Application of Raman Spectroscopy for Sorption Analysis of Functionalized Porous Materials.

Functionalized porous materials could play a key role in improving the efficiency of gas separation processes as required by applications such as carbon capture and storage (CCS) and across the hydrogen value chain. Due to the large number of different functionalizations, new experimental approaches are needed to determine if an adsorbent is suitable for a specific separation task. Here, it is shown for the first time that Raman spectroscopy is an efficient tool to characterize the adsorption capacity and selectivity of translucent functionalized porous materials at high pressures, whereby translucence is the precondition to study mass transport inside of a material. As a proof of function, the performance of three silica ionogels to separate an equimolar (hydrogen + carbon dioxide) gas mixture is determined by both accurate gravimetric sorption measurements and Raman spectroscopy, with the observed consistency establishing the latter as a novel measurement technique for the determination of adsorption capacity. These results encourage the use of the spectroscopic approach as a rapid screening method for translucent porous materials, particularly since only very small amounts of sample are required.

What influences the information exchange during self-medication consultations in community pharmacies? A non-participant observation study.

BACKGROUND: Although pharmaceutical staff consider guideline-compliant information exchange during self-medication consultations as crucial, they have not fully integrated it into practice. It is unclear what prevents pharmaceutical staff from implementing their positive intentions into their actual everyday practice. To improve the information exchange during the current consultation practice, a deeper understanding of its implementation is needed. OBJECTIVES: The aim was to evaluate the factors influencing information exchange during self-medication consultations in German community pharmacies. METHODS: We performed a non-participant observation of real-life consultations with post-consultation interviews of pharmaceutical staff in 10 pharmacies. The information exchanged during self-medication consultations was evaluated via 7 guideline-recommended information parameters in 2 stages of information exchange: (a) 'information gathering' and (b) 'provision of information'. Directly after each observed consultation, pharmaceutical staff's opinion about the consultation, the customer's interest and their own performance was questioned. Factors associated with the observed extent of information exchange were analysed by Poisson regression analysis. RESULTS: In the 379 self-medication consultations with 46 pharmaceutical staff members, 454 different customer enquiries were addressed, and 483 medications were dispensed. In median, 2 predefined information parameters (First-/Third quartile: 1/4) were fulfilled during an enquiry and 2 parameters (First-/Third quartile: 1/3) were fulfilled for a dispensed medication. Pharmaceutical staff were satisfied with 85% of their consultations and perceived 76% of them as easy to handle. In both information stages, information exchange increased when 'customers were perceived to be interested in counselling' (p < 0.001) and decreased when customers had a 'specific medication request' (p < 0.001). CONCLUSIONS: Information exchange in pharmacies needs to be better integrated into daily practice. Strategies to encourage information exchange should also include pharmaceutical staffs' perception of their own counselling technique. Ongoing patient-centred trainings should facilitate strategies to engage uninterested customers or costumers with specific medication requests in consultations.

Are evidence-based criteria addressed during counseling on over-the-counter products? An observational study in community pharmacies.

OBJECTIVE: We examined whether evidence-based criteria were addressed during counseling on over-the-counter products (OTCs) in community pharmacies. METHODS: Consultations were observed in 10 community pharmacies. We analyzed communications about OTCs to determine if any information on three evidence-based criteria (outcome variables: scientific evidence such as clinical study results, pharmaceutical staff's experience, and customer's experience) was mentioned. Two groups of communications were compared with Pearson's chi-square and Fisher's exact test, as appropriate: The communications about OTCs recommended by the pharmaceutical staff vs. the communications about OTCs requested by customers. RESULTS: In 379 observed consultations, 300 OTCs were recommended by staff and 390 OTCs were requested by customers. The least included criterion was scientific evidence (in OTCs recommended by pharmaceutical staff - 1% vs. requested by customers - 0%), followed by pharmaceutical staff's experience (5% vs. 1%). The customer's experience was addressed more frequently (14% vs. 41%). Statistically significant differences between the two groups were found for all criteria (p < 0.05). CONCLUSION: Evidence-based criteria were rarely addressed during counseling on OTCs. PRACTICE IMPLICATIONS: Pharmaceutical staff should be encouraged to include the three evidence-based criteria more frequently. Additionally, customers should be encouraged to request such information from the staff in community pharmacies.

