Search results for: tissues

Authors: Mahesh Kumar Farejiya, Anil Kumar Dikshit

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The toxic metal contamination and their biomagnification in marine fishes is a serious public health concern specially, in the coastal areas and the small islands. In the present study, concentration of toxic heavy metals like zinc (Zn), cadmium (Cd), lead (Pb), nickel (Ni), cobalt (Co), chromium (Cr) and mercury (Hg) were determined in the tissues of tunas (T. albacores) caught from the area near to Lakshdweep Islands. The heavy metals are one of the indicators for the marine water pollution. Geochemical weathering, industrialization, agriculture run off, fishing, shipping and oil spills are the major pollutants. The presence of heavy toxic metals in the near coastal water fishes at both western coast and eastern coast of India has been well established. The present study was conducted assuming that the distant island will not have the metals presence in a way it is at the near main land coast. However, our study shows that there is a significant amount of the toxic metals present in the tissues of tuna samples. The gill, lever and flash samples were collected in waters around Lakshdweep Islands. They were analyzed using ICP–AES for the toxic metals after microwave digestion. The concentrations of the toxic metals were found in all fish samples and the general trend of presence was in decreasing order as Zn > Al > Cd > Pb > Cr > Ni > Hg. The amount of metals was found to higher in fish having more weight.

Keywords: toxic metals, marine tuna fish, bioaccumulation, biomagnifications

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Authors: Cheng-Cao Sun, Shu-Jun Li, Cuili Yang, Yongyong Xi, Liang Wang, Feng Zhang, De-Jia Li

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MicroRNAs (miRNAs) act as key regulators of multiple cancers. Hsa-miR-329 (miR-329) functions as a tumor suppressor in some malignancies. However, its role on lung cancer remains poorly understood. In this study, we investigated the role of miR-329 on the development of lung cancer. The results indicated that miR-329 was decreased in primary lung cancer tissues compared with matched adjacent normal lung tissues and very low levels were found in a non-small cell lung cancer (NSCLC) cell lines. Ectopic expression of miR-329 in lung cancer cell lines substantially repressed cell growth as evidenced by cell viability assay, colony formation assay and BrdU staining, through inhibiting cyclin D1, cyclin D2, and up-regulatiing p57(Kip2) and p21(WAF1/CIP1). In addition, miR-329 promoted NSCLC cell apoptosis, as indicated by up-regulation of key apoptosis gene cleaved caspase-3, and down-regulation of anti-apoptosis gene Bcl2. Moreover, miR-329 inhibited cellular migration and invasiveness through inhibiting matrix metalloproteinases (MMP)-7 and MMP-9. Further, oncogene MET was revealed to be a putative target of miR-329, which was inversely correlated with miR-329 expression. Furthermore, down-regulation of MET by siRNA performed similar effects to over-expression of miR-329. Collectively, our results demonstrated that miR-329 played a pivotal role in lung cancer through inhibiting cell proliferation, migration, invasion, and promoting apoptosis by targeting oncogenic MET.

Keywords: hsa-miRNA-329(miR-329), MET, non-small cell lung cancer (NSCLC), proliferation, apoptosis

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Authors: Gina Zavaleta-Espejo, Segundo R. Jáuregui-Rosas, Fanny V. Samanamud-Moreno, José Saldaña Jiménez, Anibal Felix-Quintero, Víctor Montero-Del Aguila, Elsi Mejía-Uriarte

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Vicugnapacos "alpaca" fabrics are considered special for their finesse, and the garments in the textile market are very luxurious. It has many special characteristics such as antiallergic, soft, hygroscopic, among others. In this sense, the research aimed to evaluate the antimicrobial activity of alpaca fabrics functionalized with silver nanoparticles on the bacteria Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923. For the functionalization of the fabrics, AgNO3 and different concentrations of trisodium citrate (TSC) 2, 6, and 10 mg. Tissue characterization was performed using Raman spectroscopy, Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). The determination of the antimicrobial activity of the alpaca tissues was made by the Kirby-Bauer method with alpaca tissue discs functionalized with silver nanoparticles, an experimental design was made in completely randomized blocks with three treatments and a negative control with three repetitions. The results showed that inhibition halos were formed for both bacteria, therefore, the functionalized tissues have a high antimicrobial activity, whose mechanism of action is attributed to the free radicals (ROS) generated by the nanoparticles that cause oxidative damage to the bacteria. proteins and lipids of the bacterial cell wall.

Keywords: antimicrobial, animal fibers, fabrics, functionalization, trisodium citrate

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Authors: Tayyaba Bari, Muhammad Hamza Anjum, Samra Kanwal, Fakhera Ikram

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Osteoarthritis, a degenerative condition, affects more than 213 million individuals globally. Since articular cartilage has no or limited vessels, therefore, after deteriorating, it is unable to rejuvenate. Traditional approaches for cartilage repair, like autologous chondrocyte implantation, microfracture and cartilage transplantation are often associated with postoperative complications and lead to further degradation. Decellularized human umbilical cord has gained interest as a viable treatment for cartilage repair. Decellularization removes all cellular contents as well as debris, leaving a biologically active 3D network known as extracellular matrix (ECM). This matrix is biodegradable, non-immunogenic and provides a microenvironment for homeostasis, growth and repair. UC derived bioink function as 3D scaffolding material, not only mediates cell-matrix interactions but also adherence, proliferation and propagation of cells for 3D organoids. This study comprises different physical, chemical and biological approaches to optimize the decellularization of human umbilical cord (UC) tissues followed by the solubilization of these tissues to bioink formation. The decellularization process consisted of two cycles of freeze thaw where the umbilical cord at -20˚C was thawed at room temperature followed by dissection in small sections from 0.5 to 1cm. Similarly decellularization with ionic and non-ionic detergents Sodium dodecyl sulfate (SDS) and Triton-X 100 revealed that both concentrations of SDS i.e 0.1% and 1% were effective in complete removal of cells from the small UC tissues. The results of decellularization was further confirmed by running them on 1% agarose gel. Histological analysis revealed the efficacy of decellularization, which involves paraffin embedded samples of 4μm processed for Hematoxylin-eosin-safran and 4,6-diamidino-2-phenylindole (DAPI). ECM preservation was confirmed by Alcian Blue, and Masson’s trichrome staining on consecutive sections and images were obtained. Sulfated GAG’s content were determined by 1,9-dimethyl-methylene blue (DMMB) assay, similarly collagen quantification was done by hydroxy proline assay. This 3D bioengineered scaffold will provide a typical atmosphere as in the extracellular matrix of the tissue, which would be seeded with the mesenchymal cells to generate the desired 3D ink for in vitro and in vivo cartilage regeneration applications.

