Search results for: ex vivo bovine bone

Authors: Ruth Goldschmidt, Vyacheslav Kalchenko, Lilah Agemy, Rachel Elmoalem, Avigdor Scherz

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Vascular Targeted Photodynamic therapy (VTP) is a new modality for selective cancer treatment that leads to the complete tumor ablation. A photosensitizer, a bacteriochlorophyll derivative in our case, is first administered to the patient and followed by the illumination of the tumor area, by a near-IR laser for its photoactivation. The photoactivated drug releases reactive oxygen species (ROS) in the circulation, which reacts with blood cells and the endothelium leading to the occlusion of the blood vasculature. If the blood vessels are only partially closed, the tumor may recover, and cancer cells could survive. On the other hand, excessive treatment may lead to toxicity of healthy tissues nearby. Simultaneous VTP monitoring and image processing independent of the photoexcitation laser has not yet been reported, to our knowledge. Here we present a method for blood flow monitoring, using a real-time laser speckle imaging (RTLSI) in the tumor during VTP. We have synthesized over the years a library of bacteriochlorophyll derivatives, among them WST11 and STL-6014. Both are water soluble derivatives that are retained in the blood vasculature through their partial binding to HSA. WST11 has been approved in Mexico for VTP treatment of prostate cancer at a certain drug dose, and time/intensity of illumination. Application to other bacteriochlorophyll derivatives or other cancers may require different treatment parameters (such as light/drug administration). VTP parameters for STL-6014 are still under study. This new derivative mainly differs from WST11 by its lack of the central Palladium, and its conjugation to an Arg-Gly-Asp (RGD) sequence. RGD is a tumor-specific ligand that is used for targeting the necrotic tumor domains through its affinity to αVβ3 integrin receptors. This enables the study of cell-targeted VTP. We developed a special RTLSI module, based on Labview software environment for data processing. The new module enables to acquire raw laser speckle images and calculate the values of the laser temporal statistics of time-integrated speckles in real time, without additional off-line processing. Using RTLSI, we could monitor the tumor’s blood flow following VTP in a CT26 colon carcinoma ear model. VTP with WST11 induced an immediate slow down of the blood flow within the tumor and a complete final flow arrest, after some sporadic reperfusions. If the irradiation continued further, the blood flow stopped also in the blood vessels of the surrounding healthy tissue. This emphasizes the significance of light dose control. Using our RTLSI system, we could prevent any additional healthy tissue damage by controlling the illumination time and restrict blood flow arrest within the tumor only. In addition, we found that VTP with STL-6014 was the most effective when the photoactivation was conducted 4h post-injection, in terms of tumor ablation success in-vivo and blood vessel flow arrest. In conclusion, RTSLI application should allow to optimize VTP efficacy vs. toxicity in both the preclinical and clinical arenas.

Keywords: blood vessel occlusion, cancer treatment, photodynamic therapy, real time imaging

Procedia PDF Downloads 223

Authors: Parul Kulshreshtha, Subrata Sinha, Rakesh Bhatnagar

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This study was conducted by taking B. anthracis as a model pathogen. On infecting a host, B. anthracis secretes three proteins, namely, protective antigen (PA, 83kDa), edema factor (EF, 89 kDa) and lethal factor (LF, 90 kDa). These three proteins are the components of two anthrax toxins. PA binds to the cell surface receptors, namely, tumor endothelial marker (TEM) 8 and capillary morphogenesis protein (CMG) 2. TEM8 and CMG2 interact with LDL-receptor related protein (LRP) 6 for endocytosis of EF and LF. On entering the cell, EF acts as a calmodulin-dependent adenylate cyclase that causes a prolonged increase of cytosolic cyclic adenosine monophosphate (cAMP). LF is a metalloprotease that cleaves most isoforms of mitogen-activated protein kinase kinases (MAPKK/MEK) close to their N-terminus. By secreting these two toxins, B.anthracis ascertains death of the host. Once the systemic levels of the toxins rise, antibiotics alone cannot save the host. Therefore, toxin-specific inhibitors have to be developed. In this wake, monoclonal antibodies have been developed for the neutralization of toxic effects of anthrax toxins. We created hybridomas by using spleen of mice that were actively immunized with rLFn (recombinant N-terminal domain of lethal factor of B. anthracis) to obtain anti-toxin antibodies. Later on, separate group of mice were immunized with rLFn to obtain a polyclonal control for passive immunization studies of monoclonal antibodies. This led to the identification of one cohort of rLFn-immunized mice that harboured disease-enhancing polyclonal antibodies. At the same time, the monoclonal antibodies from all the hybridomas were being tested. Two hybridomas secreted monoclonal antibodies (H8 and H10) that were cross-reactive with EF (edema factor) and LF (lethal factor), while the other two hybridomas secreted LF-specific antibodies (H7 and H11). The protective efficacy of H7, H8, H10 and H11 was investigated. H7, H8 and H10 were found to be protective. H11 was found to have disease enhancing characteristics in-vitro and in mouse model of challenge with B. anthracis. In this study the disease enhancing character of H11 monoclonal antibody and anti-rLFn polyclonal sera was investigated. Combination of H11 with protective monoclonal antibodies (H8 and H10) reduced its disease enhancing nature both in-vitro and in-vivo. But combination of H11 with LETscFv (an scFv with VH and VL identical to H10 but lacking Fc region) could not abrogate the disease-enhancing character of H11 mAb. Therefore it was concluded that for suppression of disease enhancement, Fc portion was absolutely essential for interaction of H10 with H11. Our study indicates that the protective potential of an antibody depends equally on its idiotype/ antigen specificity and its isotype. A number of monoclonal and engineered antibodies are being explored as immunotherapeutics but it is absolutely essential to characterize each one for their individual and combined protective potential. Although new in the sphere of toxin-based diseases, it is extremely important to characterize the disease-enhancing nature of polyclonal as well as monoclonal antibodies. This is because several anti-viral therapeutics and vaccines have failed in the face of this phenomenon. The passive –immunotherapy thus needs to be well formulated to avoid any contraindications.

Keywords: immunotherapy, polyclonal, monoclonal, antibody-dependent disease enhancement

Procedia PDF Downloads 386

Authors: Anna Slupik, Anna Mosiolek, Sebastian Wojtowicz, Dariusz Bialoszewski

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Introduction: Whole-body vibration (WBV) uses high frequency mechanical stimuli generated by a vibration plate and transmitted through bone, muscle and connective tissues to the whole body. Research has shown that long-term vibration-plate training improves neuromuscular facilitation, especially in afferent neural pathways, responsible for the conduction of vibration and proprioceptive stimuli, muscle function, balance and proprioception. Some researchers suggest that the vibration stimulus briefly inhibits the conduction of afferent signals from proprioceptors and can interfere with the maintenance of body balance. The aim of this study was to evaluate the influence of a single set of exercises associated with whole-body vibration on the joint position sense and body balance. Material and methods: The study enrolled 55 people aged 19-24 years. These individuals were randomly divided into a test group (30 persons) and a control group (25 persons). Both groups performed the same set of exercises on a vibration plate. The following vibration parameters: frequency of 20Hz and amplitude of 3mm, were used in the test group. The control group performed exercises on the vibration plate while it was off. All participants were instructed to perform six dynamic exercises lasting 30 seconds each with a 60-second period of rest between them. The exercises involved large muscle groups of the trunk, pelvis and lower limbs. Measurements were carried out before and immediately after exercise. Joint position sense (JPS) was measured in the knee joint for the starting position at 45° in an open kinematic chain. JPS error was measured using a digital inclinometer. Balance was assessed in a standing position with both feet on the ground with the eyes open and closed (each test lasting 30 sec). Balance was assessed using Matscan with FootMat 7.0 SAM software. The surface of the ellipse of confidence and front-back as well as right-left swing were measured to assess balance. Statistical analysis was performed using Statistica 10.0 PL software. Results: There were no significant differences between the groups, both before and after the exercise (p> 0.05). JPS did not change in both the test (10.7° vs. 8.4°) and control groups (9.0° vs. 8.4°). No significant differences were shown in any of the test parameters during balance tests with the eyes open or closed in both the test and control groups (p> 0.05). Conclusions. 1. Deterioration in proprioception or balance was not observed immediately after the vibration stimulus. This suggests that vibration-induced blockage of proprioceptive stimuli conduction can have only a short-lasting effect that occurs only as long as a vibration stimulus is present. 2. Short-term use of vibration in treatment does not impair proprioception and seems to be safe for patients with proprioceptive impairment. 3. These results need to be supplemented with an assessment of proprioception during the application of vibration stimuli. Additionally, the impact of vibration parameters used in the exercises should be evaluated.

Keywords: balance, joint position sense, proprioception, whole body vibration

Procedia PDF Downloads 328

Authors: Petr Daniel Kovarik, Matt Kennedy, James Adams, Ajay Wilson, Andy Burns, Charles Kelly, Malcolm Jackson, Rahul Patil, Shahid Iqbal

