Search results for: inhibition of cancer cells

Authors: Jienny Lee, In-Soo Cho, Sang-Ho Cha

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Adult mesenchymal stem cells (MSCs) have been investigated using preclinical approaches for tissue regeneration. Porcine MSCs (pMSCs) are capable of growing and attaching to plastic with a fibroblast-like morphology and then differentiating into bone, adipose, and cartilage tissues in vitro. This study was conducted to investigate the proliferating abilities, differentiation potentials, and multipotency of miniature pig adipose tissue-derived MSCs (mpAD-MSCs) with or without long-term cryopreservation, considering that cryostorage has the potential for use in clinical applications. After confirming the characteristics of the mpAD-MSCs, we examined the effect of long-term cryopreservation (> 2 years) on expression of cell surface markers (CD34, CD90 and CD105), proliferating abilities (cumulative population doubling level, doubling time, colony-forming unit, and MTT assay) and differentiation potentials into mesodermal cell lineages. As a result, the expression of cell surface markers is similar between thawed and fresh mpAD-MSCs. However, long-term cryopreservation significantly lowered the differentiation potentials (adipogenic, chondrogenic, and osteogenic) of mpAD-MSCs. When compared with fresh mpAD-MSCs, thawed mpAD-MSCs exhibited lower expression of mesodermal cell lineage-related genes such as peroxisome proliferator-activated receptor-g2, lipoprotein lipase, collagen Type II alpha 1, osteonectin, and osteocalcin. Interestingly, long-term cryostoraged mpAD-MSCs exhibited significantly higher cell viability than the fresh mpAD-MSCs. Long-term cryopreservation induced a 30% increase in the cell viability of mpAD-MSCs when compared with the fresh mpAD-MSCs at 5 days after thawing. However, long-term cryopreservation significantly lowered expression of stemness markers such as Oct3/4, Sox2, and Nanog. Furthermore, long-term cryopreservation negatively affected expression of senescence-associated genes such as telomerase reverse transcriptase and heat shock protein 90 of mpAD-MSCs when compared with the fresh mpAD-MSCs. The results from this study might be important for the successful application of MSCs in clinical trials after long-term cryopreservation.

Keywords: mesenchymal stem cells, cryopreservation, stemness, senescence

Procedia PDF Downloads 227

Authors: Ildiko Fekete-Kertesz, Jade Chaker, Sylvain Berthelot, Viktoria Feigl, Monika Molnar, Lidia Favier

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There has been growing concern about emerging micropollutants in recent years, because of the possible environmental and health risk posed by these substances, which are released into the environment as a consequence of anthropogenic activities. Among them pharmaceuticals are currently not considered under water quality regulations; however, their potential effect on the environment have become more frequent in recent years. Due to the fact that these compounds can be detected in natural water matrices, it can be concluded, that the currently applied water treatment processes are not efficient enough for their effective elimination. To date, advanced oxidation processes (AOPs) are considered as highly competitive water treatment technologies for the removal of those organic micropollutants not treatable by conventional techniques due to their high chemical stability and/or low biodegradability. AOPs such as (photo)chemical oxidation and heterogeneous photocatalysis have proven their potential in degrading harmful organic compounds from aqueous matrices. However, some of these technologies generate reaction by-products, which can even be more toxic to aquatic organisms than the parent compounds. Thus, target compound removal does not necessarily result in the removal of toxicity. Therefore, to evaluate process efficiency the determination of the toxicity and ecotoxicity of the reaction intermediates is crucial to estimate the environmental risk of such techniques. In this context, the present study investigates the effectiveness of TiO2 assisted photodegradation for the removal of emerging water contaminants. Two drugs named losartan (used in high blood pressure medication) and levetiracetam (used to treat epilepsy) were considered in this work. The photocatalytic reactions were carried out with a commercial catalyst usually employed in photocatalysis. Moreover, the toxicity of the by-products generated during the process was assessed with various ecotoxicological methods applying aquatic test organisms from different trophic levels. A series of experiments were performed to evaluate the toxicity of untreated and treated solutions applying the Aliivibrio fischeri bioluminescence inhibition test, the Tetrahymena pyriformis proliferation inhibition test, the Daphnia magna lethality and immobilization tests and the Lemna minor growth inhibition test. The applied ecotoxicological methodology indicated sensitively the toxic effects of the treated and untreated water samples, hence the applied test battery is suitable for the ecotoxicological characterization of TiO2 based photocatalytic water treatment technologies and the indication of the formation of toxic by-products from the parent chemical compounds. Obtained results clearly showed that the TiO2 assisted photodegradation was more efficient in the elimination of losartan than levetiracetam. It was also observed that the treated levetiracetam solutions had more severe effect on the applied test organisms. A possible explanation would be the production of levetiracetam by-products, which are more toxic than the parent compound. The increased toxicity and the risk of formation of toxic metabolites represent one possible limitation to the implementation of photocatalytic treatment using TiO2 for the removal of losartan and levetiracetam. Our results proved that, the battery of ecotoxicity tests used in this work can be a promising investigation tool for the environmental risk assessment of photocatalytic processes.

Keywords: aquatic micropollutants, ecotoxicology, nano titanium dioxide, photocatalysis, water treatment

Procedia PDF Downloads 183

Authors: N. Rassoul-ibrahim, E. Siahmed, L. Tadrist

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Water management plays a crucial role in the performance and durability of PEM fuel cells. Whereas the membrane must be hydrated enough, liquid droplets formed by water in excess can block the flow in the gas distribution channels and hinder the fuel cell performance. The main purpose of this work is to increase the understanding of liquid transport and mixing through mini- or micro-channels for various engineering or medical process applications including cool-ing of equipment according to the operations considered. For that purpose and as a first step, a technique was devel-oped to automatically detect and characterize two-phase flow patterns that may appear in such. The investigation, mainly experimental, was conducted on transparent channel with a 1mm x 1mm square cross section and a 0.3mm x 0.3 mm water injection normal to the gas channel. Three main flow patterns were identified liquid slug, bubble flow and annular flow. A flow map has been built accord-ing to the flow rate of both phases. As a sample the follow-ing figures show representative images of the flow struc-tures observed. An analysis and discussion of the flow pattern, in mini-channel, will be provided and compared to the case old micro-channel. . Keywords: Two phase flow, Clean Energy, Minichannels, Fuel Cells. Flow patterns, Maps.

Keywords: two phase flox, T-juncion, Micro and minichannels, clean energy, flow patterns, maps

Procedia PDF Downloads 70

Authors: Seid Yimer Abate, Ding-Chi Huang, Yu-Tai Tao

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In this study, charge extraction characteristics at the perovskite/TiO2 interface in the conventional perovskite solar cell is studied by interface engineering. Self-assembled monolayers of phosphonic acids with different chain length and terminal functional group were used to modify mesoporous TiO2 surface to modulate the surface property and interfacial energy barrier to investigate their effect on charge extraction and transport from the perovskite to the mp-TiO2 and then the electrode. The chain length introduces a tunnelling distance and the end group modulate the energy level alignment at the mp-TiO2 and perovskite interface. The work function of these SAM-modified mp-TiO2 varied from −3.89 eV to −4.61 eV, with that of the pristine mp-TiO2 at −4.19 eV. A correlation of charge extraction and transport with respect to the modification was attempted. The study serves as a guide to engineer ETL interfaces with simple SAMs to improve the charge extraction, carrier balance and device long term stability. In this study, a maximum PCE of ~16.09% with insignificant hysteresis was obtained, which is 17% higher than the standard device.

