Search results for: small cell

Authors: M. Hosseinnejad, K. Gharanjig

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In the present study, metal free organic dyes were prepared and used as photo-sensitizers in dye-sensitized solar cells. Double rhodanine was utilized as the fundamental electron acceptor group to which electron donor aldehyde with varying substituents was attached to produce new organic dye. This dye was first purified and then characterized by analytical techniques. Spectrophotometric evaluations of the prepared dye in solution and on a nano anatase TiO₂ substrate were carried out in order to assess possible changes in the status of the dyes in different environments. The results show that the dye form j-type aggregates on the nano TiO₂. Additionally, oxidation potential measurements were also carried out. Finally, dye sensitized solar cell based on synthesized dye was fabricated in order to determine the photovoltaic behavior and conversion efficiency of individual dye.

Keywords: conversion efficiency, dye-sensitized solar cell, photovoltaic behavior, sensitizer

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Authors: B. Fazekas, I. Korolov, K. Kutasi

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Plasma medicine, which involves the use of gas discharge plasmas for medical applications is a rapidly growing research field. The use of non-thermal atmospheric pressure plasmas in dermatology to assist tissue regeneration by improving the healing of infected and/or chronic wounds is a promising application. It is believed that plasma can activate cells, which are involved in the wound closure. Non-thermal atmospheric plasmas are rich in chemically active species (such as O and N-atoms, O2(a) molecules) and radiative species such as the NO, N2+ and N2 excited molecules, which dominantly radiate in the 200-500 nm spectral range. In order to understand the effect of plasma species, both of chemically active and radiative species on wound healing process, the interaction of physical plasma with the human skin cells is necessary. In order to clarify the effect of plasma radiation on the wound healing process we treated keratinocyte cells – that are one of the main cell types in human skin epidermis – covered with a layer of phosphate-buffered saline (PBS) with a low power atmospheric pressure plasma. For the generation of such plasma we have applied a plasma needle. Here, the plasma is ignited at the tip of the needle in flowing helium gas in contact with the ambient air. To study the effect of plasma radiation we used a plasma needle configuration, where the plasma species – chemically active radicals and charged species – could not reach the treated cells, but only the radiation. For the comparison purposes, we also irradiated the cells using a UV-B light source (FS20 lamp) with a 20 and 40 mJ cm-2 dose of 312 nm. After treatment the viability and the proliferation of the cells have been examined. The proliferation of cells has been studied with a real time monitoring system called Xcelligence. The results have indicated, that the 20 mJ cm-2 dose did not affect cell viability, whereas the 40 mJ cm-2 dose resulted a decrease in cell viability. The results have shown that the plasma radiation have no quantifiable effect on the cell proliferation as compared to the non-treated cells.

Keywords: UV radiation, non-equilibrium gas discharges (non-thermal plasmas), plasma emission, keratinocyte cells

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Authors: Bhavya Tripathi, Bhupendra Kumar Sharma

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In blood, the concentration of red blood cell varies with the arterial diameter. In the case of narrow arteries, red blood cells concentrate around the center of the artery and there exists a cell-free plasma layer near the arterial wall due to Fahraeus-Lindqvist effect. Due to non- uniformity of the fluid in the narrow arteries, it is preferable to consider the two-phase model of the blood flow. In the present article, coupled nonlinear differential equations have been developed for momentum, energy and concentration of two phase model of the blood flow assuming the Newtonian fluid in both central core and cell free plasma layer and the exact solutions have been found for the problem. For having an adequate insight into the stenosed arterial two-phase blood flow, major components of the flow as flow resistance, total flow rate, and wall shear stress have been estimated for different values of magnetic and radiation parameter. Results show that the increase in the effects of magnetic field decreases the velocity of both cores as well as plasma regions. This result can be helpful to control the blood flow in narrow arteries during surgical process. Temperature of core as well plasma regions decrease as value of radiation parameter increases. The present result is implemented in the form of radiation therapy which is very helpful for cancer patients.

Keywords: two phase blood flow, radiation, magnetohydrodynamics (MHD), stenosis

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Authors: Chien Y. Lin, Jung Y. Huang, Leu-Wei Lo

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A growing body of data reveals that the membrane cholesterol molecules can alter the signaling pathways of living cells. However, the understanding about how membrane cholesterol modulates receptor proteins is still lacking. Single-molecule tracking can effectively probe into the microscopic environments and thermal fluctuations of receptor proteins in a living cell. In this study we applies single-molecule optical tracking on ligand-induced dimerization process of EGFRs in the plasma membranes of two cancer cell lines (HeLa and A431) and one normal endothelial cell line (MCF12A). We tracked individual EGFR and dual receptors, diffusing in a correlated manner in the plasma membranes of live cells. We developed an energetic model by integrating the generalized Langevin equation with the Cahn-Hilliard equation to help extracting important information from single-molecule trajectories. From the study, we discovered that ligand-bound EGFRs move from non-raft areas into lipid raft domains. This ligand-induced motion is a common behavior in both cancer and normal cells. By manipulating the total amount of membrane cholesterol with methyl-β-cyclodextrin and the local concentration of membrane cholesterol with nystatin, we further found that the amount of cholesterol can affect the stability of EGFR dimers. The EGFR dimers in the plasma membrane of normal cells are more sensitive to the local concentration changes of cholesterol than EGFR dimers in the cancer cells. Our method successfully captures dynamic interactions of receptors at the single-molecule level and provides insight into the functional architecture of both the diffusing EGFR molecules and their local cellular environment.

