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Josef Skopalik,1 Katerina Polakova,2 Marketa Havrdova,2 Ivan Justan,1 Massimiliano Magro,3 David Milde,2 Lucia Knopfova,4 Jan Smarda,4 Helena Polakova,1 Eva Gabrielova,5 Fabio Vianello,2,3 Jaroslav Michalek,1 Radek Zboril21Department of Pharmacology, Masaryk University, Brno, Czech Republic; 2Regional Centre of Advanced Technologies and Materials, Department of Physical Chemistry and Analytical Chemistry, Faculty of Science, Palacky University, Olomouc, Czech Republic; 3Department of Comparative Biomedicine and Food Science, University of Padua, Padova, Italy; 4Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic; 5Department of Medical Chemistry and Biochemistry, Faculty of Medicine, Palacky University, Olomouc, Czech RepublicObjective: Cell therapies have emerged as a promising approach in medicine. The basis of each therapy is the injection of 1–100×106 cells with regenerative potential into some part of the body. Mesenchymal stromal cells (MSCs) are the most used cell type in the cell therapy nowadays, but no gold standard for the labeling of the MSCs for magnetic resonance imaging (MRI) is available yet. This work evaluates our newly synthesized uncoated superparamagnetic maghemite nanoparticles (surface-active maghemite nanoparticles – SAMNs) as an MRI contrast intracellular probe usable in a clinical 1.5 T MRI system.Methods: MSCs from rat and human donors were isolated, and then incubated at different concentrations (10–200 µg/mL) of SAMN maghemite nanoparticles for 48 hours. Viability, proliferation, and nanoparticle uptake efficiency were tested (using fluorescence microscopy, xCELLigence analysis, atomic absorption spectroscopy, and advanced microscopy techniques). Migration capacity, cluster of differentiation markers, effect of nanoparticles on long-term viability, contrast properties in MRI, and cocultivation of labeled cells with myocytes were also studied.Results: SAMNs do not affect MSC viability if the concentration does not exceed 100 µg ferumoxide/mL, and this concentration does not alter their cell phenotype and long-term proliferation profile. After 48 hours of incubation, MSCs labeled with SAMNs show more than double the amount of iron per cell compared to Resovist-labeled cells, which correlates well with the better contrast properties of the SAMN cell sample in T2-weighted MRI. SAMN-labeled MSCs display strong adherence and excellent elasticity in a beating myocyte culture for a minimum of 7 days.Conclusion: Detailed in vitro tests and phantom tests on ex vivo tissue show that the new SAMNs are efficient MRI contrast agent probes with exclusive intracellular uptake and high biological safety.Keywords: mesenchymal stromal cells, stem cell tracking, magnetic resonance imaging, superparamagnetic iron oxide nanoparticles, stem cell labelling

2-Ethylhexyl methoxycinnamate (EHMC) is one of the most used ultraviolet filters in personal care products. It undergoes cis/trans isomerization in sunlight, and there is limited toxicological understanding of the effects of the cis-isomer. It is known that two geometric isomers of one compound can have different physico-chemical properties and effects. However, there are no studies focusing on toxicokinetics of EHMC isomerization products to compare their potential difference in dermal exposure to cis-EHMC and trans-EHMC due to the difference in their dermatotoxicokinetics. In this study, dermal absorption of the parental trans-EHMC and its cis isomer was studied. A commercially available sunscreen lotion containing trans-EHMC and spiked with laboratory-prepared cis-EHMC was locally applied on the forearm skin of two volunteers. After 8 h of skin exposure, the stratum corneum (SC) layer was removed by tape stripping. The removed thickness of the SC was determined spectrophotometrically using a total protein assay. The concentration of both isomers in the removed SC was measured by HPLC-DAD. A new diffusion and permeability coefficient of both EHMC isomers in SC were determined by Fick's second law of diffusion in vivo. The difference in dermatotoxicokinetic parameters between the two isomers was not statistically significant. However, separate toxicological studies of isomeric forms and the determination of their dermatotoxicokinetic parameters are crucial for refinement of human risk assessment.