Keywords: Energy level alignment, Interface engineering, Perovskite solar cells, Phosphonic acid monolayer, Tunnelling distance

Procedia PDF Downloads 130

Authors: Ali Bahkali, Rayan Yousef Booq, Mohammad Khiyami

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Soil samples were collected from five different regions in the Kingdom of Saudi Arabia. Microbiological methods included dilution methods and pour plates to isolate and purify bacteria soil. The ability of isolates to develop biopolymer was investigated on petri dishes containing elements and substance concentrations stimulating developing biopolymer. Fluorescent stains, Nile red and Nile blue were used to stain the bacterial cells developing biopolymers. In addition, Sudan black was used to detect biopolymers in bacterial cells. The isolates which developed biopolymers were identified based on their gene sequence of 1 6sRNA and their ability to grow and synthesize PHAs on mineral medium supplemented with 1% dates molasses as the only carbon source under nitrogen limitation. During the study 293 bacterial isolates were isolated and detected. Through the initial survey on the petri dishes, 84 isolates showed the ability to develop biopolymers. These bacterial colonies developed a pink color due to accumulation of the biopolymers in the cells. Twenty-three isolates were able to grow on dates molasses, three strains of which showed the ability to accumulate biopolymers. These strains included Bacillus sp., Ralstonia sp. and Microbacterium sp. They were detected by Nile blue A stain with fluorescence microscopy (OLYMPUS IX 51). Among the isolated strains Ralstonia sp. was selected after its ability to grow on molasses dates in the presence of a limited nitrogen source was detected. The optimum conditions for formation of biopolymers by isolated strains were investigated. Conditions studied included, best incubation duration (2 days), temperature (30°C) and pH (7-8). The maximum PHB production was raised by 1% (v1v) when using concentrations of dates molasses 1, 2, 3, 4 and 5% in MSM. The best inoculated with 1% old inoculum (1= OD). The ideal extraction method of PHA and PHB proved to be 0.4% sodium hypochlorite solution, producing a quantity of polymer 98.79% of the cell's dry weight. The maximum PHB production was 1.79 g/L recorded by Ralstonia sp. after 48 h, while it was 1.40 g/L produced by R.eutropha ATCC 17697 after 48 h.

Keywords: bacteria forming polyhydroxyalkanoate, detection, molecular, Saudi Arabia

Procedia PDF Downloads 341

Authors: Jakub Młoźniak, Adam Szymański, Gabriela Stondzik, Dagny Krankowska, Tomasz Mikuła

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Nontuberculous mycobacteria (NTM) are bacterial species that cause diversely manifesting diseases mainly in immunocompromised patients. In people with HIV, NTM infection is an AIDS-defining disease and usually appears when the lymphocyte T CD4 count is below 50 cells/μl. The usage of antiretroviral therapy has decreased the prevalence of NTM among people with HIV, but the disease can still be observed especially among patients with late HIV diagnosis. Common presence in environment, human colonization, clinical similarity with tuberculosis and slow growth on culture makes NTM especially hard to diagnose. The study aimed to analyze the epidemiology and clinical course of NTM among patients with HIV. This study included patients with NTM and HIV admitted to our department between 2017 and 2023. Medical records of patients were analyzed and data on age, sex, median time from HIV diagnosis to identification of NTM infection, median CD4 count at NTM diagnosis, methods of determining NTM infection, type of species of mycobacteria identified, clinical symptoms and treatment course were gathered. Twenty-four patients (20 men, 4 women) with identified NTM were included in this study. Among them, 20 were HIV late presenters. The patients' median age was 40. The main symptoms which patients presented were fever, weight loss and cough. Pulmonary disease confirmed with positive cultures from sputum/bronchoalveolar lavage was present in 18 patients. M. avium was the most common species identified. M. marinum caused disseminated skin lesions in 1 patient. Out of all, 5 people were not treated for NTM caused by lack of symptoms and suspicion of colonization with mycobacterium. Concomitant tuberculosis was present in 6 patients. The median diagnostic time from HIV to NTM infections was 3.5 months. The median CD4 count at NTM identification was 69.5 cells/μl. Median NTM treatment time was 16 months but 7 patients haven’t finished their treatment yet. The most commonly used medications were ethambutol and clarithromycin. Among analyzed patients, 4 of them have died. NTM infections are still an important disease among patients who are HIV late presenters. This disease should be taken into consideration during the differential diagnosis of fever, weight loss and cough in people with HIV with lymphocyte T CD4 count <100 cells/μl. Presence of tuberculosis does not exclude nontuberculous mycobacterium coinfection.

Keywords: mycobacteriosis, HIV, late presenter, epidemiology

Procedia PDF Downloads 37

Authors: Simon Grossemy, Peggy P. Y. Chan, Pauline M. Doran

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Nerve tissue engineering is the main field of research aimed at finding an alternative to autografts as a treatment for nerve injuries. Scaffolds are used as a support to enhance nerve regeneration. In order to successfully design novel scaffolds and in vitro cell culture systems, a deep understanding of the factors affecting nerve regeneration processes is needed. Physical and biological parameters associated with the culture environment have been identified as potentially influential in nerve cell differentiation, including electrical stimulation, exposure to extracellular-matrix (ECM) proteins, dynamic medium conditions and co-culture with glial cells. The mechanisms involved in driving the cell to differentiation in the presence of these factors are poorly understood; the complexity of each of them raises the possibility that they may strongly influence each other. Some questions that arise in investigating nerve regeneration include: What are the best protein coatings to promote neural cell attachment? Is the scaffold design suitable for providing all the required factors combined? What is the influence of dynamic stimulation on cell viability and differentiation? In order to study these effects, scaffolds adaptable to bioreactor culture conditions were designed to allow electrical stimulation of cells exposed to ECM proteins, all within a dynamic medium environment. Gold coatings were used to make the surface of viscose rayon microfiber scaffolds (VRMS) conductive, and poly-L-lysine (PLL) and laminin (LN) surface coatings were used to mimic the ECM environment and allow the attachment of rat PC12 neural cells. The robustness of the coatings was analyzed by surface resistivity measurements, scanning electron microscope (SEM) observation and immunocytochemistry. Cell attachment to protein coatings of PLL, LN and PLL+LN was studied using DNA quantification with Hoechst. The double coating of PLL+LN was selected based on high levels of PC12 cell attachment and the reported advantages of laminin for neural differentiation. The underlying gold coatings were shown to be biocompatible using cell proliferation and live/dead staining assays. Coatings exhibiting stable properties over time under dynamic fluid conditions were developed; indeed, cell attachment and the conductive power of the scaffolds were maintained over 2 weeks of bioreactor operation. These scaffolds are promising research tools for understanding complex neural cell behavior. They have been used to investigate major factors in the physical culture environment that affect nerve cell viability and differentiation, including electrical stimulation, bioreactor hydrodynamic conditions, and combinations of these parameters. The cell and tissue differentiation response was evaluated using DNA quantification, immunocytochemistry, RT-qPCR and functional analyses.

Keywords: bioreactor, electrical stimulation, nerve differentiation, PC12 cells, scaffold

Procedia PDF Downloads 236

Authors: Gizem Buldum, Alexander Bismarck, Athanasios Mantalaris

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Cellulose is the most abundant biopolymer on earth and it is currently being utilised in a multitude of industrial applications. Over the last 30 years, attention has been paid to the bacterial cellulose (BC), since BC exhibits unique physical, chemical and mechanical properties when compared to plant-based cellulose, including high purity and biocompatibility. Although Acetobacter xylinum is the most efficient producer of BC, it’s long doubling time results in insufficient yields of the cellulose production. This limits widespread and continued use of BC. In this study, E. coli BL21 (DE3) or E. coli HMS cells are selected as host organisms for the expression of bacterial cellulose synthase operon (bcs) of A.xylinum. The expression system is created based on pET-Duet1 and pCDF plasmid vectors, which carry bcs operon. The results showed that all bcs genes were successfully transferred and expressed in E.coli strains. The expressions of bcs proteins were shown by SDS and Native page analyses. The functionality of the bcs operon was proved by congo red binding assay. The effect of culturing temperature and the inducer concentration (IPTG) on cell growth and plasmid stability were monitored. The percentage of plasmid harboring cells induced with 0.025 mM IPTG was obtained as 85% at 22˚C in the end of 10-hr culturing period. It was confirmed that the high output cellulose production machinery of A.xylinum can be transferred into other organisms.

