Response by B. Eberlein, I. León Suárez, U. Darsow, F. Ruëff, H. Behrendt, J. Ring

Abstract

We appreciate the comments by Dr Monneret about the staining and gating strategies in optimizing the identification of basophils to our study on the newly developed basophil activation protocol using CD63 and CCR3 [1]. He points out that CCR3 as an identification marker for basophils is also expressed in T cells (Th2 lymphocytes and regulatory T cells), thus reducing the purity of basophils and lowering their sensitivity. According to the information of the manufacturer [2] a contamination of CD3 cells in the CCR3 population occurs with a mean of 3.85% (95% CI = 2.52-5.18%; n = 8). Furthermore, in CD4 1 CD25 1 T cells, CCR3 expression appears not to exceed 5% ( fig. 1 of [3]). If we would have these 5% T cells in the CCR3 population, they would not express CD63. Therefore an activation of, e.g., 90% CD63 would be in fact only 85.5% and an activation of 15% would be 14.25% activation. These differences might be negligible for clinical purposes. Therefore we are not in accordance with the argument that staining only with CCR3 is an insufficient step for the identification of basophils. Nevertheless, we agree with Monneret, that an additional antibody labelling T cells based on CD3 expression would be an optimal staining strategy. As the Flow2 CAST was developed for clinical routine testing, the existing protocol with two antibodies is easier to handle, particularly if studies of different labs are compared. Staining with a third antibody might be the source of additional errors and cannot be performed with older flow cytometers still in use. References1 Eberlein B, Suárez IL, Darsow U, Ruëff H, Behrendt H, Ring J. A new basophil activation test using CD63 and CCR3 in allergy to antibiotics. Clin Exp Allergy 2010; 40:411-8. 2 Test Brochure Flow2 CAST s , BÜHLMANN Laboratories AG, Schönenbuch, Switzerland. Available at http://www.buhlmann labs.ch/core/allergy/flow2-cast/ 3 Ahern D, Lloyd CM, Robinson DS. Chemokine responsiveness of CD41CD251 regulatory and CD41CD25À T cells from atopic and nontopic donors. Allergy 2009; 64:1121-9. c 2010 Blackwell Publishing Ltd, Clinical & Experimental Allergy, 40 : 953-954 954 B. Eberlein et al

Highly Skin‐Conformal Microhairy Sensor for Pulse Signal Amplification

“…In addition to the cross‐sectional analysis, the structural properties of a pig skin were investigated using AFM and optical microscopy (see Figure S4, Supporting Information). In general, skin is known to have a morphologically dual‐level rough surface with micro‐ and nano levels . It was measured that the roughness of the examined pig skin ranged from minimum 1.9 nm to maximum of 90 to 240 μm (average roughness = 18 μm, moisture‐dependent) in Figure S4 (Supporting Information, via both AFM and optical microscopy).…”

Expression of γ-H2AX, 53BP1 and Micronuclei as Genome Damage Biomarker of Population in Keang and Salumati Village, Mamuju West Sulawesi Province

“…The observation was verified by an experienced investigator (IKHB) using a fluorescence microscope (Nikon) equipped with red, green and blue fluorescence filters and a 100x lens under immersion oil. Generally, 50 γ-H2AX and 53BP1 foci were counted per individual [11][12][13][14].…”

Targeting the gut‐skin axis—Probiotics as new tools for skin disorder management?

“…The negative effect of the use of soaps and detergents on skin is most possibly due to the consequent marked increase (3 U) of skin surface pH that can last for almost two hours, and an elevated pH is detrimental on epidermal barrier function . It is of note that AD‐affected patients have a higher skin surface pH than individuals with normal skin . Whether the prevalent cutaneous dysbiosis of AD patients is a cause or a consequence of the elevation of skin pH is not yet clarified.…”