Keywords: umbilical cord, 3d printing, bioink, tissue engineering, cartilage regeneration

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Authors: Ieva Bruzauskaite, Jovile Raudoniute, Karina Poliakovaite, Danguole Zabulyte, Daiva Bironaite, Ruta Aldonyte

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Introduction: Air pollution is a growing global problem which has been shown to be responsible for various adverse health outcomes. Immunotoxicity, such as dysregulated inflammation, has been proposed as one of the main mechanisms in air pollution-associated diseases. Chronic obstructive pulmonary disease (COPD) is among major morbidity and mortality causes worldwide and is characterized by persistent airflow limitation caused by the small airways disease (obstructive bronchiolitis) and irreversible parenchymal destruction (emphysema). Exact pathways explaining the air pollution induced and mediated disease states are still not clear. However, modern societies understand dangers of polluted air, seek to mitigate such effects and are in need for reliable biomarkers of air pollution. We hypothesise that post-translational modifications of structural proteins, e.g. citrullination, might be a good candidate biomarker. Thus, we have designed this study, where mice were exposed to diesel exhaust and the ongoing protein modifications and inflammation in lungs and other tissues were assessed. Materials And Methods: To assess the effects of diesel exhaust a in vivo study was designed. Mice (n=10) were subjected to everyday 2-hour exposure to diesel exhaust for 14 days. Control mice were treated the same way without diesel exhaust. The effects within lung and other tissues were assessed by immunohistochemistry of formalin-fixed and paraffin-embedded tissues. Levels of inflammation and citrullination related markers were investigated. Levels of parenchymal damage were also measured. Results: In vivo study corroborates our own data from in vitro and reveals diesel exhaust initiated inflammatory shift and modulation of lung peptidyl arginine deiminase 4 (PAD4), citrullination associated enzyme, levels. In addition, high levels of citrulline were observed in exposed lung tissue sections co-localising with increased parenchymal destruction. Conclusions: Subacute exposure to diesel exhaust renders mice lungs inflammatory and modifies certain structural proteins. Such structural changes of proteins may pave a pathways to lost/gain function of affected molecules and also propagate autoimmune processes within the lung and systemically.

Keywords: air pollution, citrullination, in vivo, lungs

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Authors: Nunthawan Nowwarote, Waleerat Sukarawan, Prasit Pavasant, Thanaphum Osathanon

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Interleukin 6 (IL-6) is a multifunctional cytokine, regulating various biological responses in several tissues. A Recent study shows that IL-6 plays a role in stemness maintenance in stem cells isolated from human exfoliated deciduous teeth (SHEDs). However, the role of IL-6 on cell differentiation in SHEDs remains unknown. The present study investigated the effect of IL-6 on SHEDs differentiation. Cells were isolated from dental pulp tissues of human deciduous teeth. Flow cytometry was used to determined mesenchymal stem cell marker expression, and the multipotential differentiation (osteogenic, adipogenic and neurogenic lineage ) was also determined. The mRNA was determined using real-time quantitative polymerase chain reaction, and the phenotypes were confirmed by chemical and immunofluorescence staining. Results demonstrated that SHEDs expressed CD44, CD73, CD90, CD105 but not CD45. Further, the up-regulation of osteogenic, adipogenic and neurogenic marker genes was observed upon maintaining cells in osteogenic, adipogenic and neurogenic induction medium, respectively. The addition of IL-6 induced osteogenic by up-regulated osteogenic marker gene also increased in vitro mineralization. Under neurogenic medium supplement with IL-6, up-regulated neurogenic marker. Whereas, an addition of IL-6 attenuated adipogenic differentiation by SHEDs. In conclusion, this evidence implies that IL-6 may participate in cells differentiation ability of SHEDs.

Keywords: SHEDs, IL-6, cell differentiations, dental pulp

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Authors: Kyu Rae Lee

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The aim of the study is to investigate the clinical efficacy of combination therapeutic modalities using liraglutide (1.2mg/d) add on low-dose phentermine (7.5 mg/d)/topiramate (50mg/d) medication on the obese patient in the bariatric clinic. We assessed the retrospective cohort clinical analyses to the clinical efficacy of medication and combination in the patients who visited the bariatric clinic. We measured all participants’ body fat (bioelectric impedance analysis), weight, height, and the cross-sectional areas of adipose tissues (umbilicus level) after keep fasting for 8 hours at 0, 4, 12 weeks. The design of the study was opened, paired t-test and Wilcoxon test were performed using SPSS for windows (ver.18, IL, USA) for comparison of weight, body fat, and adipose tissues. The participants were one hundred twenty-eight subjects aged 44.67 (1.18) years, 28.95 (0.39) kg/m², and female (82.7%). Their body fat was 40.57 (2.23%), and waist to hip ratio was 0.96 (0.01). The mean cross-sectional area of visceral adipose tissue was 142.59 (7.06) mm², and that of subcutaneous adipose was 274.37 (9.18) mm². 73 of them (57.5%) took medication only, 54 of them took medication with liraglutide for 12 weeks. The subjects in the medication group lost 5.4165 kg, 6.8069%, and those of the combination group did 6.2481 kg, 3.564%. The mean cross-sectional areas of visceral, subcutaneous adipose tissue in the medication group significantly decreased (p=.043), even more in the combination group. (p=.028). Further controlled clinical trials should be considered in the future. We conclude that the low dose of phentermine/topiramate with liraglutide therapeutic modalities would be more effective than phentermine/topiramate medication only in obesity treatment for 12 weeks.

Keywords: low dose phentermine, topiramate, liraglutide, obesity, efficacy

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Authors: Vishwanath Mali, Hemant Warhatkar, Raju Pawade

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Drilling of bone has been always challenging for surgeons due to the adverse effect it may impart to bone tissues. Force has to be applied manually by the surgeon while performing conventional bone drilling which may lead to permanent death of bone tissues and nerves. During bone drilling the temperature of the bone tissues increases to higher values above 47 ⁰C that causes thermal osteonecrosis resulting into screw loosening and subsequent implant failures. An attempt has been made here to study the input drilling parameters and surgical drill bit geometry affecting bone health during bone drilling. A One Factor At a Time (OFAT) method is used to plan the experiments. Input drilling parameters studied include spindle speed and feed rate. The drill bit geometry parameter studied include point angle and helix angle. The output variables are drilling thrust force and bone temperature. The experiments were conducted on goat femur bone at room temperature 30 ⁰C. For measurement of thrust forces KISTLER cutting force dynamometer Type 9257BA was used. For continuous data acquisition of temperature NI LabVIEW software was used. Fixture was made on RPT machine for holding the bone specimen while performing drilling operation. Bone specimen were preserved in deep freezer (LABTOP make) under -40 ⁰C. In case of drilling parameters, it is observed that at constant feed rate when spindle speed increases, thrust force as well as temperature decreases and at constant spindle speed when feed rate increases thrust force as well as temperature increases. The effect of drill bit geometry shows that at constant helix angle when point angle increases thrust force as well as temperature increases and at constant point angle when helix angle increase thrust force as well as temperature decreases. Hence it is concluded that as the thrust force increases temperature increases. In case of drilling parameter, the lowest thrust force and temperature i.e. 35.55 N and 36.04 ⁰C respectively were recorded at spindle speed 2000 rpm and feed rate 0.04 mm/rev. In case of drill bit geometry parameter, the lowest thrust force and temperature i.e. 40.81 N and 34 ⁰C respectively were recorded at point angle 70⁰ and helix angle 25⁰ Hence to avoid thermal necrosis of bone it is recommended to use higher spindle speed, lower feed rate, low point angle and high helix angle. The hard nature of cortical bone contributes to a greater rise in temperature whereas a considerable drop in temperature is observed during cancellous bone drilling.

