Search results for: red blood cells

Authors: A. Armin, R. Fotouhi, W. Szyszkowski

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In this paper a 3-D finite element (FE) investigation of soil-blade interaction is described. The effects of blade’s shape and rake angle are examined both numerically and experimentally. The soil is considered as an elastic-plastic granular material with non-associated Drucker-Prager material model. Contact elements with different properties are used to mimic soil-blade sliding and soil-soil cutting phenomena. A separation criterion is presented and a procedure to evaluate the forces acting on the blade is given and discussed in detail. Experimental results were derived from tests using soil bin facility and instruments at the University of Saskatchewan. During motion of the blade, load cells collect data and send them to a computer. The measured forces using load cells had noisy signals which are needed to be filtered. The FE results are compared with experimental results for verification. This technique can be used in blade shape optimization and design of more complicated blade’s shape.

Keywords: finite element analysis, experimental results, blade force, soil-blade contact modeling

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Authors: E. Carraro, Sa. Bonetta, Si. Bonetta, E. Ceretti, G. C. V. Viola, C. Pignata, S. Levorato, T. Salvatori, S. Vannini, V. Romanazzi, A. Carducci, G. Donzelli, T. Schilirò, A. De Donno, T. Grassi, S. Bonizzoni, A. Bonetti, G. Gilli, U. Gelatti

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In 2013, air pollution and particulate matter were classified as carcinogenic to human by the IARC. At present, PM is Europe's most problematic pollutant in terms of harm to health, as reported by European Environmental Agency (EEA) in the EEA Technical Report on Air quality in Europe, 2015. A percentage between 17-30 of the EU urban population lives in areas where the EU air quality 24-hour limit value for PM10 is exceeded. Many studies have found a consistent association between exposure to PM and the incidence and mortality for some chronic diseases (i.e. lung cancer, cardiovascular diseases). Among the mechanisms responsible for these adverse effects, genotoxic damage is of particular concern. Children are a high-risk group in terms of the health effects of air pollution and early exposure during childhood can increase the risk of developing chronic diseases in adulthood. The MAPEC_LIFE (Monitoring Air Pollution Effects on Children for supporting public health policy) is a project founded by EU Life+ Programme (LIFE12 ENV/IT/000614) which intends to evaluate the associations between air pollution and early biological effects in children and to propose a model for estimating the global risk of early biological effects due to air pollutants and other factors in children. This work is focused on the micronuclei frequency in child buccal cells in association with airborne PM levels taking into account the influence of other factors associated with the lifestyle of children. The micronucleus test was performed in exfoliated buccal cells of 6–8 years old children from 5 Italian towns with different air pollution levels. Data on air quality during the study period were obtained from the Regional Agency for Environmental Protection. A questionnaire administered to children’s parents was used to obtain details on family socio-economic status, children health condition, exposures to other indoor and outdoor pollutants (i.e. passive smoke) and life-style, with particular reference to eating habits. During the first sampling campaign (winter 2014-15) 1315 children were recruited and sampled for Micronuclei test in buccal cells. In the sampling period the levels of the main pollutants and PM10 were, as expected, higher in the North of Italy (PM10 mean values 62 μg/m3 in Torino and 40 μg/m3 in Brescia) than in the other towns (Pisa, Perugia, Lecce). A higher Micronucleus frequency in buccal cells of children was found in Brescia (0.6/1000 cells) than in the other towns (range 0.3-0.5/1000 cells). The statistical analysis underlines a relation of the micronuclei frequency with PM concentrations, traffic level near child residence, and level of education of parents. The results suggest that, in addition to air pollution exposure, some other factors, related to lifestyle or further exposures, may influence micronucleus frequency and cellular response to air pollutants.

Keywords: air pollution, buccal cells, children, micronucleus cytome assay

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Authors: Kang Jian Xian, Huda Abdullah, Md. Akhtaruzzaman, Iskandar Yahya, Mohd Hafiz Dzarfan Othman, Brian Yulianto

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The CZTS absorber layer doped with a silver (Ag) is one of the candidates that suggest improving the efficiency of thin films. Silver element functions to reduce antisite defects, increase grain size and create the plasmonic effect. In this work, an experimental study has been done to investigate the electrical and physical properties of CZTS, ACZTS, and AZTS. Ag replaces the Cu in (Cu1-xAgx)2ZnSnS4 (ACZTS) is up to x ≤1. ACZTS thin-films solar cells have been deposited by sol–the gel spin coating method. There are a total of 19 samples done with 11 significant percentages (0%, 10%, 20%… 100%) to show the whole phenomena of efficiency rate and nine specific percentages to find out the best concentration rate for Ag-doped. The obtained results can be helpful for better understanding ACZTS layers.