In the last few years, magnetically labeled cells have been intensively explored, and non-invasive cell tracking and magnetic manipulation methods have been tested in preclinical studies focused on cell transplantation. For clinical applications, it is desirable to know the intracellular pathway of nanoparticles, which can predict their biocompatibility with cells and the long-term imaging properties of labeled cells. Here, we quantified labeling efficiency, localization, and fluorescence properties of Rhodamine derivatized superparamagnetic maghemite nanoparticles (SAMN-R) in mesenchymal stromal cells (MSC). We investigated the stability of SAMN-R in the intracellular space during a long culture (20 days). Analyses were based on advanced confocal microscopy accompanied by atomic absorption spectroscopy (AAS) and magnetic resonance imaging. SAMN-R displayed excellent cellular uptake (24 h of labeling), and no toxicity of SAMN-R labeling was found. 83% of SAMN-R nanoparticles were localized in lysosomes, only 4.8% were found in mitochondria, and no particles were localized in the nucleus. On the basis of the MSC fluorescence measurement every 6 days, we also quantified the continual decrease of SAMN-R fluorescence in the average single MSC during 18 days. An additional set of analyses showed that the intracellular SAMN-R signal decrease was minimally caused by fluorophore degradation or nanoparticles extraction from the cells, main reason is a cell division. The fluorescence of SAMN-R nanoparticles within the cells was detectable minimally for 20 days. These observations indicate that SAMN-R nanoparticles have a potential for application in transplantation medicine.

Due to their widespread usage, people are exposed to pesticides on a daily basis. Although these compounds may have adverse effects on their health, there is a gap in the data and the methodology needed to reliably quantify the risks of non-occupational human dermal exposure to pesticides. We used Franz cells and human skin in order to measure the dermal absorption kinetics (steady-state flux, lag time and permeability coefficient) of Carbendazim and Simazine. These parameters were then used to refine the dermal exposure model and a probabilistic simulation was used to quantify risks resulting from exposure to pesticide-polluted waters. The experimentally derived permeability coefficient was 0.0034 cm h(-1) for Carbendazim and 0.0047 cm h(-1) for Simazine. Two scenarios (varying exposure duration and concentration, i.e. environmentally relevant and maximum solubility) were used to quantify the human health risks (hazard quotients) for Carbendazim and Simazine. While no risks were determined in the case of either scenario, the permeability coefficient, which is concentration independent and donor, formulation, compound and membrane specific, may be used in other scenarios and exposure models to quantify more precisely the dermally absorbed dose during exposure to polluted water. To the best of our knowledge, the dermal absorption kinetics parameters defined here are being published for the first time. The usage of experimental permeability parameters in combination with probabilistic risk assessment thus provides a new tool for quantifying the risks of human dermal exposure to pesticides., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Objectives: The aim of the study was to develop a clinical prediction model for assessing the probability of having invasive cancer in the definitive surgical resection specimen in patients with biopsy diagnosis of ductal carcinoma in situ (DCIS) of the breast, to facilitate decision making regarding axillary surgery., Methods: In 349 women with DCIS, predictors of invasion in the definitive resection specimen were identified. A model to predict the probability of invasion was developed and subsequently simplified to divide patients into two risk categories. The model's performance was validated on another patient population., Results: Multivariate logistic regression revealed four independent predictors of invasion: (i) suspicious (micro)invasion in the biopsy specimen; (ii) visibility of the lesion on ultrasonography; (iii) size of the lesion on mammography>30 mm; (iv) clinical palpability of the lesion. The actual frequency of invasion in the high-risk patient group in the test and validation population was 52.6% and 48.3%, respectively; in the low-risk group it was 16.8% and 7.1%, respectively., Conclusion: The model proved to have good performance. In patients with a low probability of invasion, an axillary procedure can be omitted without a substantial risk of additional surgery.