Keywords: bone drilling, helix angle, point angle, thrust force, temperature, thermal necrosis

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Authors: Asim Abbasi, Muhammad Sufyan, Muhammad Kamran, Iqra

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Development of resistance in insect pests against various groups of insecticides has prompted the use of alternative integrated pest management approaches. Among these induced host plant resistance represents an important strategy as it offers a practical, cheap and long lasting solution to keep pests populations below economic threshold level (ETL). Silicon (Si) has a major role in regulating plant eco-relationship by providing strength to the plant in the form of anti-stress mechanism which was utilized in coping with the environmental extremes to get a better yield and quality end produce. Among biotic stresses, insect herbivore signifies one class against which Si provide defense. Silicon in its neutral form (H₄SiO₄) is absorbed by the plants via roots through an active process accompanied by the help of different transporters which were located in the plasma membrane of root cells or by a passive process mostly regulated by transpiration stream, which occurs via the xylem cells along with the water. Plants tissues mainly the epidermal cell walls are the sinks of absorbed silicon where it polymerizes in the form of amorphous silica or monosilicic acid. The noteworthy function of this absorbed silicon is to provide structural rigidity to the tissues and strength to the cell walls. Silicon has both direct and indirect effects on insect herbivores. Increased abrasiveness and hardness of epidermal plant tissues and reduced digestibility as a result of deposition of Si primarily as phytoliths within cuticle layer is now the most authenticated mechanisms of Si in enhancing plant resistance to insect herbivores. Moreover, increased Si content in the diet also impedes the efficiency by which insects transformed consumed food into the body mass. The palatability of food material has also been changed by Si application, and it also deters herbivore feeding for food. The production of defensive compounds of plants like silica and phenols have also been amplified by the exogenous application of silicon sources which results in reduction of the probing time of certain insects. Some studies also highlighted the role of silicon at the third trophic level as it also attracts natural enemies of insects attacking the crop. Hence, the inclusion of Si in pest management approaches can be a healthy and eco-friendly tool in future.

Keywords: defensive, phytoliths, resistance, stresses

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Authors: Jimmy Jiun-Ming Su, Yuan-Min Lin

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Bioprinting has been applied to produce 3D cellular constructs for tissue engineering. Microextrusion printing is the most common used method. However, printing low viscosity bioink is a challenge for this method. Herein, we developed a new 3D printing system to fabricate cell-laden hydrogels via a DLP-based projector. The bioprinter is assembled from affordable equipment including a stepper motor, screw, LED-based DLP projector, open source computer hardware and software. The system can use low viscosity and photo-polymerized bioink to fabricate 3D tissue mimics in a layer-by-layer manner. In this study, we used gelatin methylacrylate (GelMA) as bioink for stem cell encapsulation. In order to reinforce the printed construct, surface modified hydroxyapatite has been added in the bioink. We demonstrated the silanization of hydroxyapatite could improve the crosslinking between the interface of hydroxyapatite and GelMA. The results showed that the incorporation of silanized hydroxyapatite into the bioink had an enhancing effect on the mechanical properties of printed hydrogel, in addition, the hydrogel had low cytotoxicity and promoted the differentiation of embedded human bone marrow stem cells (hBMSCs) and retinal pigment epithelium (RPE) cells. Moreover, this bioprinting system has the ability to generate microchannels inside the engineered tissues to facilitate diffusion of nutrients. We believe this 3D bioprinting system has potential to fabricate various tissues for clinical applications and regenerative medicine in the future.

Keywords: bioprinting, cell encapsulation, digital light processing, GelMA hydrogel

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Authors: J. C. Akan, F. I. Abdulrahman, Z. M. Chellube

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The aim of this study was to determine the levels of some organochlorine (o, p-DDE, p,p’-DDD, o,p’-DDD, p,p’-DDT, p,p’-DDT, α-BHC, γ-BHC, lindane, Endosulfan sulphate, dieldrin and aldrin and organophosphorus (Dichlorvos, Diazinon, Chlorpyrifos, fenitrothion and Fenitrothion) pesticide residues in the gills, liver, stomach, kidney and flesh of four fish species (Tilapia zilli, Clarias anguillaris Hetrotis niloticus and Oreochronmis niloticus) between the periods of September 2010 to October, 2011. Samples were collected from Kwantan turare in Lake Chad, Baga, Borno State, Nigeria. Extraction of the fish samples and de-fattening of the fish sample extracts were performed using standard procedures. Analysis of the fish samples for pesticide residues were carried out using Shimadzu GC/MS (GC – 17A), equipped with fluorescence detector. Large differences in the levels of pesticide residues were observed between tissues within each fish. The concentrations of all the organophosphorus pesticides were higher in the organs of Oreochronmis niloticus, while Hetrotis niloticus shows the lowest. For organochlorine pesticides, the organs of Tilapia zilli showed the highest concentrations, while Hetrotis niloticus shows the lowest. The highest pesticide concentrations were observed in gills and liver tissues of all the species of fish study, while the lowest concentrations were observed in flesh. Based on the above results, it can therefore be concluded that the concentrations of pesticide in the four fish species study did exceed the permissible limits set by FAO and FEPA.

Keywords: organochlorine, organophosphorus, pesticides, accumulation, fish, lake chad

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Authors: Yong Zhang

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Light has proven to be useful in a wide range of biomedical applications such as fluorescence imaging, photoacoustic imaging, optogenetics, photodynamic therapy, photothermal therapy, and light controlled drug/gene delivery. Taking photodynamic therapy (PDT) as an example, PDT has been proven clinically effective in early lung cancer, bladder cancer, head, and neck cancer and is the primary treatment for skin cancer as well. However, clinical use of PDT is severely constrained by the low penetration depth of visible light through thick tissue, limiting its use to target regions only a few millimeters deep. One way to enhance the range is to use invisible near-infrared (NIR) light within the optical window (700–1100nm) for biological tissues, extending the depth up to 1cm with no observable damage to the intervening tissue. We have demonstrated use of NIR-to-visible upconversion fluorescent nanoparticles (UCNPs), emitting visible fluorescence when excited by a NIR light at 980nm, as a nanotransducer for PDT to convert deep tissue-penetrating NIR light to visible light suitable for activating photosensitizers. The unique optical properties of UCNPs enable the upconversion wavelength to be tuned and matched to the activation absorption wavelength of the photosensitizer. At depths beyond 1cm, however, tissue remains inaccessible to light even within the NIR window, and this critical depth limitation renders existing phototherapy ineffective against most deep-seated cancers. We have demonstrated some new treatment modalities for deep-seated cancers based on UCNP hydrogel implants and miniaturized, wirelessly powered optoelectronic devices for light delivery to deep tissues.