Keywords: membrane proteins, single-molecule tracking, Cahn-Hilliard equation, EGFR dimers

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Authors: Chinedu Peter, Dashrath Singh

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Membrane computing is a computability model which abstracts its structures and functions from the biological cell. The main ingredient of membrane computing is the notion of a membrane structure, which consists of several cell-like membranes recurrently placed inside a unique skin membrane. The emergence of several variants of membrane computing gives rise to the notion of a P system. The paper presents a variant of P systems for arithmetic operations on non-negative integers based on the weak priorities for rule application. Consequently, we obtain deterministic P systems. Two membranes suffice. There are at most four objects for multiplication and five objects for division throughout the computation processes. The model is simple and has a potential for possible extension to non-negative integers and real numbers in general.

Keywords: P system, binary operation, determinism, weak rule priority

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Authors: Shen Jian, Hu Jie, Ma Jin, Peng Ying Hong, Fang Yi, Liu Wen Hai

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Biologically inspired design inspires inventions and new technologies in the field of engineering by mimicking functions, principles, and structures in the biological domain. To deal with the obstacles of cross-domain knowledge acquisition in the existing biologically inspired design process, functional semantic clustering based on functional feature semantic correlation and environmental constraint clustering composition based on environmental characteristic constraining adaptability are proposed. A knowledge cell clustering algorithm and the corresponding prototype system is developed. Finally, the effectiveness of the method is verified by the visual prosthetic device design.

Keywords: knowledge clustering, knowledge acquisition, knowledge based engineering, knowledge cell, biologically inspired design

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Authors: Sanja Pfeifer, Nataša Šarlija, Marina Jeger, Ana Bilandžić

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The purpose of this study is to explore the different facets of growth among micro, small and medium-sized firms in Croatia and to analyze the differences between models designed for all micro, small and medium-sized firms and those in creative industries. Three growth prediction models were designed and tested using the growth of sales, employment and assets of the company as dependent variables. The key drivers of sales growth are: prudent use of cash, industry affiliation and higher share of intangible assets. Growth of assets depends on retained profits, internal and external sources of financing, as well as industry affiliation. Growth in employment is closely related to sources of financing, in particular, debt and it occurs less frequently than growth in sales and assets. The findings confirm the assumption that growth strategies of small and medium-sized enterprises (SMEs) in creative industries have specific differences in comparison to SMEs in general. Interestingly, only 2.2% of growing enterprises achieve growth in employment, assets and sales simultaneously.

Keywords: creative industries, growth prediction model, growth determinants, growth measures

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Authors: Vipulkumar Jagodana

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Recently the use of renewable sources has increased, these sources include fuel cell, photo voltaic, and wind turbine. Islanding occurs when one portion of grid is isolated from remaining grid. Use of the renewable sources can provide continuous power to isolated portion in islanding condition. One of the common renewable sources is wind generation using wind turbine. The efficiency of wind generation can be increased in combination with conventional sources. When islanding occurs, few parameters change which may be frequency, voltage, active power, and harmonics. According to large change in one of these parameters islanding is detected. In this paper, two passive methods Rate of Change of Frequency (ROCOF) and Rate of change of Power (ROCOP) have been implemented for islanding detection of small wind-turbine. Islanding detection of both methods have been simulated in PSCAD. Simulation results show at different islanding inception angle response of ROCOF and ROCOP.

Keywords: islanding, adopted methods, PSCAD simulation, comparison

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Authors: Mohamed Dhouibi, Bogdan Stirbu, Chabotier André, Marc Pirlot

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Internal barrel wear is a major factor affecting the performance of small caliber guns in their different life phases. Wear analysis is, therefore, a very important process for understanding how wear occurs, where it takes place, and how it spreads with the aim on improving the accuracy and effectiveness of small caliber weapons. This paper discusses the measurement and analysis of combustion chamber wear for a small-caliber gun using a Coordinate Measuring Machine (CMM). Initially, two different NATO small caliber guns: 5.56x45mm and 7.62x51mm, are considered. A Micura Zeiss Coordinate Measuring Machine (CMM) equipped with the VAST XTR gold high-end sensor is used to measure the inner profile of the two guns every 300-shot cycle. The CMM parameters, such us (i) the measuring force, (ii) the measured points, (iii) the time of masking, and (iv) the scanning velocity, are investigated. In order to ensure minimum measurement error, a statistical analysis is adopted to select the reliable CMM parameters combination. Next, two measurement strategies are developed to capture the shape and the volume of each gun chamber. Thus, a task-specific measurement uncertainty (TSMU) analysis is carried out for each measurement plan. Different approaches of TSMU evaluation have been proposed in the literature. This paper discusses two different techniques. The first is the substitution method described in ISO 15530 part 3. This approach is based on the use of calibrated workpieces with similar shape and size as the measured part. The second is the Monte Carlo simulation method presented in ISO 15530 part 4. Uncertainty evaluation software (UES), also known as the Virtual Coordinate Measuring Machine (VCMM), is utilized in this technique to perform a point-by-point simulation of the measurements. To conclude, a comparison between both approaches is performed. Finally, the results of the measurements are verified through calibrated gauges of several dimensions specially designed for the two barrels. On this basis, an experimental database is developed for further analysis aiming to quantify the relationship between the volume of wear and the muzzle velocity of small caliber guns.