Keywords: CZTS, ACZTS, AZTS, silver, antisite, efficiency, thin-film solar cell

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Authors: Michelle C. S. Azevedo, Priscila M. Colavite, Carolina F. Francisconi, Ana P. Trombone, Gustavo P. Garlet

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VIP (vasoactive intestinal peptide) know as a potential protective factor in the view of its marked immunosuppressive properties. In this work, we investigated a possible association of VIP with the clinical status of experimental periapical granulomas and the association with expression markers in the lesions potentially associated with periapical lesions pathogenesis. C57BL/6WT mice were treated or not with recombinant VIP. Animals with active/progressive (N=40), inactive/stable (N=70) periapical granulomas and controls (N=50) were anesthetized and the right mandibular first molar was surgically opened, allowing exposure of dental pulp. Endodontic pathogenic bacterial strains were inoculated: Porphyromonas gingivalis, Prevotella nigrescens, Actinomyces viscosus, and Fusobacterium nucleatum subsp. polymorphum. The cavity was not sealed after bacterial inoculation. During lesion development, animals were treated or not with recombinant VIP 3 days post infection. Animals were killed after 3, 7, 14, and 21 days of infection and the jaws were dissected. The extraction of total RNA from periodontal tissues was performed and the integrity of samples was checked. qPCR reaction using TaqMan chemistry with inventoried primers were performed in ViiA7 equipment. The results, depicted as the relative levels of gene expression, were calculated in reference to GAPDH and β-actin expression. Periodontal tissues from upper molars were vested and incubated supplemented RPMI, followed by processing with 0.05% DNase. Cell viability and couting were determined by Neubauer chamber analysis. For flow cytometry analysis, after cell counting the cells were stained with the optimal dilution of each antibody; (PE)-conjugated and (FITC)-conjugated antibodies against CD4, CD25, FOXP3, IL-4, IL-17 and IFN-γ antibodies, as well their respective isotype controls. Cells were analyzed by FACScan and CellQuest software. Results are presented as the number of cells in the periodontal tissues or the number of positive cells for each marker in the CD4+FOXp3+, CD4+IL-4+, CD4+IFNg+ and CD4+IL-17+ subpopulations. The levels mRNA were measured by qPCR. The VIP expression was predominated in inactive lesions, as well part of the clusters of cytokine/Th markers identified as protective factors and a negative correlation between VIP expression and lesion evolution was observed. A quantitative analysis of IL1β, IL17, TNF, IFN, MMP2, RANKL, OPG, IL10, TGFβ, CTLA4, COL5A1, CTGF, CXCL11, FGF7, ITGA4, ITGA5, SERP1 and VTN expression was measured in experimental periapical lesions treated with VIP 7 and 14 days after lesion induction and healthy animals. After 7 days, all targets presented a significate increase in comparison to untreated animals. About migration kinetics, profile of chemokine receptors expression of TCD4+ subsets and phenotypic analysis of Tregs, Th1, Th2 and Th17 cells during the course of experimental periodontal disease evaluated by flow cytometry and depicted as the number of positive cells for each marker. CD4+IFNg+ and CD4+FOXp3+ cells migration were significate increased 7 days post VIP treatment. CD4+IL17+ cells migration were significate increased 7 and 14 days post VIP treatment, CD4+IL4+ cells migration were significate increased 14 and 21 days post VIP treatment compared to the control group. In conclusion, our experimental data support VIP involvement in determining the inactivity of periapical lesions. Financial support: FAPESP #2015/25618-2.

Keywords: chronic inflammation, cytokines, osteolytic lesions, VIP (Vasoactive Intestinal Peptide)

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Authors: Parsa Sheikhzadeh

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Polar bodies are cells that form by oogenesis in meiosis which differentiate and develop from oocytes. Although in many animals, these cells often die following meiotic maturation of the oocyte. Oocyte activation is during mammalian fertilization, sperm is fused with the oocyte's membrane, triggering the resumption of meiosis from the metaphase II arrest, the extrusion of the second polar body, and the exocytosis of cortical granules. The origin recognition complex proteins 4 (ORC4) forms a cage around the set of chromosomes that will be extruded during polar body formation before it binds to the chromatin shortly before zygotic DNA replication. One unique feature of the female gamete is that the polar bodies can provide beneficial information about the genetic background of the oocyte without potentially destroying it. Testing at the polar body (PB) stage was the least accurate, mainly due to the high incidence of post-zygotic events. On the other hand, the results from PB1-MII oocyte pair validated that PB1 contains nearly the same methylome (average Pearson correlation is 0.92) with sibling MII oocyte. In this article, we comprehensively examine the role of polar bodies in female human gametes.