Keywords: bacterial cellulose, biopolymer, recombinant expression system, production

Procedia PDF Downloads 392

Authors: Jagruti Barot

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The textile industry plays a major role in the economy of India and on the other side of the coin it is the major source for water pollution. As azo dyes is the largest dye class they are extensively used in many fields such as textile industry, leather tanning industry, paper production, food, colour photography, pharmaceuticals and medicine, cosmetic, hair colourings, wood staining, agricultural, biological and chemical research etc. In addition to these, they can have acute and/or chronic effects on organisms depending on their concentration and length of exposure when they discharged as effluent in the environment. The aim of this study was to assess the genotoxic and histotoxic potentials of environmentally relevant concentrations of RR 120 on Catla catla, important edible freshwater fingerlings. For this, healthy Catla catla fingerlings were procured from the Government Fish Farm and acclimatized in 100 L capacity and continuously aerated glass aquarium in laboratory for 15 days. According to APHA some physic-chemical parameters were measured and maintained such as temperature, pH, dissolve oxygen, alkalinity, total hardness. Water along with excreta had been changed every 24 hrs. All fingerlings were fed artificial food palates once a day @ body weight. After 15 days fingerlings were grouped in 5 (10 in each) and exposed to various concentrations of RR 120 (Control, 10, 20, 30 and 40 mg/L) and samples (peripheral blood and gills, kidney) were collected and analyzed at 96 hrs. of interval. All results were compared with the control. Micronuclei (MN), nuclear buds (NB), fragmented-apoptotic (FA) and bi-nucleated (BN) cells in blood cells and in tissues (gills and kidney cells) were observed. Prominent histopathological alterations were noticed in gills such as aneurism, hyperplasia, degenerated central axis, lifting of gill epithelium, curved secondary gill lamellae etc. Similarly kidney showed some detrimental changes like shrunken glomeruli with increased periglomerular space, degenerated renal tubules etc. Both haematological and histopathological changes clearly reveal the toxic potential of RR 120. This work concludes that water pollution assessment can be done by these two biomarkers which provide baseline to the further chromosomal or molecular work.

Keywords: micronuclei, genotoxicity, RR 120, Catla catla

Procedia PDF Downloads 204

Authors: Aftab Ali

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Cryopreservation of semen is one of the key factors for a successful breeding business along with other factors. To achieve high fertility in boar, one should know about spermatozoa response to different treatments proceeds during cryopreservation. The running project is highly focused on cryopreservation and its effects on sperm quality parameters in both boar and bull semen. Semen sample from A, B, C, and D, were subjected to different thawing conditions and were analyzed upon different treatments in the study. Parameters like sperm cell motility, viability, acrosome, DNA integrity, and phospholipase C zeta were detected by different established methods. Different techniques were used to assess different parameters. Motility was detected using computer assisted sperm analysis, phospholipase C zeta using luminometry while viability, acrosome integrity, and DNA integrity were analyzed using flow cytometry. Thawing conditions were noted to have an effect on sperm quality parameters with motility being the most critical parameter. The results further indicated that the most critical step during cryopreservation of boar semen is when sperm cells are subjected to freezing and thawing. The findings of the present study provide insight that; boar semen cryopreservation is still suboptimal in comparison to bull semen cryopreservation. Thus, there is a need to conduct more research to improve the fertilizing potential of cryopreserved boar semen.

Keywords: cryopreservation, computer assisted sperm, flow cytometry, luminometry

Procedia PDF Downloads 142

Authors: Jerman Madonsela, Wallace Matizamhuka, Akiko Yamamoto, Ronald Machaka, Brendon Shongwe

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In this study, biocompatibility evaluation of nanostructured near beta Ti-24Nb-4Zr-8Sn (Ti2448) alloy with non-toxic elements produced utilizing Spark plasma sintering (SPS) of very fine microsized powders attained through mechanical alloying was performed. The results were compared with pure titanium and Ti-6Al-4V (Ti64) alloy. Cell proliferation test was performed using murine osteoblastic cells, MC3T3-E1 at two cell densities; 400 and 4000 cells/mL for 7 days incubation. Pure titanium took a lead under both conditions suggesting that the presence of other oxide layers influence cell proliferation. No significant difference in cell proliferation was observed between Ti64 and Ti2448. Potentiodynamic measurement in Hanks, 0.9% NaCl and cell culture medium showed no distinct difference on the anodic polarization curves of the three alloys, indicating that the same anodic reaction occurred on their surface but with different rates. However, Ti2448 showed better corrosion resistance in cell culture medium with a slightly lower corrosion rate of 2.96 nA/cm2 compared to 4.86 nA/cm2 and 5.62 nA/cm2 of Ti and Ti64 respectively. Ti2448 adsorbed less protein as compared to Ti and Ti64 though no notable difference in surface wettability was observed.

Keywords: biocompatibility, osteoblast, corrosion, surface wettability, protein adsorption

Procedia PDF Downloads 216

Authors: A. P. Attanayake, K. A. P. W. Jayatilaka, L. K. B. Mudduwa, C. Pathirana

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Triggering of β-cell regeneration is a recognized therapeutic strategy for the treatment of type 1 diabetes mellitus. One such approach to foster restoration and regeneration of β-cells is from exogenous natural extracts. The aim of the present study was to investigate and compare the β-cell regenerative potentials of the extracts of Spondias pinnata (Linn. f.) Kurz, Coccinia grandis (L.) Voigt and Gmelina arborea Roxb. in alloxan induced diabetic rats. Wistar rats were divided in to six groups (n=6); healthy untreated rats, alloxan induced diabetic untreated rats (150 mg/kg, ip), diabetic rats receiving the extracts of S. pinnata (1.0 g/kg), C. grandis (0.75 g/kg), G. arobrea (1.00 g/kg) and diabetic rats receiving glibenclamide (0.5 mg/kg) for 30 days. The assessment of selected biochemical parameters, histopathology and immunohistochemistry in the pancreatic tissue were done on the 30th day. The reduction in the percentage of HbA1C was in the decreasing order of C. grandis (35%), G. arborea (31%) and S. pinnata (29%) in alloxan induced diabetic rats (p< 0.05). The concentration of serum fructosamine, insulin and C-peptide were decreased significantly in a decreasing order of C. grandis (30%, 72%, 51%), G. arborea (25%, 44%, 44%) and S. pinnata (27%, 34%, 24%) in alloxan induced diabetic rats (p < 0.05). The extent of β-cell regeneration was in the decreasing order of C. grandis, G. arborea, S. pinnata reflected through the increased percentage of insulin secreting β-cells in alloxan induced diabetic rats. The extract of C. grandis produced the highest degree of β-cell regeneration demonstrated through an increase in the number of islets and percentage of the insulin secreting β-cells (75%) in the pancreas of diabetic rats (p < 0.05). Further the C. grandis extract produced a significant increase in mean profile diameter in small (118%), average (10%), and large (13%) islets as compared with diabetic control rats respectively. However, statistically significant increase in the islet profile diameter was shown only in average (2%) and large (5%) islets in the G. arborea extract treated rats and large islets (5%) in S. pinnata extract treated diabetic rats (p < 0.05). The β-cell regeneration potency was in the decreasing order of C. grandis (0.75 g/kg), G. arborea (1.00 g/kg) and S. pinnata (1.00 g/kg) in alloxan induced diabetic rats. The three plant extracts may be useful as natural agents of triggering the β-cell regeneration in the management of type 1 diabetes mellitus.

