3D integration of micro- and nanostructures into bio-analytical devices

This study aims to develop a process which allows 3D integration of micro and nanostructures in microchannels. A fabrication process was established for the large area integration of hierarchical micro and nanostructures in microchannels. This novel process, which is called 3D molding, takes advantage of an intermediate thin flexible stamp such as PDMS from soft lithography and a hard mold such as brass from hot embossing process. However, the use of a thin intermediate polydimethylsiloxane (PDMS) stamp inevitably causes dimensional changes in the 3D molded channel, with respect to those in the brass mold protrusion and the intermediate PDMS stamp structures. We have investigated the deformation behavior of the 3D molded poly(methyl methacrylate) (PMMA) substrate and the intermediate PDMS stamp in 3D molding through both experimentation and numerical simulation. It was found that for high aspect ratio brass mold protrusion, the maximum strain of the intermediate layer occurs in the bottom center of the 3D channels. However, with decreasing the aspect ratio of brass mold protrusion the highest elongation occurs at the bottom corners of the channel causing less elongation of the intermediate PDMS stamp and imprinted structures on the bottom surface of the 3D channel. A modified 3D molding process which is called 3D nanomolding is developed which allows nanopatterning the surface of small microfeatures. Using 3D nanomolding process and solvent assisted bonding microdevices with no side, one side, three sides and four sides patterned were fabricated. To characterize 3D flow patterns induced by the surface structures on microdevices, confocal microscopy was used as dyed water and undyed water injected from separate inlets of micromixer were mixed along the microchannel at flow rates of 10 and 40 μL/min. The standard deviation of the normalized intensity measured in the confocal image of the cross section of the channel was used for quantifying the degree of mixing and evaluating the mixing performance of all four different microdevices. Experimental and simulation results show that by patterning the surface of the micromixer, flow patterns can be manipulated, which can improve mixing through stretching and folding of fluid elements and therefore increasing the interfacial area between fluids and cutting down the diffusion length. The effect of increasing velocity on increasing standard deviation (decreasing mixing) was also found to be less for the micromixers whose surfaces are patterned compared to the plain channel

Roles of novel biomarkers in progression of cutaneous squamous cell carcinoma

Roles of novel biomarkers was studied in progression of cutaneous squamous cell carcinoma (cSCC) as the most common metastatic skin cancer. The incidence of cSCC is increasing worldwide due to lifestyle changes such as recreational exposure to sunlight and the aging of the population. Because of an emerging need for molecular markers for the progression of cSCC, we set our goal to characterize three distinct novel markers overexpressed in cSCC cells. Our results identified overexpression of serpin peptidase inhibitor clade A member 1 (SerpinA1), EphB2 and absent in melanoma 2 (AIM2) in cSCC cell lines compared with normal human epidermal keratinocytes (NHEKs). Immunohistochemical analysis of SerpinA1, EphB2 and AIM2 revealed abundant tumor cell-specific expression of cytoplasmic SerpinA1 and AIM2 and cytoplasmic and membranous EphB2 in cSCC tumors in vivo. The staining intensity of SerpinA1, EphB2 and AIM2 was significantly stronger in cSCC as compared with carcinoma in situ (cSCCIS) and actinic keratosis (AK). Tumor cell-associated SerpinA1 and EphB2 was noted in chemically induced mouse skin SCC, and the staining intensity was stronger in mouse cSCCs than in untreated skin. AIM2 staining intensity was significantly more abundant in cSCC of organ transplant recipients (OTR) than in sporadic cSCC in vivo. EphB2 knockdown resulted in inhibition of migration in cSCC cells. In addition, knockdown of EphB2 and AIM2 was found to inhibit the proliferation and invasion of cSCC cells and to delay the growth and vascularization of cSCC xenografts in vivo. Altogether, these findings identify SerpinA1 as a novel biomarker for cSCC. In addition, characterization of the roles of EphB2 and AIM2 in the progression of cSCC was implicated them as possible therapeutic targets for the treatment of cSCC particularly in unresectable and metastatic tumors.Ihon levyepiteelisyövän kehittymiseen liittyvät uudet merkkitekijät Keratinosyyttiperäinen ihon okasolusyöpä on yleisin metastasoituva ihosyöpä. Sen ilmaantuvuus kasvaa maailmanlaajuisesti elintapojen muutoksen kuten auringonvalolle altistumisen sekä väestön ikääntymisen takia. Tarvitaankin uusia merkkitekijöitä tämän syövän etenemisen ennustamiseksi. Työssämme löysimme kolme uutta merkkitekijää ja selvitimme niiden roolia ihon okasolusyövässä. Havaitsimme, että seriiniproteaasin estäjä A1 (SerpinA1), EphB2 ja absent in melanoma 2 (AIM2) ovat voimakkaasti koholla ihon okasolusyövästä eristetyissä solulinjoissa verrattuna ihmisen normaaleihin epidermaalisiin keratinosyytteihin. Tämän ohella havaitsimme immunohistokemiallisessa analyysissä SerpinA1:n, EphB2:n ja AIM2:n ilmentyvän spesifisti kasvaimen soluissa sekä ilmentyvän merkitsevästi enemmän okasolusyövissä verrattuna in situ karsinoomaan ja aktiiniseen keratoosiin in vivo. Lisäksi SerpinA1:n ja EphB2:n havaittiin olevan yliekspressoituja hiiren ihoon kemiallisesti aiheutetussa okasolusyövässä. AIM2:n värjäytymisintensiteetti oli merkitsevästi voimakkaampi elinsiirtopotilaista peräisin olevissa ihon okasolusyövissä kuin sporadisissa okasolusyövissä. EphB2:n hiljentäminen esti merkitsevästi okasolusyövästä eristettyjen solujen migraatiota. Lisäksi EphB2:n ja AIM2:n hiljentäminen syöpäsoluissa vähensi merkitsevästi solujen jakaantumista ja invaasiota sekä tuumorien vaskularisaatiota ja kasvua xenograftimallissa in vivo. Yhdessä nämä havainnot antavat aiheen otaksua, että SerpinA1 voisi toimia merkkitekijänä ihon okasolusyövässä. Lisäksi havaitsimme, että EphB2:lla ja AIM2:lla on tärkeä rooli ihon okasolusyövän kehittymisessä, joten ne voisivat mahdollisesti toimia uusina hoidon kohteina erityisesti metastaattisten ja vaikeasti kirurgisesti poistettavien ihon okasolusyöpien hoidossa.Siirretty Doriast