Keywords: upconversion, fluorescent, nanoparticle, bioimaging, photodynamic therapy

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Authors: Anurag Gaur Nidhi

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Targeted drug delivery is a method of delivering medication to a patient in a manner that increases the concentration of the medication in some parts of the body relative to others. Targeted drug delivery seeks to concentrate the medication in the tissues of interest while reducing the relative concentration of the medication in the remaining tissues. This improves efficacy of the while reducing side effects. In the present work, we investigate the effect of magnetic field, flow rate and particle concentration on the capturing of magnetic particles transported in a stent implanted fluidic channel. Iron oxide magnetic nanoparticles (Fe3O4) nanoparticles were synthesized via co-precipitation method. The synthesized Fe3O4 nanoparticles were added in the de-ionized (DI) water to prepare the Fe3O4 magnetic particle suspended fluid. This fluid is transported in a cylindrical tube of diameter 8 mm with help of a peristaltic pump at different flow rate (25-40 ml/min). A ferromagnetic coil of SS 430 has been implanted inside the cylindrical tube to enhance the capturing of magnetic nanoparticles under magnetic field. The capturing of magnetic nanoparticles was observed at different magnetic magnetic field, flow rate and particle concentration. It is observed that capture efficiency increases from 47-67 % at magnetic field 2-5kG, respectively at particle concentration 0.6 mg/ml and at flow rate 30 ml/min. However, the capture efficiency decreases from 65 to 44 % by increasing the flow rate from 25 to 40 ml/min, respectively. Furthermore, it is observed that capture efficiency increases from 51 to 67 % by increasing the particle concentration from 0.3 to 0.6 mg/ml, respectively.

Keywords: capture efficiency, implant assisted-Magnetic drug targeting (IA-MDT), magnetic nanoparticles, In-vitro study

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Authors: A. M. Malagon Escandon, J. A. Arenas Alatorre, C. P. Chaires Rosas, N. A. Vazquez Torres, B. Hernandez Tellez, G. Pinon Zarate, M. Herrera Enriquez, A. E. Castell Rodriguez

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The current approach to the treatment of bone defects involves the use of scaffolds that provide a biological and mechanically stable niche to favor tissue repair. Despite the significant progress in the field of bone tissue engineering, several main problems associated are attributed to giving a low biodegradation degree, does not promote osseointegration and regeneration, if the bone is not healing as well as expected or fails to heal, will not be given a proper ossification or new bone formation. The actual approaches of bone tissue regeneration are directed to the use of decellularized native extracellular matrices, which are able of retain their own architecture, mechanic properties, biodegradability and promote new bone formation because they are capable of conserving proteins and other factors that are founded in physiological concentrations. Therefore, we propose an extracellular matrix-based bioscaffolds derived from bovine cancellous bone, which is processed by decellularization, demineralization, and hydrolysis of the collagen protein, these protocols have been successfully carried out in other organs and tissues; the effectiveness of its biosafety has also been previously evaluated in vivo and Food and Drug Administration (FDA) approved. In the specific case of bone, a more complex treatment is needed in comparison with other organs and tissues because is necessary demineralization and collagen denaturalization. The present work was made in order to obtain a temporal scaffold that succeed in degradation in an inversely proportional way to the synthesis of extracellular matrix and the maturation of the bone by the cells of the host.

Keywords: bioactive, biodegradable, bone, extracellular matrix-based bioscaffolds, stem cells, tissue engineering

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Authors: Salama A. Ouf, Amera A. El-Adly, Abdelaleam H. Mohamed

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Dermatophytosis is the infection of keratinized tissues such as hair, nail and the stratum corneum of the skin by dermatophytic fungi. Infection is generally cutaneous and restricted to the non-living cornified layers because of the inability of the fungi to penetrate the deeper tissues or organs of immunocompetent hosts. In Saudi Arabia, Onychomycosis is the most frequent infection (40.3%), followed by tinea capitis (21.9%), tinea pedis (16%), tinea cruris (15.1%), and tinea corporis (6.7%). Several azole compounds have been tried to control dermatophytic infection, however, the azole-containing medicines may interfere with the activity of hepatic microsomal enzymes, sex and thyroid hormones, and testosterone biosynthesis. In this research, antibody-conjugated nanoparticles stimulated by cold plasma and laser were evaluated in vitro against some dermatophytes isolated from the common types of tinea. Different types of nanomaterials were tested but silver nanoparticles (AgNPs) were proved to be most effective against the dermatophytes under test. The use of cold plasma coupled with antibody-conjugated nano-particles has severe impact on dermatophytes where the inhibition of growth, spore germination keratinase activity was more than 88% in the case of Trichophyton rubrum, T. violaceum, Microsprum canis and M. gypseum. Complete inhibition of growth for all dermatophytes was brought about by the interaction of conjugated nanoparticles, with cold plasma and laser treatment. The in vivo test with inoculated guinea pigs achieved promising results where the recovery from the infection reached 95% in the case of M. canis –inoculated pigs treated with AgNPs pretreated with cold plasma and laser.

Keywords: cold plasma, dermatophytes, laser, silver nanoparticles

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Authors: Kazushi Murata, Fabian Watermann, O. B. Herve Gonroudobou, Le Thuy Hang, Toshiro Yamanaka, M. Larry Lopez C.

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Aims: Black pine coastal forests play an important role as a windbreak and as a natural barrier to sand and salt spray inland in Japan. The recent invasion of N₂-fxing black locust (Robinia pseudoacacia) trees in these forests is expected to have a nutritional contribution to black pine trees growth. Thus, the effect of this new source of N on black pine trees' N assimilation needs to be assessed. Methods: In order to evaluate this contribution, tree-ring isotopic composition (δ¹⁵N) and nitrogen content (%N) of black pine (Pinus thunbergii) trees in a pure stand (BPP) and a mixed stand (BPM) with black locust (BL) trees were measured for the period 2000–2019 for BPP and BL and 1990–2019 for BPM. The same measurements were conducted in plant tissues and in soil samples. Results: The tree ring δ15N values showed that for the last 30 years, BPM trees gradually switched from BPP to BL-derived soil N starting in the 1990s, becoming the dominant N source from 2000 as no significant diference was found between BPM and BL tree ring δ¹⁵N values from 2000 to 2019. No difference in root and sapwood BPM and BL δ¹⁵N values were found, but BPM foliage (−2.1‰) was different to BPP (−4.4‰) and BL (−0.3‰), which is related to the different N assimilation pathways between BP and BL. Conclusions: Based on the results of this study, the assimilation of BL-derived N inferred from the BPM tissues' δ¹⁵N values is the result of an increase in soil bioavailable N with a higher δ¹⁵N value.

