Nonhost versus host resistance to the grapevine downy mildew, Plasmopara viticola, studied at the tissue level.

Following inoculation of host and nonhost plants with Plasmopara viticola, the grapevine downy mildew, a histological survey was undertaken to identify the stage where its development is contained in nonhosts and in resistant host plants. Three herbaceous nonhost species, Beta vulgaris, Lactuca sativa, and Capsicum annuum, and three grapevine species displaying different level of resistance (Vitis vinifera [susceptible], Vitis riparia [partially resistant] and Muscadinia rotundifolia [totally resistant]) where inoculated by P. viticola using a controlled leaf disk inoculation bioassay. During the early steps of infection, defined as encystment of zoospores on stomata, penetration of the germ tube, and production of the vesicle with the primary hypha, there was no evidence of a clear-cut preference to grapevine tissues that could attest to host specificity. The main difference between host grapevine species and nonhosts was observed during the haustorium formation stage. In nonhost tissues, the infection was stopped by cell wall-associated defense responses before any mature haustorium could appear. Defense responses in resistant grapevines were triggered when haustoria were fully visible and corresponded to hypersensitive responses. These observations illustrate that, for P. viticola, haustorium formation is not only a key stage for the establishment of biotrophy but also for the host specificity and the recognition by grapevine resistance factors.

Assessment of topical corticosteroid activity using the vasoconstriction assay in healthy volunteers.

OBJECTIVE AND DESIGN: The aim of the study was to evaluate the vasoconstrictive activity of four new galenic preparations of hydrocortisone and to compare it with concentration-equivalent reference preparations. The study comprised two study phases: the pilot study phase and the main study phase. During open, nonrandomized pilot study, the optimal administration period was tested. The main study was performed in a randomized, double-blind intraindividual comparative design. SUBJECTS: Twenty male and female volunteers with healthy skin who responded to topically applied clobetasol-17-propionate before entering the trial participated in this study. TREATMENT: All subjects received the same treatments. The test preparations new galenic formulation (NGF) hydrocortisone 0.25% cream, NGF hydrocortisone acetate 0.25% cream, NGF hydrocortisone 0.5% cream, and NGF hydrocortisone 1.0% cream were compared with the respective reference preparations Soventol hydrocortisone (hydrocortisone acetate 0.25%), Hydroderm HC 0.5% cream (hydrocortisone 0.5%), Hydrogalen cream (hydrocortisone 1.0%) and placebo (vehicle of test preparations). METHOD: The topical preparations were applied occlusively for 2 h (pilot study) or 24 h (main study) on outlined areas (5 x 5 cm with a distance of 3 cm) of both forearms (4 areas for each). Assessment of vasoconstriction was performed before treatment, and 0.5, 4, 6 and 24 h after treatment (observation period) using a subjective rating scale (OLSEN vasoconstriction score) and measuring the colorimetric parameter a* (redness) by use of the Chroma-Meter (Minolta company, Ahrensburg, Germany). RESULTS: A significant vasoconstriction (positive blanching effect) was measured by use of chromametry for test preparations hydrocortisone 0.25% cream, hydrocortisone 0.5% cream, hydrocortisone 1.0% cream and for the reference preparation Hydrogalen cream compared to placebo 30 min after the end of treatment. In contrast, the reference preparations Soventol hydrocortisone and Hydroderm HC 0.5% did not differ significantly from placebo 30 min after treatment. No statistically significant effect of all formulations was observed 4-24 h after treatment in comparison with placebo. CONCLUSIONS: The vasoconstrictive efficacy of test preparations was mostly stronger than the concentration-equivalent reference preparations. This effect was achieved by use of new galenics of test preparations resulting in enhanced skin penetration and improved efficiency. No unwanted side effects were observed during the course of the study despite increased efficacy of the topically applied test preparations.

Transepidermal water loss dynamics of human vulvar and thigh skin.