Public's perspectives on guideline-recommended self-medication consultations in German community pharmacies.

The aim of this study was to assess the public's opinions and expectations of self-medication consultations in German community pharmacies with special emphasis on the acceptance of guideline-recommended consultation. In a cross-sectional study in the city centre of Leipzig, Germany, we conducted a questionnaire-based survey administered via an interview with passers-by from June to September 2018. The structured questionnaire contained questions assessing (I) previous experience with self-medication consultations, (II) possible reasons for declining self-medication consultations, (III) the attitude towards information gathering and (IV) expectations of self-medication consultations. (I) 92% of the 963 respondents stated they were generally satisfied with self-medication consultations in community pharmacies. Around one-fifth of all respondents claimed that they would like to be asked more health-related questions (22%) and receive more information on non-prescription drugs (20%). (II) Privacy issues (39%) and reluctance to talk about some medical conditions (43%) were the most frequent reasons for declining self-medication consultation. (III) Respondents understood the need for answering guideline-recommended questions (85-96%) and did not mind being asked these questions (70-96%). (IV) Most of the respondents expected to be counselled even if they did not ask for it directly (69%). Pharmacies were further expected to recommend the best drug, even if it was not what the customer initially intended to buy (87%). However, more than half of the respondents would consider counselling as unimportant if they knew exactly which medication they wanted to buy (56%) or if they had used the non-prescription drug before (70%). The majority also expected to receive guideline-recommended drug information (each item at least 52%). Thus, our study shows that respondents were mostly in line with the required standards of self-medication counselling guidelines. Customers expect high-quality counselling on self-medication. These findings support pharmaceutical staff's understanding of customers' barriers and expectations during self-medication consultations.

Evidence-based self-medication: development and evaluation of a professional newsletter concept for community pharmacies.

Background Providing evidence-based care is recognized as a key competence for all healthcare professionals. In order to support community pharmacists in evidence-based self-medication counseling, the umbrella organization of German pharmacists initiated the development of a nationwide concept. The key element of the concept was a professional newsletter that should help pharmacists incorporate research findings into their daily counseling practice. Objective To develop, implement and evaluate the professional newsletter concept. Setting German community pharmacies. Method Clinical pharmacists from a German university compiled and synthesized clinical trial data in a professional newsletter that would supply community pharmacists with evidence-based information on common over-the-counter medicines as well as instructions for searching and appraising scientific literature. The electronic newsletter was offered to interested community pharmacists free of charge, once or twice a month, after they signed up for a subscription. About one year after the publication of the first newsletter issue, the subscribers were invited to take part in an anonymous cross-sectional online survey. In all, 21 newsletter issues were published through the end of the survey period. Main outcome measure Perceived value of the professional newsletter with 10 predefined objectives. Results A total of 1975 persons subscribed to the professional newsletter. Of those, 150 persons working in community pharmacies completed the survey. Most of them perceived the synthesized information as 'useful' (81-95%). They attributed positive changes in knowledge (89%), skills (87-91%), awareness (85%), and motivation (67-77%) to the newsletter. However, almost half of them (43%) found it difficult to incorporate reading the newsletter in their everyday working life. Free-text feedback suggested that further modifications should be considered to facilitate a better integration of the newsletter into everyday pharmacy practice. Conclusion A nationwide provided professional newsletter can play a vital part in supporting pharmacists in evidence-based self-medication counseling. However, the practicability of such a newsletter needs to be further improved and the newsletter should be accompanied by additional measures.

What is the attitude towards and the current practice of information exchange during self-medication counselling in German community pharmacies? An assessment through self-report and non-participant observation.