Keywords: nitrogen-15, N₂-fxing species, mixed stand, soil, tree rings

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Authors: Saad M. Howladar

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Although, there are a little data on the use of littoral plants as heavy metals bioaccumulators over large areas of the wetlands environment. So, soil samples and biomass of the five plant species: Pluchea dioscroides, Pulicaria crispa, Lavandula pubescens, Tarchononthus comporatus and Argemone ochroleuca were collected from two different sites (basin and mouth) of four dams at Baha province, KSA. Nutrients and heavy metals were extracted from plant samples (leaves and stems) for analyzing elements (Na, K, Ca, P and N) and heavy metals (Pb, Cu and Ni). The soils of the mouth of the dam had the highest concentrations of all elements, while that of basin had the highest ones of most heavy metals except Pb. The soil elements in relation to the two sites arranged as: Ca > K > P > Na > N; and the heavy metals as: Cu > Ni > Pb. The present study indicated that Pluchea dioscroides had the highest values of most elements and heavy metals, while Lavandula pubescens had the lowest. In general, leaves attain the highest concentrations of all nutrients and heavy metals in most studied species as compared with stem. It was indicated that Pluchea dioscroides showed a high transfer factor for almost elements and heavy metals such as K, Na, Cu, Ni and Pb, while Pulicaria crispa showed the highest translocation factor of N, P, Ca-Na ratio and Cu. All studied species growing in the basin had almost the highest concentrations of elements and heavy metals as compared with that in the mouth of dam except K in Pluchea dioscroides, Tarchononthus comporatus and Argemone ochroleuca tissues. Otherwise tissues of Tarchononthus comporatus growing in the basin had the lowest concentrations of K and Ni, while that growing in the mouth had the highest of P and N.

Keywords: Baha Region, bioindicators, plant, pollution, dams, heavy metals

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Authors: Sayed Ali Garossi, Azar Heidarizadi, Shahla Mohammad Ganji

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Context: Colorectal cancer is the second type of cancer-related mortality globally. Expression of cas8 (caspase 8) is closely connected to growth and metastasis of colorectal cancer.Cas8/Rip1 plays a vital role in the apoptosis pathway and resistance to chemotherapy. The aim of the present study is to investigate the pattern of gene expression in colorectal cancer and compare the differences using Real-Time PCR to find a potential biomarker candidate for colorectal cancer. Methodology: This study conducted real-time PCR to evaluate gene expression of Cas8 in colorectal cancer patients. The gene-specific primer sequences exon–exon junction was designed by OLIGO7 software for the expression of the gene under investigation. Forty-six patient samples without any chemotherapy were selected, including tumoral tissue and adjacent normal tissue samples. The age of the patients was 50 and the size of the tumors was 5.5 cm. The categories were before and after age 50. Findings: Here, we found that Caspase 8 was overexpressed in CRC tissues compared to corresponding adjacent colon tissues (Cas8: 5.2 vs. 1 ratio); high expression of Cas8 was associated with poor overall survival and independent risk factors for the prognosis of CRC patients. Conclusion: In conclusion, our study pioneered the reporting of high Casp8 expression as a predictor of poor prognosis and chemical resistance in CRC patients.Cas8 overexpression suppressed Cas 8 / Rip1-dependent apoptosis and activated the proliferation of tumor cells by activating necroptosis. The necroptosis pathway has also emerged as a new approach to anti-tumor in cancer treatment.

Keywords: Cas8, necroptosis, apoptosis, Real-Time PCR

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Authors: MyungGu Yeo, JongHan Ha, Gi-Hoon Yang, JaeYoon Lee, SeungHyun Ahn, Hyeongjin Lee, HoJun Jeon, YongBok Kim, Minseong Kim, GeunHyung Kim

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Tissue engineering is a rapidly growing interdisciplinary research area that may provide options for treating damaged tissues and organs. As a promising technique for regenerating various tissues, this technology requires biomedical scaffolds, which serve as an artificial extracellular matrix (ECM) to support neotissue growth. Electrospun micro/nanofibers have been used widely in tissue engineering because of their high surface-area-to-volume ratio and structural similarity to extracellular matrix. However, low mechanical sustainability, low 3D shape-ability, and low cell infiltration have been major limitations to their use. In this work, we propose new hybrid scaffolds interlayered with cell-laden electrospun micro/nano fibers and poly(caprolactone) microstructures. Also, we applied various concentrations of alginate and electric field strengths to determine optimal conditions for the cell-electrospinning process. The combination of cell-laden bioink (2 ⅹ 10^5 osteoblast-like MG63 cells/mL, 2 wt% alginate, 2 wt% poly(ethylene oxide), and 0.7 wt% lecithin) and a 0.16 kV/mm electric field showed the highest cell viability and fiber formation in this process. Using these conditions and PCL microstructures, we achieved mechanically stable hybrid scaffolds. In addition, the cells embedded in the fibrous structure were viable and proliferated. We suggest that the cell-embedded hybrid scaffolds fabricated using the cell-electrospinning process may be useful for various soft- and hard-tissue regeneration applications.

Keywords: bioink, cell-laden scaffold, micro/nanofibers, poly(caprolactone)

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Authors: Marek Pawlikowski, Krzysztof Jankowski, Konstanty Skalski, Anna Makuch

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Nanoindentation or depth-sensing indentation (DSI) technique has proven to be very useful to measure mechanical properties of various tissues at a micro-scale. Bone tissue, both trabecular and cortical one, is one of the most commonly tested tissues by means of DSI. Most often such tests on bone samples are carried out to compare the mechanical properties of lamellar and interlamellar bone, osteonal bone as well as compact and cancellous bone. In the paper, a relation between stress and strain for human trabecular bone is presented. The relation is based on the results of nanoindentation tests. The formulation of a constitutive model for human trabecular bone is based on nanoindentation tests. In the study, the approach proposed by Olivier-Pharr is adapted. The tests were carried out on samples of trabecular tissue extracted from human femoral heads. The heads were harvested during surgeries of artificial hip joint implantation. Before samples preparation, the heads were kept in 95% alcohol in temperature 4 Celsius degrees. The cubic samples cut out of the heads were stored in the same conditions. The dimensions of the specimens were 25 mm x 25 mm x 20 mm. The number of 20 samples have been tested. The age range of donors was between 56 and 83 years old. The tests were conducted with the indenter spherical tip of the diameter 0.200 mm. The maximum load was P = 500 mN and the loading rate 500 mN/min. The data obtained from the DSI tests allows one only to determine bone behoviour in terms of nanoindentation force vs. nanoindentation depth. However, it is more interesting and useful to know the characteristics of trabecular bone in the stress-strain domain. This allows one to simulate trabecular bone behaviour in a more realistic way. The stress-strain curves obtained in the study show relation between the age and the mechanical behaviour of trabecular bone. It was also observed that the bone matrix of trabecular tissue indicates an ability of energy absorption.