Keywords: polar bodies, ORC4, oocyte, genetic, methylome, gamete, female

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Authors: Elena Petersen, Inna Kornienko, Svetlana Guryeva, Sergey Dobdin, Anatoly Skripal, Andrey Usanov, Dmitry Usanov

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One of the most important unsolved problems in modern medicine is the increase of chronic diseases that lead to organ dysfunction or even complete loss of function. Current methods of treatment do not result in decreased mortality and disability statistics. Currently, the best treatment for many patients is still transplantation of organs and/or tissues. Therefore, finding a way of correct artificial matrix biofabrication in case of limited number of natural organs for transplantation is a critical task. One important problem that needs to be solved is development of a nondestructive and noninvasive method to analyze dynamic changes of mechanical characteristics of a matrix with minimal side effects on the growing cells. This research was focused on investigating the properties of matrix as a marker of graft condition. In this study, the collagen gel with human primary dermal fibroblasts in suspension (60, 120, 240*103 cells/mL) and collagen gel with cell spheroids were used as model objects. The stiffness and elasticity characteristics were evaluated by a semiconductor laser autodyne. The time and cell concentration dependency of the stiffness and elasticity were investigated. It was shown that these properties changed in a non-linear manner with respect to cell concentration. The maximum matrix stiffness was observed in the collagen gel with the cell concentration of 120*103 cells/mL. This study proved the opportunity to use the mechanical properties of matrix as a marker of graft condition, which can be measured by noninvasive semiconductor laser autodyne technique.

Keywords: graft, matrix, noninvasive method, regenerative medicine, semiconductor laser autodyne

Procedia PDF Downloads 344

Authors: Mohammed Auwal Ibrahim, Isa Yunusa, Nafisa Kabir, Shazali Ali Baba, Amina Muhammad Yushau, Suraj Suraj Ibrahim, Zaharaddeen Idris Bello, Suleiman Haruna Suleiman, Murtala Bindawa Isah

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Background: Inhibition of intestinal maltase and sucrase prevents postprandial blood glucose excursions which are beneficial in ameliorating diabetes-associated complications. Objective: In this study, the inhibitory effects of fruit extracts of Parinari macrophylla, Detarium microcarpum, Ziziphus spina-christi, Z. mairei and Parkia biglobosa were investigated against intestinal maltase and sucrase. Methods: Rats were given co-administration of the fruit extracts with maltose or sucrose and blood glucose levels were measured at 0, 30, 90 and 120 min. Results: The glucose-time curves indicated that all the fruits had the most potent inhibitory effects on both maltase and sucrase within the first 30 min. The computed Area Under the Curves (AUC0-120)for all the fruits indicated more potent inhibitory effects against intestinal maltase than sucrase.The ED50 range for the fruits extract against maltase and sucrase were 647.15-1118.35 and 942.44-1851.94 mg/kg bw respectively. Conclusion: The data suggests that the fruits could prevent postprandial hyperglycemia via inhibition of intestinal maltase and sucrase.

Keywords: diabetes mellitus, fruits, α-glucosidases, maltase, sucrase

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Authors: Majid Afkhami, Maryam Ehsanpour

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The present study provides information about the nature of adverse effects on fish and the ecological impact that polycyclic aromatic hydrocarbons (PAHs) pollutant are having in the northern coast of Hormuz Strait. The glucose and cholesterol levels were higher in fish from the St3 than in Walton's mudskipper from other stations however St3 samples had lowest total proteins levels. There was a significant positive correlation between glucose and cholesterol with PAHs concentrations in sediment and tissue samples (P<0.05). However, total proteins had adverse significant correlation with PAHs concentrations (P>0.05). The adverse correlation was seen between length and body weight of fish samples with PAHs concentrations. According to the results of this study, the monitoring of contaminants bioaccumulation in the northern part of Hormuz Strait is necessary, because this will give an indication of the temporal and spatial extent of the process, as well as an assessment of the potential impact on aquatic organisms health.