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Introduction: Osteoradionecrosis (ORN) is a recognised toxicity of radiotherapy (RT) for head and neck cancer (HNC). Existing literature lacks any generally accepted definition and staging system for this toxicity. Objective: The objective is to analyse the outcome of the surgical and nonsurgical treatments of ORN. Material and Method: Data on 2303 patients treated for HNC with radical or adjuvant RT or RT-chemotherapy from January 2010 - December 2021 were retrospectively analysed. Median follow-up to the whole group of patients was 37 months (range 0–148 months). Results: ORN developed in 185 patients (8.1%). The location of ORN was as follows; mandible=170, maxilla=10, and extra oral cavity=5. Multiple ORNs developed in 7 patients. 5 patients with extra oral cavity ORN were excluded from treatment analysis as the management is different. In 180 patients with oral cavity ORN, median follow-up was 59 months (range 5–148 months). ORN healed in 106 patients, treatment failed in 74 patients (improving=10, stable=43, and deteriorating=21). Median healing time was 14 months (range 3-86 months). Notani staging is available in 158 patients with jaw ORN with no previous surgery to the mandible (Notani class I=56, Notani class II=27, and Notani class III=76). 28 ORN (mandible=27, maxilla=1; Notani class I=23, Notani II=3, Notani III=1) healed spontaneously with a median healing time 7 months (range 3–46 months). In 20 patients, ORN developed after dental extraction, in 1 patient in the neomandible after radical surgery as a part of the primary treatment. In 7 patients, ORN developed and spontaneously healed in irradiated bone with no previous surgical/dental intervention. Radical resection of the ORN (segmentectomy, hemi-mandibulectomy with fibula flap) was performed in 43 patients (all mandible; Notani II=1, Notani III=39, Notani class was not established in 3 patients as ORN developed in the neomandible). 27 patients healed (63%); 15 patients failed (improving=2, stable=5, deteriorating=8). The median time from resection to healing was 6 months (range 2–30 months). 109 patients (mandible=100, maxilla=9; Notani I=3, Notani II=23, Notani III=35, Notani class was not established in 9 patients as ORN developed in the maxilla/neomandible) were treated conservatively using a combination of debridement, antibiotics and Pentoclo. 50 patients healed (46%) with a median healing time 14 months (range 3–70 months), 59 patients are recorded with persistent ORN (improving=8, stable=38, deteriorating=13). Out of 109 patients treated conservatively, 13 patients were treated with Pentoclo only (all mandible; Notani I=6, Notani II=3, Notani III=3, 1 patient with neomandible). In total, 8 patients healed (61.5%), treatment failed in 5 patients (stable=4, deteriorating=1). Median healing time was 14 months (range 4–24 months). Extra orally (n=5), 3 cases of ORN were in the auditory canal and 2 in mastoid. ORN healed in one patient (auditory canal after 32 months. Treatment failed in 4 patients (improving=3, stable=1). Conclusion: The outcome of the treatment of ORN remains in general, poor. Every effort should therefore be made to minimise the risk of development of this devastating toxicity.

Keywords: head and neck cancer, radiotherapy, osteoradionecrosis, treatment outcome

Procedia PDF Downloads 92

Authors: Sunitha Sampathi, Pankaj Kumar Tiriya, Sonia Gera, Sravanthi Reddy Pailla, V. Likhitha, A. J. Maruthi

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Surgical site infections (SSIs) are the most common nosocomial infections localized at the incision site. With an estimated 27 million surgical procedures each year in USA, approximately 2-5% rate of SSIs are predicted to occur annually. SSIs are treated with antibiotic medication. Current trend suggest that the direct drug delivery from the suture to the scared tissue can improve patient comfort and wound recovery. For that reason coating the surface of the medical device such as suture and catguts with broad spectrum antibiotics can prevent the formation of bactierial colonies with out comprimising the mechanical properties of the sutures.Hence, the present study was aimed to develop and evaluate a surgical suture coated with an antibiotic Ciprofloxacin hydrochloride loaded on gold nanoparticles. Gold nanoparticles were synthesized by chemical reduction method and conjugated with ciprofloxacin using Polyvinylpyrolidone as stabilizer and gold as carrier. Ciprofloxacin conjugated gold nanoparticles were coated over an absorbable surgical suture made of Polyglactan using sodium alginate as an immobilising agent by slurry dipping technique. The average particle size and Polydispersity Index of drug conjugated gold NPs were found to be 129±2.35 nm and 0.243±0.36 respectively. Gold nanoparticles are characterized by UV-Vis absorption spectroscopy, Fourier Transform Infrared Spectroscopy (FT-IR), Scanning electron microscopy and Transmission electron microscopy. FT-IR revealed that there is no chemical interaction between drug and polymer. Antimicrobial activity for coated sutures was evaluated by disc diffusion method on culture plates of both gram negative (E-coli) and gram positive bacteria (Staphylococcus aureus) and results found to be satisfactory. In vivo studies for coated sutures was performed on Swiss albino mice and histological evaluation of intestinal wound healing parameters such as wound edges in mucosa, muscularis, presence of necrosis, exudates, granulation tissue, granulocytes, macrophages, restoration, and repair of mucosal epithelium and muscularis propria on day 7 after surgery were studied. The control animal group, sutured with plain suture (uncoated suture) showed signs of restoration and repair, but presence of necrosis, heamorraghic infiltration and granulation tissue was still noticed. Whereas the animal group treated with ciprofloxacin and ciprofloxacin gold nanoparticle coated sutures has shown promising decrease in terms of haemorraghic infiltration, granulation tissue, necrosis and better repaired muscularis layers on comparision with plain coated sutures indicating faster rate of repair and less chance of sepsis. Hence coating of sutures with broad spectrum antibiotics can be an alternate technique to reduce SSIs.

Keywords: ciprofloxacin hydrochloride, gold nanoparticles, surgical site infections, sutures

Procedia PDF Downloads 256

Authors: Sofie Verstraete, Stijn Debruyne, Frederik Desplentere

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Considering environmental issues, the interest to apply sustainable materials in industry increases. Specifically for composites, there is an emerging need for suitable materials and bonding techniques. As an alternative to traditional composites, short bio-fiber (cellulose-based flax) reinforced Polylactic Acid (PLA) is gaining popularity. However, these thermoplastic based composites show issues in adhesive bonding. This research focusses on analyzing the effects of the fibers near the bonding interphase. The research applies injection molded plate structures. A first important parameter concerns the fiber volume fraction, which directly affects adhesion characteristics of the surface. This parameter is varied between 0 (pure PLA) and 30%. Next to fiber volume fraction, the orientation of fibers near the bonding surface governs the adhesion characteristics of the injection molded parts. This parameter is not directly controlled in this work, but its effects are analyzed. Surface roughness also greatly determines surface wettability, thus adhesion. Therefore, this research work considers three different roughness conditions. Different mechanical treatments yield values up to 0.5 mm. In this preliminary research, only one adhesive type is considered. This is a two-part epoxy which is cured at 23 °C for 48 hours. In order to assure a dedicated parametric study, simple and reproduceable adhesive bonds are manufactured. Both single lap (substrate width 25 mm, thickness 3 mm, overlap length 10 mm) and double lap tests are considered since these are well documented and quite straightforward to conduct. These tests are conducted for the different substrate and surface conditions. Dog bone tensile testing is applied to retrieve the stiffness and strength characteristics of the substrates (with different fiber volume fractions). Numerical modelling (non-linear FEA) relates the effects of the considered parameters on the stiffness and strength of the different joints, obtained through the abovementioned tests. Ongoing work deals with developing dedicated numerical models, incorporating the different considered adhesion parameters. Although this work is the start of an extensive research project on the bonding characteristics of thermoplastic bio-fiber reinforced composites, some interesting results are already prominent. Firstly, a clear correlation between the surface roughness and the wettability of the substrates is observed. Given the adhesive type (and viscosity), it is noticed that an increase in surface energy is proportional to the surface roughness, to some extent. This becomes more pronounced when fiber volume fraction increases. Secondly, ultimate bond strength (single lap) also increases with increasing fiber volume fraction. On a macroscopic level, this confirms the positive effect of fibers near the adhesive bond line.

Keywords: adhesive bonding, bio-fiber reinforced composite, flax fibers, lap joint

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Authors: Geum Yeon Sim, Jasal Patel, Anand Kumthekar, Stanley Wainapel

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A 65-year-old right-hand dominant African-American man, formerly employed as a security guard, was referred to Rehabilitation Medicine with bilateral hand stiffness and weakness. His past medical history was only significant for myelofibrosis, diagnosed 4 years earlier, for which he was receiving scheduled blood transfusions. Approximately 2 years ago, he began to notice stiffness and swelling in his non-dominant hand that progressed to pain and decreased strength, limiting his hand function. Similar but milder symptoms developed in his right hand several months later. There was no history of prior injury or exposure to cold. Physical examination showed enlargement of metacarpophalangeal (MCP) and proximal interphalangeal (PIP) joints with finger flexion contractures, Swan-neck and Boutonniere deformities, and associated joint tenderness. Changes were more prominent in the left hand. X-rays showed mild osteoarthritis of several bilateral PIP joints. Anti-nuclear antibodies, rheumatoid factor, and cyclic citrullinated peptide antibodies were negative. MRI of the hand showed no erosions or synovitis. A rheumatology consultation was obtained, and the cause of his symptoms was attributed to myelofibrosis-related arthropathy with secondary osteoarthritis. The patient was tried on diclofenac cream and received a few courses of Occupational Therapy with limited functional improvement. Primary myelofibrosis (PMF) is a rare myeloproliferative neoplasm characterized by clonal proliferation of myeloid cells with variable morphologic maturity and hematopoietic efficiency. Rheumatic manifestations of malignancies include direct invasion, paraneoplastic presentations, secondary gout, or hypertrophic osteoarthropathy. PMF causes gradual bone marrow fibrosis with extramedullary metaplastic hematopoiesis in the liver, spleen, or lymph nodes. Musculoskeletal symptoms are not common and are not well described in the literature. The first reported case of myelofibrosis related arthritis was seronegative arthritis due to synovial invasion of myeloproliferative elements. Myelofibrosis has been associated with autoimmune diseases such as systemic lupus erythematosus, progressive systemic sclerosis, and rheumatoid arthritis. Gout has been reported in patients with myelofibrosis, and the underlying mechanism is thought to be related to the high turnover of nucleic acids that is greatly augmented in this disease. X-ray findings in these patients usually include erosive arthritis with synovitis. Treatment of underlying PMF is the treatment of choice, along with anti-inflammatory medications. Physicians should be cognizant of recognizing this rare entity in patients with PMF while maintaining clinical suspicion for more common causes of joint deformities, such as rheumatic diseases.

