Modeling the Simultaneous Dynamics of Proteins in Blood Plasma and the Cerebrospinal Fluid in Human In Vivo.

The analysis of protein dynamics or turnover in patients has the potential to reveal altered protein recycling, such as in Alzheimer's disease, and to provide informative data regarding drug efficacy or certain biological processes. The observed protein dynamics in a solid tissue or a fluid is the net result of not only protein synthesis and degradation but also transport across biological compartments. We report an accurate 3-biological compartment model able to simultaneously account for the protein dynamics observed in blood plasma and the cerebrospinal fluid (CSF) including a hidden central nervous system (CNS) compartment. We successfully applied this model to 69 proteins of a single individual displaying similar or very different dynamics in plasma and CSF. This study puts a strong emphasis on the methods and tools needed to develop this type of model. We believe that it will be useful to any researcher dealing with protein dynamics data modeling.

miRViz: a novel webserver application to visualize and interpret microRNA datasets.

MicroRNAs (miRNAs) are small non-coding RNAs that are involved in the regulation of major pathways in eukaryotic cells through their binding to and repression of multiple mRNAs. With high-throughput methodologies, various outcomes can be measured that produce long lists of miRNAs that are often difficult to interpret. A common question is: after differential expression or phenotypic screening of miRNA mimics, which miRNA should be chosen for further investigation? Here, we present miRViz (http://mirviz.prabi.fr/), a webserver application designed to visualize and interpret large miRNA datasets, with no need for programming skills. MiRViz has two main goals: (i) to help biologists to raise data-driven hypotheses and (ii) to share miRNA datasets in a straightforward way through publishable quality data representation, with emphasis on relevant groups of miRNAs. MiRViz can currently handle datasets from 11 eukaryotic species. We present real-case applications of miRViz, and provide both datasets and procedures to reproduce the corresponding figures. MiRViz offers rapid identification of miRNA families, as demonstrated here for the miRNA-320 family, which is significantly exported in exosomes of colon cancer cells. We also visually highlight a group of miRNAs associated with pluripotency that is particularly active in control of a breast cancer stem-cell population in culture.

The Outcomes Assessment of the Plasma Blade Technology in Upper Blepharoplasties: A Prospective Study on a Series of 25 Patients.

BACKGROUND: The Dermo Ablation Surgery (DAS) Medical® (Technolux, Italy) device is a plasma blade which induces a plasma voltaic arc causing a retraction in the epidermis and superficial dermis. OBJECTIVE: The aim of our study is to prove the efficacy and safety of the DAS Medical® device in dermatochalasis size reduction. METHODS: Our prospective study included 25 adult patients presenting with upper eyelid dermatochalasis undergoing a two-session treatment protocol with the DAS Medical® device (with a month treatment-free interval). The primary end point was the reduction in the size of the dermatochalasis. The secondary end points were patient satisfaction, and a blinded assessment of the outcomes was carried out by 15 plastic surgery specialists on post-procedural pictures. RESULTS: The mean reduction in the size of the dermatochalasis was estimated at 2.47 mm on a 6-month follow-up (13.5 mm at T0 vs. 11.03 mm at 6 months, p = 0.0002) and 1.97 mm on a 12-month follow-up ((13.5 mm at T0 vs. 11.53 mm at 12 months, p = 0.0055). Eighty per cent of the patients and 78% of the assessing clinicians were globally satisfied with the results on a 12-month follow-up. The mean visual analogue pain score reported during the treatment was 4.5/10; MEOPA® was used in 23% of cases. No irreversible post-procedural sequelae (complications) were observed. CONCLUSION: Voltaic plasma arc treatment with DAS Medical® is an effective technique for non-invasive blepharoplasty on moderate dermatochalasis patients not suffering from palpebral lipoptosis and is very well tolerated. It can be usefully and successfully associated with surgery. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Cooperative activation of the T-type CaV3.2 channel: interaction between Domains II and III.