Keywords: alloxan-induced diabetic rats, β-cell regeneration, histopathology, immunohistochemistry

Procedia PDF Downloads 236

Authors: Carlos Y. Soto, Camila A. Lota, G. Astrid Garzón

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Bacterial biofilms cause an ongoing problem for food safety. They are formed when microorganisms aggregate to form a community that attaches to solid surfaces. Biofilms increase the resistance of pathogens to cleaning, disinfection and antibacterial products. This resistance gives rise to problems for human health, industry, and agriculture. At present, plant extracts rich in polyphenolics are being investigated as natural alternatives to degrade bacterial biofilms. The pomace of the tropical Berry Vaccinium meridionale S. contains high amounts of phenolic compounds. Therefore, in the current study, the antimicrobial and antibiofilm effects of extracts from the pomace of Vaccinium meridionale S. were tested on three foodborne pathogens: Enterohaemorrhagic Escherichia coli O157:H7 (ATCC®700728TM), Staphylococcus aureus subsp. aureus (ATCC® 6538TM), and Salmonella enterica serovar Enteritidis (ATCC® 13076TM). Microwave-assisted extraction was used to extract polyphenols with aqueous methanol (80% v/v) at a solid to solvent ratio of 1:10 (w/v) for 20 min. The magnetic stirring was set at 400 rpm, and the microwave power was adjusted to 400 W. The antimicrobial effect of the extract was assessed by determining the half maximal inhibitory concentration (IC50) against the three food poisoning pathogens at concentrations ranging from 50 to 2,850 μg gallic acid equivalents (GAE)/mL of the extract. Biofilm inhibition was assessed using a crystal violet assay applying the same range of concentration. Three replications of the experiments were carried out, and all analyses were run in triplicate. IC50 values were determined using the GraphPad Prism8® program. Significant differences (P<0.05) among means were identified using one-factor analysis of variance (ANOVA) and the post-hoc least significant difference (LSD) test using the Statgraphics plus program, version 2.1.There was significant difference among the mean IC50 values for the tested bacteria. The IC50 for S. aureus was 48 ± 9 μg GAE/mL, followed by 123 ± 49 μg GAE/mL for Salmonella and 376 ± 32 μg GAE/mL for E. coli. The percent inhibition of the extract on biofilm formation was significantly higher for S. aureus (85.8  0.3), followed by E. coli (74.5  1.0) and Salmonella (53.6  9.7). These findings suggest that polyphenolic extracts obtained from the pomace of V. meridionale S. might be used as natural antimicrobial and anti-biofilm natural agents, effective against S. aureus, E. coli and Salmonella enterica.

Keywords: antibiofilm, antimicrobial, E. coli, S. aureus, salmonella, IC50, pomace, V. meridionale

Procedia PDF Downloads 56

Authors: Yen-Ni Teng, Hsing-Yi Chen, Hsien-An Pan, Yung-Ming Lin, Hany A. Omar, Jui-Hsiang Hung

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Leucine-rich repeats and WD repeat domain containing 1 (LRWD1) is highly expressed in the testes of healthy males. On the other hand, LRWD1 is significantly down-regulated in the testicular tissues of patients with severe spermatogenic defects. In our study, the downregulation of LRWD1 expression by shRNA caused a significant reduction of cell growth and mitosis and a noteworthy increase in the cell microtubule atrophy rate. Here, we used EMBOSS CpG plot analysis to explore the promoter region of LRWD1 gene. We found that CpG islands are located between positions -253 to +5 nucleotides upstream from the LRWD1 transcription start site. Luciferase reporter assay revealed that the hypermethylation of the LRWD1 promoter reduced the transcription activity in cells. In addition, quantitative methylation-specific PCR and immunostaining showed that the methylation inhibitor, 5-Aza-2'-deoxycytidine, increased LRWD1 promoter activity, LRWD1 mRNA, protein expression and cell viability. Whereas, the methylation activator, S-adenosylmethionine, caused opposite effects. The overexpression of p53 and Nrf2 in NT2/D1 cells increased LRWD1 promoter activity while 5-fluorodeoxyuridine decreased it. In conclusion, this study highlights evidence that the methylation status of LRWD1 promoter is associated with LRWD1 expression. Since the expression level of LRWD1 plays an important role in spermatogenesis, the methylation status of LRWD1 may serve as a novel molecular diagnostic or therapeutic approach in male's infertility.

Keywords: LRWD1, DNA methylation, p53, Nrf2

Procedia PDF Downloads 141

Authors: Mohamad Bolandmartabe, Nafiseh Hassani, Saeed Abdi Darake, Maryam Asghari

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Dogs are highly susceptible to diseases, nutritional problems, toxins, and parasites, with parasitic infections being common and causing hardship in their lives. Some important internal parasites include worms (such as roundworms and tapeworms) and protozoa, which can lead to anemia in dogs. Important bloodborne parasites in dogs include microfilariae and adult forms of Dirofilaria immitis, Dipetalonema reconditum, Babesia, Trypanosoma, Hepatozoon, Leishmania, Ehrlichia, and Hemobartonella. Babesia and Hemobartonella are parasites that reside inside red blood cells and cause regenerative anemia by directly destroying the red blood cells. Hepatozoon, Leishmania, and Ehrlichia are also parasites that reside within white blood cells and can infiltrate other tissues, such as the liver and lymph nodes. Since intermediate hosts are more commonly found in the open environment, the prevalence of parasites in stray and free-roaming dogs is higher compared to pet dogs. Furthermore, pet dogs are less exposed to internal and external parasites due to better care, hygiene, and being predominantly indoors. Therefore, they are less likely to be affected by them. Among the parasites, Leishmania carries significant importance as it is shared between dogs and humans, causing a dangerous disease known as visceral Leishmaniasis or kala-azar and cutaneous Leishmaniasis. Furthermore, dogs can act as reservoirs and spread the disease agent within human communities. Therefore, timely and accurate diagnosis of these diseases in dogs can be highly beneficial in preventing their occurrence in humans. In this article, we employed the Giemsa staining technique under a light microscope for the identification of bloodborne parasites in dogs. However, considering the negative impact of these parasites on the natural life of dogs, the development of chronic diseases, and the gradual loss of the animal's well-being, rapid and timely diagnosis is essential. Serological methods and PCR are available for the diagnosis of certain parasites, which have high sensitivity and desirable characteristics. Therefore, this research aims to investigate the molecular aspects of bloodborne parasites in dogs referred to veterinary hospitals in Tehran city.