BACKGROUND: Guidelines encourage relevant information exchange between pharmaceutical staff and patients during self-medication consultation. Thereby, assessing the patient's situation and providing information is crucial for patient safety. So far, limited studies have investigated this information exchange, particularly in Germany. We aimed to assess the attitude towards and the current practice of guideline-recommended information exchange in German community pharmacies. METHODS: In total, twelve guideline-recommended parameters were predefined for gathering patient-related information and for the provision of information. These information exchange parameters were evaluated in two parts: Firstly, in a self-report of pharmaceutical staff via an online questionnaire to assess the reported importance, difficulty and frequency of the parameters as well as barriers to their implementation; secondly, in a non-participant observation in five pharmacies to evaluate the actual consultation practice. RESULTS: In the self-report, all parameters were rated by more than 76% of 1068 participants as important. 'Concurrent medication' was determined to be the most difficult parameter to address (54%). All parameters of information gathering were rated to be addressed during routine counselling by at least 70% of the respondents. Parameters of information provision were all rated to be addressed by at least 45%. 'Lack of patient's interest' was identified as the most frequent barrier to appropriate counselling (84%). During the observation, the information gathering parameters were each addressed between 8 to 63% in the consultations, parameters of information provision between 3 to 34%. CONCLUSION: Despite broad acceptance, the guideline parameters of information exchange were comparatively little addressed during the actual routine care. This might be due to a perceived 'lack of patient's interest' in counselling. Our results suggest to scrutinize whether patients are in fact not interested in counselling and to further explore how the positive intention of pharmaceutical staff towards information exchange can be further translated into everyday practice.

Boosting Escherichia coli's heterologous production rate of ectoines by exploiting the non-halophilic gene cluster from Acidiphilium cryptum.

The compatible solutes ectoine and hydroxyectoine are synthesized by many microorganisms as potent osmostress and desiccation protectants. Besides their successful implementation into various skincare products, they are of increasing biotechnological interest due to new applications in the healthcare sector. To meet this growing demand, efficient heterologous overproduction solutions for ectoines need to be found. This study is the first report on the utilization of the non-halophilic biosynthesis enzymes from Acidiphilium cryptum DSM 2389T for efficient heterologous production of ectoines in Escherichia coli. When grown at low salt conditions (≤ 0.5% NaCl) and utilizing the cheap carbon source glycerol, the production was characterized by the highest specific production of ectoine [2.9 g/g dry cell weight (dcw)] and hydroxyectoine (2.2 g/g dcw) reported so far and occurred at rapid specific production rates of up to 345 mg/(g dcw × h). This efficiency in production was related to an unprecedented carbon source conversion rate of approx. 60% of the theoretical maximum. These findings confirm the unique potential of the here implemented non-halophilic enzymes for ectoine production processes in E. coli and demonstrate the first efficient heterologous solution for hydroxyectoine production, as well as an extraordinary efficient low-salt ectoine production.

The impact of evidence from clinical trials on counselling for over-the-counter drugs: A national survey of pharmaceutical staff in German pharmacies.

WHAT IS KNOWN AND OBJECTIVE: Evidence from clinical trials should form the foundation of healthcare advice. Comparatively little has been investigated about the implementation of evidence in over-the-counter (OTC) counselling in community pharmacies. The objective of this study was to explore the current impact of clinical trials on OTC counselling from the perspective of pharmaceutical staff. METHODS: Pharmaceutical staff in German community pharmacies were invited to participate in a nationwide cross-sectional survey using SoSci Survey. Within the questionnaire, the following items were queried: (i) attitudes towards influencing factors on OTC drug recommendation, (ii) practical procedure of OTC drug recommendations, (iii) experiences and challenges in handling clinical trials, and (iv) current and desired continuing education on self-medication. Multiple logistic regression was employed to identify predictors for OTC recommendations based on clinical trials in routine. RESULTS AND DISCUSSION: In total, 1068 participants completed the survey. (i) Clinical trial data were rated as 'very important' for drug recommendations by 33% and as 'rather important' by 50%. (ii) 8% reported that they base their recommendations on clinical trial data 'in almost all consultations', 40% 'in most consultations'. (iii) 69% faced difficulties in including clinical trial data into their counselling. (iv) 7% read original clinical trials at least once a month. Predictors for routine OTC recommendations based on clinical trials were as follows: male gender, university degree, clinical trials considered to be important for recommendations, inclusion of clinical trials in counselling assessed as easy, higher frequency of reading specialized literature per year, increasing number of working pharmacists in the pharmacy, and more casual than regular customers in the pharmacy. WHAT IS NEW AND CONCLUSION: Most of the pharmaceutical staff recognize the importance of clinical studies for self-medication advice. Based on self-report, however, clinical trial data are not taken routinely into account in practice. Only a minor proportion of staff routinely deal with studies. Therefore, appropriate instruments are required to make knowledge from clinical trials available in community pharmacies.