Keywords: PAHs, blood metabolic factors, Periophthalmus waltoni, Hormuz Strait

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Authors: Attahiru Sifawa Ahmad

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The media; electronic, print, and social, is playing very significant roles towards promoting political awareness and stability of any nation. However, for the media to play its role effectively, a clear and sound grasp of the language of communication is necessary. Otherwise, there is the tendency of the media spreading wrong and, or, misinterpreted information to the public, capable of generating rancour and political instability. One such clear misinterpretation or misrepresentation of information was the Hausa metaphorical expression, Kare Jinni Biri Jinni quoted from the statement made by Rtd. General Muhammadu Buhari, sometimes in April, 2013, while addressing his supporters from Niger State. In the political presentation of the term Kare - Jini Biri – Jini, quoted and translated by many print media in Nigeria, it was interpreted to mean; ‘The Dog and the Baboon will be soaked in blood’, denoting bloodshed and declaration of war. However, the term Kare - jini Biri - Jini, literally; the Dog with blood and the Baboon with blood, or, the Dog is bleeding the Baboon is bleeding, or, both the Dog and the Baboon sustained injuries. It is a metaphorical expression denoting a hot competition, and serious struggle, between two competing parties that are closer in strength and stamina. The expression got its origin among the hunting communities in traditional Hausa Societies. From experience, it was always not easy to wrestle and hunt Baboon by the Hunter’s Dog. In many instances, it ended a futile exercise, and even at instances whereby the latter hunted the former, it would be after a serious struggle with both two sustaining injuries. This paper seeks to highlight the poverty of vocabulary, and poor grasp of Nigerian languages among Journalists and young citizens in the country. The paper, therefore, advocated for the retention and effective teaching of the indigenous languages in primary and secondary school’s curriculums in Nigeria. The paper equally analysed the political origin of the print media in Nigeria, how since its first appearance, the print Media is being assigned very important political role by political elites in the country.

Keywords: Baboon, dog, media, politics

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Authors: Farhan Ahmad Atif, Abd Ur Rehman, Muhammad Babir

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Dietary cation anion difference (DCAD) is an important aspect of dairy nutrition, especially in the transition period. Sahiwal cattle is the highest milk producing breed among Zebu cattle. We planned first study on transition Sahiwal cattle to determine the effects of feeding varying levels of negative DCAD. For this purpose, twenty pregnant cows (at the 250th day of gestation) were selected and randomly divided into 5 groups comprising four animals each. Five iso-caloric (2100 Kcal) and iso-nitrogenous (12%) diets were formulated and each diet was allotted to each group. The animals received positive DCAD diet served as control. Diets were supplemented with NutriCAB® to attain 0, -15, -30 and -45 DCAD levels. Experimental diets were fed at ad-libitum upto parturition and data regarding feed intake were recorded on daily. Post-partum incidence of milk fever, dystocia, retention of placenta (RP), mastitis as well as milk production, milk fat percentage and serum Ca levels were recorded. Urine and blood pH were determined weekly during the last month of pregnancy. Results showed that prepartum feed intake and blood pH were not affected (P > 0.05), while urine pH was significantly reduced (P < 0.05) by lowering DCAD levels. Post parturient blood calcium level linearly increased (P < 0.05) with decreasing DCAD. Pre-partum negative DCAD feeding had no effect (P > 0.05) on post-parturient milk production and fat percentage. However, parturient related problems decreased with decreasing DCAD feeding. It was concluded that negative DCAD feeding raised serum calcium level and reduced the incidence of post-parturient problems in Sahiwal cattle.

Keywords: dairy cattle, transition, metabolic diseases, reproductive disorders, incidence

Procedia PDF Downloads 186

Authors: Achim Kampker, Heiner Hans Heimes, Nemanja Sarovic, Jan-Philip Ganser, Saskia Wessel, Christoph Lienemann

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The electrification of the power train is a fundamental technical transition in the automotive industry and poses a major challenge for established car companies. Providing the traction energy, requiring an ever greater amount of space within the car and having a high share of value-add the lithium-ion battery is a central component of the electric power train and a completely new component to car manufacturers at the same time. Being relatively new to the automotive industry, the current design of the product architecture and production process (including manufacturing and assembling processes) of lithium-ion battery modules do not allow for an easy and cost-efficient disassembly or product design change. Yet these two requirements will increase in importance with rising sales volumes of electric cars in the near future and need to be addressed for the electric car to be competitive with conventional power train systems. This paper focuses on the current product architecture and production process of common automotive battery modules from prismatic lithium-ion battery cells to derive impacts for a remanufacturing concept. The information necessary for this purpose were gathered by literature research, patent inquiries, industry expert interviews and first-hand experiences of the authors. On the basis of these results, the underlying causes for the design´s lack of remanufacturability and flexibility with regards to product design changes are examined. In all, this paper gives an extensive and detailed overview of the state of the art of the product architecture and production process of lithium-ion battery modules from prismatic battery cells, identifies its deficiencies and derives improvement measures.