Keywords: myelofibrosis, arthritis, arthralgia, malignancy

Procedia PDF Downloads 99

Authors: Scott M. Black, Craig Maclean, Pauline Hall Barrientos, Konstantinos Ritos, Asimina Kazakidi

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Aortic aneurysms and dissections are leading causes of death in cardiovascular disease. Both inevitably lead to hemodynamic instability without surgical intervention in the form of vascular stent-graft deployment. An accurate description of the aortic geometry and blood flow in patient-specific cases is vital for treatment planning and long-term success of such grafts, as they must generate physiological branch perfusion and in-stent hemodynamics. The aim of this study was to create patient-specific computational fluid dynamics (CFD) models through a multi-modality, multi-dimensional approach with boundary condition optimization to predict branch flow rates and in-stent hemodynamics in custom stent-graft configurations. Three-dimensional (3D) thoracoabdominal aortae were reconstructed from four-dimensional flow-magnetic resonance imaging (4D Flow-MRI) and computed tomography (CT) medical images. The former employed a novel approach to generate and enhance vessel lumen contrast via through-plane velocity at discrete, user defined cardiac time steps post-hoc. To produce patient-specific boundary conditions (BCs), the aortic geometry was reduced to a one-dimensional (1D) model. Thereafter, a zero-dimensional (0D) 3-Element Windkessel model (3EWM) was coupled to each terminal branch to represent the distal vasculature. In this coupled 0D-1D model, the 3EWM parameters were optimized to yield branch flow waveforms which are representative of the 4D Flow-MRI-derived in-vivo data. Thereafter, a 0D-3D CFD model was created, utilizing the optimized 3EWM BCs and a 4D Flow-MRI-obtained inlet velocity profile. A sensitivity analysis on the effects of stent-graft configuration and BC parameters was then undertaken using multiple stent-graft configurations and a range of distal vasculature conditions. 4D Flow-MRI granted unparalleled visualization of blood flow throughout the cardiac cycle in both the pre- and postsurgical states. Segmentation and reconstruction of healthy and stented regions from retrospective 4D Flow-MRI images also generated 3D models with geometries which were successfully validated against their CT-derived counterparts. 0D-1D coupling efficiently captured branch flow and pressure waveforms, while 0D-3D models also enabled 3D flow visualization and quantification of clinically relevant hemodynamic parameters for in-stent thrombosis and graft limb occlusion. It was apparent that changes in 3EWM BC parameters had a pronounced effect on perfusion distribution and near-wall hemodynamics. Results show that the 3EWM parameters could be iteratively changed to simulate a range of graft limb diameters and distal vasculature conditions for a given stent-graft to determine the optimal configuration prior to surgery. To conclude, this study outlined a methodology to aid in the prediction post-surgical branch perfusion and in-stent hemodynamics in patient specific cases for the implementation of custom stent-grafts.

Keywords: 4D flow-MRI, computational fluid dynamics, vascular stent-grafts, windkessel

Procedia PDF Downloads 181

Authors: Shrestha Sharma, Jasjeet K. Sahni, Javed Ali, Sanjula Baboota

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Introduction: Rutin is a naturally occurring strong antioxidant molecule belonging to bioflavonoid category. Due to its free radical scavenging properties, it has been found to be beneficial in the treatment of various diseases including inflammation, cancer, diabetes, allergy, cardiovascular disorders and various types of microbial infections. Despite its beneficial effects, it suffers from the problem of low aqueous solubility which is responsible for low oral bioavailability. The aim of our study was to optimize and characterize self-nanoemulsifying drug delivery system (SNEDDS) of rutin using Box-Behnken design (BBD) combined with a desirability function. Further various antioxidant, pharmacokinetic and pharmacodynamic studies were performed for the optimized rutin SNEDDS formulation. Methodologies: Selection of oil, surfactant and co-surfactant was done on the basis of solubility/miscibility studies. Sefsol+ Vitamin E, Solutol HS 15 and Transcutol P were selected as oil phase, surfactant and co-surfactant respectively. Optimization of SNEDDS formulations was done by a three-factor, three-level (33)BBD. The independent factors were Sefsol+ Vitamin E, Solutol HS15, and Transcutol P. The dependent variables were globule size, self emulsification time (SEF), % transmittance and cumulative percentage drug released. Various response surface graphs and contour plots were constructed to understand the effect of different factor, their levels and combinations on the responses. The optimized Rutin SNEDDS formulation was characterized for various parameters such as globule size, zeta potential, viscosity, refractive index , % Transmittance and in vitro drug release. Ex vivo permeation studies and pharmacokinetic studies were performed for optimized formulation. Antioxidant activity was determined by DPPH and reducing power assays. Anti-inflammatory activity was determined by using carrageenan induced rat paw oedema method. Permeation of rutin across small intestine was assessed using confocal laser scanning microscopy (CLSM). Major findings:The optimized SNEDDS formulation consisting of Sefsol+ Vitamin E - Solutol HS15 -Transcutol HP at proportions of 25:35:17.5 (w/w) was prepared and a comparison of the predicted values and experimental values were found to be in close agreement. The globule size and PDI of optimized SNEDDS formulation was found to be 16.08 ± 0.02 nm and 0.124±0.01 respectively. Significant (p˂0.05) increase in percentage drug release was achieved in the case of optimized SNEDDS formulation (98.8 %) as compared to rutin suspension. Furthermore, pharmacokinetic study showed a 2.3-fold increase in relative oral bioavailability compared with that of the suspension. Antioxidant assay results indicated better efficacy of the developed formulation than the pure drug and it was found to be comparable with ascorbic acid. The results of anti-inflammatory studies showed 72.93 % inhibition for the SNEDDS formulation which was significantly higher than the drug suspension 46.56%. The results of CLSM indicated that the absorption of SNEDDS formulation was considerably higher than that from rutin suspension. Conclusion: Rutin SNEDDS have been successfully prepared and they can serve as an effective tool in enhancing oral bioavailability and efficacy of Rutin.

Keywords: rutin, oral bioavilability, pharamacokinetics, pharmacodynamics

Procedia PDF Downloads 500

Authors: T. Vijay Bhasker, N. K. S Gowda, P. Krishnamoorthy, D. T. Pal, A. K. Pattanaik, A. K. Verma

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Boron (B) is a newer trace element and its biological importance and dietary essentiality is unclear in animals. The available literature suggests its putative role in bone mineralization, antioxidant status and steroid hormone synthesis. A feeding trial was conducted in Wister strain (Rattus norvegicus) albino rats for duration of 90 days. A total of 84 healthy weaned (3-4 weeks) experimental rats were randomly divided into 7 dietary groups (4 replicates of three each) viz., A (Basal diet/ Control), B (Basal diet + 5 ppm B), C (Basal diet + 10 ppm B), D (Basal diet + 20 ppm B), E (Basal diet + 40 ppm B), F (Basal diet-Ca 50%), G (Basal diet-Ca 50% + 40 ppm B). Dietary level of calcium (Ca) was maintained at two levels, 100% and 50% of requirement. Sodium borate was used as source of boron along with other ingredients of basal diet while preparing the pelletized diets. All the rats were kept in proper ventilated laboratory animal house maintained at temperature (23±2º C) and humidity (50 to 70%). At the end of experiment digestibility trial was conducted for 5 days to estimate nutrient digestibility and gut absorption of minerals. Eight rats from each group were sacrificed to collect the vital organs (liver, kidney and spleen) to study histopathology. Blood sample was drawn by heart puncture to determine biochemical profile. The average daily feed intake (g/rat/day), water intake (ml/rat/day) and body weight gain (g/rat/day) were similar among the dietary groups. The digestibility (%) of organic matter and crude fat were significantly improved (P < 0.05) was by B supplementation. The gut absorption (%) Ca was significantly increased (P < 0.01) in B supplemented groups compared to control. However, digestibility of dry matter and crude protein, gut absorption of magnesium and phosphorus showed a non-significant increasing trend with B supplementation. The gut absorption (%) of B (P < 0.01) was significantly lowered (P<0.05) in supplemented groups compared to un-supplemented ones. The serum level of triglycerides (mg/dL), HDL-cholesterol (mg/dL) and alanine transaminase (IU/L) were significantly lowered (P < 0.05) in B supplemented groups. While serum level of glucose (mg/dL) and alkaline phosphatase (KA units) showed a non-significant decreasing trend with B supplementation. However the serum levels of total cholesterol (mg/dL) and aspartate transaminase (IU/L) were similar among dietary groups. The histology sections of kidney and spleen revealed no significant changes among the dietary groups and were observed to be normal in anatomical architecture. However, the liver histology revealed cell degenerative changes with vacuolar degeneration and nuclear condensation in Ca deficient groups. But the comparative degenerative changes were mild in 40 ppm B supplemented Ca deficient group. In conclusion, dietary supplementation of graded levels of boron in rats had a positive effect on metabolism and health by improving nutrient digestibility and gut absorption of Ca. This indicates the beneficial role of dietary boron supplementation.

Keywords: boron, calcium, nutrient utilization, histopathology

Procedia PDF Downloads 318

Authors: Ernest Moles, Patricia Urbán, María Belén Jiménez-Díaz, Sara Viera-Morilla, Iñigo Angulo-Barturen, Maria Antònia Busquets, Xavier Fernàndez-Busquets

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Bearing in mind the absence of an effective vaccine against malaria and its severe clinical manifestations causing nearly half a million deaths every year, this disease represents nowadays a major threat to life. Besides, the basic rationale followed by currently marketed antimalarial approaches is based on the administration of drugs on their own, promoting the emergence of drug-resistant parasites owing to the limitation in delivering drug payloads into the parasitized erythrocyte high enough to kill the intracellular pathogen while minimizing the risk of causing toxic side effects to the patient. Such dichotomy has been successfully addressed through the specific delivery of immunoliposome (iLP)-encapsulated antimalarials to Plasmodium falciparum-infected red blood cells (pRBCs). Unfortunately, this strategy has not progressed towards clinical applications, whereas in vitro assays rarely reach drug efficacy improvements above 10-fold. Here, we show that encapsulation efficiencies reaching >96% can be achieved for the weakly basic drugs chloroquine (CQ) and primaquine using the pH gradient active loading method in liposomes composed of neutrally charged, saturated phospholipids. Targeting antibodies are best conjugated through their primary amino groups, adjusting chemical crosslinker concentration to retain significant antigen recognition. Antigens from non-parasitized RBCs have also been considered as targets for the intracellular delivery of drugs not affecting the erythrocytic metabolism. Using this strategy, we have obtained unprecedented nanocarrier targeting to early intraerythrocytic stages of the malaria parasite for which there is a lack of specific extracellular molecular tags. Polyethylene glycol-coated liposomes conjugated with monoclonal antibodies specific for the erythrocyte surface protein glycophorin A (anti-GPA iLP) were capable of targeting 100% RBCs and pRBCs at the low concentration of 0.5 μM total lipid in the culture, with >95% of added iLPs retained into the cells. When exposed for only 15 min to P. falciparum in vitro cultures synchronized at early stages, free CQ had no significant effect over parasite viability up to 200 nM drug, whereas iLP-encapsulated 50 nM CQ completely arrested its growth. Furthermore, when assayed in vivo in P. falciparum-infected humanized mice, anti-GPA iLPs cleared the pathogen below detectable levels at a CQ dose of 0.5 mg/kg. In comparison, free CQ administered at 1.75 mg/kg was, at most, 40-fold less efficient. Our data suggest that this significant improvement in drug antimalarial efficacy is in part due to a prophylactic effect of CQ found by the pathogen in its host cell right at the very moment of invasion.