Keywords: coordinate measuring machine, measurement uncertainty, erosion and wear volume, small caliber guns

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Authors: Gunnar Oskarsson, Gudjon Helgi Egilsson

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Due to globalization, small and medium-sized enterprises (SME) increasingly offer their products and services in foreign markets. The main reasons are either to compensate for a decreased market share in their home market or to exploit opportunities in foreign markets, which are becoming less distant and better accessible than before. International markets are particularly important for companies located in a small economy and offering specialized products. Although more accessible, entering international markets is both expensive and difficult. In order to select the most appropriate markets, it is, therefore, important to gain an insight into the factors that have an impact on success, or potential failure. Although there has been a reasonable volume of research into the theory of internationalization, there is still a need to gain further understanding of the decision-making process of SMEs in small economies and the most important characteristics that distinguish between success and failure. The main objective of this research is to enhance knowledge on the internationalization of SMEs, including the drivers for the decision to internationalize, and the most important factors contributing to success in their internationalization activities. A qualitative approach was found to be most appropriate for this kind of research, with the objective of gaining a deeper understanding and discovering factors which impact a company’s decision-making and potential success. In-depth interviews were conducted with 14 companies in different industries located in Iceland, a country extensively dependent on export revenues. The interviews revealed several factors as drivers of internationalization and, not surprisingly, the most frequently mentioned source of motivation was that the local market is inadequate to maintain a sustainable operation. Good networking relationships were seen as a particular priority for potential success, searching for new markets was mainly carried out through the internet, although sales exhibitions and sales trips were also considered important. When it comes to the final decision as to whether a market should be considered for further analysis, economy, labor cost, legal environment, and cultural barriers were the most common factors to be weighted. The ultimate answer to successful internationalization, however, is largely dependent on a coordinated and experienced management team. The main contribution of this research is offering an insight into factors affecting decision-making in the internationalization process of SMEs, based on the opinion and experience of managers of SMEs in a small economy.

Keywords: internationalization, success factors, small and medium-sized enterprises (SMEs), drivers, decision making

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Authors: E. Kaygan, A. Gatto

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An investigation of adaptable winglets for morphing aircraft control and performance is described in this paper. The concepts investigated consist of various winglet configurations fundamentally centred on a baseline swept wing. The impetus for the work was to identify and optimize winglets to enhance controllability and the aerodynamic efficiency of a small unmanned aerial vehicle. All computations were performed with Athena Vortex Lattice modelling with varying degrees of twist, swept, and dihedral angle considered. The results from this work indicate that if adaptable winglets were employed on small scale UAV’s improvements in both aircraft control and performance could be achieved.

Keywords: aircraft, rolling, wing, winglet

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Authors: Felipa De Mello-Sampayo

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Background: The aim of this study is to undertake, from a patient perspective, an economic analysis of presurgical teledermatology, comparing it with a conventional referral system. Store-and-forward teledermatology allows surgical planning, saving both time and number of visits involving travel, thereby reducing patients’ out-of-pocket expenses, i.e., costs that patients incur when traveling to and from health providers for treatment, visits’ fees, and the opportunity cost of time spent in visits. Method: Patients’ out-of-pocket expenses-effectiveness of presurgical teledermatology were analyzed in the setting of a public hospital during two years. The mean delay in surgery was used to measure effectiveness. The teledermatology network covering the area served by the Hospital Garcia da Horta (HGO), Portugal, linked the primary care centers of 24 health districts with the hospital’s dermatology department. The patients’ opportunity cost of visits, travel costs, and visits’ fee of each presurgical modality (teledermatology and conventional referral), the cost ratio between the most and least expensive alternative, and the incremental cost-effectiveness ratio were calculated from initial primary care visit until surgical intervention. Two groups of patients: those with squamous cell carcinoma and those with basal cell carcinoma were distinguished in order to compare the effectiveness according to the dermatoses. Results: From a patient perspective, the conventional system was 2.15 times more expensive than presurgical teledermatology. Teledermatology had an incremental out-of-pocket expenses-effectiveness ratio of €1.22 per patient and per day of delay avoided. This saving was greater in patients with squamous cell carcinoma than in patients with basal cell carcinoma. Conclusion: From a patient economic perspective, teledermatology used for presurgical planning and preparation is the dominant strategy in terms of out-of-pocket expenses-effectiveness than the conventional referral system, especially for patients with severe dermatoses.

Keywords: economic analysis, out-of-pocket expenses, opportunity cost, teledermatology, waiting time

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Authors: Rojith K. Balakrishnan

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Plasmablastic lymphoma (PBL) is an uncommon, recently described B-cell derived lymphoma that is most commonly seen in patients with Human Immunodeficiency Virus (HIV) infection. Here we report a case of PBL in a 35 year old man with HIV who presented with multiple subcutaneous swellings all over the body and oral mucosal lesions.The biopsy report was suggestive of Diffuse Large B Cell Lymphoma. Immunohistochemistry was done which showed, lymphoma cells, positive for MUM1, CD 138, and VS 38. The proliferation index (MIB) was 95%. Final report was consistent with the diagnosis of Plasmablastic Lymphoma. The lesion completely regressed after treatment with systemic chemotherapy. Up to date, only a few cases of plasmablastic lymphoma have been reported from India. Increased frequency of this lymphoma in HIV patients and rarity of the tumour, along with rapid response of the same to chemotherapy, make this case a unique one. Hence the knowledge about this new entity is important for clinicians who deal with HIV patients.

Keywords: human immunodeficiency virus (HIV), oral cavity lesion, plasmablastic lymphoma, subcutaneous swelling

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Authors: Yuriko Takayama, Norihiro Kato