Keywords: constitutive model, mechanical behaviour, nanoindentation, trabecular bone

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Authors: Abdul Soofi, Katherine Wolf, Egon Ranghini, Gregory Dressler

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Obesity and its associated complications, such as insulin resistance and non-alcoholic fatty liver disease, are reaching epidemic proportions. In mice, the TGF-β superfamily is implicated in the regulation of white and brown adipose tissues differentiation. The Kielin/Chordin-like Protein (KCP) is a secreted regulator of the TGF-β superfamily pathways that can inhibit both TGF-β and Activin signals while enhancing the Bone Morphogenetic protein (BMP) signaling. However, the effects of KCP on metabolism and obesity have not been studied in animal models. Thus, we examined the effects of KCP loss or gain of function in mice that were maintained on either a regular or a high fat diet. Loss of KCP sensitized mice to obesity and associated complications such as hepatic steatosis and glucose intolerance. In contrast, transgenic mice that expressed KCP in the kidney, liver and adipose tissues were resistant to developing high fat diet induced obesity and had significantly reduced white adipose tissue. KCP over-expression was able to shift the pattern of Smad signaling in vivo, to increase the levels of P-Smad1 and decrease P-Smad3, resulting in resistance to high fat diet induced hepatic steatosis and glucose intolerance. In aging mice, loss of KCP promoted liver pathology even when mice were fed a normal diet. The data demonstrate that shifting the TGF-β superfamily signaling with a secreted inhibitor or enhancer can alter the physiology of adipose tissue to reduce obesity and can inhibit the initiation and progression of hepatic steatosis to significantly reduce the effects of high fat diet induced metabolic disease.

Keywords: adipose tissue, KCP, obesity, TGF-β, BMP, hepatic steatosis, metabolic syndrome

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Authors: Azza El-Medany, Jamila El-Medany

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Residential and agricultural pesticide use is widespread in the world. Their extensive and indiscriminative use, in addition with their ability to interact with biological systems other than their primary targets constitute a health hazards to both humans and animals. The toxic effects of pesticides include alterations in metabolism; there is a lack of knowledge that organophosphates can cause pancreatic toxicity. The primary goal of this work is to study the effects of chronic exposure to Diazinon an organophosphate used in agriculture on pancreatic tissues and evaluate the ameliorating effect of Mesna as antioxidant on the toxicity of Diazinon on pancreatic tissues.40 adult male rats, their weight ranged between 300-350 g. The rats were classified into three groups; control (10 rats) was received corn oil at a dose of 1 0 mg/kg/day by gavage once a day for 2 months. Diazinon (15 rats) was received Diazinon at a dose of 10 mg/kg/day dissolved in corn oil by gavage once a day for 2 months. Treated group (15 rats), were received Mesna 180mg/kg once a week by gavage 15 minutes before administration of Diazinon for 2 months. At the end of the experiment, animals were anesthetized, blood samples were taken by cardiac puncture for glucose and insulin assays and pancreas was removed and divided into 3 portions; first portion for histopathological study; second portion for ultrastructural study; third portion for biochemical study using Elisa Kits including determination of malondialdehyde (MDA), tumor necrosis factor α (TNF-α), myeloperoxidase activity (MPO), interleukin 1β (IL-1β). A significant increase in the levels of MDA, TNF-α, MPO activity, IL-1β, serum glucose levels in the toxicated group with Diazinon were observed, while a significant reduction was noticed in GSH in serum insulin levels. After treatment with Mesna a significant reduction was observed in the previously mentioned parameters except that there was a significant rise in GSH in insulin levels. Histopathological and ultra-structural studies showed destruction in pancreatic tissues and β cells were the most affected cells among the injured islets as compared with the control group. The current study try to spot light about the effects of chronic exposure to pesticides on vital organs as pancreas also the role of oxidative stress that may be induced by them in evoking their toxicity. This study shows the role of antioxidant drugs in ameliorating or preventing the toxicity. This appears to be a promising approach that may be considered as a complementary treatment of pesticide toxicity.

Keywords: Diazinon, reduced glutathione, myeloperoxidase activity, tumor necrosis factor α, Mesna

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Authors: Naeim Akbari Shahkhosravi, Reza Kakavand, Helen M. S. Davies, Amin Komeili

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Equine locomotion and performance are significantly affected by hoof health. One of the most critical diseases of the hoof is laminitis, which can lead to horse lameness in a severe condition. This disease exhibits the mechanical properties degradation of the laminar junction tissue within the hoof. Therefore, it is essential to investigate the biomechanics of the hoof, focusing specifically on excessive and cumulatively accumulated stresses within the laminar junction tissue. For this aim, the current study generated a novel equine hoof Finite Element (FE) model under dynamic physiological loading conditions and employing a hyperelastic material model. Associated tissues of the equine hoof were segmented from computed tomography scans of an equine forelimb, including the navicular bone, third phalanx, sole, frog, laminar junction, digital cushion, and medial- dorsal- lateral wall areas. The inner tissues were connected based on the hoof anatomy, and the hoof was under a dynamic loading over cyclic strides at the trot. The strain distribution on the hoof wall of the model was compared with the published in vivo strain measurements to validate the model. Then the validated model was used to study the development of laminitis. The ultimate stress tolerated by the laminar junction before rupture was considered as a stress threshold. The tissue damage was simulated through iterative reduction of the tissue’s mechanical properties in the presence of excessive maximum principal stresses. The findings of this investigation revealed how damage initiates from the medial and lateral sides of the tissue and propagates through the hoof dorsal area.

Keywords: horse hoof, laminitis, finite element model, continuous damage

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Authors: Martyna Ekiert, Andrzej Mlyniec

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Tendons are the biological structures, which primary function is to transfer force generated by muscles to the bones. Unfortunately, damages of tendons are also one of the most common injuries of the human musculoskeletal system. For the most severe cases of tendon rupture, such as the tear of calcaneus tendon or anterior cruciate ligament of the knee, a surgical procedure is the only possible way of full recovery. Tendons used as biological grafts are usually subjected to the process of deep freezing and subsequent thawing. This, in particular for multiple freezing/thawing cycles, may result in changes of tendon internal structure causing deterioration of mechanical properties of the tissue. Therefore, studies on the influence of freezing on tendons biomechanics, including internal water content in soft tissue, seems to be greatly needed. An experimental study of the influence of freezing on mechanical properties of the tendon was performed on fascicles samples dissected form bovine flexor tendons. The preparation procedure was performed with the presence of 0.9% saline solution in order to prevent an excessive tissue drying. All prepared samples were subjected to the different number of freezing/thawing cycles. For freezing part of the protocol we used -80°C temperature while for slow thawing we used fridge temperature (4°C) combined with equalizing temperatures in the standard state (25°C). After final thawing, the mechanical properties of each sample was examined using cyclic loading test. Our results may contribute for better understanding of negative effects of soft tissues freezing, resulting from abnormal thermal expansion of water. This also may help to determine the limit of freezing/thawing cycles disqualifying tissue for surgical purposes and thus help optimize tissues storage conditions.