Refinement in procedures to assess skin surface water loss (SSWL) dynamics of the vulvar skin on a large sample of subjects (60) is described and compared to another semi-occluded skin site, the inner thigh. Vulvar SSWL significantly decreased over a 30-min period from 46.2 +/- 2.6 (SE) to 24.7 +/- 1.6 g m(-2) h (p < 0.001). The inner thigh, another semi-occluded region, showed no similar pattern for SSWL (6.2 +/- 0.3 to 6.6 +/- 0.5 g m(-2) h), and the values were significantly less than those for vulvar skin. There was no significant effect of age, body mass index or atopic status on vulvar SSWL.

Vulvar dermatoses--irritant and allergic contact dermatitis of the vulva.

Irritant and allergic contact dermatitis are commonly seen in patients complaining about itching, burning and irritation in the vulvar area. Irritation often precedes allergic sensitization. Clinically, irritant and allergic contact dermatitis can be difficult to distinguish. Diagnosis is made by history, clinical investigation and patch testing. Recommended patch test series are the standard series, a medicament series, the patient's own topical medicaments, popular remedies and other suspected products. A skin biopsy may be useful to establish the diagnosis of contact dermatitis, but it is usually not helpful for the differential diagnosis between irritant and allergic dermatitis.

Percutaneous penetration of topically applied melatonin in a cream and an alcoholic solution.

In a clinical study, the skin penetration properties of melatonin 0.01% in a cream and 0.01 and 0.03% in a solution were investigated by evaluation of the serum melatonin levels over a 24-hour time course in 15 healthy volunteers. Blood samples for melatonin measurements were taken at 9.00 a.m. before applying the test preparations and 1, 4, 8 and 24 h after application. The measurements were carried out by radioimmunoassay for melatonin. In 15 volunteers, the serum levels of melatonin before application of the topical preparations were between 0.6 and 15.9 pg/ml. After application of the 0.01% melatonin cream, there was a steady increase starting from 9.00 a.m. up to a mean serum value of 9.0 pg/ml at 9.00 a.m. the next day. The solution of 0.01% melatonin also showed an increase, starting from 5.00 p.m., up to a mean melatonin level of 12.7 pg/ml 24 h after application. The solution containing 0.03% melatonin resulted in elevated melatonin levels 1 and 8 h after application. The values were 18.1 and 19.0 pg/ml. The cumulative melatonin values for each preparation were 7.1, 8.6 and 15.7 pg/ml, respectively. This study shows that the strongly lipophilic substance melatonin is able to penetrate through the skin and leads to dose- and galenic-dependent melatonin levels in the blood. No increase of melatonin above the physiological range was observed.

Vulvar pain. Psychological profiles and treatment responses.

OBJECTIVE: To compare the psychological and quality of life profiles of women with different subsets of vulvodynia and identify differential response patterns to a combined gynecologic-dermatologic treatment program. STUDY DESIGN: Fifty-three women with vulvar dermatoses, dysesthetic vulvodynia and vulvar vestibulitis were recruited from a combined dermatologic and gynecologic vulvar clinic and completed instruments related to quality of life (QoL) and psychologic symptomatology. A repeated measurement design was employed. However, only 33 women could be assessed at the second point of measurement. RESULTS: Women with different subsets of vulvodynia showed (1) distinct quality of life and psychological profiles, and (2) different response patterns to treatment. There were significant differences on most QoL scales, with patients with dysesthetic vulvodynia and with vulvar dermatoses showing greater impairment than patients with vestibulitis. However, patients with vestibulitis displayed a highly specific pattern of psychologic symptomatology. Patients with vulvar dermatoses had an improvement in problems related to intimacy than patients with vulvar pain syndromes. CONCLUSION: Treatment intervention studies seem to be indispensable in order to gain a better understanding of the complex interplay between different subsets of vulvodynia and psychological symptoms concurrently with differential treatment reactions.

Vulval diseases need an interdisciplinary approach.