FROM REPRESENTATIVE TO TRANSFORMATIVE USER PARTICIPATION – A CASE STUDY OF PUBLIC HEALTHCARE DIGITALIZATION

User participation in IS implementation is a core topic for the IS community. While most of our current participation theories emerged in the 1990s and 2000s, recent developments such as the emergence of large half-built products and the increased emphasis on digital transformation necessitate revisiting current understanding of participation. User participation is not anymore about merely representing the organization\u27s domain knowledge - what we call representative participation. Users participating in modern IS implementation projects need to possess additional types of knowledge and skills. This includes knowledge about the products, knowledge about the intended transformation, and leadership skills, to name a few. We investigate this emerging type of user participation - what we call transformative participation through a case study of an IS implementation project within healthcare. We discuss the knowledge and skills needed to function as transformative users as well as the challenges faced by these users

Drug-induced skin reactions: A 2-year study

Background: The aim of this study was to analyze the clinical characteristics of patients with adverse cutaneous drug reactions, which occur when a medicinal product results in cutaneous morbidity. Methods: The study included 308 patients who were diagnosed as having an adverse cutaneous drug reaction during the study period (2007�2009). In 84 cases, histopathologic examination of skin biopsies were also performed. Results: Patients with drug reactions were found to be more commonly female (63) than male (37). Beta-lactam antibiotics were found to be the most frequent cause of adverse cutaneous drug reactions (42.7), followed by non-steroidal anti-infammatory drugs (16.5). Acute urticaria was the most common clinical presentation (59.2) followed by fxed drug eruptions (18.5), and maculopapular eruptions (14.9). Conclusion: Adverse cutaneous drug reactions in our study population were mainly induced by beta-lactam antibiotics and non-steroidal anti-inflammatory drugs. The most common forms of cutaneous adverse drug reactions were found to be acute urticaria, fxed drug eruptions, and maculopapular rashes. © 2015 Farshchian et al

Comparative analysis of morphometric parameters of intercondylar notch in patients with and without anterior cruciate ligament tears