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Quorum sensing (QS) is a cell-to-cell communication system in bacteria to regulate expression of target genes. In gram-negative bacteria, activation on QS is controlled by a concentration increase of N-acylhomoserine lactone (AHL), which can diffuse in and out of the cell. Effective control of QS is expected to avoid virulence factor production in infectious pathogens, biofilm formation, and antibiotic production because various cell functions in gram-negative bacteria are controlled by AHL-mediated QS. In this research, we applied cyclodextrins (CDs) as artificial hosts for the AHL signal to reduce the AHL concentration in the culture broth below its threshold for QS activation. The AHL-receptor complex induced under the high AHL concentration activates transcription of the QS-target gene. Accordingly, artificial reduction of the AHL concentration is one of the effective strategies to inhibit the QS. A hydrophobic cavity of the CD can interact with the acyl-chain of the AHL due to hydrophobic interaction in aqueous media. We studied N-hexanoylhomoserine lactone (C6HSL)-mediated QS in Serratia marcescens; accumulation of C6HSL is responsible for regulation of the expression of pig cluster. Inhibitory effects of added CDs on QS were demonstrated by determination of prodigiosin amount inside cells after reaching stationary phase, because production of prodigiosin depends on the C6HSL-mediated QS. By adding approximately 6 wt% hydroxypropyl-β-CD (HP-β-CD) in Luria-Bertani (LB) medium prior to inoculation of S. maecescens AS-1, the intracellularly accumulated prodigiosin was drastically reduced to 7-10%, which was determined after the extraction of prodigiosin in acidified ethanol. The AHL retention ability of HP-β-CD was also demonstrated by Chromobacterium violacuem CV026 bioassay. The CV026 strain is an AHL-synthase defective mutant that activates QS solely by adding AHLs from outside of cells. A purple pigment violacein is induced by activation of the AHL-mediated QS. We demonstrated that the violacein production was effectively suppressed when the C6HSL standard solution was spotted on a LB agar plate dispersing CV026 cells and HP-β-CD. Physico-chemical analysis was performed to study the affinity between the immobilized CD and added C6HSL using a quartz crystal microbalance (QCM) sensor. The COOH-terminated self-assembled monolayer was prepared on a gold electrode of 27-MHz AT-cut quartz crystal. Mono(6-deoxy-6-N, N-diethylamino)-β-CD was immobilized on the electrode using water-soluble carbodiimide. The C6HSL interaction with the β-CD cavity was studied by injecting the C6HSL solution to a cup-type sensor cell filled with buffer solution. A decrement of resonant frequency (ΔFs) clearly showed the effective C6HSL complexation with immobilized β-CD and its stability constant for MBP-SpnR-C6HSL complex was on the order of 102 M-1. The CD has high potential for engineered control of QS because it is safe for human use.

Keywords: acylhomoserine lactone, cyclodextrin, intracellular signaling, quorum sensing

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Authors: Edward Hage, Pieter Lietaert, Gabriel Abedrabbo

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Musculoskeletal disorders are an important category of work-related diseases. They are often caused by working in non-ergonomic postures and are preventable with proper workplace design, possibly including human-machine collaboration. This paper presents a methodology and a supporting software prototype to design a simple assembly cell with minimal ergonomic risk. The methodology helps to determine the optimal position and orientation of workpieces and workplace resources for specific operator assembly actions. The methodology is tested on an industrial use case: a collaborative robot (cobot) assisted assembly of a clamping device. It is shown that the automated methodology results in a workplace design with significantly reduced ergonomic risk to the operator compared to a manual design of the cell.

Keywords: ergonomics optimization, design for ergonomics, workplace design, pose generation

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Authors: Varsha Salian, Shama Rao, N. Narendra, B. Mohana Kumar

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Evolving evidence proposes the existence of a highly tumorigenic subpopulation of undifferentiated, self-renewing cancer stem cells, responsible for exhibiting resistance to conventional anti-cancer therapy, recurrence, metastasis and heterogeneous tumor formation. Importantly, the mechanisms exploited by cancer stem cells to resist chemotherapy are very less understood. Oral squamous cell carcinoma (OSCC) is one of the most regularly diagnosed cancer types in India and is associated commonly with alcohol and tobacco use. Therefore, the isolation and in vitro characterization of cancer stem-like cells from patients with OSCC is a critical step to advance the understanding of the chemoresistance processes and for designing therapeutic strategies. With this, the present study aimed to establish and characterize cancer stem-like cells in vitro from OSCC. The primary cultures of cancer stem-like cell lines were established from the tissue biopsies of patients with clinical evidence of an ulceroproliferative lesion and histopathological confirmation of OSCC. The viability of cells assessed by trypan blue exclusion assay showed more than 95% at passage 1 (P1), P2 and P3. Replication rate was performed by plating cells in 12-well plate and counting them at various time points of culture. Cells had a more marked proliferative activity and the average doubling time was less than 20 hrs. After being cultured for 10 to 14 days, cancer stem-like cells gradually aggregated and formed sphere-like bodies. More spheroid bodies were observed when cultured in DMEM/F-12 under low serum conditions. Interestingly, cells with higher proliferative activity had a tendency to form more sphere-like bodies. Expression of specific markers, including membrane proteins or cell enzymes, such as CD24, CD29, CD44, CD133, and aldehyde dehydrogenase 1 (ALDH1) is being explored for further characterization of cancer stem-like cells. To summarize the findings, the establishment of OSCC derived cancer stem-like cells may provide scope for better understanding the cause for recurrence and metastasis in oral epithelial malignancies. Particularly, identification and characterization studies on cancer stem-like cells in Indian population seem to be lacking thus provoking the need for such studies in a population where alcohol consumption and tobacco chewing are major risk habits.

Keywords: cancer stem-like cells, characterization, in vitro, oral squamous cell carcinoma

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Authors: Jun Hong Lee

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Background: Alzheimer`s disease (AD) I: creases with age and is characterized by the premature progressive loss of neuronal cell. In contrast, cancer cells have inappropriate cell proliferation and resistance to cell death. Objective: We evaluated the association between cancer and AD and also examined the specific types of cancer. Patients and Methods/Material and Methods: This retrospective, nationwide, longitudinal study used National Health Insurance Service – Senior cohort (NHIS-Senior) 2002-2013, which was released by the KNHIS in 2016, comprising 550,000 random subjects who were selected from over than 60. The study included a cohort of 4,408 patients who were first diagnoses as AD between 2003 and 2005. To match each dementia patient, 19,150 subjects were selected from the database by Propensity Score Matching. Results: We enrolled 4,790 patients for analysis in this cohort and the prevalence of AD was higher in female (19.29%) than in male (17.71%). A higher prevalence of AD was observed in the 70-84 year age group and in the higher income status group. A total of 540 cancers occurred within the observation interval. Overall cancer was less frequent in those with AD (12.25%) than in the control (18.46%), with HR 0.704 (95% Confidence Intervals (CIs)=0.0.64-0.775, p-Value < 0.0001). Conclusion: Our data showed a decreased incidence of overall cancers in patients with AD similar to previous studies. Patients with AD had a significantly decreased risk of colon & rectum, lung and stomach cancer. This finding lower than but consistent with Western countries. We need further investigation of genetic evidence linking AD to cancer.