Background: For most breast cancer patients in the Czech Republic, breast reconstruction is available only in a delayed manner. In the Masaryk Memorial Cancer Institute (MMCI), suitable candidates are offered immediate breast reconstruction using tissue expander with later exchange to a permanent silicone implant. The aim of this study was to assess patient satisfaction with this type of reconstruction., Patients and Methods: Sixty-two women who had undergone surgery at the MMCI from 2007 through 2013 were sent a simple questionnaire developed by our working team. Fifty-seven patients completed the questionnaire. The data were evaluated by description methods and statistical tests., Results: Patient response was 92%. The absolute majority of patients (56/57) would opt for this method again. The vast majority of patients (48/57) are generally satisfied with their reconstruction. Most women (8/14) younger than 50 years after the unilateral surgery would prefer synchronous contralateral prophylactic mastectomy and bilateral reconstruction if they could choose again. After bilateral surgery, reconstructed breasts are more often regarded as a part of the patients body. Dressed women rate their look substantially better than when they are undressed. As for self-esteem, these women are feeling excellent or good. Their psychosocial well-being in common situations is predominantly excellent. Their sexual well-being is significantly worse, and almost half of these women indicate occasional pain in their reconstructed breasts. The patients emphasize the need for appropriate information before the surgery., Conclusion: Immediate two-stage implant-based breast reconstruction is a suitable option for some breast cancer patients. With regard to the less natural cosmetic result and feeling of the implant-reconstructed breast, appropriate selection of women for this type of surgery is necessary and potential candidates must be thoroughly informed in the preoperative setting.

Background: The finger range of motion (ROM) was evaluated in patients with hand flexor tendon replacement using a tendon graft and the effect of smoking was investigated. The first hypothesis was: a tendon graft increases the post-operative ROM. The second hypothesis was: smoking has a negative effect on the final finger ROM., Patients and Methods: Fifty seven autologous tendon grafts in 48 patients were included. The patients were grouped as non-smokers or smokers ("light" or "heavy"). The modified Strickland system and Total Active Motion (TAM) system, were chosen for the evaluation., Results: The first hypothesis was proved by all the measurements. The post-operative status assessed by the Strickland method was different between the non-smokers and smokers with a better score in the smokers., Conclusion: A significant ROM improvement occurs after reconstruction and is even slightly better in smokers than in non-smokers.

Background: The purpose of our study was to indentify the mechanical characteristics of various suture materials. We created an in-vitro experimental flexor tendon model., Material and Methods: Materials were divided into four groups: monofilament polypropylene non-absorbable material (group 1); monofilament long-term absorbable material (group 2); polyester multifilament non-absorbable coated material (group 3) and polyester multifilament non-absorbable uncoated material (group 4). We performed 135 tests. The mean maximal tensile strength was 62.92 N in group 1, 75.20 N in group 2, 36.38 N in group 3 and 72.4 N in group 4. Elasticity in millimetres was adjusted at the 35N level: group 1:2.01 mm, group 2:2.18 mm, group 3:2.14 and group 4:1.51 mm., Discussion: With regard to its elasticity and favourable SD for tensile strength measurements, polyester multifilament non-absorbable uncoated material was considered to be the most suitable material.

Background: Currently, several types of plasma discharge devices have become established for performing surgical procedures or superficial sterilization. Our goal is to introduce a brand new type of plasma discharge device with different characteristics, which we have used for two experiments involving thermal damage to biological tissues., Material and Methods: A plasma discharge device with the following characteristics was used for all incisions in plasma group: radio frequency barrier discharger at atmospheric pressure with a working frequency of 13.56 MHz. The working gas was Argon. We used a Cesar 136 generator with typical power 100-150W and with a working gas flow rate between 0.5 and 3 litres per minute. The inner diameter of the end of the nozzle was 0.4mm. A standard Martin electrotome was used for all comparative incisions in the electrotome group. We used two experimental fresh rat cadavers, employing their skin, tails, lungs, livers, kidneys and spleens to make a comparative study between incisions made with a plasma discharge knife and those made with a standard electrotome device. All samples underwent histological verification., Discussion: Our experiment showed slight differences in the impact on incision edges made by the plasma knife and the electrotome. The most interesting effect is on the skin: the plasma knife does not damage hair bulbs, and it results in less thermal damage. The plasma discharge knife is a promising device for cutting various biological tissues. Its effect can be compared with a standard electrotome, while in some tissues it causes less harm then an electrotome. Moreover, we can adjust its therapeutic effect. However, we do not consider the plasma knife a better choice at present, because maneuvering it is unwieldy.