BACKGROUND: Anterior cruciate ligament (ACL) injury is the most common type of ligament injury whose prevalence is higher in athletes. There are different external risk factors for this injury. However, it is important to find its physiological risk factors, as well. This study assessed the relationship between morphometric parameters of intercondylar notch and ACL tears in patients suffering from knee complications. METHODS: Patients with or without ACL tears who had undergone knee magnetic resonance imaging for any reason were recruited based on inclusion criteria. Intercondylar notch width, femoral bicondylar width, and intercondylar notch index were calculated in both groups. Then, significant variables from univariate analysis were entered in multiple regression analysis with intercondylar notch width, femoral bicondylar width, and intercondylar notch index being assumed as dependent variables. RESULTS: Overall, 199 participants, including 81 patients with ACL tear and 118 without ACL tear, were evaluated. Multiple regression analysis revealed intercondylar notch width and intercondylar notch index to be less common among women and subjects with ACL tears (P < 0.001). CONCLUSION: Based on the results of this study, there are relationships between ACL tears and being female and intercondylar notch width. Therefore, intercondylar notch index can be used for screening athletes and people at risk of ACL tears

Optimization Model for Sustainable Renovations in Buildings

Buildings consume a substantial amount of energy and adversely affect the global climate and environment. According to the US Department of Energy (DOE), buildings account for 39% of total primary energy consumption and 71% of the electricity consumption. The construction and operation phases constitute the largest proportion of the total energy end-use worldwide (Ma et al. 2012). An innovative and comprehensive set of sustainable materials aiming at the envelope of buildings excluding the roofs is employed to define the renovation alternatives in order to ameliorate the sustainability status of the buildings. The model is comprised of a NSGA-II multi-objective optimization algorithm integrated into a simulation engine. Simulation runs are performed to compute the objective function values and transfer them to the optimization algorithm. A hybrid fuzzy simulation-based optimization model is developed to select the optimum renovation alternatives. The model simultaneously minimizes annual energy consumption and capital cost of an existing office building based on a multi-objective optimization problem. Fuzzy set theory is assigned to the objective functions to address the uncertainty associated with calculation of energy consumption and capital cost values. Conclusively, the model is implemented on a sample case to substantiate the capabilities of the developed model. The case study is a one-story office building with a double skin facade on the south facing facade in Montreal. The results illustrate nine Pareto optimal points and demonstrate that the generated optimum solutions are capable of causing an average of 35% decrease in the annual energy consumption compared to the conventional building scenario

Evaluation and Optimization of Lipofectamine 3000 Reagents for Transient Gene Expression in KYSE-30 Esophagus Cancer Cell Line

Background and Aim: Transfection of DNA/RNA sequence into eukaryotic cells has a major effect on scientific studies. Various methods are used to transfer the DNA/RNA sequence into cells, such as lipid-based carriers as the available and easy procedure. Transfection with cationic lipid liposome is introduced as a simple and efficient procedure for monitoring the DNA/RNA sequence through gene function analysis, including fluorescence imaging RNA and protein expression. This study aimed to investigate the transfection efficiency and cell death through GFP expression in human esophageal squamous cell carcinoma (ESCC) cell line KYSE-30 using Lipofectamine 3000 reagent. Methods: The pCDH-513b plasmid DNA was transfected into KYSE-30 cells using Lipofectamine 3000 in different concentrations of the plasmid DNA and reagent. The transfection efficiency was evaluated by fluorescence microscope and flow cytometry analysis to determine the percentage of GFP-expressing cells. Moreover, the viability and death of transfected KYSE-30 cells were evaluated using a trypan blue exclusion assay. Results: The transfection efficiency of KYSE-30 with Lipofectamine 3000 was increased with higher plasmid DNA concentration and a lower amount of Lipofectamine 3000 reagent. The Optimized concentration of 1.5 µg plasmid DNA and volume of one µl of lipofectamine 3000 reagents were identified for 95% transfection efficiency in the KYSE-30 cell line. The viability and death of transfected cells were 43% and 58% after transfection, respectively. Conclusion: The results indicated that Lipofectamine 3000 might not be suitable for transfection in KYSE-30 cells due to increased cell death. *Corresponding Author: Mohammad Reza Abbaszadegan; Email: [email protected] Please cite this article as: Mahmoudian RA, Farshchian M, Abbaszadegan MR. Evaluation and Optimization of Lipofectamine 3000 Reagents for Transient Gene Expression in KYSE-30 Esophagus Cancer Cell Line. Arch Med Lab Sci. 2019;5(4):1-9. https://doi.org/10.22037/amls.v5i4.3108