Keywords: Alzheimer, cancer, nationwide, longitudinal study

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Authors: R. Stalin, D. Karthick, H. Haseena Banu, T. P. Sachidanandam, P. Shanthi

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Background: Breast cancer is emerging as one of the leading cause of cancer related deaths and hence there arises the need to look out for drugs which are more targets specific with minimal side effects. In recent times, there is a shift towards alternative medicine due to low cost and less side effects. Siddha system of medicine is one the oldest system of medicine practiced against various ailments. Tridham (TD) is a herbal formulation prepared in our laboratory consisting of Terminalia chebula, Elaeocarpus ganitrus and Prosopis cineraria in a definite ratio (TD) and its anticancer potential is evaluated in terms of induction of apoptosis. Objective: The present study was designed to investigate the anti proliferative effect of TD and 1,2,3,4,6-penta-O-galloyl-b-D-glucose (PGG), a pure compound isolated from TD on human mammary carcinoma cell line (MCF-7). Materials and Methods: Cell viability was studied using MTT analysis and trypan blue staining. Mitochondrial membrane potential was studied using DAPI staining. The protein and mRNA expressions of pro-apoptotic and anti- apoptotic markers namely Bax, Bad, Bcl-2 and caspases were also assessed by Western Blotting and RT PCR. Results: Viability studies of TD and PGG treated MCF-7 cells showed an inhibition in cell growth in time and dose dependent manner. The alteration in mitochondrial membrane potential was restored through treatment with TD and PGG which was confirmed by DAPI staining. The protein and mRNA expression of pro-apoptotic markers was found to be significantly increased in TD and PGG treated cells with a concomitant decrease in anti-apoptotic markers. Conclusion: The results of the study suggest that TD and PGG exhibit their anticancer effect through its membrane stabilizing property and activation of apoptotic cascade in MCF-7 cells.

Keywords: apoptosis, mammary carcinoma, MCF-7, penta galloyl glucose, Tridham

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Authors: Anamarija Kuhar, Veronika Kloboves Prevodnik, Nataša Nolde, Ulrika Klopčič

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The decision for immunocytochemistry (ICC) is usually made in the basis of the findings in Giemsa- and/or Papanicolaou- smears. More demanding diagnostic cases require preparation of additional cytological preparations. Therefore, it is convenient to suspend cytological samples in a liquid based medium (LBM) that preserve antigen and morphological properties. However, the duration of these properties being preserved in the medium is usually unknown. Eventually, cell morphology becomes impaired and altered, as well as antigen properties may be lost or become diffused. In this study, the influence of cytological sample storage length in in-house liquid based medium on antigen properties and cell morphology is evaluated. The question is how long the cytological samples in this medium can be stored so that the results of immunocytochemical reactions are still reliable and can be safely used in routine cytopathological diagnostics. The stability of 6 ICC markers that are most frequently used in everyday routine work were tested; Cytokeratin AE1/AE3, Calretinin, Epithelial specific antigen Ep-CAM (MOC-31), CD 45, Oestrogen receptor (ER), and Melanoma triple cocktail were tested on methanol fixed cytospins prepared from fresh fine needle aspiration biopsies, effusion samples, and disintegrated lymph nodes suspended in in-house cell medium. Cytospins were prepared on the day of the sampling as well as on the second, fourth, fifth, and eight day after sample collection. Next, they were fixed in methanol and immunocytochemically stained. Finally, the percentage of positive stained cells, reaction intensity, counterstaining, and cell morphology were assessed using two assessment methods: the internal assessment and the UK NEQAS ICC scheme assessment. Results show that the antigen properties for Cytokeratin AE1/AE3, MOC-31, CD 45, ER, and Melanoma triple cocktail were preserved even after 8 days of storage in in-house LBM, while the antigen properties for Calretinin remained unchanged only for 4 days. The key parameters for assessing detection of antigen are the proportion of cells with a positive reaction and intensity of staining. Well preserved cell morphology is highly important for reliable interpretation of ICC reaction. Therefore, it would be valuable to perform a similar analysis for other ICC markers to determine the duration in which the antigen and morphological properties are preserved in LBM.

Keywords: cytology samples, cytospins, immunocytochemistry, liquid-based cytology

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Authors: Priyanka Bhowmik, Subrata Majumdar, Debprasad Chattopadhyay