Keywords: leishmaniasis, babesiosis, ehrlichiosis, dirofilariasis, hepatozoonosis

Procedia PDF Downloads 95

Authors: Orapan Paecharoenchai, Tanasait Ngawhirunpat, Theerasak Rojanarata, Auayporn Apirakaramwong, Praneet Opanasopit

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Cationic niosomes formulated with Span20, cholesterol and novel synthesized spermine-cationic lipids (2-hydrocarbon tail and 4- hydrocarbon tail) in a molar ratio of 2.5:2.5:1 can mediate high gene transfection in vitro. However, the uptake mechanisms of these systems are not well clarified. In the present study, effect of endocytic inhibitors on the transfection efficiency of niosomes/DNA complexes was determined on a human cervical carcinoma cell line (HeLa cells) using the inhibitors of macropinocytosis (wortmannin), clathrin- and caveolae-mediated endocytosis (methyl-β-cyclodextrin), clathrin-mediated endocytosis (chlorpromazine), caveolae-mediated endocytosis (genistein and filipin), cytosolic transfer (ammonium chloride) and microtubules polymerization (nocodazole). The transfection of niosomes with 2-hydrocarbon tail lipid was blocked by nocodazole, genistein, ammonium chloride and filipin, respectively, whereas, the transfection of niosomes with 4-hydrocarbon tail lipid was blocked by nocodazole, genistein, ammonium chloride, methyl-β-cyclodextrin and filipin, respectively. It can be concluded that these niosomes/DNA complexes were internalized predominantly by endocytosis via clathrin and caveolae-independent pathway.

Keywords: cellular internalization, cationic niosomes, gene carriers, spermine-cationic lipids

Procedia PDF Downloads 447

Authors: Abderahim Ahmadou, Alfredo Napoli, Noel Durand, Didier Montet

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Biochar is a microporous and adsorbent solid carbon product obtained from the pyrolysis of various organic materials (biomass, agricultural waste). Biochar is distinguished from vegetable charcoal by its manufacture methods. Biochar is used as the amendment in soils to give them favorable characteristics under certain conditions, i.e., absorption of water and its release at low speed. Cashew nuts shell from Mali is usually discarded on land by local processors or burnt as a mean for waste management. The burning of this biomass poses serious socio-environmental problems including greenhouse gas emission and accumulation of tars and soot on houses closed to factories, leading to neighbor complaints. Some mycotoxins as aflatoxins are carcinogenic compounds resulting from the secondary metabolism of molds that develop on plants in the field and during their conservation. They are found at high level on some seeds and nuts in Africa. Ochratoxin A, member of mycotoxins, is produced by various species of Aspergillus and Penicillium. Human exposure to Ochratoxin A can occur through consumption of contaminated food products, particularly contaminated grain, as well as coffee, wine grapes. We showed that cashew shell biochars produced at 400, 600 and 800°C adsorbed aflatoxins (B1, B2, G1, G2) at 100% by filtration (rapid contact) as well as by stirring (long contact). The average percentage of adsorption of Ochratoxin A was 35% by filtration and 80% by stirring. The duration of the biochar-mycotoxin contact was a significant parameter. The effect of biochar was also tested on two strains of toxigenic molds: Aspergillus parasiticus (producers of Aflatoxins) and Aspergillus carbonarius (producers of Ochratoxins). The growth of the strain Aspergillus carbonarius was inhibited at up to 60% by the biochar at 600°C. An opposite effect to the inhibition was observed on Aspergillus parasiticus using the same biochar. In conclusion, we observed that biochar adsorbs mycotoxins: Aflatoxins and Ochratoxin A to different degrees; 100% adsorption of aflatoxins under all conditions (filtration and stirring) and adsorption of Ochratoxin A varied depending on the type of biochar and the experiment conditions (35% by filtration and 85% by stirring). The effects of biochar at 600 °C on the toxigenic molds: Aspergillus parasiticus and Aspergillus carbonarius, varied according to the experimental conditions and the strains. We observed an opposite effect on the growth with an inhibition of Aspergillus carbonarius up to 60% and a stimulated growth of Aspergillus parasiticus.

Keywords: biochar, cashew nut shell, mycotoxins, toxicogenic molds

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Authors: Hager Elsheikh

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Context: Pancreatitis is a condition characterized by inflammation in the pancreas, which can lead to serious complications if untreated. Both acute and chronic pancreatitis are associated with immune reactions and fibrosis, which further damage the pancreas. The type 2 immune response, primarily driven by alternative activated macrophages (AAMs), plays a significant role in the development of fibrosis. The IL-4/STAT6 pathway is a crucial signaling pathway for the activation of M2 macrophages. Pancreatic fibrosis is induced by dysregulated inflammatory responses and can result in the autodigestion and necrosis of pancreatic acinar cells. Research Aim: The aim of this study is to investigate the impact of STAT6, a crucial molecule in the IL-4/STAT6 pathway, on the severity and development of fibrosis during acute and chronic pancreatitis. The research also aims to understand the influence of the JAK/STAT6 signaling pathway on the balance between fibrosis and regeneration in the presence of different macrophage populations. Methodology: The research utilizes murine models of acute and chronic pancreatitis induced by cerulean injection. Animal models will be employed to study the effect of STAT6 knockout on disease severity and fibrosis. Isolation of acinar cells and cell culture techniques will be used to assess the impact of different macrophage populations on wound healing and regeneration. Various techniques such as PCR, histology, immunofluorescence, and transcriptomics will be employed to analyze the tissues and cells. Findings: The research aims to provide insights into the mechanisms underlying tissue fibrosis and wound healing during acute and chronic pancreatitis. By investigating the influence of the JAK/STAT6 signaling pathway and different macrophage populations, the study aims to understand their impact on tissue fibrosis, disease severity, and pancreatic regeneration. Theoretical Importance: This research contributes to our understanding of the role of specific signaling pathways, macrophage polarization, and the type 2 immune response in pancreatitis. It provides insights into the molecular mechanisms underlying tissue fibrosis and the potential for targeted therapies. Data Collection and Analysis Procedures: Data will be collected through the use of murine models, isolation and culture of acinar cells, and various experimental techniques such as PCR, histology, immunofluorescence, and transcriptomics. Data will be analyzed using appropriate statistical methods and techniques, and the findings will be interpreted in the context of the research objectives. Conclusion: By investigating the mechanisms of tissue fibrosis and wound healing during acute and chronic pancreatitis, this research aims to enhance our understanding of the disease progression and potential therapeutic targets. The findings have theoretical importance in expanding our knowledge of pancreatic fibrosis and the role of macrophage polarization in the context of the type 2 immune response.

Keywords: immunity in chronic diseases, pancreatitis, macrophages, immune response

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Authors: Dike H. Ogbuagu, Etsede J. Oritsematosan

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This work investigated possible inductions of CaC₂, often misused by fruit vendors to stimulate artificial ripening, on mammalian sperm morphology and viability. Thirty isogenic strains of male albino mice, Mus musculus (age≈ 8weeks; weight= 32.5±2.0g) were acclimatized (ambient temperature 28.0±1.0°C) for 2 weeks and fed standard growers mash and water ad libutum. They were later exposed to graded toxicant concentrations (w/w) of 2.5000, 1.2500, 0.6250, and 0.3125% in 4 cages. A control cage was also established. After 5 weeks, 3 animals from each cage were sacrificed by cervical dislocation and the cauda epididymis excised. Sperm morphology and viability were determined by microscopic procedures. The ANOVA, means plots, Student’s t-test and variation plots were used to analyze data. The common abnormalities observed included Double Head, Pin Head, Knobbed Head, No Tail and With Hook. The higher toxicant concentrations induced significantly lower body weights [F(829.899) ˃ Fcrit(4.19)] and more abnormalities [F(26.52) ˃ Fcrit(4.00)] at P˂0.05. Sperm cells in the control setup were significantly more viable than those in the 0.625% (t=0.005) and 2.500% toxicant doses (t=0.018) at the 95% confidence limit. CaC₂ appeared to induced morphological abnormalities and reduced viability in sperm cells of M. musculus.