Keywords: immunoliposomal nanoparticles, malaria, prophylactic-therapeutic polyvalent activity, targeted drug delivery

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Authors: Iman Farasat, Howard M. Salis

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Engineered genetic circuits reprogram cellular behavior to act as living computers with applications in detecting cancer, creating self-controlling artificial tissues, and dynamically regulating metabolic pathways. Phenemenological models are often used to simulate and design genetic circuit behavior towards a desired behavior. While such models assume that each circuit component’s function is modular and independent, even small changes in a circuit (e.g. a new promoter, a change in transcription factor expression level, or even a new media) can have significant effects on the circuit’s function. Here, we use statistical thermodynamics to account for the several factors that control transcriptional regulation in bacteria, and experimentally demonstrate the model’s accuracy across 825 measurements in several genetic contexts and hosts. We then employ our first principles model to design, experimentally construct, and characterize a family of signal amplifying genetic circuits (genetic OpAmps) that expand the dynamic range of cell sensors. To develop these models, we needed a new approach to measuring the in vivo binding free energies of transcription factors (TFs), a key ingredient of statistical thermodynamic models of gene regulation. We developed a new high-throughput assay to measure RNA polymerase and TF binding free energies, requiring the construction and characterization of only a few constructs and data analysis (Figure 1A). We experimentally verified the assay on 6 TetR-homolog repressors and a CRISPR/dCas9 guide RNA. We found that our binding free energy measurements quantitatively explains why changing TF expression levels alters circuit function. Altogether, by combining these measurements with our biophysical model of translation (the RBS Calculator) as well as other measurements (Figure 1B), our model can account for changes in TF binding sites, TF expression levels, circuit copy number, host genome size, and host growth rate (Figure 1C). Model predictions correctly accounted for how these 8 factors control a promoter’s transcription rate (Figure 1D). Using the model, we developed a design framework for engineering multi-promoter genetic circuits that greatly reduces the number of degrees of freedom (8 factors per promoter) to a single dimensionless unit. We propose the Ptashne (Pt) number to encapsulate the 8 co-dependent factors that control transcriptional regulation into a single number. Therefore, a single number controls a promoter’s output rather than these 8 co-dependent factors, and designing a genetic circuit with N promoters requires specification of only N Pt numbers. We demonstrate how to design genetic circuits in Pt number space by constructing and characterizing 15 2-repressor OpAmp circuits that act as signal amplifiers when within an optimal Pt region. We experimentally show that OpAmp circuits using different TFs and TF expression levels will only amplify the dynamic range of input signals when their corresponding Pt numbers are within the optimal region. Thus, the use of the Pt number greatly simplifies the genetic circuit design, particularly important as circuits employ more TFs to perform increasingly complex functions.

Keywords: transcription factor, synthetic biology, genetic circuit, biophysical model, binding energy measurement

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Authors: Vrushali Guhe, Shailza Singh

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Cutaneous leishmaniasis is neglected tropical disease present worldwide caused by the protozoan parasite Leishmania major, the therapeutic armamentarium for leishmaniasis are showing several limitations as drugs are showing toxic effects with increasing resistance by a parasite. Thus identification of novel therapeutic targets is of paramount importance. Previous studies have shown that autophagy, a cellular process, can either facilitate infection or aid in the elimination of the parasite, depending on the specific parasite species and host background in leishmaniasis. In the present study, our objective was to target the essential autophagy protein ATG8, which plays a crucial role in the survival, infection dynamics, and differentiation of the Leishmania parasite. ATG8 in Leishmania major and its homologue, LC3, in Homo sapiens, act as autophagic markers. Present study manifested the crucial role of ATG8 protein as a potential target for combating Leishmania major infection. Through bioinformatics analysis, we identified non-conserved motifs within the ATG8 protein of Leishmania major, which are not present in LC3 of Homo sapiens. Against these two non-conserved motifs, we generated a peptide library of 60 peptides on the basis of physicochemical properties. These peptides underwent a filtering process based on various parameters, including feasibility of synthesis and purification, compatibility with Selective Reaction Monitoring (SRM)/Multiple reaction monitoring (MRM), hydrophobicity, hydropathy index, average molecular weight (Mw average), monoisotopic molecular weight (Mw monoisotopic), theoretical isoelectric point (pI), and half-life. Further filtering criterion shortlisted three peptides by using molecular docking and molecular dynamics simulations. The direct interaction between ATG8 and the shortlisted peptides was confirmed through Surface Plasmon Resonance (SPR) experiments. Notably, these peptides exhibited the remarkable ability to penetrate the parasite membrane and exert profound effects on Leishmania major. The treatment with these peptides significantly impacted parasite survival, leading to alterations in the cell cycle and morphology. Furthermore, the peptides were found to modulate autophagosome formation, particularly under starved conditions, suggesting their involvement in disrupting the regulation of autophagy within Leishmania major. In vitro, studies demonstrated that the selected peptides effectively reduced the parasite load within infected host cells. Encouragingly, these findings were corroborated by in vivo experiments, which showed a reduction in parasite burden upon peptide administration. Additionally, the peptides were observed to affect the levels of LC3II within host cells. In conclusion, our findings highlight the efficacy of these novel peptides in targeting Leishmania major’s ATG8 and disrupting parasite survival. These results provide valuable insights into the development of innovative therapeutic strategies against leishmaniasis via targeting autophagy protein ATG8 of Leishmania major.

Keywords: ATG8, leishmaniasis, surface plasmon resonance, MD simulation, molecular docking, peptide designing, therapeutics

Procedia PDF Downloads 82

Authors: Aurica P. Chiriac, Vera Balan, Mihai Asandulesa, Elena Butnaru, Nita Tudorachi, Elena Stoleru, Loredana E. Nita, Iordana Neamtu, Alina Diaconu, Liliana Mititelu-Tartau

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The interest for studying ‘smart’ materials is entirely justified and in this context were realized investigations on poly(2-hydroxyethylmethacrylate-co-3, 9-divinyl-2, 4, 8, 10-tetraoxaspiro (5.5) undecane), which is a macromolecular compound with sensibility at pH and temperature, gel formation capacity, binding properties, amphilicity, good oxidative and thermal stability. Physico-chemical characteristics in terms of the molecular weight, temperature-sensitive abilities and thermal stability, as well rheological, dielectric and spectroscopic properties were evaluated in correlation with further coupling capabilities. Differential scanning calorimetry investigation indicated Tg at 36.6 °C and a melting point at Tm=72.8°C, for the studied copolymer, and up to 200oC two exothermic processes (at 99.7°C and 148.8°C) were registered with losing weight of about 4 %, respective 19.27%, which indicate just processes of thermal decomposition (and not phenomena of thermal transition) owing to scission of the functional groups and breakage of the macromolecular chains. At the same time, the rheological studies (rotational tests) confirmed the non-Newtonian shear-thinning fluid behavior of the copolymer solution. The dielectric properties of the copolymer have been evaluated in order to investigate the relaxation processes and two relaxation processes under Tg value were registered and attributed to localized motions of polar groups from side chain macromolecules, or parts of them, without disturbing the main chains. According to literature and confirmed as well by our investigations, β-relaxation is assigned with the rotation of the ester side group and the γ-relaxation corresponds to the rotation of hydroxy- methyl side groups. The fluorescence spectroscopy confirmed the copolymer structure, the spiroacetal moiety getting an axial conformation, more stable, with lower energy, able for specific interactions with molecules from environment, phenomena underlined by different shapes of the emission spectra of the copolymer. Also, the copolymer was used as template for indomethacin incorporation as model drug, and the biocompatible character of the complex was confirmed. The release behavior of the bioactive compound was dependent by the copolymer matrix composition, the increasing of 3, 9-divinyl-2, 4, 8, 10-tetraoxaspiro (5.5) undecane comonomer amount attenuating the drug release. At the same time, the in vivo studies did not show significant differences of leucocyte formula elements, GOT, GPT and LDH levels, nor immune parameters (OC, PC, and BC) between control mice group and groups treated just with copolymer samples, with or without drug, data attesting the biocompatibility of the polymer samples. The investigation of the physico-chemical characteristics of poly(2-hydrxyethyl methacrylate-co-3, 9-divinyl-2, 4, 8, 10-tetraoxaspiro (5.5) undecane) in terms of temperature-sensitive abilities, rheological and dielectrical properties, are bringing useful information for further specific use of this polymeric compound.

Keywords: bioapplications, dielectric and spectroscopic properties, dual sensitivity at pH and temperature, smart materials

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Authors: Nabil Omar, Farah Jibril, Oraib Amjad

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Purpose: Trabecatine is a is a potent marine-derived antineoplastic drug which binds to the minor groove of the DNA, bending DNA towards the major groove resulting in a changed conformation that interferes with several DNA transcription factors, repair pathways and cell proliferation. Trabectedin was approved by the European Medicines Agency (EMA; London, UK) for the treatment of adult patients with advanced stage soft tissue sarcomas in whom treatment with anthracyclines and ifosfamide has failed, or for those who are not candidates for these therapies. The recommended dosing regimen is 1.5 mg/m2 IV over 24 hours every 3 weeks. The purpose of this study was to comprehensively review available data on the safety and efficacy of trabectedin used as indicated for patients at a Tertiary Cancer Center at Qatar. Methods: A medication administration report generated in the electronic health record identified all patients who received trabectedin between November 1, 2015 and November 1, 2017. This retrospective chart review evaluated the indication of trabectedin use, compliance to administration protocol and the recommended monitoring parameters, number of patients improved on the drug and continued treatment, number of patients discontinued treatment due to side-effects and the reported side effects. Progress and discharged notes were utilized to report experienced side effects during trabectedin therapy. A total of 3 patients were reviewed. Results: Total of 2 out of 3 patients who received trabectedin were receiving it for non-FDA and non-EMA, approved indications; metastatic rhabdomyosarcoma and ovarian cancer stage IV with poor prognosis. And only one patient received it as indicated for leiomyosarcoma of left ureter with metastases to liver, lungs and bone. None of the patients has continued the therapy due to development of serious side effects. One patient had stopped the medication after one cycle due to disease progression and transient hepatic toxicity, the other one had disease progression and developed 12 % reduction in LVEF after 12 cycles of trabectedin, and the third patient deceased, had disease progression on trabectedin after the 10th cycle that was received through peripheral line which resulted in developing extravasation and left arm cellulitis requiring debridement. Regarding monitoring parameters, at baseline the three patients had ECHO, and Creatine Phosphokinase (CPK) but it was not monitored during treatment as recommended. Conclusion: Utilizing this medication as indicated with performing the appropriate monitoring parameters as recommended can benefit patients who are receiving it. It is important to reinforce the intravenous administration via central intravenous line, the re-assessment of left ventricular ejection fraction (LVEF) by echocardiogram or multigated acquisition (MUGA) scan at 2- to 3-month intervals thereafter until therapy is discontinued, and CPK and LFTs levels prior to each administration of trabectedin.