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Background: Cervical cancer is the third major cause of cancer in women and the second most frequent cause of cancer related deaths causing 300,000 deaths annually worldwide. Evasion of immune response by Human Papilloma Virus (HPV), the key contributing factor behind cancer and pre-cancerous lesions of the uterine cervix, makes immunotherapy a necessity to treat this disease. Objective: A Heat killed fraction of Mycobacterium indicus pranii (MIP), a non-pathogenic Mycobacterium has been shown to exhibit cytotoxic effects on different cancer cells, including human cervical carcinoma cell line HeLa. However, the underlying mechanisms remain unknown. The aim of this study is to decipher the mechanism of MIP induced HeLa cell death. Methods: The cytotoxicity of Mycobacterium indicus pranii against HeLa cells was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis was detected by annexin V and Propidium iodide (PI) staining. The assessment of reactive oxygen species (ROS) generation and cell cycle analysis were measured by flow cytometry. The expression of apoptosis associated genes was analyzed by real time PCR. Result: MIP could inhibit the proliferation of HeLa cell in a time and dose dependent manner but caused minor damage to normal cells. The induction of apoptosis was confirmed by the cell surface presentation of phosphatidyl serine, DNA fragmentation, and mitochondrial damage. MIP caused very early (as early as 30 minutes) transcriptional activation of p53, followed by a higher activation (32 fold) at 24 hours suggesting prime importance of p53 in MIP-induced apoptosis in HeLa cell. The up regulation of p53 dependent pro-apoptotic genes Bax, Bak, PUMA, and Noxa followed a lag phase that was required for the transcriptional p53 program. MIP also caused the transcriptional up regulation of Toll like receptor 2 and 4 after 30 minutes of MIP treatment suggesting recognition of MIP by toll like receptors. Moreover, MIP caused the inhibition of expression of HPV anti apoptotic gene E6, which is known to interfere with p53/PUMA/Bax apoptotic cascade. This inhibition might have played a role in transcriptional up regulation of PUMA and subsequently apoptosis. ROS was generated transiently which was concomitant with the highest transcription activation of p53 suggesting a plausible feedback loop network of p53 and ROS in the apoptosis of HeLa cells. Scavenger of ROS, such as N-acetyl-L-cysteine, decreased apoptosis suggesting ROS is an important effector of MIP induced apoptosis. Conclusion: Taken together, MIP possesses full potential to be a novel therapeutic agent in the clinical treatment of cervical cancer.

Keywords: cancer, mycobacterium, immunity, immunotherapy.

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Authors: Z. Adnan, M. M. Habib

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The laboratory tests for measuring the effects of small strain to the shear strength and maximum stiffness development of post-cyclic degradation of hemic peat are reviewed in this paper. A series of laboratory testing has been conducted to fulfil the objective of this research to study the post-cyclic behaviour of peat soil and focuses on the small strain characteristics. For this purpose, a number of strain-controlled static, cyclic and post-cyclic triaxial tests were carried out in undrained condition on hemic peat soil. The shear strength and maximum stiffness of hemic peat are evaluated immediately after post-cyclic monotonic testing. There are two soil samples taken from West Johor and East Malaysia peat soil. Based on these laboratories and field testing data, it was found that the shear strength and maximum stiffness of peat soil decreased in post-cyclic monotonic loading than its initial shear strength and stiffness. In particular, degradation in shear strength and stiffness is more sensitive for peat soil due to fragile and uniform fibre structures. Shear strength of peat soil, τ_max = 12.53 kPa (Beaufort peat, BFpt) and 36.61 kPa (Parit Nipah peat, PNpt) decreased than its initial 58.46 kPa and 91.67 kPa. The maximum stiffness, G_max = 0.23 and 0.25 decreased markedly with post-cyclic, G_max = 0.04 and 0.09. Simple correlations between the G_max and the τ_max effects due to small strain, ε = 0.1, the G_max values for post-cyclic are relatively low compared to its initial G_max. As a consequence, the reported values and patterns of both the West Johor and East Malaysia peat soil are generally the same.

Keywords: post-cyclic, strain, maximum stiffness, shear strength

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Authors: Débora P. Jaeschke, Eduardo A. Merlo, Rosane Rech, Giovana D. Mercali, Ligia D. F. Marczak

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Carotenoids are high value added pigments that can be alternatively extracted from some microalgae species. However, the application of carotenoids synthetized by microalgae is still limited due to the utilization of organic toxic solvents. In this context, studies involving alternative extraction methods have been conducted with more sustainable solvents to replace and reduce the solvent volume and the extraction time. The aim of the present work was to evaluate the extraction time of carotenoids from the microalgae Heterochlorella luteoviridis using moderate electric field (MEF) as a pre-treatment to the extraction. The extraction methodology consisted of a pre-treatment in the presence of MEF (180 V) and ethanol (25 %, v/v) for 10 min, followed by a diffusive step performed for 50 min using a higher ethanol concentration (75 %, v/v). The extraction experiments were conducted at 30 °C and, to keep the temperature at this value, it was used an extraction cell with a water jacket that was connected to a water bath. Also, to enable the evaluation of MEF effect on the extraction, control experiments were performed using the same cell and conditions without voltage application. During the extraction experiments, samples were withdrawn at 1, 5 and 10 min of the pre-treatment and at 1, 5, 30, 40 and 50 min of the diffusive step. Samples were, then, centrifuged and carotenoids analyses were performed in the supernatant. Furthermore, an exhaustive extraction with ethyl acetate and methanol was performed, and the carotenoids content found for this analyses was considered as the total carotenoids content of the microalgae. The results showed that the application of MEF as a pre-treatment to the extraction influenced the extraction yield and the extraction time during the diffusive step; after the MEF pre-treatment and 50 min of the diffusive step, it was possible to extract up to 60 % of the total carotenoids content. Also, results found for carotenoids concentration of the extracts withdrawn at 5 and 30 min of the diffusive step did not presented statistical difference, meaning that carotenoids diffusion occurs mainly in the very beginning of the extraction. On the other hand, the results for control experiments showed that carotenoids diffusion occurs mostly during 30 min of the diffusive step, which evidenced MEF effect on the extraction time. Moreover, carotenoids concentration on samples withdrawn during the pre-treatment (1, 5 and 10 min) were below the quantification limit of the analyses, indicating that the extraction occurred in the diffusive step, when ethanol (75 %, v/v) was added to the medium. It is possible that MEF promoted cell membrane permeabilization and, when ethanol (75 %) was added, carotenoids interacted with the solvent and the diffusion occurred easily. Based on the results, it is possible to infer that MEF promoted the decrease of carotenoids extraction time due to the increasing of the permeability of the cell membrane which facilitates the diffusion from the cell to the medium.