Keywords: battery module, prismatic lithium-ion battery cell, product architecture, production process, remanufacturing, flexibility

Procedia PDF Downloads 267

Authors: Zair Mohammed El Amine, Chemouri Hafida, Derbal Habak Hassina

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Organic photovoltaic cells are electronic devices that convert sunlight into electricity. To this end, the number of studies on organic photovoltaic cells (OVCs) is growing, and this trend is expected to continue. Computational studies are still needed to verify and prove the capability of CVOs, specifically the nanometer molecule PCBM, based on successful experimental results. In this paper, we present a theoretical and computational investigation of PCBM and PC71BM derivatives using the DFT method. On this basis, we employ independent and time-dependent density theories. HOMO, LUMO and GAPH-L energies, ionization potentials and electronic affinity are determined and found to be in agreement with experiments. Using DFT theory based on B3LYP and M062X methods with bases 6-31G (d,p) and 6-311G (d), calculations show that the most efficient acceptors are presented in the group of PC71BM derivatives and are in substantial agreement with experiments. The geometries of the structures are optimized by Gaussian 09.

Keywords: PCBM, P3HT, organic cell solar, DFT, TD-DFT

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Authors: Cheng-Cao Sun, Shu-Jun Li, De-Jia Li

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Determinants of growth and metastasis in cancer remain of great interest to define. MicroRNAs (miRNAs) have frequently emerged as tumor metastatic regulator by acting on multiple signaling pathways. Here, we report the definition of miR-346 as an oncogenic microRNA that facilitates non-small cell lung cancer (NSCLC) cell growth and metastasis. XPC, an important DNA damage recognition factor in nucleotide excision repair was defined as a target for down-regulation by miR-346, functioning through direct interaction with the 3'-UTR of XPC mRNA. Blocking miR-346 by an antagomiR was sufficient to inhibit NSCLC cell growth and metastasis, an effect that could be phenol-copied by RNAi-mediated silencing of XPC. In vivo studies established that miR-346 overexpression was sufficient to promote tumor growth by A549 cells in xenografts mice, relative to control cells. Overall, our results defined miR-346 as an oncogenic miRNA in NSCLC, the levels of which contributed to tumor growth and invasive aggressiveness.

Keywords: microRNA-346, miR-346, XPC, non-small cell lung cancer, oncogenesis

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Authors: Renata Rank Miranda, Arandi Ginane Bezerra, Ciro Alberto Oliveira Ribeiro, Marco AntôNio Ferreira Randi, Carmen Lúcia Voigt, Lilian Skytte, Kaare Lund Rasmussen, Francisco Filipak Neto, Frank Kjeldsen

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Synergetic and antagonistic effects of drugs are well-known concerns in pharmacological assessments of dose and toxicity. Similar approach should be used in assessing cellular uptake and cytotoxicity of nanoparticles. Since nanoparticles are released into the aquatic environment they may interact with existing xenobiotics. Here we used biochemical assays and quantitative proteomics to assess the cytotoxicity of silver nanoparticles (AgNP) when human hepatoma HepG2 cells were co-exposed to 2 nm AgNP together with either Cd2+ or Hg2+ ions. Time-course experiments (2h, 4h, and 24h) were conducted to assess the first response to the exposure studies. The general trend was that a synergetic toxicological response was observed in cells exposed to both AgNP and Cd2+ or Hg2+, with AgNP and Cd2+ being more toxic. This was observed by a significant increase in the ROS and superoxide level of >35% in the case of AgNP+Cd2+ compared to the sum of responses of AgNP and Cd2+, individually. Metabolic activity and viability also dropped more for AgNP+Cd2+ (>10%) than for AgNP and Cd2+ combined. We used inductively coupled plasma mass spectrometry to investigate if AgNP facilitates larger influx of toxic metal ions into HepG2 cells. Only Hg2+ ions was found to be more efficiently engulfed as the concentration of Hg2+ was found 2.8 times larger compared to exposure experiments with only Hg2+. This effect was not observed for Cd2+. We now continue with deep proteomics studies to obtain wider details on the mechanism of the toxicity related to AgNP, Cd2+, and AgNP+Cd2+, respectively.

Keywords: nanotoxicology, silver nanoparticles, proteomics, human cell line

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Authors: N. F. Hamid, A. Kipar, J. Stewart, D. J. Antoine, B. K. Park, D. P. Williams