T-type CaV3 channels are important mediators of Ca(2+) entry near the resting membrane potential. Little is known about the molecular mechanisms responsible for channel activation. Homology models based upon the high-resolution structure of bacterial NaV channels predict interaction between the S4-S5 helix of Domain II (IIS4-S5) and the distal S6 pore region of Domain II (IIS6) and Domain III (IIIS6). Functional intra- and inter-domain interactions were investigated with a double mutant cycle analysis. Activation gating and channel kinetics were measured for 47 single mutants and 20 pairs of mutants. Significant coupling energies (ΔΔG(interact) ≥ 1.5 kcal mol(-1)) were measured for 4 specific pairs of mutants introduced between IIS4-S5 and IIS6 and between IIS4-S5 and IIIS6. In agreement with the computer based models, Thr-911 in IIS4-S5 was functionally coupled with Ile-1013 in IIS6 during channel activation. The interaction energy was, however, found to be stronger between Val-907 in IIS4-S5 and Ile-1013 in IIS6. In addition Val-907 was significantly coupled with Asn-1548 in IIIS6 but not with Asn-1853 in IVS6. Altogether, our results demonstrate that the S4-S5 and S6 helices from adjacent domains are energetically coupled during the activation of a low voltage-gated T-type CaV3 channel.

A quartet of leucine residues in the guanylate kinase domain of CaVß determines the plasma membrane density of the CaV2.3 channel.

Ca(V)ß subunits are formed by a Src homology 3 domain and a guanylate kinase-like (GK) domain connected through a variable HOOK domain. Complete deletion of the Src homology 3 domain (75 residues) as well as deletion of the HOOK domain (47 residues) did not alter plasma membrane density of Ca(V)2.3 nor its typical activation gating. In contrast, six-residue deletions in the GK domain disrupted cell surface trafficking and functional expression of Ca(V)2.3. Mutations of residues known to carry nanomolar affinity binding in the GK domain of Ca(V)ß (P175A, P179A, M195A, M196A, K198A, S295A, R302G, R307A, E339G, N340G, and A345G) did not significantly alter cell surface targeting or gating modulation of Ca(V)2.3. Nonetheless, mutations of a quartet of leucine residues (either single or multiple mutants) in the α3, α6, ß10, and α9 regions of the GK domain were found to significantly impair cell surface density of Ca(V)2.3 channels. Furthermore, the normalized protein density of Ca(V)2.3 was nearly abolished with the quadruple Ca(V)ß3 Leu mutant L200G/L303G/L337G/L342G. Altogether, our observations suggest that the four leucine residues in Ca(V)ß3 form a hydrophobic pocket surrounding key residues in the α-interacting domain of Ca(V)2.3. This interaction appears to play an essential role in conferring Ca(V)ß-induced modulation of the protein density of Ca(V)α1 subunits in Ca(V)2 channels.

Internal pudendal perforator artery flap harvesting without pre-operative imaging: Reliability and approach.

Since its initial description in 1996 by Yii and Niranjan, the internal pudendal perforator flap (also known as the Singapore flap, the gluteal fold flap, and the lotus petal flap) has become a workhorse in perineal soft tissue reconstruction. In 2001, Hashimoto described the presence of three to five perforators in the perineal anogenital triangle. The ischial tuberosity has thus become a useful anatomic landmark for the safe boundary of medial dissection during flap elevation, in order to avoid damaging the perforator vessels. The objective of the present study was to evaluate the perforators' positions within the anogenital triangle by using color Doppler ultrasound. In a study of 15 subjects in the lithotomy position, we identified a total of 24 perforator vessels with a diameter greater than 5 mm. We noted the vessels' positions using orthonormal measurements, according to the distance from the midline and the distance on a straight line between the two ischial tuberosities (i.e. consistent bony anatomic landmarks that are independent of the patient's height and body mass index). The mean distance between the ischial tuberosity and the internal pudendal perforator was 27.3 mm. Based on our present results, we consider that routine ultrasound identification and dissection of the perforators is not always required before pudendal flap harvesting. This decreases the operating time and simplifies the flap harvesting procedure.