Keywords: moderate electric field (MEF), pigments, microalgae, ethanol

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Authors: Ki-Yeo Kim

Abstract:

Trends in genetics are transforming in order to identify differential coexpressions of correlated gene expression rather than the significant individual gene. Moreover, it is known that a combined biomarker pattern improves the discrimination of a specific cancer. The identification of the combined biomarker is also necessary for the early detection of invasive oral squamous cell carcinoma (OSCC). To identify the combined biomarker that could improve the discrimination of OSCC, we explored an appropriate number of genes in a combined gene set in order to attain the highest level of accuracy. After detecting a significant gene set, including the pre-defined number of genes, a combined expression was identified using the weights of genes in a gene set. We used the Principal Component Analysis (PCA) for the weight calculation. In this process, we used three public microarray datasets. One dataset was used for identifying the combined biomarker, and the other two datasets were used for validation. The discrimination accuracy was measured by the out-of-bag (OOB) error. There was no relation between the significance and the discrimination accuracy in each individual gene. The identified gene set included both significant and insignificant genes. One of the most significant gene sets in the classification of normal and OSCC included MMP1, SOCS3 and ACOX1. Furthermore, in the case of oral dysplasia and OSCC discrimination, two combined biomarkers were identified. The combined genomic expression achieved better performance in the discrimination of different conditions than in a single significant gene. Therefore, it could be expected that accurate diagnosis for cancer could be possible with a combined biomarker.

Keywords: oral squamous cell carcinoma, combined biomarker, microarray dataset, correlated genes

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Authors: Brian M. Mehling, Louis Quartararo, Marine Manvelyan, Paul Wang, Dong-Cheng Wu

Abstract:

Numerous investigations suggest that Mesenchymal Stem Cells (MSCs) in general represent a valuable tool for therapy of symptoms related to chronic inflammatory diseases. Blue Horizon Stem Cell Therapy Program is a leading provider of adult and children’s stem cell therapies. Uniquely we have safely and efficiently treated more than 600 patients with documenting each procedure. The purpose of our study is primarily to monitor the immune response in order to validate the safety of intravenous infusion of human umbilical cord blood derived MSCs (UC-MSCs), and secondly, to evaluate effects on biomarkers associated with chronic inflammation. Nine patients were treated for conditions associated with chronic inflammation and for the purpose of anti-aging. They have been given one intravenous infusion of UC-MSCs. Our study of blood test markers of 9 patients with chronic inflammation before and within three months after MSCs treatment demonstrates that there is no significant changes and MSCs treatment was safe for the patients. Analysis of different indicators of chronic inflammation and aging included in initial, 24-hours, two weeks and three months protocols showed that stem cell treatment was safe for the patients; there were no adverse reactions. Moreover data from follow up protocols demonstrates significant improvement in energy level, hair, nails growth and skin conditions. Intravenously administered UC-MSCs were safe and effective in the improvement of symptoms related to chronic inflammation. Further close monitoring and inclusion of more patients are necessary to fully characterize the advantages of UC-MSCs application in treatment of symptoms related to chronic inflammation.

Keywords: chronic inflammatory diseases, intravenous infusion, stem cell therapy, umbilical cord blood derived mesenchymal stem cells (UC-MSCs)

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Authors: Pratthana Ammaraphitak, Piyachon Ketsuwan, Rattapoom Prommana

Abstract:

Electricity production from vermicompost liquid was investigated in microbial fuel cells (MFCs). The aim of this study was to determine the performance of vermicompost liquid as a biocatalyst for electricity production by MFCs. Chemical and physical parameters of vermicompost liquid as total nitrogen, ammonia-nitrogen, nitrate, nitrite, total phosphorus, potassium, organic matter, C:N ratio, pH, and electrical conductivity in MFCs were studied. The performance of MFCs was operated in open circuit mode for 7 days. The maximum open circuit voltage (OCV) was 0.45 V. The maximum power density of 5.29 ± 0.75 W/m² corresponding to a current density of 0.024 2 ± 0.0017 A/m² was achieved by the 1000 Ω on day 2. Vermicompost liquid has efficiency to generate electricity from organic waste.

Keywords: vermicompost liquid, microbial fuel cell, nutrient, electricity production

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Authors: Haiyan Wang, Dongyun Wang, Yongyong Yan, Richard T. Jaspers, Gang Wu

Abstract:

Mesenchymal stem cell and 3D printing-based bone tissue engineering present a promising technique to repair large-volume bone defects. Its success is highly dependent on cell attachment, spreading, osteogenic differentiation, and in vivo survival of stem cells on 3D-printed scaffolds. In this study, human salivary histatin-1 (Hst1) was utilized to enhance the interactions between human adipose-derived stem cells (hASCs) and 3D-printed β-tricalcium phosphate (β-TCP) bioceramic scaffolds. Fluorescent images showed that Hst1 significantly enhanced the adhesion of hASCs to both bioinert glass and 3D-printed β-TCP scaffold. In addition, Hst1 was associated with significantly higher proliferation and osteogenic differentiation of hASCs on 3D-printed β-TCP scaffolds. Moreover, coating 3D-printed β-TCP scaffolds with histatin significantly promotes the survival of hASCs in vivo. The ERK and p38 but not JNK signaling was found to be involved in the superior adhesion of hASCs to β-TCP scaffolds with the aid of Hst1. In conclusion, Hst1 could significantly promote the adhesion, spreading, osteogenic differentiation, and in vivo survival of hASCs on 3D-printed β-TCP scaffolds, bearing a promising application in stem cell/3D printing-based constructs for bone tissue engineering.