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Acetaminophen (APAP) is a widely used analgesic that is safe at therapeutic doses. The mouse model of APAP has been extensively used for studies on pathogenesis and intervention of drug induced liver injury based on the CytP450 mediated formation of N-acetyl-p-benzo-quinoneimine and, more recently, as model for mechanism based biomarkers. Delay of the fasted CD1 mice to rebound to the basal level of hepatic GSH compare to fed mice is reported in this study. Histologically, 15 hours fasted mice prior to APAP treatment leading to overall more intense cell loss with no evidence of apoptosis as compared to non-fasted mice, where the apoptotic cells were clearly seen on cleaved caspase-3 immunostaining. After 15 hours post APAP administration, hepatocytes underwent stage of recovery with evidence of mitotic figures in fed mice and return to completely no histological difference to control at 24 hours. On the contrary, the evidence of ongoing cells damage and inflammatory cells infiltration are still present on fasted mice until the end of the study. To further measure the regenerative capacity of the hepatocytes, the inflammatory mediators of cytokines that involved in the progression or regression of the toxicity like TNF-α and IL-6 in liver and spleen using RT-qPCR were also included. Yet, quantification of proliferating cell nuclear antigen (PCNA) has demonstrated the time for hepatic regenerative in fasted is longer than that to fed mice. Together, these data would probably confirm that fasting prior to APAP treatment does not only modulate liver injury, but could have further effects to delay subsequent regeneration of the hepatocytes.

Keywords: acetaminophen, liver, proliferating cell nuclear antigen, regeneration, apoptosis

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Authors: Fangfang Wang, Fan Qu

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Polycystic ovary syndrome (PCOS), the most common endocrinopathy among women of reproductive age, is characterized by ovarian dysfunction, hyperandrogenism and reduced fecundity. The aim of this study is to investigate whether the circadian disruption is involved in pathogenesis of PCOS in androgen-induced animal model. We established a rat model of PCOS using single subcutaneous injection with testosterone propionate on the ninth day after birth, and confirmed their PCOS-like phenotypes with vaginal smears, ovarian hematoxylin and eosin (HE) staining and serum androgen measurement. The control group rats received the vehicle only. Gene expression was detected by real-time quantitative PCR. (1) Compared with control group, PCOS model rats of 10-week group showed persistently keratinized vaginal cells, while all the control rats showed at least two consecutive estrous cycles. (2) Ovarian HE staining and histological examination showed that PCOS model rats of 10-week group presented many cystic follicles with decreased numbers of granulosa cells and corpora lutea in their ovaries, while the control rats had follicles with normal layers of granulosa cells at various stages of development and several generations of corpora lutea. (3) In the 10-week group, serum free androgen index was notably higher in PCOS model rats than controls. (4) Disturbed mRNA expression patterns of core clock genes were found in ovaries of PCOS model rats of 10-week group. Abnormal expression of key genes associated with circadian rhythm in ovary may be one of the mechanisms for ovarian dysfunction in PCOS model rats induced by androgen.

Keywords: polycystic ovary syndrome, androgen, animal model, circadian disruption

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Authors: Gyati Shilakari Asthana, Abhay Asthana, Dharm Veer Kohli, Suresh Prasad Vyas

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Purpose: The therapeutic potential of oligonucleotide (ODN) is primarily dependent upon its safe and efficient delivery to specific cells overcoming degradation and maximizing cellular uptake in vivo. The present study is focused to design low molecular weight chitosan nanoconstructs to meet the requirements of safe and effectual delivery of ODNs. LMW-chitosan is a biodegradable, water soluble, biocompatible polymer and is useful as a non-viral vector for gene delivery due to its better stability in water. Methods: LMW chitosan ODN nanoparticles (CHODN NPs) were formulated by self-assembled method using various N/P ratios (moles ratio of amine groups of CH to phosphate moieties of ODNs; 0.5:1, 1:1, 3:1, 5:1, and 7:1) of CH to ODN. The developed CHODN NPs were evaluated with respect to gel retardation assay, particle size, zeta potential and cytotoxicity and transfection efficiency. Results: Complete complexation of CH/ODN was achieved at the charge ratio of 0.5:1 or above and CHODN NPs displayed resistance against DNase I. On increasing the N/P ratio of CH/ODN, the particle size of the NPs decreased whereas zeta potential (ZV) value increased. No significant toxicity was observed at all CH concentrations. The transfection efficiency was increased on increasing N/P ratio from 1:1 to 3:1, whereas it was decreased with further increment in N/P ratio upto 7:1. Maximum transfection of CHODN NPs with both the cell lines (Raw 267.4 cells and Hela cells) was achieved at N/P ratio of 3:1. The results suggest that transfection efficiency of CHODN NPs is dependent on N/P ratio. Conclusion: Thus the present study states that LMW chitosan nanoparticulate carriers would be acceptable choice to improve transfection efficiency in vitro as well as in vivo delivery of oligonucleotide.

