Validation of questionnaire algorithm based on repeated open application testing with the constituents of fragrance mix II:the EDEN Fragrance Study

Background: In a European study on contact allergy in the general population, it has been hypothesized that the combination of contact allergy to a fragrance together with a history indicating dermatitis at exposure and thereafter subsequent avoidance of scented products implied a diagnosis of allergic contact dermatitis. Objectives: The primary aim of this study was to validate this hypothesis/algorithm. The secondary aim was to investigate whether there was any association between the outcome of the recent repeated open application test (ROAT) and the patch test reactivity. Methods: One hundred nine subjects with and without contact allergy to fragrance mix II (FM II) were recruited. Volunteers from six European dermatology clinics participated in the study including a patch test and a ROAT. Results: Twenty-four positive ROAT reactions were noted in total including 20 of those 32 with contact allergy to FM II. None of the volunteers reacted to the vehicle (P < 0.001). More individuals with a positive algorithm had positive ROATs when compared with those with a negative algorithm. However, the difference was not statistically significant (P = 0.12). The lower the patch test concentration eliciting a positive test reaction, the more likely was a positive ROAT and the more likely that the positive ROAT appeared early during the investigative period. Conclusions: The algorithm used in this study was not validated but it was indicated in this ROAT setup. The stronger the patch test reactivity the more likely was a positive ROAT and the more likely it was that the positive ROAT appeared early during the application period

CD8+ T cell concentration determines their efficiency in killing cognate antigen–expressing syngeneic mammalian cells in vitro and in mouse tissues

We describe a quantitative model for assessing the cytolytic activity of antigen-specific CD8+ T cells in vitro and in vivo in which the concentration of antigen-specific CD8+ T cells determines the efficiency with which these cells kill cognate antigen–expressing melanoma cells in packed cell pellets, in three-dimensional collagen-fibrin gels in vitro, and in established melanomas in vivo. In combination with a clonogenic assay for melanoma cells, collagen-fibrin gels are 4,500–5,500-fold more sensitive than the packed cell pellet–type assays generally used to measure CD8+ T cell cytolytic activity. An equation previously used to describe neutrophil bactericidal activity in vitro and in vivo also describes antigen-specific CD8+ T cell–mediated cytolysis of cognate antigen-expressing melanoma cells in collagen-fibrin gels in vitro and in transplanted tumors in vivo. We have used this equation to calculate the critical concentration of antigen-specific CD8+ T cells, which is the concentration of these cells required to hold constant the concentration of a growing population of cognate antigen-expressing melanoma cells. It is ∼3.5 × 105/ml collagen-fibrin gel in vitro and ∼3 × 106/ml or /g melanoma for previously published studies of ex vivo–activated adoptively transferred tumor antigen–specific CD8+ T cell killing of cognate antigen–expressing melanoma cells in established tumors in vivo. The antigen-specific CD8+ T cell concentration required to kill 100% of 2 × 107/ml cognate antigen-expressing melanoma cells in collagen fibrin gels is ≥107/ml of gel

In Vivo Methods for the Assessment of Topical Drug Bioavailability

This paper reviews some current methods for the in vivo assessment of local cutaneous bioavailability in humans after topical drug application. After an introduction discussing the importance of local drug bioavailability assessment and the limitations of model-based predictions, the focus turns to the relevance of experimental studies. The available techniques are then reviewed in detail, with particular emphasis on the tape stripping and microdialysis methodologies. Other less developed techniques, including the skin biopsy, suction blister, follicle removal and confocal Raman spectroscopy techniques are also described

Prevalencija infekcije Neisseriom gonorrhoeae ili Chlamydiom trachomatis kod akutnoga mukopurulentnog cervicitisa

The aim of this study was to determine the incidence of N. gonorrhoae (NG) and/or C. trachomatis (CT) in acute mucopurulent cervicitis (MPC). The study included 617 non-pregnant women with MPC, who had not been receiving any antimicrobial treatment. The average age of patients was 22.2 years. There were no statistically significant differences according to place of residence, education, and marital status. Samples for laboratory analysis were collected using a routine procedure; NG was identified using the cytochrome oxidase test and Gram staining. CT was isolated on McCoy cell culture and stained with Lugol solution. NG was isolated in three women (0.8 %) and CT in 58 women (9.4 %). Fifty-six of the CT-positive patients were nullipara and only two were unipara. All NG-positive patients were also nullipara. The mean number of sexual partners was 2.2 in all study subjects, 2.4 in CT-positive subjects, and 2.9 in NG-positive subjects. Vaginal discharge purity according to Schröder was significantly deteriorated in CT-positive patients (p=0.011). When asked about the use of contraceptives, as many as 32.7 % patients answered that they did not use any protection, 39 % women used the rhythm method and coitus interruptus, 20 % were taking oral contraceptives, 6.1 % used mechanical devices, and 1.9 % used chemical protection. Previous acute and chronic pelvic infl ammatory diseases correlated with MPC (p<0.01). Our statistical analysis suggests that chlamydial infection significantly reduces the purity of vaginal discharge, which is more pronounced in nulliparae. Pap smear was not specific enough to demonstrate chlamydial infection. In view of the MPC findings, the prevalence of CT and NG infection is low.Cilj istraživanja jest utvrditi koincidenciju N. gonorrhoae (NG) i/ili C. trachomatis (CT) u akutnim mukopurulentnim cervicitisima (MPC). Istraživanje je provedeno na populaciji 617 žena izvan trudnoće koje prethodno nisu uzimale antimikrobnu terapiju. NG i CT su izolirane iz obrisaka vrata maternice primjenom standardnih metoda. NG je transportirana na selektivnoj hranjivoj podlozi i identificirana citokrom-oksidaznim testom bojenjem po Gramu. CT je izolirana McCoyevom staničnom kulturom nakon bojenja Lugolovom otopinom. NG je izolirana u tri ispitanice (0,8 %), a CT u 58 ispitanica (9,4 %). Srednja dob bolesnica iznosila je 22,2 godine. Školovanje i bračno stanje nisu statistički značajno utjecali na rezultate istraživanja. Među bolesnicama s izoliranom CT prevladavaju nulipare, a u slučaju NG sve su pozitivne bolesnice također bile nulipare. Srednja vrijednost broja partnera za čitavu populaciju iznosi 2,2. Bolesnice s potvrđenom CT imale su prosječno 2,4, a one s potvrđenom NG 2,9 partnera. Vrijednost određivanja stupnja čistoće rodničkog iscjetka po Schroderu značajno je povišena u CT-pozitivnih bolesnica (p=0.011). Utvrđeno je da 32.7 % ispitanica uopće nije koristilo kontracepciju, dok je većina (39 %) rabila ritmičke metode i prekinuti snošaj. Preostale ispitanice uzimale su oralne kontraceptive (20 %), mehaničku zaštitu (6,1 %) ili lokalne kemijske kontraceptive (1,9 %). Također je dokazano da prethodna akutna i kronična zdjelična upalna bolest korelira s MPC-om (p<0.01). Zaključeno je da klamidijska infekcija statistički značajno korelira sa stupnjem čistoće rodničkog iscjetka, posebice u nulipara. Papanicolaouov razmaz nije specifičan u određivanju klamidijske infekcije. Rezultati istraživanja pokazuju da je učestalost klamidijske i gonokokne infekcije ipak relativno niska u odnosu na nalaz MPC-a