Keywords: artificial ripening, calcium carbide, fruit vendors, sperm morphology, sperm viability

Procedia PDF Downloads 217

Authors: Boukli Hacene Faiza, Soufi Wassila, Ghalem Said

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The oral antidiabetics drugs such as alpha glucosidase inhibitors present undesirable effects like acarbose. Flavonoids are class of molecules widely distributed in plants, for this reason we are interested in our work to study the inhibition in silico of alpha glucosidase by natural ligands ( flavonoids analogues) using molecular modeling methods using MOE (Molecular Operating Environment) software to predict their interaction with this enzyme with score energy, ADME /T tests and druglikeness properties experiments. Two flavonoids Beicalein and Apigenin have high binding affinity with alpha glucosidase with lower IC50 supposed potent inhibitors.

Keywords: alpha glucosidase, flavonoides analogues, drug research, molecular modeling

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Authors: Mounir Baccar, Imen Jmal

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Because of the rise in energy costs, thermal storage systems designed for the heating and cooling of buildings are becoming increasingly important. Energy storage can not only reduce the time or rate mismatch between energy supply and demand but also plays an important role in energy conservation. One of the most preferable storage techniques is the Latent Heat Thermal Energy Storage (LHTES) by Phase Change Materials (PCM) due to its important energy storage density and isothermal storage process. This paper presents a numerical study of the solidification of a PCM (paraffin RT27) in a rectangular thermal storage exchanger for air conditioning systems taking into account the presence of natural convection. Resolution of continuity, momentum and thermal energy equations are treated by the finite volume method. The main objective of this numerical approach is to study the effect of natural convection on the PCM solidification time and the impact of fins number on heat transfer enhancement. It also aims at investigating the temporal evolution of PCM solidification, as well as the longitudinal profiles of the HTF circling in the duct. The present research undertakes the study of two cases: the first one treats the solidification of PCM in a PCM-air heat exchanger without fins, while the second focuses on the solidification of PCM in a heat exchanger of the same type with the addition of fins (3 fins, 5 fins, and 9 fins). Without fins, the stratification of the PCM from colder to hotter during the heat transfer process has been noted. This behavior prevents the formation of thermo-convective cells in PCM area and then makes transferring almost conductive. In the presence of fins, energy extraction from PCM to airflow occurs at a faster rate, which contributes to the reduction of the discharging time and the increase of the outlet air temperature (HTF). However, for a great number of fins (9 fins), the enhancement of the solidification process is not significant because of the effect of confinement of PCM liquid spaces for the development of thermo-convective flow. Hence, it can be concluded that the effect of natural convection is not very significant for a high number of fins. In the optimum case, using 3 fins, the increasing temperature of the HTF exceeds approximately 10°C during the first 30 minutes. When solidification progresses from the surfaces of the PCM-container and propagates to the central liquid phase, an insulating layer will be created in the vicinity of the container surfaces and the fins, causing a low heat exchange rate between PCM and air. As the solid PCM layer gets thicker, a progressive regression of the field of movements is induced in the liquid phase, thus leading to the inhibition of heat extraction process. After about 2 hours, 68% of the PCM became solid, and heat transfer was almost dominated by conduction mechanism.

Keywords: heat transfer enhancement, front solidification, PCM, natural convection

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Authors: Hassan Mohammadi Khujin

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Tissue engineering is the science of tissues and complex organs creation using scaffolds, cells and biologically active components. Most cells require scaffolds to grow and proliferate. These temporary support structures for tissue regeneration are later replaced with extracellular matrix produced inside the body. Recent advances in additive manufacturing methods allow production of highly porous, complex three dimensional scaffolds suitable for cell growth and proliferation. The current paper investigates the mechanical properties, including elastic modulus and compressive strength, as well as fluid flow dynamics, including permeability and flow-induced shear stress of scaffolds with four triply periodic minimal surface (TPMS) configurations, namely the Schwarz primitive, the Schwarz diamond, the gyroid, and the Neovius structures. Higher porosity in all scaffold types resulted in lower mechanical properties. The permeability of the scaffolds was determined using Darcy's law with reference to geometrical parameters and the pressure drop derived from the computational fluid dynamics (CFD) analysis. Higher porosity enhanced permeability and reduced wall shear stress in all scaffold designs.

Keywords: highly porous scaffolds, tissue engineering, finite elements analysis, CFD analysis

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Authors: Yulistiani Yulistiani, Zamrotul Izzah, Lintang Bismantara, Wenny Putri Nilamsari, Arif Bachtiar, Budi Suprapti

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Interferon-gamma (IFN-γ) and interleukin-6 (IL-6) are pro-inflammatory cytokines, which have the protective immune response against Tuberculosis (TB). Indeed, pro-inflammatory cytokines Mycobacterium tuberculosis antigen-specific CD4+ and CD8+ T cells and NK cells increase the level of production of IFN-γ, a cytokine critical for augmenting the microbicidal activity of phagocytes. On the other hand, M. tuberculosis reduces the effects of IFN-γ by inhibiting the transcription of IFN-γ- responsive genes and by inducing the secretion of IL-6, which inhibits IFN-γ signaling. Probiotics Lactobacillus sp. and Bifidobacterium sp. were known to increase IFN-γ production in vivo, while vitamin B1, B6, and B12 worked on macrophages and releasing cytokines. Therefore, the present study was to evaluate the effect of probiotics and vitamin B supplement on changes of plasma cytokine levels in active pulmonary TB. From October to November 2016, twelve M. tuberculosis-infected patients starting anti-TB drugs were recruited, then divided into two groups. Seven patients were given a combination of probiotics and vitamin B, while five patients were in the control group. Plasma IFN-γ and IL-6 levels were measured by the ELISA kit before and a month after treatment. IFN-γ levels raised in four patients receiving the supplement (P = 0.743), while IL-6 increased in three patients in this group until day 30 of treatment (P = 0.298). Taken together, these results show the promising effect of probiotics and vitamin B on stimulation of IFN-γ and IL-6 production during intensive therapy of TB.

Keywords: interferon-gamma, interleukin-6, probiotic, tuberculosis

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Authors: Wissam Saliba, Mandy Nicholson

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Secretory carcinoma (SC) is a rare type of salivary gland cancer. It was first realized as a distinct type of malignancy in 2010and wasinitially termed “mammary analogue secretory carcinoma” because of similarities with secretory breast cancer. The name was later changed to SC. Most SCs originate in parotid glands, and most harbour a rare gene mutation: ETV6-NTRK3. This mutation is rare in common cancers and common in rare cancers; it is present in most secretory carcinomas. Disease outcomes for SC are usually described as favourable as many cases of SC are lowgrade (LG), and cancer growth is slow. In early stages, localized therapy is usually indicated (surgery and/or radiation). Despitea favourable prognosis, a sub-set of casescan be much more aggressive.These cases tend to be of high-grade(HG).HG casesare associated with a poorer prognosis.Management of such cases can be challenging due to limited evidence for effective systemic therapy options. This case report describes the clinical management of a 46-year-oldmale patient with a unique case of SC. He was initially diagnosed with a low/intermediate grade carcinoma of the left parotid gland in 2009; he was treated with surgery and adjuvant radiation. Surgical pathology favoured primary salivary adenocarcinoma, and 2 lymph nodes were positive for malignancy. SC was not yet realized as a distinct type of cancerat the time of diagnosis, and the pathology reportvalidated this gap by stating that the specimen lacked features of the defined types of salivary carcinoma.Slow-growing pulmonary nodules were identified in 2017. In 2020, approximately 11 years after the initial diagnosis, the patient presented with malignant pleural effusion. Pathology from a pleural biopsy was consistent with metastatic poorly differentiated cancer of likely parotid origin, likely mammary analogue secretory carcinoma. The specimen was sent for Next Generation Sequencing (NGS); ETV6-NTRK3 gene fusion was confirmed, and systemic therapy was initiated.One cycle ofcarboplatin/paclitaxel was given in June 2020. He was switched to Larotrectinib (NTRK inhibitor (NTRKi)) later that month. Larotrectinib continued for approximately 9 months, with discontinuation in March 2021 due to disease progression. A second-generation NTRKi (Selitrectinib) was accessed and prescribedthrough a single patient study. Selitrectinib was well tolerated. The patient experienced a complete radiological response within~4 months. Disease progression occurred once again in October 2021. Progression was slow, and Selitrectinib continuedwhile the medical team performed a thorough search for additional treatment options. In January 2022, a liver lesion biopsy was performed, and NGS showed an NTRKG623R solvent-front resistance mutation. Various treatment pathways were considered. The patient pursuedanother investigational NTRKi through a clinical trial, and Selitrectinib was discontinued in July 2022. Excellent performance status was maintained throughout the entire course of treatment.It can be concluded that NTRK inhibitors provided satisfactory treatment efficacy and tolerance for this patient with high-grade transformation and NTRK gene fusion cancer. In the future, more clinical research is needed on systemic treatment options for high-grade transformations in NTRK gene fusion SCs.