Keywords: LMW-chitosan, chitosan nanoparticles, biocompatibility, cytotoxicity study, transfection efficiency, oligonucleotide

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Authors: N. Rajendra Prasad

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Resveratrol (RSV), a grape phytochemical, has drawn greater attention because of its beneficial ef-fects against cancer. However, RSV has some draw-backs such as unstabilization, poor water solubility and short biological half time, which limit the utili-zation of RSV in medicine, food and pharmaceutical industries. In this study, we have encapsulated RSV in gelatin nanoparticles (GNPs) and studied its anti-cancer efficacy in NCI-H460 lung cancer cells. SEM and DLS studies have revealed that the prepared RSV-GNPs possess spherical shape with a mean diameter of 294 nm. The successful encapsulation of RSV in GNPs has been achieved by the cross-linker glutaraldehyde probably through Schiff base reaction and hydrogen bond interaction. Spectrophotometric analysis revealed that the max-imum of 93.6% of RSV has been entrapped in GNPs. In vitro drug release kinetics indicated that there was an initial burst release followed by a slow and sustained release of RSV from GNPs. The prepared RSV-GNPs exhibited very rapid and more efficient cellular uptake than free RSV. Further, RSV-GNPs treatment showed greater antiproliferative efficacy than free RSV treatment in NCI-H460 cells. It has been found that greater ROS generation, DNA damage and apoptotic incidence in RSV-GNPs treated cells than free RSV treatment. Erythrocyte aggregation assay showed that the prepared RSV-GNPs formulation elicit no toxic response. HPLC analysis revealed that RSV-GNPs was more bioavailable and had a longer half-life than free RSV. Hence, GNPs carrier system might be a promising mode for controlled delivery and for improved therapeutic index of poorly water soluble RSV.

Keywords: resveratrol, coacervation, anticancer gelatin nanoparticles, lung cancer, controlled release

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Authors: Nima Samie, Batoul Sadat Haerian, Sekaran Muniandy, M. S. Kanthimathi

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Colon and breast cancers are categorized as the most prevalent types of cancer worldwide. Recently, the broad clinical application of metal complex compounds has led to the discovery of potential therapeutic drugs. The aim of this study was to evaluate the cytotoxic action of a selected nickel complex compound (NCC) against human colon and breast cancer cells. In this context, we determined the potency of the compound in the induction of apoptosis, cell cycle arrest, and cytoskeleton rearrangement. HT-29, WiDr, CCD-18Co, MCF-7 and Hs 190.T cell lines were used to determine the IC50 of the compound using the MTT assay. Analysis of apoptosis was carried out using immunofluorescence, acridine orange/ propidium iodide double staining, Annexin-V-FITC assay, evaluation of the translocation of NF-kB, oxygen radical antioxidant capacity, quenching of reactive oxygen species content , measurement of LDH release, caspase-3/-7, -8 and -9 assays and western blotting. The cell cycle arrest was examined using flowcytometry and gene expression was assessed using qPCR array. Results showed that our nickel complex compound displayed a potent suppressive effect on HT-29, WiDr, MCF-7 and Hs 190.T after 24 h of treatment with IC50 value of 2.02±0.54, 2.13±0.65, 3.76±015 and 3.14±0.45 µM respectively. This cytotoxic effect on normal cells was insignificant. Dipping in the mitochondrial membrane potential and increased release of cytochrome c from the mitochondria indicated induction of the intrinsic apoptosis pathway by the nickel complex compound. Activation of this pathway was further evidenced by significant activation of caspase 9 and 3/7.The nickel complex compound (NCC) was also shown activate the extrinsic pathways of apoptosis by activation of caspase-8 which is linked to the suppression of NF-kB translocation to the nucleus. Cell cycle arrest in the G1 phase and up-regulation of glutathione reductase, based on excessive ROS production were also observed. The results of this study suggest that the nickel complex compound is a potent anti-cancer agent inducing both intrinsic and extrinsic pathways as well as cell cycle arrest in colon and breast cancer cells.

Keywords: nickel complex, apoptosis, cytoskeletal rearrangement, colon cancer, breast cancer

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Authors: Badawia Ibrahim, Iman Hussein, Samar El Sheikh, Fatma Abou Elkasem, Hazem Abo Ismael