BACKGROUND: Vulval diseases are common, but their frequency and importance are often underestimated. OBJECTIVE: The study intended to evaluate the number of patients with vulval diseases seen in medical practice. Furthermore, the acceptance of an interdisciplinary vulval disease clinic, managed by a dermatologist in cooperation with a gynaecologist and a psychologist, was evaluated. METHODS: In April and Mai 1998, a mailing survey of all gynaecologists (n = 239), general practitioners (n = 244) and dermatologists (n = 96) in Thuringia, Germany, containing 7 questions concerning patients with vulval diseases was performed. RESULTS: 49% (n = 286) of the questionnaires were returned. On average, 97% of the physicians regularly treated females with genital diseases. The most common diseases mentioned by all specialities were local fungal and bacterial infections. The further ranking of diagnosis was different between the specialities. It was estimated that 83% of patients with chronic disorders would develop additional psychological problems. The necessity of an interdisciplinary vulval disease clinic in Thuringia was stated by 71% of the interviewed physicians. 76% of the gynaecologists and 75% of the general practitioners welcomed the introduction of the clinic. Interestingly, only 51% of the dermatologists felt that a vulval disease clinic was necessary. CONCLUSION: The majority of the physicians in Thuringia welcomed an interdisciplinary vulval disease clinic to improve the conditions for women suffering from genital diseases.

[A severe epicutaneous test reaction to the bufexamac in a hemorrhoidal therapeutic preparation]. / Schwere Epikutantestreaktion auf Bufexamac in einem Hämorrhoidal-Therapeutikum.

HISTORY AND FINDINGS: A 49-year-old woman presented with acute perianal vesicular/bullous contact dermatitis. Other areas were over the trunk, face, neck and wrists. She reported occasional application of an ointment (Mastu S) to treat her hemorrhoids. INVESTIGATION: Patch tests (basic series, anal block, own ointment, local anesthetic, cosmetics) provoked strong vesicular and bullous reactions of persisting crescendo type, spreading far beyond the site of application, to Bufexamac, to a derivative of hydroxxamine acid, and to local applied ointment with mild or moderate antiinflammatory action. TREATMENT AND COURSE: A week after the patch tests there was a flare-up of the previous foci of dermatitis. These reactions subsided two days later after intravenous injection of prednisolone. The skin lesions healed after rapid reduction of the systemic treatment and local application of corticosteroids, bathing with tanning substances and basic preparations. CONCLUSION: While Bufexamac is not absorbed when applied rectally, perianal contamination may not be avoidable on intra-anal application and can produce sensitization.

[Giant condylomata acuminata (Buschke-Löwenstein tumor)]. / Condylomata gigantea Buschke-Löwenstein.

HISTORY AND ADMISSION FINDINGS: A 59-year-old woman had for 3 weeks been suffering from painful, moist skin changes on the external genitals, for 3 months having noticed vegetations in the anogenital and perineal region. She had been fatigued and lacking in energy since then. Her last gynecological examination had been 20 years ago. Examination of the skin on admission revealed extensive plaque and cauliflower-like warts and large areas of maceration in the anogenital region, the labia and perineum, with large areas of maceration. Nodules were clearly palpable in the left labium majora: the inner aspects were markedly reddened and had some cutaneous erosions. These findings suggested giant condyloma (GC; Buschke-Löwenstein tumour). INVESTIGATIONS: Biopsy showed marked condylomatous epithelial proliferations which, with the formation of markedly thickened epithelial cones, had reached the stage of a verrucous carcinoma. TREATMENT AND COURSE: Under local anti-inflammatory and adstringent medications the inflammatory reaction quickly subsided. Vulvectomy was performed to remove the tumour. CONCLUSION: Giant condyloma (Buschke-Löwenstein tumour) is a rare pseudocancereous lesion. If human papilloma viruses of type 16 or 18 is demonstrated in the lesions, malignant degeneration should be considered. Histological examination is essential to differentiate it from squamous cell carcinoma.

Effects of various grit-containing cleansers on skin barrier function.

Products intended for individuals in contact with strongly adhering dirt often contain grit. Various clinical test methods have been developed for evaluating the potential of personal washing products to induce skin irritation. In the present study, differences in the irritant effects of washing products containing naturally-derived grit and synthetic grit were investigated in a forearm wash test. The forearms of 16 test subjects were washed in a total of 18 treatments (4 per day for 4 days, with 2 treatments on the 5th day). Treatment consisted of continuous washing for 2 min by a technician, who gently slid his fingertips with the lather up and down the forearm. Non-invasive instrumental measurements of skin barrier function were performed. Repetitive washing for 1 week lead to increased TEWL values, skin redness and decreased stratum corneum hydration. Results indicate differences in irritancy potential due to different types of grit, their surface and concentration. It is concluded that the repeated wash test seems to be adequate for rating personal washing products that contain grit.