Keywords: secretory carcinoma, high-grade transformations, NTRK gene fusion, NTRK inhibitor

Procedia PDF Downloads 106

Authors: Mitesh Rathod, Jungho Ahn, Noo Li Jeon, Junghoon Lee

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Endothelial cells respond to cues from both biochemical as well as micro mechanical environment. Significant effort has been directed to understand the effects of biochemical signaling, however, relatively little is known about regulation of endothelial cell biology by the micro mechanical environment. Numerous studies have been performed to understand how physical forces regulate endothelial cell behavior. In this regard, past studies have majorly focused on exploring how fluid shear stress governs endothelial cell behavior. Parallel plate flow chambers and rectangular microchannels are routinely employed for applying fluid shear force on endothelial cells. However, these studies fall short in mimicking the in vivo like micro environment from topological aspects. Few studies have only used circular microchannels to replicate in vivo like condition. Seldom efforts have been directed to elucidate the combined effect of topology, substrate rigidity and fluid shear stress on endothelial cell response. In this regard, we demonstrate a facile fabrication process to develop a hybrid polydimethylsiloxane microfluidic platform to study endothelial cell biology. On a single chip microchannels with different cross sections i.e., circular, rectangular and square have been fabricated. In addition, our fabrication approach allows variation in the substrate rigidity along the channel length. Two different variants of polydimethylsiloxane, namely Sylgard 184 and Sylgard 527, were utilized to achieve the variation in rigidity. Moreover, our approach also enables in creating Y bifurcation circular microchannels. Our microfluidic platform thus facilitates for conducting studies pertaining to endothelial cell morphology with respect to change in topology, substrate rigidity and fluid flow on a single chip. The hybrid platform was tested by culturing Human Umbilical Vein Endothelial Cells in circular microchannels with varying substrate rigidity, and exposed to fluid shear stress of 12 dynes/cm² and static conditions. Results indicate the cell area response to flow induced shear stress was governed by the underlying substrate mechanics.

Keywords: hybrid, microfluidic platform, PDMS, shear flow, substrate rigidity

Procedia PDF Downloads 268

Authors: Wiam El Kheir, Anais Dumais, Maude Beaudoin, Bernard Marcos, Nick Virgilio, Benoit Paquette, Nathalie Faucheux, Marc-Antoine Lauzon

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The high infiltrative pattern of glioblastoma multiforme cells (GBM) is the main cause responsible for the actual standard treatments failure. The tumor high heterogeneity, the interstitial fluid flow (IFF) and chemokines guides GBM cells migration in the brain parenchyma resulting in tumor recurrence. Drug delivery systems emerged as an alternative approach to develop effective treatments for the disease. Some recent studies have proposed to harness the effect CXC-lchemokine-ligand-12 to direct and control the cancer cell migration through delivery system. However, the dynamics of the brain environment on the delivery system remains poorly understood. Nanoparticles (NPs) and hydrogels are known as good carriers for the encapsulation of different agents and control their release. We studied the release of CXCL12 (free or loaded into NPs) from an alginate-based hydrogel under static and indirect perfusion (IP) conditions. Under static conditions, the main phenomena driving CXCL12 release from the hydrogel was diffusion with the presence of strong interactions between the positively charged CXCL12 and the negatively charge alginate. CXCL12 release profiles were independent from the initial mass loadings. Afterwards, we demonstrated that the release could tuned by loading CXCL12 into Alginate/Chitosan-Nanoparticles (Alg/Chit-NPs) and embedded them into alginate-hydrogel. The initial burst release was substantially attenuated and the overall cumulative release percentages of 21%, 16% and 7% were observed for initial mass loadings of 0.07, 0.13 and 0.26 µg, respectively, suggesting stronger electrostatic interactions. Results were mathematically modeled based on Fick’s second law of diffusion framework developed previously to estimate the effective diffusion coefficient (Deff) and the mass transfer coefficient. Embedding the CXCL12 into NPs decreased the Deff an order of magnitude, which was coherent with experimental data. Thereafter, we developed an in-vitro 3D model that takes into consideration the convective contribution of the brain IFF to study CXCL12 release in an in-vitro microenvironment that mimics as faithfully as possible the human brain. From is unique design, the model also allowed us to understand the effect of IP on CXCL12 release in respect to time and space. Four flow rates (0.5, 3, 6.5 and 10 µL/min) which may increase CXCL12 release in-vivo depending on the tumor location were assessed. Under IP, cumulative percentages varying between 4.5-7.3%, 23-58.5%, 77.8-92.5% and 89.2-95.9% were released for the three initial mass loadings of 0.08, 0.16 and 0.33 µg, respectively. As the flow rate increase, IP culture conditions resulted in a higher release of CXCL12 compared to static conditions as the convection contribution became the main driving mass transport phenomena. Further, depending on the flow rate, IP had a direct impact on CXCL12 distribution within the simulated brain tissue, which illustrates the importance of developing such 3D in-vitro models to assess the efficiency of a delivery system targeting the brain. In future work, using this very model, we aim to understand the impact of the different phenomenon occurring on GBM cell behaviors in response to the resulting chemokine gradient subjected to various flow while allowing them to express their invasive characteristics in an in-vitro microenvironment that mimics the in-vivo brain parenchyma.

Keywords: 3D culture system, chemokines gradient, glioblastoma multiforme, kinetic release, mathematical modeling

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Authors: Islam Nowisser, Noha Farag, Mohamed El Azizi

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Helicobacter pylori is a highly important human pathogen which inhabits about 50% of the population worldwide. Infection with this bacteria is very hard to treat, with high probability of recurrence. H. pylori causes severe gastric diseases, including peptic ulcer, gastritis, and gastric cancer, which has been linked to chronic inflammation. The infection has been reported to be associated with high levels of pro-inflammatory cytokines, especially IL-1β and TNF-α. The aim of the current study is to investigate the molecular mechanisms by which H. pylori activates NLRP3 inflammasome and its contribution to Il-1 β production in an innate cellular model. H. pylori PMSS1 and G27 standard strains, as well as the PMSS1 isogenic mutant strain PMSS1ΔVacA and G27ΔVacA, G27ΔCagA in addition to clinical isolates obtained from biopsy samples from the antrum and corpus mucosa of chronic gastritis patients, were used to establish infection in RAW-264.7 macrophages. The production levels of TNF-α and IL-1β was assessed using ELISA. Since expression of these cytokines is often regulated by the transcription factor complex, nuclear factor-kB (NF-kB), the activation of NF-κB in H. pylori infected cells was also evaluated by luciferase assay. Genomic DNA was extracted from bacterial cultures of H. pylori clinical isolates as well as the standard strains and their corresponding mutants, where they were evaluated for the cagA pathogenicity island and vacA expression. The correlation between these findings and expression of the cagA Pathogenicity Island and vacA in the bacteria was also investigated. The results showed IL-1β, and TNF-α production significantly increased in raw macrophages following H. pylori infection. The cagA+ and vacA+ H. pylori strains induced significant production of IL-1β compared to cagA- and vacA- strains. The activation pattern of NF-κB was correlated in the isolates to their cagA and vacA expression profiles. A similar finding could not be confirmed for TNF-α production. Our study shows the ability of H. pylori to activate NF-kB and induce significant IL-1β production as a possible mechanism for the augmented inflammatory response seen in subjects infected with cagA+ and vacA+ H. pylori strains that would lead to the progression to more severe form of the disease.