Keywords: freezing, soft tissue, tendon, bovine fascicles

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Authors: Paola Modesto, Floriana Fruscione, Isabella Martini, Simona Perga, Federica Riccardo, Mariateresa Camerino, Davide Giacobino, Cecilia Gola, Luca Licenziato, Elisabetta Razzuoli, Katia Varello, Lorella Maniscalco, Elena Bozzetta, Angelo Ferrari

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Canine and human melanoma, osteosarcoma (OSA), and mammary carcinomas are aggressive tumors with common characteristics making dogs a good model for comparative oncology. Novel therapeutic strategies against these tumors could be useful to both species. In humans, chondroitin sulphate proteoglycan 4 (CSPG4) is a marker involved in tumor progression and could be a candidate target for immunotherapy. The anti-CSPG4 DNA electrovaccination has shown to be an effective approach for canine malignant melanoma (CMM) [1]. An immunohistochemistry evaluation of CSPG4 expression in tumour tissue is generally performed prior to electrovaccination. To assess the possibility to perform a rapid molecular evaluation and in order to validate these spontaneous canine tumors as the model for human studies, we investigate the CSPG4 gene expression by RT qPCR in CMM, OSA, and canine mammary tumors (CMT). The total RNA was extracted from RNAlater stored tissue samples (CMM n=16; OSA n=13; CMT n=6; five paired normal tissues for CMM, five paired normal tissues for OSA and one paired normal tissue for CMT), retro-transcribed and then analyzed by duplex RT-qPCR using two different TaqMan assays for the target gene CSPG4 and the internal reference gene (RG) Ribosomal Protein S19 (RPS19). RPS19 was selected from a panel of 9 candidate RGs, according to NormFinder analysis following the protocol already described [2]. Relative expression was analyzed by CFX Maestro™ Software. Student t-test and ANOVA were performed (significance set at P<0.05). Results showed that gene expression of CSPG4 in OSA tissues is significantly increased by 3-4 folds when compared to controls. In CMT, gene expression of the target was increased from 1.5 to 19.9 folds. In melanoma, although an increasing trend was observed, no significant differences between the two groups were highlighted. Immunohistochemistry analysis of the two cancer types showed that the expression of CSPG4 within CMM is concentrated in isles of cells compared to OSA, where the distribution of positive cells is homogeneous. This evidence could explain the differences in gene expression results.CSPG4 immunohistochemistry evaluation in mammary carcinoma is in progress. The evidence of CSPG4 expression in a different type of canine tumors opens the way to the possibility of extending the CSPG4 immunotherapy marker in CMM, OSA, and CMT and may have an impact to translate this strategy modality to human oncology.

Keywords: canine melanoma, canine mammary carcinomas, canine osteosarcoma, CSPG4, gene expression, immunotherapy

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Authors: Lola T. Alimkhodjaeva, Lola T. Zakirova, Soniya S. Ziyavidenova

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Background: Breast cancer occupies the first place among the cancer in women in the world. It is urgent today to study the role of molecular markers which are capable of predicting the dynamics and outcome of the disease. The aim of this study is to define the prognostic value of the content of estrogen receptor (ER), progesterone receptor (PgR), and amplification of HER-2 / neu oncoprotein by studying 3 and 5-year overall and relapse-free survival in 470 patients with primary operable and 280 patients with locally–advanced breast cancer. Materials and methods: Study results of 3 and 5-year overall and relapse-free survival, depending on the content of RE, PgR in primary operable patients showed that ER positive (+) and PgR (+) survival was 100 (96.2%) and 97.3 (94.6%), for ER negative (-) and PgR (-) - 69.2 (60.3%) and 65.4 (57.7%), for ER positive (+) and negative PgR (-) 87.4 (80.1%) and 81.5 (79.3%), for ER negative (-) and positive PgR (+) - 97.4 (93.4%) and 90.4 (88.5%), respectively. Survival results depended also on the level of HER-2 / neu expression. In patients with HER-2 / neu negative the survival rates were as follows: 98.6 (94.7%) and 96.2 (92.3%). In group of patients with the level of HER-2 / neu (2+) expression these figures were: 45.3 (44.3%) and 45.1 (40.2%), and in group of patients with the level of HER-2 / neu (3+) expression - 41.2 (33.1%) and 34.3 (29.4%). The combination of ER negative (-), PgR (-), HER-2 / neu (-) they were 27.2 (25.4%) and 19.5 (15.3%), respectively. In patients with locally-advanced breast cancer the results of 3 and 5-year OS and RFS for ER (+) and PgR (+) were 76.3 (69.3%) and 62.2 (61.4%), for ER (-) and RP (-) 29.1 (23.7%) and 18.3 (12.6%), for ER (+) and PgR (-) 61.2 (47.2%) and 39.4 (25.6%), for ER (-) and PgR (+) 54.3 (43.1%) and 41.3 (18.3%), respectively. The level of HER-2 / neu expression also affected the survival results. Therefore, in HER-2/ neu negative patients the survival rate was 74.1 (67.6%) and 65.1 (57.3%), with the level of expression (2+) 20.4 (14.2%) and 8.6 (6.4%), with the level of expression (3+) 6.2 (3.1%) and 1.2 (1.5%), respectively. The combination for ER, PgR, HER-2 / neu negative was 22.1 (14.3%) and 8.4 (1.2%). Conclusion: Thus, the presence of steroid hormone receptors in breast tumor tissues at primary operable and locally- advanced process as the lack of HER-2/neu oncoprotein correlates with the highest rates of 3- and 5-year overall and relapse-free survival. The absence of steroid hormone receptors as well as of HER-2/neu overexpression in malignant breast tissues significantly degrades the 3- and 5-year overall and relapse-free survival. Tumors with ER, PgR and HER-2/neu negative have the most unfavorable prognostics.

