Development and validation of a minimally invasive and image-guided tape stripping method to sample atopic skin in children.

BACKGROUND: Molecular skin profiling techniques, typically performed on skin samples taken by punch biopsy, have enhanced the understanding of the pathophysiology of atopic dermatitis (AD), thereby enabling the development of novel targeted therapeutics. However, punch biopsies are not always feasible or desirable, and novel minimally invasive methods such as skin tape stripping have been developed. AIM: To develop, optimize and validate a novel tape stripping method guided by noninvasive in vivo skin imaging to sample atopic skin in children. METHODS: Skin tape stripping-based procedures were compared and optimized using data from 30 healthy controls (HCs: 5 adults, 25 children) and 39 atopic children. Evaluations were guided by high-resolution photography, reflectance confocal microscopy, optical coherence tomography and transepidermal water loss measurements. We assessed and compared adverse events (AEs), the time needed to perform the sampling and the cDNA levels obtained from the tapes. RESULTS: Tape stripping methods based on previously described protocols resulted in erosions in all participants and required a median time of 65 min to perform (range 60-70 min), but provided good cDNA yield. Shorter durations appeared less invasive but provided lower cDNA yield. The final optimized tape stripping protocol, using 11 tapes of 22 mm in diameter, each applied twice for 5 s with 90° rotation, did not produce significant AEs, was completed within a median time of 7 min (range 5-15 min) and provided good cDNA yield both in HCs and atopic children. CONCLUSION: Our minimally invasive method is safe and reliable, and provides reproducible acquisition of cDNA in atopic children. In addition, it enables rapid sample collection, a crucial factor in clinical practice.

Microarray Gene Expression Analysis to Evaluate Cell Type Specific Expression of Targets Relevant for Immunotherapy of Hematological Malignancies.

Cellular immunotherapy has proven to be effective in the treatment of hematological cancers by donor lymphocyte infusion after allogeneic hematopoietic stem cell transplantation and more recently by targeted therapy with chimeric antigen or T-cell receptor-engineered T cells. However, dependent on the tissue distribution of the antigens that are targeted, anti-tumor responses can be accompanied by undesired side effects. Therefore, detailed tissue distribution analysis is essential to estimate potential efficacy and toxicity of candidate targets for immunotherapy of hematological malignancies. We performed microarray gene expression analysis of hematological malignancies of different origins, healthy hematopoietic cells and various non-hematopoietic cell types from organs that are often targeted in detrimental immune responses after allogeneic stem cell transplantation leading to graft-versus-host disease. Non-hematopoietic cells were also cultured in the presence of IFN-γ to analyze gene expression under inflammatory circumstances. Gene expression was investigated by Illumina HT12.0 microarrays and quality control analysis was performed to confirm the cell-type origin and exclude contamination of non-hematopoietic cell samples with peripheral blood cells. Microarray data were validated by quantitative RT-PCR showing strong correlations between both platforms. Detailed gene expression profiles were generated for various minor histocompatibility antigens and B-cell surface antigens to illustrate the value of the microarray dataset to estimate efficacy and toxicity of candidate targets for immunotherapy. In conclusion, our microarray database provides a relevant platform to analyze and select candidate antigens with hematopoietic (lineage)-restricted expression as potential targets for immunotherapy of hematological cancers.

Multidisciplinary rehabilitation treatment versus cognitive behavioural therapy for patients with chronic fatigue syndrome: a randomized controlled trial.

OBJECTIVES: The aim of this trial was to evaluate the difference in treatment effect, at 26 and 52 weeks after the start of treatment, between cognitive behavioural therapy (CBT) and multidisciplinary rehabilitation treatment (MRT) for patients with chronic fatigue syndrome (CFS). DESIGN: Multicentre, randomized controlled trial of patients with CFS. Participants were randomly assigned to MRT or CBT. SETTING: Four rehabilitation centres in the Netherlands. SUBJECTS: A total of 122 patients participated in the trial. MAIN OUTCOME MEASURES: Primary outcomes were fatigue measured by the fatigue subscale of the Checklist Individual Strength and health-related quality of life measured by the Short-Form 36. Outcomes were assessed prior to treatment and at 26 and 52 weeks after treatment initiation. RESULTS: A total of 114 participants completed the assessment at 26 weeks, and 112 completed the assessment at 52 weeks. MRT was significantly more effective than CBT in reducing fatigue at 52 weeks. The estimated difference in fatigue between the two treatments was -3.02 [95% confidence interval (CI) -8.07 to 2.03; P = 0.24] at 26 weeks and -5.69 (95% CI -10.62 to -0.76; P = 0.02) at 52 weeks. Patients showed an improvement in quality of life over time, but between-group differences were not significant. CONCLUSION: This study provides evidence that MRT is more effective in reducing long-term fatigue severity than CBT in patients with CFS. Although implementation in comparable populations can be recommended based on clinical effectiveness, it is advisable to analyse the cost-effectiveness and replicate these findings in another multicentre trial.

