylate) and MMA (methyl methacrylate) were most frequent. 9 of the 24 were positive only to (meth)acrylates, the remaining 15 also had reactions to allergens in the standard series. 23 of these had hand eczema and 1 facial eczema. In 17 of the 24 allergic to (meth)acrylates, the dermatitis had started in 1995 or later, in 15 within the previous 12 months. Of 8 who had been sick-listed, 7 also had a history of atopy and 6 were allergic to nickel. The most frequent allergens besides (meth)acrylates were nickel, cobalt, palladium, fragrance mix, colophonium and thiuram mix. Allergy to natural rubber latex was diagnosed in 14/137 (10%). In conclusion, irritant hand dermatitis was the dominant diagnosis. Contact allergy to (meth)acrylate was seen in 22% of the patch tested patients, with reactions to 3 predominant test substances. 1/3 of the patients with allergy to (meth)acrylates had been sick-listed for dermatitis, but in all these cases the (meth)acrylate allergy was seen together with atopy and/or further contact allergies.

Adult↗

Meibomian gland disease. Classification and grading of lid changes.

In recent years attention has been paid to meibomian gland dysfunction (MGD) as a distinct clinical entity responsible for chronic symptoms and signs and occurring independently or in association with atopy, cicatrising mucosal disorders and rosacea. Attempts to correlate MGD with microbiological and lipid biochemical changes are confounded by the absence of a clear descriptive language for the disorder and its associated changes. Such a language is crucial for the conduct of cross-sectional and natural history studies and therapeutic clinical trials. We present a comprehensive classification and grading scheme of meibomian gland disease, supporting our observations with illustrations.

Adolescent↗

Recurrent corneal erosion: clinical features.

The clinical features of a group of 30 patients with recalcitrant recurrent corneal erosions (i.e. those who failed to respond to conventional therapy) were evaluated. Associated ocular and facial abnormalities were documented. Meibomian gland dysfunction was present in all patients as manifest by dropout and inspissation of the meibomian glands, reduced tear film break-up time and debris in the tear film. Dropout of meibomian glands was present in 25 (83%) patients and was maximum in the medial half of the lid in 21 (84%) of these 25 patients. Tear film break-up time was reduced in all patients, being instant in 7 (23%), between 1 and 5 seconds in 22 (74%) and between 10 and 15 seconds in 1 (3%) patient. Superficial corneal abnormalities were present in 28 (93%) patients as manifest by maps, dots and fingerprints. Facial abnormalities such as telangiectasia, rhinophyma and acne rosacea were present in 22 (73%) patients. The findings of our study suggest an association between recalcitrant recurrent corneal erosions and meibomian gland dysfunction.

Adult↗

Superior forniceal conjunctival advancement pedicles (SFCAP) in the management of acute and impending corneal perforations.

OBJECTIVE/AIM: Corneal perforations can result from a wide variety of disorders and can lead to devastating visual sequelae. Various surgical procedures have been described to manage nontraumatic corneal perforation. Conjunctival flaps offer an important technique in dealing with such corneal emergencies. We report a modified conjunctival flap procedure referred to as superior forniceal conjunctival advancement pedicle (SFCAP) in the successful management of corneal perforation and impending corneal perforation. PATIENTS AND METHODS: Out of 20 patients who underwent SFCAP for non-traumatic corneal perforation and impending perforation, 16 had corneal perforations. Perforations were secondary to acne rosacea (2), advanced bacterial keratitis (6), corneal anaesthesia(2), multiple retinal procedures (2), previous corneal grafts with a compromised ocular surface (3), and advanced Mooren's ulcer (1). RESULTS: The globe was preserved in all patients. In 14 of the 16 eyes with perforated corneas the pedicle stabilised. One patient, who was a chronic alcoholic, rubbed the eye during sleep detaching the pedicle a week after surgery. However, the ulcer healed and the integrity of the globe was restored. None of these patients developed secondary glaucoma or ptosis. CONCLUSION: SFCAP is an appropriate procedure to manage corneal perforations and impending corneal perforations where donor material is not available and transplantation of such tissue is not suitable.

Acute Disease↗

Pathogenesis, clinical features and management of recurrent corneal erosions.