Keywords: trabectedin, drug-use evaluation, safety, effectiveness, adverse drug reaction, monitoring

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Authors: N. Slepickova Kasalkova, P. Slepicka, L. Bacakova, V. Svorcik

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Tissue engineering includes combination of materials and techniques used for the improvement, repair or replacement of the tissue. Scaffolds, permanent or temporally material, are used as support for the creation of the "new cell structures". For this important component (scaffold), a variety of materials can be used. The advantage of some polymeric materials is their cytocompatibility and possibility of biodegradation. Poly(L-lactic acid) (PLLA) is a biodegradable,  semi-crystalline thermoplastic polymer. PLLA can be fully degraded into H₂O and CO₂. In this experiment, the effect of the surface modification of biodegradable polymer (performed by plasma treatment) on the various cell types was studied. The surface parameters and changes of the physicochemical properties of modified PLLA substrates were studied by different methods. Surface wettability was determined by goniometry, surface morphology and roughness study were performed with atomic force microscopy and chemical composition was determined using photoelectron spectroscopy. The physicochemical properties were studied in relation to cytocompatibility of human osteoblast (MG 63 cells), rat vascular smooth muscle cells (VSMC), and human stem cells (ASC) of the adipose tissue in vitro. A fluorescence microscopy was chosen to study and compare cell-material interaction. Important parameters of the cytocompatibility like adhesion, proliferation, viability, shape, spreading of the cells were evaluated. It was found that the modification leads to the change of the surface wettability depending on the time of modification. Short time of exposition (10-120 s) can reduce the wettability of the aged samples, exposition longer than 150 s causes to increase of contact angle of the aged PLLA. The surface morphology is significantly influenced by duration of modification, too. The plasma treatment involves the formation of the crystallites, whose number increases with increasing time of modification. On the basis of physicochemical properties evaluation, the cells were cultivated on the selected samples. Cell-material interactions are strongly affected by material chemical structure and surface morphology. It was proved that the plasma treatment of PLLA has a positive effect on the adhesion, spreading, homogeneity of distribution and viability of all cultivated cells. This effect was even more apparent for the VSMCs and ASCs which homogeneously covered almost the whole surface of the substrate after 7 days of cultivation. The viability of these cells was high (more than 98% for VSMCs, 89-96% for ASCs). This experiment is one part of the basic research, which aims to easily create scaffolds for tissue engineering with subsequent use of stem cells and their subsequent "reorientation" towards the bone cells or smooth muscle cells.

Keywords: poly(L-lactic acid), plasma treatment, surface characterization, cytocompatibility, human osteoblast, rat vascular smooth muscle cells, human stem cells

Procedia PDF Downloads 228

Authors: Tomohiko Utsuki, Kyoka Sato

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In emergency medicine, it is recognized that brain resuscitation is very important for the reduction of mortality rate and neurological sequelae. Especially, the control of brain temperature (BT), intracranial pressure (ICP), and cerebral blood flow (CBF) are most required for stabilizing brain’s physiological state in the treatment for such as brain injury, stroke, and encephalopathy. However, the manual control of BT, ICP, and CBF frequently requires the decision and operation of medical staff, relevant to medication and the setting of therapeutic apparatus. Thus, the integration and the automation of the control of those is very effective for not only improving therapeutic effect but also reducing staff burden and medical cost. For realizing such integration and automation, a mathematical model of brain physiological state is necessary as the controlled object in simulations, because the performance test of a prototype of the control system using patients is not ethically allowed. A model of cerebral blood circulation has already been constructed, which is the most basic part of brain physiological state. Also, a migration model of extracellular fluid in brain has been constructed, however the condition that the total volume of intracranial cavity is almost changeless due to the hardness of cranial bone has not been considered in that model. Therefore, in this research, the dynamic migration model of extracellular fluid in brain was constructed on the consideration of the changelessness of intracranial cavity’s total volume. This model is connectable to the cerebral blood circulation model. The constructed model consists of fourteen compartments, twelve of which corresponds to perfused area of bilateral anterior, middle and posterior cerebral arteries, the others corresponds to cerebral ventricles and subarachnoid space. This model enable to calculate the migration of tissue fluid from capillaries to gray matter and white matter, the flow of tissue fluid between compartments, the production and absorption of cerebrospinal fluid at choroid plexus and arachnoid granulation, and the production of metabolic water. Further, the volume, the colloid concentration, and the tissue pressure of/in each compartment are also calculable by solving 40-dimensional non-linear simultaneous differential equations. In this research, the obtained model was analyzed for its validation under the four condition of a normal adult, an adult with higher cerebral capillary pressure, an adult with lower cerebral capillary pressure, and an adult with lower colloid concentration in cerebral capillary. In the result, calculated fluid flow, tissue volume, colloid concentration, and tissue pressure were all converged to suitable value for the set condition within 60 minutes at a maximum. Also, because these results were not conflict with prior knowledge, it is certain that the model can enough represent physiological state of brain under such limited conditions at least. One of next challenges is to integrate this model and the already constructed cerebral blood circulation model. This modification enable to simulate CBF and ICP more precisely due to calculating the effect of blood pressure change to extracellular fluid migration and that of ICP change to CBF.

Keywords: dynamic model, cerebral extracellular migration, brain resuscitation, automatic control

Procedia PDF Downloads 156

Authors: Farrah Putri Salmanida, Mei-Li Wu, Rika Wahyuningtyas, Wen-Bin Chung, Hso-Chi Chaung, Ko-Tung Chang

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Within the field of immunotherapy, oncolytic virotherapy (OVT) employs dual approaches that directly eliminate tumor cells while preserving healthy ones and indirectly reprogram the tumor microenvironment (TME) to elicit antitumor responses. Within the TME, tumor associated macrophages (TAMs) manifest characteristics akin to those of anti-inflammatory M2 macrophages, thus earning the designation of M2-like TAMs. In prior research, two antigens denoted as A1 (g6Ld10T) and A3 (ORF6L5), derived from a complete sequence of ORF5 with partial sequence of ORF6 in Porcine Reproductive and Respiratory Syndrome Virus Genotype 2 (PRRSV-2), demonstrated the capacity to repolarize M2-type porcine alveolar macrophages (PAMs) into M1 phenotypes. In this study, we sought for utilizing OVT strategies by introducing A1 or A3 on TAMs to endow them with the anti-tumor traits of M1 macrophages while retaining their capacity to target cancer cells. Upon exposing human THP-1-derived M2 macrophages to a cross-species test with 2 µg/ml of either A1 or A3 for 24 hours, real time PCR revealed that A3, but not A1, treated cells exhibited upregulated gene expressions of M1 markers (CCR7, IL-1ß, CCL2, Cox2, CD80). These cells reacted to virus-derived antigen, as evidenced by increased expression of pattern-recognition receptors TLR3, TLR7, and TLR9, subsequently providing feedback in the form of type I interferon responses like IFNAR1, IFN-ß, IRF3, IRF7, OAS1, Mx1, and ISG15. Through an MTT assay, only after 15 µg/ml of A3 treatment could the cell viability decrease, with a predicted IC50 of 16.96 µg/ml. Interestingly, A3 caused dose-dependent toxicity to a rat C6 glial cancer cell line even at doses as low as 2.5 µg/ml and reached its IC50 at 9.419 µg/ml. Using Annexin V/7AAD staining and PCR test, we deduced that a significant proportion of C6 cells were undergoing the early apoptosis phase predominantly through the intrinsic apoptosis cascade involving Bcl-2 family proteins. Following this stage, we conducted a test on A3’s repolarization ability, which revealed a significant rise in M1 gene expression markers, such as TNF, CD80, and IL-1ß, in M2-like TAMs generated in vitro from murine RAW264.7 macrophages grown with conditioned medium of 4T1 breast cancer cells. This was corroborated by the results of transcriptome analysis, which revealed that the primary subset among the top 10 to top 30 significantly upregulated differentially expressed genes (DEGs) dominantly consisted of M1 macrophages profiles, including Ccl3, Ccl4, Csf3, TNF, Bcl6b, Stc1, and Dusp2. Our findings unveiled the remarkable potential of the PRRSV-derived antigen A3 to repolarize macrophages while also being capable of selectively inducing apoptosis in cancerous cells. While further in vivo study is needed for A3, it holds promise as an adjuvant by its dual effects in cancer therapy modalities.