Epicutaneous allergen application preferentially boosts specific T cell responses in sensitized patients.

The effects of epicutaneous allergen administration on systemic immune responses in allergic and non-allergic individuals has not been investigated with defined allergen molecules. We studied the effects of epicutaneous administration of rBet v 1 and rBet v 1 fragments on systemic immune responses in allergic and non-allergic subjects. We conducted a clinical trial in which rBet v 1 and two hypoallergenic rBet v 1 fragments were applied epicutaneously by atopy patch testing (APT) to 15 birch pollen (bp) allergic patients suffering from atopic dermatitis, 5 bp-allergic patients suffering from rhinoconjunctivitis only, 5 patients with respiratory allergy without bp allergy and 5 non-allergic individuals. Epicutaneous administration of rBet v 1 and rBet v 1 fragments led to strong and significant increases of allergen-specific T cell proliferation (CLA+ and CCR4+T cell responses) only in bp-allergic patients with a positive APT reaction. There were no relevant changes of Bet v 1-specific IgE and IgG responses. No changes were noted in allergic subjects without bp allergy and in non-allergic subjects. Epicutaneous allergen application boosts specific T cell but not antibody responses mainly in allergic, APT-positive patients suggesting IgE-facilitated allergen presentation as mechanism for its effects on systemic allergen-specific immune responses.

Development of a new densimeter for the combined investigation of dew-point densities and sorption phenomena of fluid mixtures.

For the determination of dew-point densities and pressures of fluid mixtures, a new densimeter has been developed. The new apparatus is based on the well-established two-sinker density measurement principle with the additional capability of quantifying sorption effects. In the vicinity of the dew line, such effects cause a change in composition of the gas mixture under study, which can significantly distort accurate density measurements. The new experimental technique enables the accurate measurement of dew-point densities and pressures and the quantification of sorption effects at the same time.

Standardising analgesic administration for nurses: a prospective intervention study.

Background Uncontrolled pain reduces quality of life, delays recovery from surgery and can potentially contribute to the development of chronic pain. Strategies to improve pain management are desirable in view of these detrimental outcomes, yet frequently they are predominately medically based despite nurses playing key roles in pain assessment and analgesic administration. Objective To develop an algorithm-based guidance addressing nurses' pain assessment and analgesic administration and investigating its impact on quality of pain management. Setting Two orthopaedic units in a university hospital. Method An interdisciplinary expert panel consisting of physicians, nurses and clinical pharmacists developed an algorithm-based guidance in analgesic administration for nurses. The guidance was based on current guidelines addressing the appropriate use of analgesics. In a prospective intervention study, clinical pharmacists acted as independent monitors who assessed nurses' pain assessment and analgesic administration before (control period: usual care without any further support) and after implementation of the algorithm-based guidance (intervention period). We evaluated patient-nurse contacts for guideline adherence. We predefined guideline adherence (main outcome) as fulfilling all three of the following criteria: (A) nurses' pain intensity assessment, (B) their assessment of the patients' need for analgesics, and (C) analgesic administration depending on patients' individual pain intensity (including choice of prescribed analgesics). Main outcome measure Adherence to pain management guidelines. Results We analysed 706 patient-nurse contacts with 162 patients in the control and 748 contacts with 168 patients in the intervention period. Without support, guidelines were followed in 6 % of the patient contacts. We achieved an increase to 54 % (p < 0.001) after guidance implementation (main outcome). Guideline adherence decreased with rising pain intensity (slope coefficient -0.763, p < 0.001). Conclusion Especially in patients suffering from severe pain, current guidelines were not strictly followed by nurses when administering analgesics. Guideline adherence improved eightfold by implementing an algorithm-based guidance.

Frequent occurrence of T cell-mediated late reactions revealed by atopy patch testing with hypoallergenic rBet v 1 fragments.