Recurrent corneal erosions (RCE) are common. They are characterised by repeated episodes of pain, difficulty in opening the eyes, watering, and photophobia resulting from poor epithelial adhesion. In the majority of patients with RCE, trauma is the initiating factor. Epithelial, stromal, and endothelial corneal dystrophies have all been described in association with RCE. Other causes that may lead to RCE include chemical and thermal injuries, previous herpetic keratitis, meibomian gland dysfunction, ocular rosacea, diabetes mellitus, Salzmann's nodular degeneration, band keratopathy, previous bacterial ulceration, kerato-conjunctivitis sicca, and epidermolysis bullosa. The conditions that are associated with RCE can be either primary or secondary depending on whether the basement membrane complex abnormality is intrinsic or acquired. Primary types tend to be bilateral, symmetrical and develop in multiple corneal locations. The pathogenetic mechanism of this disorder is related to poor adhesion of the corneal epithelium to the underlying stroma. Excessive matrix metalloproteinase (MMP) activity may play a role in the pathogenesis. Although the majority of patients will respond to simple measures such as padding and antibiotic ointment, RCE resistant to simple measures require approaches that are more elaborate. The common goal of these approaches is to encourage proper formation of adhesion complexes between the epithelium and the stroma. The use of long-term contact lenses, autologous serum eye drops, botulinum toxin, induced ptosis, oral MMP inhibitors, diamond burr polishing of Bowman's membrane have been reported with varying degree of success in treating RCE. Anterior stromal puncture with insulin needles or Neodymium : aluminium-yttrium-garnet may enhance the epithelial adhesion to the basement membrane by scar formation and success rates of up to 80% have been reported in the treatment of recalcitrant RCE. Excimer laser photo-therapeutic keratectomy (PTK) is now a well-established treatment modality for RCE and is being used both safely and effectively. Partial ablation of Bowman's layer with PTK gives a smooth surface for the newly generating epithelium to migrate and form adhesion complexes. The pathogenesis, clinical features, and management options of this common disorder are discussed in this review article.

Corneal Diseases↗

Molecular variation at the self-incompatibility locus in natural populations of the genera Antirrhinum and Misopates.

The self-incompatibility system of flowering plants is a classic example of extreme allelic polymorphism maintained by frequency-dependent selection. We used primers designed from three published Antirrhinum hispanicum S-allele sequences in PCR reactions with genomic DNA of plants sampled from natural populations of Antirrhinum and Misopates species. Not surprisingly, given the polymorphism of S-alleles, only a minority of individuals yielded PCR products of the expected size. These yielded 35 genomic sequences, of nine different sequence types of which eight are highly similar to the A. hispanicum S-allele sequences, and one to a very similar unpublished Antirrhinum S-like RNase sequence. The sequence types are well separated from the S-RNase sequences from Solanaceae and Rosaceae, and also from most known "S-like" RNase sequences (which encode proteins not involved in self-incompatibility). An association with incompatibility types has so far been established for only one of the putative S-alleles, but we describe evidence that the other sequences are also S-alleles. Variability in these sequences follows the pattern of conserved and hypervariable regions seen in other S-RNases, but no regions have higher replacement than silent diversity, unlike the results in some other species.

Alleles↗

Host-associated allozyme variation in tree cambium miners, Phytobia spp. (Diptera: Agromyzidae).

The larvae of the agromyzid flies that belong to the genus Phytobia Lioy feed by mining in the differentiating xylem just below the cambium of growing forest trees. The genus, which is apparently one of the most primitive groups in the Agromyzidae, comprises over 50 currently recognized species. Most of the species are mono- or oligophagous, and the host plants belong to numerous genera in about 60 families. Thus, Phytobia is an attractive candidate for studies on the evolution of insect-plant relationships. In spite of this, the taxonomy of Phytobia is currently poorly understood, mainly because the morphological differences between species are small. We used allozyme electrophoresis to investigate whether molecular markers could be used to separate and identify species in Phytobia, and to study the patterns of host use in the group. For this, we collected Phytobia larvae from eight host tree species occurring in southern Finland. An analysis of 10 variable allozyme loci showed that there are probably five species of Phytobia that feed on the hosts included in our study: one occurs on birches (Betula pubescens Ehrh. and B. pendula Roth) and alders (Alnus incana (L.) Moench and A. glutinosa (L.) Gaertn.), one on rowan (Sorbus aucuparia L.), and three species with overlapping feeding ranges on aspen (Populus tremula L.) and two willow species (Salix phylicifolia L. and S. caprea L.). Because birches and alders belong to the plant family Betulaceae, rowan to Rosaceae, and aspen and willows to Salicaceae, the host associations of the individual fly species can be explained by the taxonomic affinities of the hosts. However, our results also show that on a larger scale the evolution of host-plant associations in Phytobia cannot be explained by strict parallel cladogenesis (cospeciation) between the flies and their hosts.