Keywords: cancer cell apoptosis, interferon responses, macrophage repolarization, recombinant protein

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Authors: Dalila Chaid, Bennameur Fedilli, Mohammed Amine Bellelou

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The study focuses on the experience of lower-limb amputees, who face both physical and psychological challenges due to their disability. Chronic neuropathic pain and various types of limb pain are common in these patients. They often require orthopaedic interventions for issues such as dressings, infection, ulceration, and bone-related problems. Research Aim: The aim of this study is to determine the most suitable anaesthetic technique for lower-limb amputees, which can provide them with the greatest comfort and prolonged analgesia. The study also aims to demonstrate the effectiveness and cost-effectiveness of ultrasound-guided local regional anaesthesia (LRA) in this patient population. Methodology: The study is an observational analytical study conducted over a period of eight years, from 2010 to 2018. It includes a total of 955 cases of revisions performed on lower limb stumps. The parameters analyzed in this study include the effectiveness of the block and the use of sedation, the duration of the block, the post-operative visual analog scale (VAS) scores, and patient comfort. Findings: The study findings highlight the benefits of ultrasound-guided LRA in providing comfort by optimizing post-operative analgesia, which can contribute to psychological and bodily repair in lower-limb amputees. Additionally, the study emphasizes the use of alpha2 agonist adjuvants with sedative and analgesic properties, long-acting local anaesthetics, and larger volumes for better outcomes. Theoretical Importance: This study contributes to the existing knowledge by emphasizing the importance of choosing an appropriate anaesthetic technique for lower-limb amputees. It highlights the potential of ultrasound-guided LRA and the use of specific adjuvants and local anaesthetics in improving post-operative analgesia and overall patient outcomes. Data Collection and Analysis Procedures: Data for this study were collected through the analysis of medical records and relevant documentation related to the 955 cases included in the study. The effectiveness of the anaesthetic technique, duration of the block, post-operative pain scores, and patient comfort were analyzed using statistical methods. Question Addressed: The study addresses the question of which anaesthetic technique would be most suitable for lower-limb amputees to provide them with optimal comfort and prolonged analgesia. Conclusion: The study concludes that ultrasound-guided LRA, along with the use of alpha2 agonist adjuvants, long-acting local anaesthetics, and larger volumes, can be an effective approach in providing comfort and improving post-operative analgesia for lower-limb amputees. This technique can potentially contribute to the psychological and bodily repair of these patients. The findings of this study have implications for clinical practice in the management of lower-limb amputees, highlighting the importance of personalized anaesthetic approaches for better outcomes.

Keywords: neuropathic pain, ultrasound-guided peripheral nerve block, DN4 quiz, EMG

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Authors: Diego F. Carrillo-González, Milena Osorio, Natalia M. Cerro, Yasser Y. Lenis

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Introduction: mortality during the implantation and early embryonic development periods reach around 30% in different mammalian species. It has been described that progesterone (P4) and Arginine (Arg) play a beneficial role in establishing and maintaining early pregnancy in mammals. The combined effect between Arg and P4 on reproductive parameters in the rabbit species is not yet elucidated, to our best knowledge. Objective: to assess the effect of L-arginine and progesterone during the post-breeding period in female rabbits on the composition of the amniotic fluid, the placental structure, and the bone growth in their fetuses. Methods: crossbred female rabbits (n=16) were randomly distributed into four experimental groups (Ctrl, Arg, P4, and Arg+P4). In the control group, 0.9% saline solution was administered as a placebo, the Arg group was administered arginine (50 mg/kg BW) from day 4.5 to day 19 post-breeding, the P4 group was administered progesterone (Gestavec®, 1.5 mg/kg BW) from 24 hours to day 4 post-breeding and for the Arg+P4 group, an administration was performed under the same time and dose guidelines as the Arg and P4 treatments. Four females were sacrificed, and the amniotic fluid was collected and analyzed with rapid urine test strips, while the placenta and fetuses were processed in the laboratory to obtain histological plates. The percentage of deciduous, labyrinthine, and junctional zones was determined, and the length of the femur for each fetus was measured as an indicator of growth. Descriptive statistics were applied to identify the success rates for each of the tests. Afterwards, A one-way analysis of variance (ANOVA) was performed, and a comparison of means was conducted by Tukey's test. Results: a higher density (p<0.05) was observed in the amniotic fluid for fetuses in the control group (1022±2.5g/mL) compared to the P4 (1015±5.3g/mL) and Arg+P4 (1016±4,9g/mL) groups. Additionally, the density of amniotic fluid in the Arg group (1021±2.5g/mL) was higher (p<0.05) than in the P4 group. The concentration of protein, glucose, and ascorbic acid had no statistical difference between treatments (p>0.05). The histological analysis of the uteroplacental regions, a statistical difference (p<0,05) in the proportion of deciduous zone was found between the P4 group (9.6±2.6%) when compared with the Ctrl (28.15±12.3%), and Arg+P4 (26.3±4.9) groups. In the analysis of the fetuses, the weight was higher for the Arg group (2.69±0.18), compared to the other groups (p<0.05), while a shorter length was observed (p<0.05) in the fetuses for the Arg+P4 group (25.97±1.17). However, no difference (p>0.05) was found when comparing the length of the developing femurs between the experimental groups. Conclusion: the combination of L-arginine and progesterone allows a reduction in the density of amniotic fluid, without affecting the protein, energy, and antioxidant components. However, the use of L-arginine stimulates weight gain in fetuses, without affecting size, which could be used to improve production parameters in rabbit production systems. In addition, the modification in the deciduous zone could show a placental adaptation based on the fetal growth process, however more specific studies on the placentation process are required.

Keywords: arginine, progesterone, rabbits, reproduction

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Authors: Angelis P. Barlampas

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Learning Objectives: The purpose of this study is to inform briefly the reader about the applications of AI in chest radiology. Background: Currently, there are 190 FDA-approved radiology AI applications, with 42 (22%) pertaining specifically to thoracic radiology. Imaging findings OR Procedure details Aids of AI in chest radiology1: Detects and segments pulmonary nodules. Subtracts bone to provide an unobstructed view of the underlying lung parenchyma and provides further information on nodule characteristics, such as nodule location, nodule two-dimensional size or three dimensional (3D) volume, change in nodule size over time, attenuation data (i.e., mean, minimum, and/or maximum Hounsfield units [HU]), morphological assessments, or combinations of the above. Reclassifies indeterminate pulmonary nodules into low or high risk with higher accuracy than conventional risk models. Detects pleural effusion . Differentiates tension pneumothorax from nontension pneumothorax. Detects cardiomegaly, calcification, consolidation, mediastinal widening, atelectasis, fibrosis and pneumoperitoneum. Localises automatically vertebrae segments, labels ribs and detects rib fractures. Measures the distance from the tube tip to the carina and localizes both endotracheal tubes and central vascular lines. Detects consolidation and progression of parenchymal diseases such as pulmonary fibrosis or chronic obstructive pulmonary disease (COPD).Can evaluate lobar volumes. Identifies and labels pulmonary bronchi and vasculature and quantifies air-trapping. Offers emphysema evaluation. Provides functional respiratory imaging, whereby high-resolution CT images are post-processed to quantify airflow by lung region and may be used to quantify key biomarkers such as airway resistance, air-trapping, ventilation mapping, lung and lobar volume, and blood vessel and airway volume. Assesses the lung parenchyma by way of density evaluation. Provides percentages of tissues within defined attenuation (HU) ranges besides furnishing automated lung segmentation and lung volume information. Improves image quality for noisy images with built-in denoising function. Detects emphysema, a common condition seen in patients with history of smoking and hyperdense or opacified regions, thereby aiding in the diagnosis of certain pathologies, such as COVID-19 pneumonia. It aids in cardiac segmentation and calcium detection, aorta segmentation and diameter measurements, and vertebral body segmentation and density measurements. Conclusion: The future is yet to come, but AI already is a helpful tool for the daily practice in radiology. It is assumed, that the continuing progression of the computerized systems and the improvements in software algorithms , will redder AI into the second hand of the radiologist.

Keywords: artificial intelligence, chest imaging, nodule detection, automated diagnoses

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Authors: Iordana Neamtu, Aurica P. Chiriac, Loredana E. Nita, Mihai Asandulesa, Elena Butnaru, Nita Tudorachi, Alina Diaconu

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In the last decade, the attention of many researchers is focused on the synthesis of innovative “intelligent” copolymer structures with great potential for different uses. This considerable scientific interest is stimulated by possibility of the significant improvements in physical, mechanical, thermal and other important specific properties of these materials. Functionalization of polymer in synthesis by designing a suitable composition with the desired properties and applications is recognized as a valuable tool. In this work is presented a comparative study of the properties of the new copolymers poly(maleic anhydride maleic-co-3,9-divinyl-2,4,8,10-tetraoxaspiro[5.5]undecane) and poly(itaconic-anhydride-co-3,9-divinyl-2,4,8,10-tetraoxaspiro[5.5]undecane) obtained by radical polymerization in dioxane, using 2,2′-azobis(2-methylpropionitrile) as free-radical initiator. The comonomers are able for generating special effects as for example network formation, biodegradability and biocompatibility, gel formation capacity, binding properties, amphiphilicity, good oxidative and thermal stability, good film formers, and temperature and pH sensitivity. Maleic anhydride (MA) and also the isostructural analog itaconic anhydride (ITA) as polyfunctional monomers are widely used in the synthesis of reactive macromolecules with linear, hyperbranched and self & assembled structures to prepare high performance engineering, bioengineering and nano engineering materials. The incorporation of spiroacetal groups in polymer structures improves the solubility and the adhesive properties, induce good oxidative and thermal stability, are formers of good fiber or films with good flexibility and tensile strength. Also, the spiroacetal rings induce interactions on ether oxygen such as hydrogen bonds or coordinate bonds with other functional groups determining bulkiness and stiffness. The synthesized copolymers are analyzed by DSC, oscillatory and rotational rheological measurements and dielectric spectroscopy with the aim of underlying the heating behavior, solution viscosity as a function of shear rate and temperature and to investigate the relaxation processes and the motion of functional groups present in side chain around the main chain or bonds of the side chain. Acknowledgments This work was financially supported by the grant of the Romanian National Authority for Scientific Research, CNCS-UEFISCDI, project number PN-II-132/2014 “Magnetic biomimetic supports as alternative strategy for bone tissue engineering and repair’’ (MAGBIOTISS).