BACKGROUND: Late allergic reactions are common in the course of allergen-specific immunotherapy and even occur with allergy vaccines with reduced IgE reactivity. OBJECTIVE: We sought to study atopy patch test (APT) reactions and T-cell responses to the recombinant birch pollen allergen Bet v 1 and recombinant hypoallergenic T-cell epitope-containing Bet v 1 fragments in patients with birch pollen allergy with and without atopic dermatitis (AD). METHODS: A clinical study was conducted in 15 patients with birch pollen allergy with AD (group 1), 5 patients with birch pollen allergy without AD (group 2), 5 allergic patients without birch pollen allergy (group 3), and 5 nonallergic subjects (group 4) by performing skin prick tests and APTs with rBet v 1 and hypoallergenic rBet v 1 fragments. T-cell, cutaneous lymphocyte antigen (CLA)(+) and CCR4(+) T-cell and cytokine responses were studied by thymidine uptake, carboxyfluorescein diacetate succinimidyl ester staining, and Luminex technology, respectively. RESULTS: rBet v 1 and hypoallergenic rBet v 1 fragments induced APT reactions in not only most of the patients with birch pollen allergy with AD (11/15) but also in most of those without AD (4/5). Patients with birch pollen allergy with AD had higher Bet v 1-specific proliferation of CLA(+) and CCR4(+) T cells compared with patients with birch pollen allergy without AD. There were no differences in Bet v 1-specific CLA(+) and CCR4(+) proliferation and cytokine secretion in patients with and without APT reactions. CONCLUSION: Hypoallergenic rBet v 1 fragments induce T cell-dependent late reactions not only in patients with birch pollen allergy with AD but also in those without AD, which can be determined based on APT results but not based on in vitro parameters.

The hydroxyectoine gene cluster of the non-halophilic acidophile Acidiphilium cryptum.

Acidiphilium cryptum is an acidophilic, heterotrophic α-Proteobacterium which thrives in acidic, metal-rich environments (e.g. acid mine drainage). Recently, an ectABCDask gene cluster for biosynthesis of the compatible solutes ectoine and hydroxyectoine was detected in the genome sequence of A. cryptum JF-5. We were able to demonstrate that the type strain A. cryptum DSM 2389(T) is capable of synthesizing the compatible solute hydroxyectoine in response to moderate osmotic stress caused by sodium chloride and aluminium sulphate, respectively. Furthermore, we used the A. cryptum JF-5 sequence to amplify the ectABCDask gene cluster from strain DSM 2389(T) and achieved heterologous expression of the gene cluster in Escherichia coli. Hence, we could for the first time prove metabolic functionality of the genes responsible for hydroxyectoine biosynthesis in the acidophile A. cryptum. In addition, we present information on specific enzyme activity of A. cryptum DSM 2389(T) ectoine synthase (EctC) in vitro. In contrast to EctCs from halophilic microorganisms, the A. cryptum enzyme exhibits a higher isoelectric point, thus a lower acidity, and has maximum specific activity in the absence of sodium chloride.

R1 resection in pancreatic cancer has significant impact on long-term outcome in standardized pathology modified for routine use.

BACKGROUND: The quality of a histopathologic workup after oncologic resection of pancreatic malignancies has changed the central role of surgery substantially for radical tumor clearance over the past years. The development of standardized protocols for pathologic workup increased the rate of R1 resections from around 20% up to 80%. In the present study, we investigated the incidence of R1 and its impact on survival after oncologic pancreatic resections using a standardized pathologic routine protocol. PATIENTS AND METHODS: We performed 265 pancreatic resections from September 2003 to September 2010. Among 128 patients with malignant neoplasms, histology revealed ductal pancreatic adenocarcinoma in 97, ampullary cancer in 10, and distal bile duct cancer in 21 patients. Resected specimens were analyzed according to this improved standardized pathology protocol introduced in 2000. Follow-up data on overall and cancer-related survival, presence and site of tumor recurrence, and chemotherapy were obtained from 120 patients. RESULTS: Pancreatic resection comprised a pylorus-preserving or classical pancreaticoduodenectomy in 112, a distal pancreatectomy in 8, and a total pancreatectomy in 7 patients. In the overall series, 56 (44%) were classified R1 resections and 68 (43%) R0 resections, 3 patients with R2 resections were excluded, leaving 125 patients for analysis. In pancreatic adenocarcinoma, the rate of R1 was 51% (48/94). R1 resection involved most frequently the circumferential margin in 86% (48/125) of the total group and in 92% (44/48) in pancreatic cancer. Follow-up was performed after a median of 17 months (range, 1-85) postoperatively. Cancer-related death rate in R0 and R1-resected patients was 60% and 83% (P < .02) in all cancers (n = 117) and 66% and 80% in patients with pancreatic adenocarcinoma (n = 88). Median tumor-related survival in R0 and R1 resections was 22 (range, 4-85) vs 14 months (range, 2-48) in all cancers (P < .002), and 19 (range, 4-85) vs 14 months (range, 2-48) in pancreatic adenocarcinoma (P < .04). Kaplan-Meier survival analysis revealed a survival benefit after R0 resection in both all cancers (P = .002) and pancreatic adenocarcinoma (P < .02). The pattern of tumor recurrence had a greater rate of regional metastases in the R1 group (P < .05). CONCLUSION: Our 51% rate of R1 resections in ductal pancreatic carcinoma indicates a high quality standard of pathologic evaluation. The vast majority of R1 margins are located at the retroperitoneal dissection surface. Standardization of histopathologic analysis has a clinically relevant impact on survival after oncologic resection of pancreatic cancer and can be achieved by less extensive protocols.