Keywords: breast cancer, estrogen receptor, oncoprotein, progesterone receptor

Procedia PDF Downloads 156

Authors: Hande Yesil, Seda Aycan Altan, M. Vehbi Bal, Alper Uyar, O. Cumhur Sipahi

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Purpose: Fixed partial dentures (FPDs) must fulfill functional requirements such as phonetics, chewing efficiency and esthetics especially in the anterior region. A convex type tissue surface is usually recommended for pontics of FPDs. That pontic design also provides suitable oral hygiene and ease of cleaning. However, high esthetic requirements and correct emergence profile are not always achievable because of the convex shape of adjacent soft tissues. Therefore, the ovate type pontic which fulfills the high esthetic demands of the patients may be a good alternative to the modified ridge lap pontic design. Clinical Report: A female patient referred with the complaint of anterior upper edentulousness. In the oral examination it was determined that teeth 11, 12, 21, 22 were deficient. A thick and convex gingival tissue that may cause aesthetic problems was also observed.. Periodontal augmentation surgery was performed to ensure proper papillary configuration and gingival contour. An interim removable partial denture (IRPD) which applied pressure to operated gingival tissues was fabricated postoperatively. The IRPD was used for 4 weeks and after completion of tissue sculpting, the permanent FPD with an ovate pontic was fabricated and cemented. After a follow-up period of 6 months, not any esthetical and hygienic problem was detected and the patient was satisfied with her prosthesis. Conclusion: It was concluded that shaping of gingival contours with IRPD and use of a FPD with ovate pontic fulfills all esthetic and hygienic requirements.

Keywords: interim appliance, ovate pontic, tissue sculpturing, fixed partial denture

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Authors: Mihail Nagea, Olivera Lupescu, Nicolae Ciurea, Alexandru Dimitriu, Alina Grosu

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Introduction: As severe open injuries are more and more frequent in modern traumatology, threatening not only the integrity of the affected limb but even the life of the patients, new methods desired to cope with the consequences of these traumas were described. Vacuum therapy is one such method which has been described as enhancing healing in trauma with extensive soft-tissue injuries, included those with septic complications. Material and methods: Authors prospectively analyze 15 patients with severe lower limb trauma with MESS less than 6, with considerable soft tissue loss following initial debridement and fracture fixation. The patients needed serial debridements and vacuum therapy was applied after delayed healing due to initial severity of the trauma, for an average period of 12 days (7 - 23 days).In 7 cases vacuum therapy was applied for septic complications. Results: Within the study group, there were no local complications; secondary debridements were performed for all the patients and vacuum system was re-installed after these debridements. No amputations were needed. Medical records were reviewed in order to compare the outcome of the patients: the hospital stay, anti-microbial therapy, time to healing of the bone and soft tissues (there is no standard group to be compared with) and the result showed considerable improvements in the outcome of the patients. Conclusion: Vacuum therapy improves healing of the soft tissues, including those infected; hospital stay and the number of secondary necessary procedures are reduced. Therefore it is considered a valuable support in treating trauma of the limbs with severe soft tissue injuries.

Keywords: complex injuries, negative pressure, open fractures, wound therapy

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Authors: Maria Karnachoriti, Ellas Spyratou, Dimitrios Lykidis, Maria Lambropoulou, Yiannis S. Raptis, Ioannis Seimenis, Efstathios P. Efstathopoulos, Athanassios G. Kontos

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Colorectal cancer is the third most common cancer diagnosed in Europe, according to the latest incidence data provided by the World Health Organization (WHO), and early diagnosis has proved to be the key in reducing cancer-related mortality. In cases where surgical interventions are required for cancer treatment, the accurate discrimination between healthy and cancerous tissues is critical for the postoperative care of the patient. The current study focuses on the ex vivo handling of surgically excised colorectal specimens and the acquisition of their spectral fingerprints using Raman spectroscopy. Acquired data were analyzed in an effort to discriminate, in microscopic scale, between healthy and malignant margins. Raman spectroscopy is a spectroscopic technique with high detection sensitivity and spatial resolution of few micrometers. The spectral fingerprint which is produced during laser-tissue interaction is unique and characterizes the biostructure and its inflammatory or cancer state. Numerous published studies have demonstrated the potential of the technique as a tool for the discrimination between healthy and malignant tissues/cells either ex vivo or in vivo. However, the handling of the excised human specimens and the Raman measurement conditions remain challenging, unavoidably affecting measurement reliability and repeatability, as well as the technique’s overall accuracy and sensitivity. Therefore, tissue handling has to be optimized and standardized to ensure preservation of cell integrity and hydration level. Various strategies have been implemented in the past, including the use of balanced salt solutions, small humidifiers or pump-reservoir-pipette systems. In the current study, human colorectal specimens of 10X5 mm were collected from 5 patients up to now who underwent open surgery for colorectal cancer. A novel, non-toxic zinc-based fixative (Z7) was used for tissue preservation. Z7 demonstrates excellent protein preservation and protection against tissue autolysis. Micro-Raman spectra were recorded with a Renishaw Invia spectrometer from successive random 2 micrometers spots upon excitation at 785 nm to decrease fluorescent background and secure avoidance of tissue photodegradation. A temperature-controlled approach was adopted to stabilize the tissue at 2 °C, thus minimizing dehydration effects and consequent focus drift during measurement. A broad spectral range, 500-3200 cm-1,was covered with five consecutive full scans that lasted for 20 minutes in total. The average spectra were used for least square fitting analysis of the Raman modes.Subtle Raman differences were observed between normal and cancerous colorectal tissues mainly in the intensities of the 1556 cm-1 and 1628 cm-1 Raman modes which correspond to v(C=C) vibrations in porphyrins, as well as in the range of 2800-3000 cm-1 due to CH2 stretching of lipids and CH3 stretching of proteins. Raman spectra evaluation was supported by histological findings from twin specimens. This study demonstrates that Raman spectroscopy may constitute a promising tool for real-time verification of clear margins in colorectal cancer open surgery.

Keywords: colorectal cancer, Raman spectroscopy, malignant margins, spectral fingerprints

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Authors: Maryam Shirmohammadi, Prasad K. D. V. Yarlagadda, YuanTong Gu

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The mechanical damage on the agricultural crop during and after harvesting can create high volume of damage on tissue. Uniaxial compression and tensile loading were performed on flesh and peel samples of pumpkin. To investigate the structural changes on the tissue, Scanning Electron Microscopy (SEM) was used to capture the cellular structure change before and after loading on tissue for tensile, compression and indentation tests. To obtain required mechanical properties of tissue for the finite element analysis (FEA) model, laser measurement sensors were used to record the lateral displacement of tissue under the compression loading. Uniaxial force versus deformation data were recorded using Universal Testing Machine for both tensile and compression tests. The experimental Results were employed to develop a material model with failure criteria. The results obtained by the simulation were compared with those obtained by experiments. Note that although modelling food materials’ behaviour is not a new concept however, majority of previous studies focused on elastic behaviour and damages under linear limit, this study, however, has developed FEA models for tensile and compressive loading of pumpkin flesh and peel samples using, as the first study, both elastic and elasto-plastic material types. In addition, pumpkin peel and flesh tissues were considered as two different materials with different properties under mechanical loadings. The tensile and compression loadings were used to develop the material model for a composite structure for FEA model of mechanical peeling of pumpkin as a tough skinned vegetable.

Keywords: compressive and tensile response, finite element analysis, poisson’s ratio, elastic modulus, elastic and plastic response, rupture and bio-yielding

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