Determination of lactones in wines by headspace solid-phase microextraction and gas chromatography coupled with mass spectrometry.

Application of headspace solid-phase microextraction (HS-SPME) coupled with high-resolution gas chromatographic (HRGC) analysis was studied for determining lactones in wines. Six different SPME fibers were tested, and the influence of different factors such as temperature and time of desorption, ionic strength, time of extraction, content of sugar, ethanol, tannins and anthocyanins, and pH and influence of SO2 were studied. The proposed HS-SPME-GC method is an appropriate technique for the quantitative analysis of γ-butyrolactone, γ-hexalactone, trans-whiskey lactone, γ-octalactone, cis-whiskey lactone, γ-nonalactone, γ-decalactone, δ-decalactone, and γ-undecalactone in wines. Method reproducibility and repeatability ranged between 0.6 and 5.2% for all compounds. Detection limit for γ-butyrolactone was 0.17 mg/L and a few µg/L for the rest of the compounds. The optimized method has been applied to several wine samples.

[Female sexuality after surgical treatment of symptomatic deep pelvic endometriosis]. / Sexualité féminine après chirurgie pour endométriose pelvienne profonde.

OBJECTIVES: The aim of this study is to assess the impact on sexuality of the surgical treatment in patients with symptomatic deep pelvic endometriosis. PATIENTS AND METHODS: The design is a single-center cohort prospective study including all patients with symptomatic pelvic endometriosis and regular sexual activity who underwent surgery between October 2009 and September 2010. Sexual function was evaluated by the "Brief Index of Sexual Functioning for Women" (BISF-W) questionnaire translated and validated in French, including a global evaluation by the Composite Score (CS). Pain symptoms related to endometriosis were evaluated by the Visual Analog Scale (VAS) and the simple Verbal Rating Scale (VRS). Questionnaires were answered before surgery. A standardized mid and long-term postoperative follow-up was performed to compare sexuality and pain symptoms. RESULTS: Twenty women were included in the study. Mean follow-up was 23.3 months. When compared to a French reference population, global preoperative sexual function was significatively deteriorated (CS=14.3±10.8 vs 32.2±12.6; P<0.001), especially for arousal, frequency of sexual activity, pleasure and orgasm. Significant improvements in sex life were observed after surgery at the long-term follow-up (CS=33.0±11.7 vs 14.3±10.8; P=0.02). and sexual function was similar to the reference population (CS=33.0±11.7 vs 32.2±12.6; P=0.806). At the mid-follow-up, a significant improvement in the intensity of dysmenorrhoea, non-cyclic pelvic pain, dyspareunia and bowel symptoms were observed on the VAS. At the long-term follow-up, dysmenorrhoea and dyspareunia were significatively ameliorated. Pelvic pain recurrence related to endometriosis was 13.3%. DISCUSSION AND CONCLUSION: Surgical management of deep pelvic endometriosis in symptomatic patients improves sexual life at the long term follow-up. Deep dyspareunia pain decreases significantly, although other conditions are involved in the improvement of sexual function.

[Solar urticaria: pre-PUVA UVA desensitization]. / Urticaire solaire : désensibilisation UVA avant PUVAthérapie.