Malignancy and specific allergen immunotherapy: the results of a case series.

BACKGROUND: Specific immunotherapy with allergen is the only causative treatment for IgE-mediated allergies such as stinging insect allergy or hay fever and works by the induction of blocking antibodies and regulatory T lymphocytes. OBJECTIVE: Does a hypothetical obstruction of tumor surveillance presupposing the induction of regulatory T cells really justify detaining immunotherapy to oncologic patients as suggested by recent guidelines? METHODS: We report 6 patients (4 female, 2 male) suffering or having suffered from stage 1 cancer (4 melanomas, 1 lung cancer, 1 breast cancer) and concomitant IgE-mediated allergy. Four of them had a history of severe anaphylactic reactions to the insect yellow jacket, the 5th suffered from allergic rhinoconjunctivitis to dust mites, and the 6th to grass/rye pollen. RESULTS: Between 2004 and 2010, subcutaneous immunotherapy was safely performed in 5 patients without signs of tumor reactivation. The cancer in 2 of them was diagnosed immediately after specific immunotherapy had been initiated and in another 2 the active cancer phase had already finished years before; the 5th suffered from a relapse around the time of the initiation of immunotherapy. At the time of the writing of the manuscript, 4 of them had already concluded 3 years of treatment, another one almost 1 year. The melanoma in the 6th patient was diagnosed 5 months after reaching the maintenance dose. Immunotherapy with grass/rye pollen was aborted in this patient based on current guidelines. CONCLUSIONS: Specific immunotherapy was safely administered in patients suffering concomitantly from IgE-mediated allergy and lower stage cancer.

Persistence of antibodies in 4-8 year old Austrian children after vaccination with hexavalent DTaP-HBV-IPV/Hib and MMR vaccines.

To determine the proficiency of the Austrian childhood vaccination schedule to induce long lasting seroprotection against vaccine preventable diseases a seroepidemiological study in 348 children between four and eight years of age was conducted. Antibodies against diphtheria, tetanus, pertussis, hepatitis B, measles, mumps and rubella antigens were assessed in children, who had been vaccinated with hexavalent DTaP-HBV-IPV/Hib vaccines at three, four, five months and in the second year of life and/or MMR vaccines in the second year of life at least once, but mostly twice. High seroprotection rates (SPRs) were detected for tetanus (96%) and measles (90%). SPRs regarding diphtheria and mumps were 81% and 72%, respectively. Rubella-SPRs were 68% in females and 58% in males. Hepatitis B-antibody levels ≥10 mIU/mL were present in 52%; antibodies against pertussis were detected in 27% of the children. SPRs for measles and rubella depended on the interval since last vaccination; mumps-antibodies were significantly lower after one MMR-vaccination only. Antibodies against diphtheria, tetanus and pertussis depended on the interval since last vaccination while HBs-antibodies did not. The low levels of antibodies 1-7 years after vaccination against pertussis, rubella and mumps after only one vaccination should be considered when recommending new vaccination schedules.

Fixed drug eruption caused by mefenamic acid: a case series and diagnostic algorithms.