Keywords: Poly(maleic anhydride-co-3, 9-divinyl-2, 4, 8, 10-tetraoxaspiro (5.5)undecane); Poly(itaconic anhydride-co-3, 9-divinyl-2, 4, 8, 10-tetraoxaspiro (5.5)undecane); DSC; oscillatory and rotational rheological analysis; dielectric spectroscopy

Procedia PDF Downloads 227

Authors: Sean Yao Zu Kong, Khong Yik Chew

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Introduction: We described a case report of the successful use of advanced surgical techniques in a patient with recurrent breast cancer who underwent a wide resection including the hemi-sternum, clavicle, multiple ribs, and a lobe of the lung due to tumor involvement. This extensive resection exposed critical structures, requiring a creative approach to reconstruction. To address this complex chest wall reconstruction, a free fibula flap and a 4-zone rectus abdominis musculocutaneous flap were successfully utilized. The use of a free vascularized bone flap allowed for rapid osteointegration and resistance against osteoradionecrosis after adjuvant radiation, while a four-zone tram flap allowed for reconstruction of both the chest wall and breast mound. Although limited recipient vessels made free flaps challenging, the free fibula flap served as both a bony reconstruction and vascular conduit, supercharged with the distal peroneal artery and veins of the peroneal artery from the fibula graft. Our approach highlights the potential of advanced surgical techniques to improve outcomes in complex cases of chest wall reconstruction in patients with recurrent breast cancer, which is becoming increasingly relevant as breast cancer incidence rates increases. Case presentation: This report describes a successful reconstruction of a patient with recurrent breast cancer who required extensive resection, including the anterior chest wall, clavicle, and sternoclavicular joint. Challenges arose due to the loss of accessory muscles and the non-rigid rib cage, which could lead to compromised ventilation and instability. A free fibula osteocutaneous flap and a four-zone TRAM flap with vascular supercharging were utilized to achieve long-term stability and function. The patient has since fully recovered, and during the review, both flaps remained viable, and chest mound reconstruction was satisfactory. A planned nipple/areolar reconstruction was offered pending the patient’s decision after adjuvant radiotherapy. Conclusion: In conclusion, this case report highlights the successful use of innovative surgical techniques in addressing a complex case of recurrent breast cancer requiring extensive resection and radical reconstruction. Our approach, utilized a combination of a free fibula flap and a 4-zone rectus abdominis musculocutaneous flap, demonstrates the potential for advanced techniques in chest wall reconstruction to minimize complications and ensure long-term stability and function. As the incidence of breast cancer continues to rise, it is crucial that healthcare professionals explore and utilize innovative techniques to improve patient outcomes and quality of life.

Keywords: free fibula flap, rectus abdominis musculocutaneous flap, post-adjuvant radiotherapy, reconstructive surgery, malignancy

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Authors: Sahabettin Mete, Abdullah C. Hocagil, Hilal Hocagil, Volkan Ulker, Hasan C. Taskin

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Objectives and Goals: An emergency department is a place where people can come for a multitude of reasons 24 hours a day. As it is an easy, accessible place, thanks to self-sacrificing people who work in emergency departments. But the workload and overcrowding of emergency departments are increasing day by day. Under these circumstances, it is important to choose a quick, easily accessible and effective test for diagnosis. This results in laboratory and imaging tests being more than 40% of all emergency department costs. Despite all of the technological advances in imaging methods and available computerized tomography (CT), chest X-ray, the older imaging method, has not lost its appeal and effectiveness for nearly all emergency physicians. Progress in imaging methods are very convenient, but physicians should consider the radiation dose, cost, and effectiveness, as well as imaging methods to be carefully selected and used. The aim of the study was to investigate the effectiveness of chest X-ray in immediate diagnosis against the advancing technology by comparing chest X-ray and chest CT scan results of the patients in the emergency department. Methods: Patients who applied to Bulent Ecevit University Faculty of Medicine’s emergency department were investigated retrospectively in between 1 September 2014 and 28 February 2015. Data were obtained via MIAMED (Clear Canvas Image Server v6.2, Toronto, Canada), information management system which patients’ files are saved electronically in the clinic, and were retrospectively scanned. The study included 199 patients who were 18 or older, had both chest X-ray and chest CT imaging. Chest X-ray images were evaluated by the emergency medicine senior assistant in the emergency department, and the findings were saved to the study form. CT findings were obtained from already reported data by radiology department in the clinic. Chest X-ray was evaluated with seven questions in terms of technique and dose adequacy. Patients’ age, gender, application complaints, comorbid diseases, vital signs, physical examination findings, diagnosis, chest X-ray findings and chest CT findings were evaluated. Data saved and statistical analyses have made via using SPSS 19.0 for Windows. And the value of p < 0.05 were accepted statistically significant. Results: 199 patients were included in the study. In 38,2% (n=76) of all patients were diagnosed with pneumonia and it was the most common diagnosis. The chest X-ray imaging technique was appropriate in patients with the rate of 31% (n=62) of all patients. There was not any statistically significant difference (p > 0.05) between both imaging methods (chest X-ray and chest CT) in terms of determining the rates of displacement of the trachea, pneumothorax, parenchymal consolidation, increased cardiothoracic ratio, lymphadenopathy, diaphragmatic hernia, free air levels in the abdomen (in sections including the image), pleural thickening, parenchymal cyst, parenchymal mass, parenchymal cavity, parenchymal atelectasis and bone fractures. Conclusions: When imaging findings, showing cases that needed to be quickly diagnosed, were investigated, chest X-ray and chest CT findings were matched at a high rate in patients with an appropriate imaging technique. However, chest X-rays, evaluated in the emergency department, were frequently taken with an inappropriate technique.

Keywords: chest x-ray, chest computerized tomography, chest imaging, emergency department

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Authors: R. P. Hewawasam, M. H. A. D. de Silva, M. A. G. Iresha

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In recent years childhood obesity has increased to pan-epidemic proportions along with a concomitant increase in obesity-associated morbidity. Birth weight is an important determinant of later adult health, with neonates at both ends of the birth weight spectrum at risk of future health complications. Consequently, infants who are born large for gestational age (LGA) are more likely to be obese in childhood and adolescence and are at risk of cardiovascular and metabolic complications later in life. Adipose tissue plays a role in linking events in fetal growth to the subsequent development of adult diseases. In addition to its role as a storage depot for fat, adipose tissue produces and secrets a number of hormones of importance in modulating metabolism and energy homeostasis. Cord blood leptin level has been positively correlated with fetal adiposity at birth. It is established that Asians have lower skeletal muscle mass, low bone mineral content and excess body fat for a given body mass index indicating a genetic predisposition in the occurrence of obesity. To our knowledge, studies have never been conducted in Sri Lanka to determine the relationship between adipocytokine profile in cord blood and anthropometric parameters in newborns. Thus, the objective of this study is to establish the above relationship for the Sri Lankan population to implement awareness programs to minimize childhood obesity in the future. Umbilical cord blood was collected from 90 newborns (Male 40, Female 50; gestational age 35-42 weeks) after double clamping the umbilical cord before separation of the placenta and the concentration of leptin was measured by ELISA technique. Anthropometric parameters of the newborn such as birth weight, length, ponderal index, occipital frontal, chest, hip and calf circumferences were measured. Pearson’s correlation was used to assess the relationship between leptin and anthropometric parameters while the Mann-Whitney U test was used to assess the differences in cord blood leptin levels between small for gestational age (SGA), appropriate for gestational age (AGA) and LGA infants. There was a significant difference (P < 0.05) between the cord blood leptin concentrations of LGA infants (12.67 ng/mL ± 2.34) and AGA infants (7.10 ng/mL ± 0.90). However, a significant difference was not observed between leptin levels of SGA infants (8.86 ng/mL ± 0.70) and AGA infants. In both male and female neonates, umbilical leptin levels showed significant positive correlations (P < 0.05) with birth weight of the newborn, pre-pregnancy maternal weight and pre pregnancy BMI between the infants of large and appropriate for gestational ages. Increased concentrations of leptin levels in the cord blood of large for gestational age infants suggest that they may be involved in regulating fetal growth. Leptin concentration of Sri Lankan population was not significantly deviated from published data of Asian populations. Fetal leptin may be an important predictor of neonatal adiposity; however, interventional studies are required to assess its impact on the possible risk of childhood obesity.

Keywords: appropriate for gestational age, childhood obesity, leptin, anthropometry

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Authors: Inna Kornienko, Svetlana Guryeva, Anatoly Shekhter, Elena Petersen

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Transplantation is an effective treatment option for patients suffering from different end-stage diseases. However, it is plagued by a constant shortage of donor organs and the subsequent need of a lifelong immunosuppressive therapy for the patient. Currently some researchers look towards using of pig organs to replace human organs for transplantation since the matrix derived from porcine organs is a convenient substitute for the human matrix. As an initial step to create a new ex vivo tissue engineered model, optimized protocols have been created to obtain organ-specific acellular matrices and evaluated their potential as tissue engineered scaffolds for culture of normal cells and tumor cell lines. These protocols include decellularization by perfusion in a bioreactor system and immersion-agitation on an orbital shaker with use of various detergents (SDS, Triton X-100) and freezing. Complete decellularization – in terms of residual DNA amount – is an important predictor of probability of immune rejection of materials of natural origin. However, the signs of cellular material may still remain within the matrix even after harsh decellularization protocols. In this regard, the matrices obtained from tissues of low-immunogenic pigs with α3Galactosyl-tranferase gene knock out (GalT-KO) may be a promising alternative to native animal sources. The research included a study of induced effect of frozen and fresh fragments of GalT-KO skin on healing of full-thickness plane wounds in 80 rats. Commercially available wound dressings (Ksenoderm, Hyamatrix and Alloderm) as well as allogenic skin were used as a positive control and untreated wounds were analyzed as a negative control. The results were evaluated on the 4th day after grafting, which corresponds to the time of start of normal wound epithelization. It has been shown that a non-specific immune response in models treated with GalT-Ko pig skin was milder than in all the control groups. Research has been performed to measure technical skin characteristics: stiffness and elasticity properties, corneometry, tevametry, and cutometry. These metrics enabled the evaluation of hydratation level, corneous layer husking level, as well as skin elasticity and micro- and macro-landscape. These preliminary data may contribute to development of personalized transplantable organs from GalT-Ko pigs with significantly limited potential of immune rejection. By applying growth factors to a decellularized skin sample it is possible to achieve various regenerative effects based on the particular situation. In this particular research BMP2 and Heparin-binding EGF-like growth factor have been used. Ideally, a bioengineered organ must be biocompatible, non-immunogenic and support cell growth. Porcine organs are attractive for xenotransplantation if severe immunologic concerns can be bypassed. The results indicate that genetically modified pig tissues with knock-outed α3Galactosyl-tranferase gene may be used for production of low-immunogenic matrix suitable for transplantation.