Animals↗

Allelic diversity of S-RNase at the self-incompatibility locus in natural flowering cherry populations (Prunus lannesiana var. speciosa).

In the Rosaceae family, which includes Prunus, gametophytic self-incompatibility (GSI) is controlled by a single multiallelic locus (S-locus), and the S-locus product expressed in the pistils is a glycoprotein with ribonuclease activity (S-RNase). Two populations of flowering cherry (Prunus lannesiana var. speciosa), located on Hachijo Island in Japan's Izu Islands, were sampled, and S-allele diversity was surveyed based on the sequence polymorphism of S-RNase. A total of seven S-alleles were cloned and sequenced. The S-RNases of flowering cherry showed high homology to those of Prunus cultivars (P. avium and P. dulcis). In the phylogenetic tree, the S-RNases of flowering cherry and other Prunus cultivars formed a distinct group, but they did not form species-specific subgroups. The nucleotide substitution pattern in S-RNases of flowering cherry showed no excess of nonsynonymous substitutions relative to synonymous substitutions. However, the S-RNases of flowering cherry had a higher Ka/Ks ratio than those of other Prunus cultivars, and a subtle heterogeneity in the nucleotide substitution rates was observed among the Prunus species. The S-genotype of each individual was determined by Southern blotting of restriction enzyme-digested genomic DNA, using cDNA for S-RNase as a probe. A total of 22 S-alleles were identified. All individuals examined were heterozygous, as expected under GSI. The allele frequencies were, contrary to the expectation under GSI, significantly unequal. The two populations studied showed a high degree of overlap, with 18 shared alleles. However, the allele frequencies differed considerably between the two populations.

Alleles↗

ATPgammaS enhances the production of inflammatory mediators by a human dermal endothelial cell line via purinergic receptor signaling.

Adenosine 5'-triphosphate (ATP) affects multiple intra- and extracellular processes, including vascular tone and immune responses. Microvascular endothelial cells (EC) play a central role in inflammation by recruitment of inflammatory cells from blood to tissues. We hypothesized that ATP (secreted by neurons and/or released after perturbation of cutaneous cells) may influence secretion of inflammatory messengers by dermal microvascular EC through actions on purinergic P2 receptors. Addition of the hydrolysis-resistant ATP analogue, adenosine 5'-O-(3-thiotriphosphate) (ATPgammaS), to subconfluent cultures of the human microvascular endothelial cell-1 (HMEC-1) cell line led to a dose- and time-dependent increase in release of IL-6, IL-8, monocyte chemoattractant protein-1, and growth-regulated oncogene alpha. Both ATPgammaS-induced release and basal production of these proteins were significantly inhibited by the purinergic antagonists pyridoxal-5'-phosphate-6-azophenyl-2',5'-disulfonic acid (PPADS), pyridoxal-5'-phosphate-6(2'-naphthylazo-6-nitro-4',8'-disulfonate), and suramin. ATPgammaS increased expression of intercellular adhesion molecule-1 (ICAM-1), whereas suramin and PPADS decreased both ATPgammaS-induced and basal ICAM-1 expression. Using PCR, we found that HMEC-1 strongly express mRNA for the P2X(4), P2X(5), P2X(7), P2Y(2), and P2Y(11) receptors and weakly express mRNA for P2X(1) and P2X(3) receptors. Purinergic nucleotides may mediate acute inflammation in the skin and thus contribute to physiological and pathophysiological inflammation. For example, ATP may contribute to both the vasodilation and the inflammation associated with rosacea.

Adenosine Triphosphate↗