We present a group of 20 patients who underwent transposition of the extensor carpi radialis longus to the extensor pollicis longus between April 2003 and September 2007. The group was divided into ten patients with an early dynamic splint and ten patients with a static plaster cast following the standard rehabilitation protocols. We compared these two rehabilitation protocols. The post-rehabilitation total range of motion was 87.5 degrees in the dynamic group and 58.75 degrees in the patients treated by the static protocol. The total time of rehabilitation was 10.2 weeks in the dynamic and 14.2 weeks in the static group. The overall mean follow-up was 12.2 weeks. We conclude that early dynamic splinting brings shorter recovery time, shorter time off work and significantly better movement of the thumb.

Unlabelled: Bilateral prophylactic mastectomy without reconstruction is not accepted by the majority of patients. Successful reconstruction is therefore a mandatory condition for prophylactic mastectomy. Of the many options for autologous breast reconstruction, the deep inferior epigastric perforator (DIEP) flap best meets requirements for bilateral reconstruction in selected patients. The goal of this study is to verify the feasibility of the procedure in our conditions and to find out how it is accepted by patients. We present 55 consecutive patients who were scheduled for bilateral DIEP flap reconstruction during a 4-year period. We reviewed medical charts, performed clinical assessments and processed anonymous questionnaires. There were 77 immediate and 33 delayed breast reconstructions. There was 100% flap survival and no microanastomoses revisions. In 11 patients (10%) the surgeon preferred to convert the DIEP into a mini transverse rectus abdominis muscle (miniTRAM) flap in order to provide adequate blood supply., Complications: revision for haematoma under the flap in four patients (7.2%), excessive blood loss in four patients (7.2%) and partial mastectomy skin flap necrosis in 10 immediate breast reconstructions (12.9%). Patients' evaluation of the aesthetic result was good or excellent in 96.2% of cases. In 33.9% of patients the postoperative quality of life was considered unchanged and 50.9% of them it even improved. The DIEP flap is recommended for bilateral breast reconstruction. Occasional conversion into a miniTRAM flap can increase the total flap survival rate. Bilateral prophylactic mastectomy and DIEP flap reconstruction are very well accepted by patients.

From December 2001 to September 2005, the technique of total penile reconstruction with a reinnervated free latissimus dorsi myocutaneous flap was used in 22 patients (24-38 years old) with gender dysphoria. These patients were followed up for at least 11 months (range, 11-44 months). All flaps survived. Complications include hematoma (7 cases), vascular thrombosis (2 cases), partial necrosis (1 case), excessive swelling of the neophallus (3 cases), and skin graft loss at the donor site (1 case). Of the 19 patients included in the final evaluation, the transplanted muscle was able to obtain contraction in 18 (95%) cases and 8 patients (42%) had sexual intercourse by contracting the muscle to stiffen and move the neopenis. The described technique of neophalloplasty proved to be a reliable technique and the muscle movement in the neophallus can be expected in almost all cases. The muscle contraction in the neophallus leads to "paradox" erection-stiffening, widening, and shortening of the neopenis, which allows for sexual intercourse in some patients. Subsequent reconstruction of the urethra is possible.

Twenty two patients with gender dysphoria underwent neo-phalloplasties using a novel technique. Latissimus dorsi musculocutaneus re-innervated free flap was used to allow voluntary rigidity of the neo-penis. From the first 22 patients, 18 have obtained motoric function of reconstructed penis; the "paradox erection" was obtained. 14 patients came for examination after a follow-up period of mean 26.4 months. We evaluated the motility and shape changes of neo-phallus measuring its different size and dimension during relax and muscle contraction. The range of neo-phallus length in relaxed position was between 7 and 17 cm (mean 12.2 cm), its circumference in the same position had a range between 13 and 20 cm (mean 13.7 cm). All patients were able to contract the muscle with an average length reduction of 3.08 cm and an average circumference enlargement of 4 cm. In this study, the dimensions and motility were quantified demonstrating the neo-phallus function and size changes during sexual intercourse.