Keywords: Helicobacter pylori, IL-1β, inflammatory cytokines, nuclear factor KB, TNF-α

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Authors: Abdul Walusansa

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In this paper, paper notes and coins were collected from general public in Kampala City where ready-to-eat food can be served, in order to survey for bacterial contamination. The total bacterial number and potentially pathogenic organisms loading on currency were tested. All isolated potential pathogens were also tested for antibiotic resistance against four most commonly prescribed antibiotics. 1. The bacterial counts on one hundred paper notes sample were ranging between 6～10918/cm cm-2,the median was 141/ cm-2, according to the data it was much higher than credit cards and Australian notes which were made of polymer. The bacterial counts on sixty coin samples were ranging between 2～380/cm-2, much less than paper notes. 2. Coliform (65.6%), E. coli (45.9%), S. aureus (41.7%), B. cereus (67.7%), Salmonella (19.8%) were isolated on one hundred paper notes. Coliform (22.4%), E. coli (5.2%), S. aureus (24.1%), B. cereus (34.5%), Salmonella (10.3%) were isolated from sixty coin samples. These results suggested a high rate of potential pathogens contamination of paper notes than coins. 3. Antibiotic resistances are commonly in most of the pathogens isolated on currency. Ampicillin resistance was found in 60%of Staphylococcus aureus isolated on currency, as well as 76.6% of E. coil and 40% of Salmonella. Erythromycin resistance was detected in 56.6% of S. aureus and in 80.0% of E. coli. All the pathogens isolated were sensitive to Norfloxacin, Salmonella and S. aureus also sensitive to Cefaclor. In this paper, we also studied the antimicrobial capability of metal coins, coins collected from different countries were tested for the ability to inhibit the growth of E. sakazakii, S. aureus, E. coli, L. monocytogenes and S. typhimurium. 1) E. sakazakii appeared very sensitive to metal coins, the second is S. aureus, but E. coli, L. monocytogenes and S. typhimurium are more resistant to these metal coin samples. 2) Coins made of Nickel-brass alloy and Copper-nickel alloy showed a better effect in anti-microbe than other metal coins, especially the ability to inhibited the growth of E. sakazakii and S. aureus, all the inhibition zones produced on nutrient agar are more than 20.6 mm. Aluminium-bronze alloy revealed weak anti-microbe activity to S. aureus and no effect to kill other pathogens. Coins made of stainless steel also can’t resist bacteria growth. 3) Surprisingly, one cent coins of USA which were made of 97.5% Zinc and 2.5% Cu showed a significant antimicrobial capability, the average inhibition zone of these five pathogens is 45.5 mm.

Keywords: antibiotic sensitivity, bacteria, currency, coins, parasites

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Authors: Elham Mansouri, Asghar Mesbahi

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Purpose: In the current study, we aimed to investigate the macroscopic and microscopic dose enhancement effect of metallic nanoparticles in interstitial brachytherapy of uterus cancer by Iodin-125 source using a nano-lattice model in MCNPX (5) and MCNP6.1 codes. Materials and methods: Based on a nano-lattice simulation model containing a radiation source and a tumor tissue with cellular compartments loaded with 7mg/g spherical nanoparticles (bismuth, gold, and gadolinium), the energy deposited by the secondary electrons in microscopic and macroscopic level was estimated. Results: The results show that the values of macroscopic DEF is higher than microscopic DEF values and the macroscopic DEF values decreases as a function of distance from the brachytherapy source surface. Also, the results revealed a remarkable discrepancy between the DEF and secondary electron spectra calculated by MCNPX (5) and MCNP6.1 codes, which could be justified by the difference in energy cut-off and electron transport algorithms of two codes. Conclusion: According to the both MCNPX (5) and MCNP6.1 outputs, it could be concluded that the presence of metallic nanoparticles in the tumor tissue of uteruscancer increases the physical effectiveness of brachytherapy by I-125 source. The results presented herein give a physical view of radiosensitization potential of different metallic nanoparticles and could be considered in design of analytical and experimental radiosensitization studies in tumor regions using various radiotherapy modalities in the presence of heavy nanomaterials.

Keywords: MCNPX, MCNP6, nanoparticle, brachytherapy

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Authors: Francesco Pantano, Marta Fogolari, Michele Iuliani, Sonia Simonetti, Silvia Cavaliere, Marco Russano, Fabrizio Citarella, Bruno Vincenzi, Silvia Angeletti, Giuseppe Tonini

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Background: Although immune checkpoint inhibitors (ICIs) have changed the treatment paradigm of non–small cell lung cancer (NSCLC), these drugs fail to elicit durable responses in the majority of NSCLC patients. The gut microbiota, able to regulate immune responsiveness, is emerging as a promising, modifiable target to improve ICIs response rates. Since the oral microbiome has been demonstrated to be the primary source of bacterial microbiota in the lungs, we investigated its composition as a potential predictive biomarker to identify and select patients who could benefit from immunotherapy. Methods: Thirty-five patients with stage IV squamous and non-squamous cell NSCLC eligible for an anti-PD-1/PD-L1 as monotherapy were enrolled. Saliva samples were collected from patients prior to the start of treatment, bacterial DNA was extracted using the QIAamp® DNA Microbiome Kit (QIAGEN) and the 16S rRNA gene was sequenced on a MiSeq sequencing instrument (Illumina). Results: NSCLC patients were dichotomized as “Responders” (partial or complete response) and “Non-Responders” (progressive disease), after 12 weeks of treatment, based on RECIST criteria. A prevalence of the phylum Candidatus Saccharibacteria was found in the 10 responders compared to non-responders (abundance 5% vs 1% respectively; p-value = 1.46 x 10-7; False Discovery Rate (FDR) = 1.02 x 10-6). Moreover, a higher prevalence of Saccharibacteria Genera Incertae Sedis genus (belonging to the Candidatus Saccharibacteria phylum) was observed in "responders" (p-value = 6.01 x 10-7 and FDR = 2.46 x 10-5). Finally, the patients who benefit from immunotherapy showed a significant abundance of TM7 Phylum Sp Oral Clone FR058 strain, member of Saccharibacteria Genera Incertae Sedis genus (p-value = 6.13 x 10-7 and FDR=7.66 x 10-5). Conclusions: These preliminary results showed a significant association between oral microbiota and ICIs response in NSCLC patients. In particular, the higher prevalence of Candidatus Saccharibacteria phylum and TM7 Phylum Sp Oral Clone FR058 strain in responders suggests their potential immunomodulatory role. The study is still ongoing and updated data will be presented at the congress.

Keywords: oral microbiota, immune checkpoint inhibitors, non-small cell lung cancer, predictive biomarker

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