Keywords: 3d printing, adipose-derived stem cells, bone tissue engineering, histatin-1, osteogenesis

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Authors: Kakali Bhadra

Abstract:

Harmalol administration caused remarkable reduction in proliferation of HepG₂ cells with GI₅₀ of 14.2 mM, without showing much cytotoxicity in embryonic liver cell line, WRL-68. Data from circular dichroism and differential scanning calorimetric analysis of harmalol-CT DNA complex shows conformational changes with prominent CD perturbation and stabilization of CT DNA by 8 ^oC. Binding constant and stoichiometry was also calculated using the above biophysical techniques. Further, dose dependent apoptotic induction ability of harmalol was studied in HepG₂ cells using different biochemical assays. Generation of ROS, DNA damage, changes in cellular external and ultramorphology, alteration of membrane, formation of comet tail, decreased mitochondrial membrane potential and a significant increase in Sub G_o/G₁ population made the cancer cell, HepG₂, prone to apoptosis. Up regulation of p53 and caspase 3 further indicated the apoptotic role of harmalol.

Keywords: apoptosis, beta carboline alkaloid, comet assay, cytotoxicity, ROS

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Authors: Chakrit Thapphan, Suteeraporn Chaowattanapanit, Sorutsiri Chareonsudjai, Wisitsak Phoksawat, Supranee Phantanawiboon, Kiatichai Faksri, Steve W. Edwards, Kanin Salao

Abstract:

Background: Psoriasis is a chronic inflammatory disease of the skin that is mediated by crosstalk between keratinocytes and immune cells, especially CD4+ T helper (Th) cells. To date, psoriasis is established as a T helper 17 (Th17) cell-mediated inflammatory process driven by the over-expression of Th17. However, the role of other CD4+T helper cells is rather controversial. Objective: Our study, thereby, aimed to characterize and analyze T cell subsets in the circulating blood of psoriasis patients and compare them to healthy controls. Methods: Peripheral blood mononuclear cells were isolated from the participants and stained with fluorescent dye-conjugated monoclonal antibodies specific for intracellular cytokines, including interferon-gamma (IFN- γ), interleukin (IL-4), IL-17 and forkhead box P3 (FOXP3), that can be used to define T helper 1 (Th1) cells, T helper 2 (Th2), T helper 17 (Th17) and regulatory T cells (Treg) respectively. Results: We found that the numbers of Th2 (59.6% ± 17.0) and Th17 (4.0% ± 2.0) cells in the circulating blood of psoriasis patients were significantly higher than those of the healthy controls (p= 0.0007 and 0.0013 respectively). In contrast, the numbers of Th1 and Treg cells were not significantly different between psoriasis patients and healthy controls (p= 0.0593 and 0.8518, respectively). Additionally, when adjusting these numbers of Th cells to Treg, we observed a similar trend that the ratio of Th2/Treg and Th17/Treg also elevated (p = 0.0007 and 0.0047, respectively). Conclusion: Taken together, our results suggest an imbalanced T exhibit toward the Th2 and Th17 skewed-immune responses in psoriasis patients.

Keywords: psoriasis, Th cell subsets, Th2 cells, Th17 cells, Treg cells

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Authors: Mutadi

Abstract:

Pipes laying on an unstable slope of soft soil are prone to movement. Pipelines that are buried in unstable slope areas will move due to lateral loads from soil movement, which can cause damage to the pipeline. A small-scale laboratory model of the reinforcement system of piles supported by retaining walls was conducted to investigate the effect of lateral load on the reinforcement. In this experiment, the lateral forces of 0.3 kN, 0.35 kN, and 0.4 kN and vertical force of 0.05 kN, 0.1 kN, and 0.15 kN were used. Lateral load from the electric jack is equipped with load cell and vertical load using the cement-steel box. To validate the experimental result, a finite element program named 2-D Plaxis was used. The experimental results showed that with an increase in lateral loading, the displacement of the reinforcement system increased. For a Vertical Load, 0.1 kN and versus a lateral load of 0.3 kN causes a horizontal displacement of 0.35 mm and an increase of 2.94% for loading of 0.35 kN and an increase of 8.82% for loading 0.4 kN. The pattern is the same in the finite element method analysis, where there was a 6.52% increase for 0.35 kN loading and an increase to 23.91 % for 0.4 kN loading. In the same Load, the Reinforcement System is reliable, as shown in Safety Factor on dry conditions were 3.3, 2.824 and 2.474, and on wet conditions were 2.98, 2.522 and 2.235.

Keywords: soft soil, deflection, wall, pipeline

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Authors: J. Constanzo, B. Paquette, G. Charest, L. Masson-Côté, M. Guillot

Abstract:

Targeted and whole-brain irradiation in humans can result in significant side effects causing decreased patient quality of life. To adequately investigate structural and functional alterations after stereotactic radiosurgery, preclinical studies are needed. The first step is to establish a robust standardized method of targeted irradiation on small regions of the rat brain. Eleven euthanized male Fischer rats were imaged in a stereotactic bed, by computed tomographic (CT), to estimate positioning variations regarding to the bregma skull reference point. Using a rat brain atlas and the stereotactic bregma coordinates assessed from CT images, various regions of the brain were delimited and a treatment plan was generated. A dose of 37 Gy at 30% isodose which corresponds to 100 Gy in 100% of the target volume (X = 98.1; Y = 109.1; Z = 100.0) was set by Leksell Gamma Plan using sectors number 4, 5, 7, and 8 of the Gamma Knife unit with the 4-mm diameter collimators. Effects of positioning accuracy of the rat brain on the dose deposition were simulated by Gamma Plan and validated with dosimetric measurements. Our results showed that 90% of the target volume received 110 ± 4.7 Gy and the maximum of deposited dose was 124 ± 0.6 Gy, which corresponds to an excellent relative standard deviation of 0.5%. This dose deposition calculated with the Gamma Plan was validated with the dosimetric films resulting in a dose-profile agreement within 2%, both in X- and Z-axis,. Our results demonstrate the feasibility to standardize the irradiation procedure of a small volume in the rat brain using a Gamma Knife.

Keywords: brain irradiation, dosimetry, gamma knife, small-animal irradiation, stereotactic radiosurgery (SRS)

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