Purpose of the Study: The aim of the study was to evaluate the results in a group of patients with congenital pseudarthrosis of the tibia treated by transfer of a vascularized fibular graft from the contralateral extremity., Material: The group included three boys and two girls aged 2 to 8 years at the time of surgery. In two patients, the vascularized graft transfer was preceded by other operations. All patients but one had Crawford type IV pseudarthrosis. The signs of peripheral neurofibromatosis were found in four of the five patients., Methods: The operation was carried out by two surgical teams, i. e., orthopedic and microsurgery (plastic surgery) specialists. Deep dissection of the pseudarthrosis was performed down to healthy, well vascularized tissue; a vascularized pedicle bone graft was harvested from the contralateral fibula. The graft was inserted and anchored intramedullarily in both tibial fragments, and stability was provided with a K-wire introduced through the calcaneus. Subsequently, the vascular pedicle of the fibular graft was joined to the surrounding vessels (anterior tibial artery and anterior tibial vein). The extremity was immobilized in plaster cast and later a KAFO brace was applied., Results: Graft union partially failed in the proximal end of the graft due to bone resorption of both the graft and the proximal tibial fragment in two patients. This was successfully treated by additional spongioplasty. In all patients bony union was achieved at an average time of 9.8 months (range, 6 to 21 months). An increase by more than 100 % in the diameter of the transplanted fibula was recorded in four patients. The increase, which was of course related to follow-up time, was a clear proof of primary graft vascularization., Discussion: The use of vascularized fibular graft harvested from the contralateral extremity showed high effectiveness in comparison with other methods. This was in agreement with the relevant literature reports., Conclusions: This method can be used regardless of patients' age and our experience showed that, even in small children, union can be achieved and can thus allow for early weight-bearing and prevention of crus atrophy.

Correction of radius length after a premature complete or partial closure of the distal epiphyseal plate is important for both the correction of radial deviation of the hand and the restoration of normal function of the radiocarpal articulation. This study comprised four patients. Two children with complete closure of the epiphyseal plate of the distal radius underwent callotaxis. One patient was treated for atrophic pseudoarthrosis of the distal radial metaphysis by external fixation and subsequent spongioplasty. One patient with partial closure of the epiphyseal plate was treated by physeal distraction. In each patient an Ilizarov's external fixator was applied. The full correction of radius length was achieved in all cases. No complications were recorded with the exception of tissue irritation around Kirchner's wires, which occurred mainly on the volar side. In the therapy of injuries suffered during adolescent growth acceleration, it is necessary to perform a certain overdistraction of the radius. An alternative method includes epiphyseodesis of the distal ulna. Key words: post-traumatic closure of the epiphyseal plate, callotaxis, physeal distraction, chondrodiatasis.

The authors submit a summary of the most recent findings on the damage of the nail area of the fingers of the hand (nail matrix, surrounding soft tissues, skeleton excluding avulsion fractures and epiphyseolyses.). The reason is the fact that almost 50% of all finger injuries in children under 5 years include also damage of this complex. Although the incidence of this injury is relatively high there is no uniform therapeutic procedure and unfortunately very frequently therapeutic procedures do not lead to favourable results. This apparently trivial injury can however, in particular in relation to occupation, lead not only to cosmetic but also functional disorders and therefore it is necessary to pay adequate attention to it. The authors supplement data from the literature by their own experience with the treatment of this comprehensive injury. The objective of the work is therefore to summarize available therapeutic findings on the classification and diagnostic procedures of these apparently light injury from the layman's aspect and unfortunately also from the aspect of some professionals, these injuries being overlooked and underrated (Innis, 1995). Particular attention is however devoted to adequate treatment which must lead to restoration of the injured fingers to full function which subsequently will restore the comprehensive role of the whole hand.