BACKGROUND: Solar urticaria is an uncommon disorder sometimes difficult to treat. It is characterized by the occurrence of typical whealing reactions on exposed skin a few minutes after sun exposure. The reactions resolve 1 to 5 hours after sun exposure ceases. We report a case evolving over several years, unresponsive to antihistamines and successfully treated by PUVAtherapy performed after UVA desensitization. CASE REPORT: For 3 years, a 22-year-old man developed erythema and itchy wheals at each solar exposure. The lesions appeared on all exposed areas including those usually exposed (face and hand) even in winter. An antihistamine regimen given for several weeks (cetirizine, loratadine) was ineffective. One trial of PUVA therapy led to an urticarial reaction of the entire body. Phototesting showed the minimal whealing dose for UVA was 0.4 J/cm(2). Phototherapy was therefore started by segmentary UVA irradiation at an initial dose of 0.1 J/cm(2). Exposure was then progressively increased allowing initiation of the PUVAtherapy on the 9(th) day with a dose of 0.5 J/cm(2) without whealing reaction. Slow increment PUVA therapy was able to induce good tolerance to sun exposure. DISCUSSION: Solar urticaria may sometimes have a deleterious effect on normal daily life. Severe cases are characterized by a whealing reaction after minimal sun exposure, even on regularly exposed skin. Antihistamines can provide some symptom relief in many patients, but high doses are required. If antihistamines are ineffective, PUVA therapy is indicated. Pre-PUVA UVA desensitization is often necessary. However, exposure to UVA alone has to be repeated every 24 to 48 hours to maintain the refractory state. The advantage of PUVA therapy is a more long lasting protection allowing weekly maintenance sessions.

Wartime colon injuries: primary repair or colostomy?

A retrospective non-randomized study, comparing primary repair with colostomy, was made on a series of 102 patients with penetrating intraperitoneal colon injuries, in a war surgery programme in Cambodia. The overall case fatality rate (CFR) was 25.5%, whereas in the primary repair group CFR was 20%, compared to 30.8% in the colostomy group. The difference was not statistically significant (P = 0.30). Adjustment for possible confounding factors in the two groups did not alter the results. Considering the numerous advantages to the patient of a primary closure in the precarious situations where war surgery is often performed, this technique merits consideration.

[Possibilities and limitations of nutritional support in Crohn disease]. / Possibilità e limiti del supporto nutrizionale nella malattia di Crohn.

Crohn's disease involves a great risk of malnutrition. Malabsorption, bacterial contamination, frequent abdominal surgery, meal-related pain, protein loss through the damaged mucosa contribute to creating nutritional problems. Malnutrition can worsen the outcome, both in medical and surgical patients, and deteriorate an often already altered immune response. Weight loss, low levels of blood protein, electrolytes, micronutrients and vitamins are usually related to the extension of the mucosal damage. Nutritional assessment can be difficult due to oedema and bleeding, who interfere with both clinical and laboratory evaluation. The exact amount of nitrogen, lipids, minerals stool loss can be useful. It is widely accepted the use of nutritional support in Crohn's disease, but many Authors do not agree concerning the route (enteral or parenteral) and the kind of nutrient to be used. Still controversial is the role of nutrition: just support or real therapy? Most recent hypothesis concerning the pathogenesis of Crohn's disease indicate food and/or bacterial antigens as involved in determining the pathology. The "bowel rest", considered for many years as a fasting period necessarily supported by parenteral nutrition, can also be obtained by the temporarily reduction or stop in presenting those antigens to the bowel mucosa. This new concept can be achieved not only by parenteral nutrition, but with an enteral elemental diet as well. The elemental diet contains all nutrients in the simplest way and thus succeeds in lowering or eliminating the antigenic power. The reported results seem to indicate an equivalence of enteral and parenteral nutrition; anyway enteral is advisable when feasible, being more physiological and less expensive and involving a lower risk of serious complications.(ABSTRACT TRUNCATED AT 250 WORDS)

Superior vena caval thrombosis and recurrent pericarditis caused by a bronchogenic cyst.

A 49-year-old man with a past history of multiple episodes of acute pericarditis was admitted with a recurrence of symptoms combined with the superior vena caval syndrome. A bronchogenic cyst, located beneath the carina, was discovered. At operation, the cyst was partially resected, and superior vena caval flow was restored by a Gore-Tex prosthesis. Ten months later the superior vena caval syndrome reappeared. Angiography demonstrated subtotal stenosis of the graft, which was considered to be thrombotic due to the discontinuance of oral anticoagulants. Reoperation, however, showed no thrombosis, but kinking of the graft. The folded portion of the prosthesis was resected, since when the patient has done well.

Prenatal diagnosis of fetal lymphatic system abnormalities by ultrasound.

Antenatal ultrasound diagnoses of gross lymphatic system abnormalities were made in three fetuses. Although the diagnosis was made in only 1 fetus at a stage early enough in pregnancy to allow selective termination, knowledge of the abnormality in the remaining 2 fetuses proved to be valuable for subsequent management of those pregnancies. It is stressed that the extent of the abnormality must be carefully assessed because of the possibility of corrective surgery should the lesion be small and that the parents must be given detailed counseling before any definitive measures are taken.