Treatment of vascular lesions using the VersaPulse variable pulse width frequency doubled neodymium:YAG laser.

BACKGROUND: The flashlamp-pumped dye laser is considered the treatment of choice for vascular lesions including port-wine stains. However, this treatment is associated with an intensive postoperative purpura and considerable pain. OBJECTIVE: We tested a newly developed laser device with a 532-nm neodymium:YAG laser with variable pulse widths between 2 and 10 ms. METHODS: Forty-two consecutive patients from our laser clinics were treated with the new laser system delivering energy through a chilled tip by means of circulating water (cooling temperature was between 4 and 5.5 degreesC). Forty of them had been pre-treated with an argon laser and an argon-pumped dye laser. RESULTS: Out of the 42 patients, 1 patient had a complete clearing, 11 had an excellent result (more than 80% clearing), 21 patients had a good result (51-75% clearing), weak responses were seen in 9 patients. Significant success was already seen after 1 or 2 treatments. Side effects were rare. There was no purpura after treatment. Local anesthesia was only applied in 6 out of 42 patients. CONCLUSION: The variable pulse width frequency doubled neodymium:YAG laser seems to be an alternative to the flashlamp-pumped dye laser and deserves to be investigated in comparative trials.

Functional characterization of the proteolytic activity of the tomato black ring nepovirus RNA-1-encoded polyprotein.

Translation of tomato black ring virus (TBRV) RNA-1 in a rabbit reticulocyte lysate leads to the synthesis of a 250K polyprotein which cleaves itself into smaller proteins of 50, 60, 120, and 190K. Polypeptides synthesized from synthetic transcripts corresponding to different regions of TBRV RNA-1 are processed only when they encode the 23K protein delimited earlier by sequence homology with the cowpea mosaic virus 24K protease. The proteolytic activity of this protein is completely lost by mutating residues C170 (to I) or L188 (to H), residues which align with conserved residues of the viral serine-like proteases. The 120K protein is generated by cleavage of the dipeptide K/A localized in front of the VPg but is not further cleaved in vitro at the K/S site (at the C terminus of the VPg) or between the protease and polymerase domains. However, both the protein VPgProPol (120K) and the protein ProPol (117K) produced in vitro from synthetic transcripts can cleave in trans the RNA-2-encoded 150K polyprotein, but they cannot cleave in trans polypeptides containing a cleavage site expressed from RNA-1 transcripts in which the protease cistron is absent or modified.

Analysis of the in vitro cleavage products of the tomato black ring virus RNA-1-encoded 250K polyprotein.

Tomato black ring virus RNA-1 was translated in a rabbit reticulocyte lysate. The primary translation product of Mr 250K, which corresponds to its whole coding capacity, was synthesized within 45 min and, during further incubation in the translation medium, was proteolytically processed. Essentially, four cleavage products (P190, P120, P60 and P50) were detected and located within P250 by pulse-chase and immunoprecipitation experiments. P190 is an intermediate cleavage product which is further cleaved to form P60 and P120. P120, which contains the region that has been assigned to the virus protease and the virus polymerase, was not further cleaved in vitro.

In vitro synthesis of biologically active transcripts of tomato black ring virus satellite RNA.

Synthetic transcripts of tomato black ring virus satellite RNA (TBRV satRNA), isolate L, were prepared from cDNA cloned in the Bluescribe transcription vector. Transcripts with 49 (T49L) or two (T2GL) extra nucleotides at their 5' ends and 42 extra nucleotides at their 3' ends were able to induce, but to different extents, the synthesis in vitro of the satRNA-encoded 48K protein. However, when inoculated into Chenopodium quinoa together with TBRV L genomic RNAs, only T2GL was biologically active, in the presence or absence of a 5' cap analogue in the transcription reactions. Analysis of the 5' and 3' termini of the satRNA isolated from plants showed that nonviral extensions were not maintained in the transcript progeny.