Gramicidin derivatives as membrane-based pH sensors

Ion channels provide a means for sensitive pH measurement at membrane interfaces. Detailed knowledge of the structure and function of gramicidin channels permits the engineering of pH-sensitive derivatives. Two derivatives, gramicidin-ethylenediamine and gramicidin-histamine, are shown to exhibit pH-dependent single-channel behaviour over the pH ranges 9-11 and 6.5-8.5, respectively. Thermal isomerization of a carbamate group at the entrance of the channels leads to a pattern of steps in single-channel recordings. The size of the steps depends on the time-averaged degree of protonation of the appended group (ethylenediamine or histamine). Measurement of the size of the steps thus permits single-molecule pH sensing under symmetrical pH conditions or in the presence of a pH gradient.     

Ecological patterns strongly impact the biogeography of western Palaearctic longhorn beetles (Coleoptera: Cerambycoidea)

We aim to unravel the biogeographic structuring of western Palaearctic longhorn beetles with focus on the location of different refugia, barriers to dispersal and postglacial range expansions with their particular filters. The interaction of different ecological features with these structures is analysed. The western Palaearctic was divided into 95 geographic entities. We produced presence-only matrices for all 955 Cerambycoidea species autochthonous to this area and derived species richness distributions and extracted faunal regions and faunal elements by cluster analyses and principal component analyses. Similar analyses were performed for sub-families and ecological groups. Longhorn beetles show a strong biogeographic structuring in the western Palaearctic. Species numbers strongly decrease to the north and west. Less mobile species and root feeders mostly contribute to the fauna of the Mediterranean region, whilst mobile species are more widespread. Feeders on broad-leaved trees dominate in western Europe, whilst feeders on coniferous trees are most important in northern Europe. Our results support multiple refugia in the Mediterranean region and underline the importance of Provence, Crimea and Crete as such refugia. Crete even might be an area of old endemism. The Atlanto- and the Ponto-Mediterranean regions are more strongly structured than assumed in classical biogeography. Mediterranean assemblages are mostly composed of non-flying species, root feeders and species with small distributions not found outside their glacial refugia. Tree feeders left their glacial retreats with their host plants. These range dynamics result in biogeographic structures with several dispersal barriers and filters composed of mountains, sea straits and climatic conditions.     

In vitro Biological Activities of Seed Essential Oils from the Cameroonian Spices Afrostyrax lepidophyllus Mildbr. and Scorodophloeus zenkeri Harms Rich in Sulfur‐Containing Compounds

The chemical composition of the essential oils obtained from the seeds of bush onion (Afrostyrax lepidophyllus) and tropical garlic tree (Scorodophloeus zenkeri), plants used as spices in the traditional African cuisine, was determined by GC‐FID and GC/MS analyses. Moreover, in vitro biological properties of the oils, namely, the cytotoxic, antioxidant, and antimicrobial activities, were investigated by the MTT, the DPPH^. and ABTS^.+ scavenging, and the agar disc‐diffusion methods, respectively. Both oils were composed mainly by S‐containing compounds, accounting for 91.0–96.1% of the total oil compositions, which provided them the typical garlic‐ and onion‐like odors of spices. The predominant compound in both oils, 2,4,5,7‐tetrathiaoctane ( 1 ; 51.5–52.9%), was isolated by preparative TLC and structurally elucidated by ¹H‐ and ¹³C‐NMR data. The oils exhibited a strong inhibitory effect on the growth of human cancer cells, namely, T98G (human glioblastoma multiforme cell line), MDA‐MB 231 (human breast adenocarcinoma cell line), A375 (human malignant melanoma cell line), and HCT116 (human colon carcinoma cell line) cells, and a good DPPH^.‐ and ABTS^.+‐scavenging activity, while the antimicrobial effects were negligible. The volatile compositions of A. lepidophyllus and S. zenkeri oils supported their use as odorous spices. The significant inhibition activities detected make these oils worthy of further investigation as promising chemopreventive agents to be exploited in the African pharmaceutical market.     

Comprehensive proteomic analysis of human cervical-vaginal fluid using colposcopy samples

Background  

Cervical-vaginal fluid (CVF) plays an important role in the prevention of gynecological infections, although little is known about the contribution of CVF proteins to the immunity of the lower female genital tract. In order to analyze the protein composition of human CVF, we used CVF samples that are routinely collected during colposcopy, but are usually discarded. Since these samples are available in large quantities we aimed to analyze their usefulness for proteomics experiments. The samples were analyzed using different prefractionation techniques (ultrafiltration and C₄(RP)-LC protein separation) followed by C₁₈(RP)-LC peptide separation and identification by MALDI-TOF-TOF mass spectrometry. To determine the reproducibility of this proteomics platform we analyzed three technical replicates. Using spectral counting, protein abundances were estimated in a semiquantitative way. We also compared the results obtained in this study with those from previous studies derived from patients with different physiological conditions in order to determine an overlapping protein set.     

Estimation of anisotropy coefficient of swine pancreas,liver and muscle at 1064 nm based on goniometric technique

Optical properties of tissues are required for theoretical modeling of Laser Ablation in tumor therapy. The light scattering characteristic of tissues is described by the anisotropy coefficient, g. The relationship between the angular distribution of scattered light and g is given by the Henyey‐Greenstein (HG) phase function. This work describes the estimation of anisotropy coefficients of ex vivo swine pancreas, liver and muscle at 1064 nm. The intensities of scattered light at fixed angles were measured under repeatability conditions. Experimental data were fitted with a two‐term HG, estimating the anisotropy coefficients for the forward (e.g., 0.956 for pancreas, 0.964 for liver and 0.968 for muscle) and the backward (e.g., –0.481 for pancreas, –0.414 for liver and –0.372 for muscle) scattering.

Experimental set up employed to estimate the anisotropy coefficient of biological tissues. The image on the left depicts the holder used to house tissue, laser fiber and photodetector; on the left an example of scattered light beam is shown, as well as the effect due to Snell's law.     

Human exposure to mycotoxins in Egypt

This investigation examined the exposure of Egyptian infants to Aflatoxin M₁ (AfM₁) and of lactating mothers to Aflatoxin B₁, using AfM₁ in human milk as a biomarker for exposure to AfB₁. The presence of ochratoxin A (OA) in human milk was also investigated to determine the levels of infants exposure to OA from human milk. The results indicated that AfM₁ was found in 66 (55 %) of 120 human milk samples with a mean of 0.3 ± 0.53 ng/mL (range 0.02 to 2.09 ng/mL). OA was found in 43 (35.8 %) of 120 human milk samples with a mean of 21.1 ± 13.7 ng/mL (range 5.07 to 45.01 ng/mL), which will cause a daily intake of OA from human milk exceeding the suggested tolerable dose of 5 ng/kg_-1 of OA body weight. On the other side AfM₁ was found in 25 % of blood samples (5 out of 20 samples), at a mean of 1.18 ng/mL, but it was detected only in one urine sample (1 out of 20 samples). OA was detected only in 2 out of 13 blood samples (15.4 %) with an average 3.67 ng/mL. Whereas OA was not detected in all analyzed urine samples.     

Finding the best antibody dilution by repeated immunostaining of the same tissue section

One can find the optimal antibody dilution for immunostaining by repeated staining on the same tissue section by using a less dilute antibody for each attempt. Using secondary antibody and horseradish peroxidase conjugated to a dextran polymer, a section stained repeatedly with several dilutions of antibody appears as good as a section stained with only the last dilution.