BACKGROUND: Fixed drug eruption is a fairly common drug-induced hypersensitivity reaction of the skin and the mucous membranes, which is characterized by the re-occurrence of the lesion(s) exactly on the previously involved sites after repeated administration. The pathogenetic mechanisms of this site-specificity are not fully elucidated. PATIENTS AND METHODS: We report on three cases of fixed drug eruption, including a non-pigmenting generalized bullous fixed drug eruption, caused by mefenamic acid in its pure form. RESULTS AND CONCLUSION: Provocation tests with the assumed causative drug represent the gold standard for establishing the diagnosis and for identifying the culprit. Advantages and pitfalls of topical and systemic provocation tests as diagnostic approaches are discussed.

TNF-like weak inducer of apoptosis (TWEAK) and TNF-α cooperate in the induction of keratinocyte apoptosis.

BACKGROUND: Activation of skin keratinocytes followed by their apoptotic death leads to eczema and spongiosis formations in patients with atopic dermatitis (AD). TNF-like weak inducer of apoptosis (TWEAK) binds to its receptor, fibroblast growth factor-inducible 14 (Fn14), and controls many cellular activities, including proliferation, migration, differentiation, apoptosis, angiogenesis, and inflammation. OBJECTIVE: The aim of the study was to investigate the role of TWEAK and Fn14 in the formation of eczema in patients with AD. METHODS: Primary keratinocytes were isolated from nonlesional skin from patients with AD and psoriasis and from normal skin of healthy donors. Apoptosis analysis was performed by using annexin V/7-aminoactinomycin D and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling staining. The expression and regulation of TWEAK, TNF-α, Fn14, TNF receptor (TNFR) 1, and TNFR2 were measured by means of RT-PCR, flow cytometric analysis, and ELISA. TWEAK and Fn14 expression of lesional AD and psoriatic skin and normal control skin was analyzed by using immunohistochemistry and immunofluorescence. RESULTS: TWEAK and TNF-α cooperate in the induction of apoptosis in primary keratinocytes obtained from patients with AD, patients with psoriasis, and healthy subjects and in artificial skin equivalents. TNFR1 and Fn14 were the main receptors involved. TWEAK upregulates TNF-α expression in primary keratinocytes, whereas TNF-α did not affect the expression of TWEAK and its receptors. High TWEAK expression was observed in AD lesions but not in psoriatic lesions or normal skin. Fn14 was highly expressed in the lesional skin of patients with AD and patients with psoriasis and in healthy control skin. CONCLUSION: The high expression of TWEAK in lesional AD skin contributes to the difference in keratinocyte apoptosis and lesional formation between AD and psoriasis.

IL-32 is expressed by human primary keratinocytes and modulates keratinocyte apoptosis in atopic dermatitis.

BACKGROUND: Keratinocyte (KC) apoptosis is an important mechanism of eczema and spongiosis in patients with atopic dermatitis (AD) and is mediated by IFN-gamma, which is secreted by T(H)1 cells. IL-32 is a proinflammatory cytokine that is involved in the inflammatory processes of rheumatoid arthritis, chronic obstructive pulmonary disease, and Crohn disease. Recently, it was shown that upregulation of IL-32 induces apoptosis. OBJECTIVE: The aim of the study was to investigate the expression and function of IL-32 in patients with AD. METHODS: The expression of IL-32 in KCs was analyzed by means of RT-PCR, ELISA, and flow cytometry. Transfections of small interfering RNA were performed in primary KCs, and apoptosis was analyzed by means of terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling, annexin-V, and 7-amino actinomycin D stainings. Immunofluorescence stainings were used to detect IL-32 in skin biopsy specimens, and serum levels of IL-32 were analyzed by means of ELISA. RESULTS: We report that IL-32 is expressed in human primary KCs on stimulation with IFN-gamma, TNF-alpha, and T(H)1 cells in contrast to T(H)2, regulatory T (Treg), or T(H)17 cells, which showed no effect. Transfection of primary KCs and artificial skin equivalents with small interfering RNA to IL-32, which resulted in a clear decrease in IL-32 expression, significantly reduced KC apoptosis. Immunofluorescence staining demonstrated that IL-32 was expressed in AD lesional skin, whereas it was present in neither skin biopsy specimens from healthy donors nor in lesional skin from patients with psoriasis. Serum levels of IL-32 from patients with AD correlated with disease severity, but increased serum levels of IL-32 were also detected in asthmatic patients. CONCLUSION: The present study demonstrates KCs as a source of IL-32, which modulates KC apoptosis and contributes to the pathophysiology of AD.