Keywords: decellularization, low-immunogenic, matrix, scaffolds, transplants

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Authors: Rawan Ashraf, Ahmed E. Gomaa, Gehan Safwat, Ayman Diab

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Background: Three-dimensional (3D) bio-printing has become a versatile and powerful method for generating a variety of biological constructs, including bone or extracellular matrix scaffolds endo- or epithelial, muscle tissue, as well as organoids. Aim of the study: Fabricate a low cost DIY 3D bio-printer to produce 3D bio-printed products such as anti-microbial packaging or multi-organs on chips. We demonstrate the alignment between two types of 3D printer technology (3D Bio-printer and DLP) on Multi-organ-on-a-chip (multi-OoC) devices fabrication. Methods: First, Design and Fabrication of the Syringe Unit for Modification of an Off-the-Shelf 3D Printer, then Preparation of Hydrogel based on natural polymers Sodium Alginate and Gelatin, followed by acquisition of the cell suspension, then modeling the desired 3D structure. Preparation for 3D printing, then Cell-free and cell-laden hydrogels went through the printing process at room temperature under sterile conditions and finally post printing curing process and studying the printed structure regards physical and chemical characteristics. The hard scaffold of the Organ on chip devices was designed and fabricated using the DLP-3D printer, following similar approaches as the Microfluidics system fabrication. Results: The fabricated Bio-Ink was based onHydrogel polymer mix of sodium alginate and gelatin 15% to 0.5%, respectively. Later the 3D printing process was conducted using a higher percentage of alginate-based hydrogels because of it viscosity and the controllable crosslinking, unlike the thermal crosslinking of Gelatin. The hydrogels were colored to simulate the representation of two types of cells. The adaption of the hard scaffold, whether for the Microfluidics system or the hard-tissues, has been acquired by the DLP 3D printers with fabricated natural bioactive essential oils that contain antimicrobial activity, followed by printing in Situ three complex layers of soft-hydrogel as a cell-free Bio-Ink to simulate the real-life tissue engineering process. The final product was a proof of concept for a rapid 3D cell culturing approaches that uses an engineered hard scaffold along with soft-tissues, thus, several applications were offered as products of the current prototype, including the Organ-On-Chip as a successful integration between DLP and 3D bioprinter. Conclusion: Multiple designs for the organ-on-a-chip (multi-OoC) devices have been acquired in our study with main focus on the low cost fabrication of such technology and the potential to revolutionize human health research and development. We describe circumstances in which multi-organ models are useful after briefly examining the requirement for full multi-organ models with a systemic component. Following that, we took a look at the current multi-OoC platforms, such as integrated body-on-a-chip devices and modular techniques that use linked organ-specific modules.

Keywords: 3d bio-printer, hydrogel, multi-organ on chip, bio-inks

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Authors: Thiago E. Goto, Carla C. Lopes, Helena B. Nader, Anielle C. A. Silva, Noelio O. Dantas, José R. Siqueira Jr., Luciano Caseli

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Quantum dots (QD) are semiconductor nanocrystals that can be employed in biological research as a tool for fluorescence imagings, having the potential to expand in vivo and in vitro analysis as cancerous cell biomarkers. Particularly, cadmium selenide (CdSe) magic-sized quantum dots (MSQDs) exhibit stable luminescence that is feasible for biological applications, especially for imaging of tumor cells. For these facts, it is interesting to know the mechanisms of action of how such QDs mark biological cells. For that, simplified models are a suitable strategy. Among these models, Langmuir films of lipids formed at the air-water interface seem to be adequate since they can mimic half a membrane. They are monomolecular films formed at liquid-gas interfaces that can spontaneously form when organic solutions of amphiphilic compounds are spread on the liquid-gas interface. After solvent evaporation, the monomolecular film is formed, and a variety of techniques, including tensiometric, spectroscopic and optic can be applied. When the monolayer is formed by membrane lipids at the air-water interface, a model for half a membrane can be inferred where the aqueous subphase serve as a model for external or internal compartment of the cell. These films can be transferred to solid supports forming the so-called Langmuir-Blodgett (LB) films, and an ampler variety of techniques can be additionally used to characterize the film, allowing for the formation of devices and sensors. With these ideas in mind, the objective of this work was to investigate the specific interactions of CdSe MSQDs with tumorigenic and non-tumorigenic cells using Langmuir monolayers and LB films of lipids and specific cell extracts as membrane models for diagnosis of cancerous cells. Surface pressure-area isotherms and polarization modulation reflection-absorption spectroscopy (PM-IRRAS) showed an intrinsic interaction between the quantum dots, inserted in the aqueous subphase, and Langmuir monolayers, constructed either of selected lipids or of non-tumorigenic and tumorigenic cells extracts. The quantum dots expanded the monolayers and changed the PM-IRRAS spectra for the lipid monolayers. The mixed films were then compressed to high surface pressures and transferred from the floating monolayer to solid supports by using the LB technique. Images of the films were then obtained with atomic force microscopy (AFM) and confocal microscopy, which provided information about the morphology of the films. Similarities and differences between films with different composition representing cell membranes, with or without CdSe MSQDs, was analyzed. The results indicated that the interaction of quantum dots with the bioinspired films is modulated by the lipid composition. The properties of the normal cell monolayer were not significantly altered, whereas for the tumorigenic cell monolayer models, the films presented significant alteration. The images therefore exhibited a stronger effect of CdSe MSQDs on the models representing cancerous cells. As important implication of these findings, one may envisage for new bioinspired surfaces based on molecular recognition for biomedical applications.

Keywords: biomembrane, langmuir monolayers, quantum dots, surfaces

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Authors: Elissandro Voigt Beier, Cristiano Poleto

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The main objective of the research is to assess the loss through the quantification of material culture (archaeological fragments) in rural areas, sites explored economically by machining on seasonal crops, and also permanent, in a hydrographic subsystem Camaquã River in the state of Rio Grande do Sul, Brazil. The study area consists of different micro basins and differs in area, ranging between 1,000 m² and 10,000 m², respectively the largest and the smallest, all with a large number of occurrences and outcrop locations of archaeological material and high density in intense farm environment. In the first stage of the research aimed to identify the dispersion of points of archaeological material through field survey through plot points by the Global Positioning System (GPS), within each river basin, was made use of concise bibliography on the topic in the region, helping theoretically in understanding the old landscaping with preferences of occupation for reasons of ancient historical people through the settlements relating to the practice observed in the field. The mapping was followed by the cartographic development in the region through the development of cartographic products of the land elevation, consequently were created cartographic products were to contribute to the understanding of the distribution of the absolute materials; the definition and scope of the material dispersed; and as a result of human activities the development of revolving letter by mechanization of in situ material, it was also necessary for the preparation of materials found density maps, linking natural environments conducive to ancient historical occupation with the current human occupation. The third stage of the project it is for the systematic collection of archaeological material without alteration or interference in the subsurface of the indigenous settlements, thus, the material was prepared and treated in the laboratory to remove soil excesses, cleaning through previous communication methodology, measurement and quantification. Approximately 15,000 were identified archaeological fragments belonging to different periods of ancient history of the region, all collected outside of its environmental and historical context and it also has quite changed and modified. The material was identified and cataloged considering features such as object weight, size, type of material (lithic, ceramic, bone, Historical porcelain and their true association with the ancient history) and it was disregarded its principles as individual lithology of the object and functionality same. As observed preliminary results, we can point out the change of materials by heavy mechanization and consequent soil disturbance processes, and these processes generate loading of archaeological materials. Therefore, as a next step will be sought, an estimate of potential losses through a mathematical model. It is expected by this process, to reach a reliable model of high accuracy which can be applied to an archeological site of lower density without encountering a significant error.

Keywords: degradation of heritage, quantification in archaeology, watershed, use and occupation of land

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Authors: Alua Massalimova

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Situation analysis of Blood Service revealed that the strengths dominated over the weak 1.4 times. The possibilities dominate over the threats by 1.1 times. It follows that by using timely the possibility the Service, it is possible to strengthen its strengths and avoid threats. Priority directions of the resulting analysis are the use of subjective factors, such as personal management capacity managers of the Blood Center in the field of possibilities of legal activity of administrative decisions and the mobilization of stable staff in general market conditions. We have studied for the period 2011-2015 retrospectively indicators of Blood Service of Kazakhstan. Strengths of Blood Service of RK(Ps4,5): 1) indicators of donations for 1000 people is higher than in some countries of the CIS (in Russia 14, Kazakhstan - 17); 2) the functioning science centre of transfusiology; 3) the legal possibility of additional financing blood centers in the form of paid services; 4) the absence of competitors; 5) training on specialty Transfusiology; 6) the stable management staff of blood centers, a high level of competence; 7) increase in the incidence requiring transfusion therapy (oncohematology); 8) equipment upgrades; 9) the opening of a reference laboratory; 10) growth of the proportion of issued high-quality blood components; 11) governmental organization 'Drop of Life'; 12) the functioning bone marrow register; 13) equipped with modern equipment HLA-laboratory; 14) High categorization of average medical workers; 15) availability of own specialized scientific journal; 16) vivarium. The weaknesses (Ps = 3.5): 1) the incomplete equipping of blood centers and blood transfusion cabinets according to standards; 2) low specific weight of paid services of the CC; 3) low categorization of doctors; 4) high staff turnover; 5) the low scientific potential of industrial and clinical of transfusiology; 6) the low wages paid; 7) slight growth of harvested donor blood; 8) the weak continuity with offices blood transfusion; 9) lack of agitation work; 10) the formally functioning of Transfusion Association; 11) the absence of scientific laboratories; 12) high standard deviation from the average for donations in the republic. The possibilities (Ps = 2,7): 1): international grants; 2) organization of international seminars on clinical of transfusiology; 3) cross-sectoral cooperation; 4) to increase scientific research in the field of clinical of transfusiology; 5) reduce the share of donation unsuitable for transfusion and processing; 6) strengthening marketing management in the development of fee-based services; 7) advertising paid services; 8) strengthening the publishing of teaching aids; 9) team-building staff. The threats (Ps = 2.1): 1) an increase of staff turnover; 2) the risk of litigation; 3) reduction gemoprodukts based on evidence-based medicine; 4) regression of scientific capacity; 5) organization of marketing; 6) transfusiologist marketing; 7) reduction in the quality of the evidence base transfusions.

Keywords: blood service, healthcare, Kazakhstan, quantative swot analysis

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