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Background: Response of breast carcinoma to neoadjuvant chemotherapy (NAC) varies regarding many factors including hormonal receptor status. Breast cancer is a heterogenous disease with different outcomes, hence a need arises for new markers predicting the outcome of NAC especially for the triple negative group when estrogen, progesterone receptors and Her2/neu are negative. FOXP3 is a promising target with unclear role. Aim: To examine the value of FOXP3 expression in locally advanced triple negative breast cancer tumoral cells as well as tumor infiltrating lymphocytes (TILs) and to elucidate its relation to the extent of NAC response. Material and Methods: Forty five cases of immunohistochemically confirmed to be triple negative breast carcinoma were evaluated for NAC (Doxorubicin, Cyclophosphamide AC x 4 cycles + Paclitaxel x 12 weeks, patients with ejection fraction less than 60% received Taxotere or Cyclophosphamide, Methotrexate, Fluorouracil CMF) response in both tumour and lymph nodes status according to Miller & Payne's and Sataloff's systems. FOXP3 expression in tumor as well as TILs evaluated in the pretherapy biopsies was correlated with NAC response in breast tumor and lymph nodes as well as other clinicopathological factors. Results: Breast tumour cells showed FOXP3 positive cytoplasmic expression in (42%) of cases. High FOXP3 expression percentage was detected in (47%) of cases. High infiltration by FOXP3+TILs was detected in (49%) of cases. Positive FOXP3 expression was associated with negative lymph node metastasis. High FOXP3 expression percentage and high infiltration by FOXP3+TILs were significantly associated with complete therapy response in axillary lymph nodes. High FOXP3 expression in tumour cells was associated with high infiltration by FOXP3+TILs. Conclusion: This result may provide evidence that FOXP3 marker is a good prognostic and predictive marker for triple negative breast cancer (TNBC) indicated for neoadjuvant chemotherapy and can be used for stratifications of TNBC cases indicated for NAC. As well, this study confirmed the fact that the tumour cells and the surrounding microenvironment interact with each other and the tumour microenvironment can influence the treatment outcomes of TNBC.

Keywords: breast cancer, FOXP3 expression, prediction of neoadjuvant chemotherapy effect, triple negative

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Authors: Elaine Wong

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This research protocol aims to explore and justify the need for nutrition and physical activity intervention to improve health outcomes among SME (Small Medium Enterprise) employees. It was found that the worksite is an ideal and convenient setting for employees to take charge of their health thru active participation in health programmes since they spent a great deal of time at their workplace. This study will examine the impact of both general or/and targeted health interventions in both SME and non-SME companies utilizing the Workplace Health Promotion (WHP) grant over a 12 months period and assessed the improvement in chronic health disease outcomes in Singapore. Random sampling of both non-SME and SME companies will be conducted to undergo health intervention and statistical packages such as Statistical Package for Social Science (SPSS) 25 will be used to examine the impact of both general and targeted interventions on employees who participate and those who do not participate in the intervention and their effects on blood glucose (BG), blood lipid, blood pressure (BP), body mass index (BMI), and body fat percentage. Using focus groups and interviews, the data results will be transcribed to investigate enablers and barriers to workplace health intervention revealed by employees and WHP coordinators that could explain the variation in the health screening results across the organisations. Dietary habits and physical activity levels of the employees participating and not participating in the intervention will be collected before and after intervention to assess any changes in their lifestyle practices. It makes economic sense to study the impact of these interventions on health screening outcomes across various organizations that are existing grant recipients to justify the sustainability of these programmes by the local government. Healthcare policy makers and employers can then tailor appropriate and relevant programmes to manage these escalating chronic health disease conditions which is integral to the competitiveness and productivity of the nation’s workforce.

Keywords: chronic diseases, health screening, nutrition and fitness intervention , workplace health

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Authors: Shashank Patidar, Sumit Kumar, Atul Srivastava, Suneet Singh

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Present work is concerned with the numerical investigation of thermal response of biological tissues during laser-based photo-thermal therapy for destroying cancerous/abnormal cells with minimal damage to the surrounding normal cells. Light propagation through the biological sample is mathematically modelled by transient radiative transfer equation. In the present work, application of the Lattice Boltzmann Method is extended to analyze transport of short-pulse radiation in a participating medium.In order to determine the two-dimensional temperature distribution inside the tissue medium, the RTE has been coupled with Penne’s bio-heat transfer equation based on Fourier’s law by several researchers in last few years.

Keywords: lattice Boltzmann method, transient radiation transfer equation, dual phase lag model

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Authors: Karla E. Sanchez-Cazares, Kim H. Parker, Jennifer H. Tweedy

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The purpose of this work is to develop a simple compartmental model of cerebral fluid balance including blood and cerebrospinal-fluid (CSF). At the first level the cerebral arteries and veins are modelled as bifurcating trees with constant scaling factors between generations which are connected through a homogeneous microcirculation. The arteries and veins are assumed to be non-rigid and the cross-sectional area, resistance and mean pressure in each generation are determined as a function of blood volume flow rate. From the mean pressure and further assumptions about the variation of wall permeability, the transmural fluid flux can be calculated. The results suggest the next level of modelling where the cerebral vasculature is divided into three compartments; the large arteries, the small arteries, the capillaries and the veins with effective compliances and permeabilities derived from the detailed vascular model. These vascular compartments are then linked to other compartments describing the different CSF spaces, the cerebral ventricles and the subarachnoid space. This compartmental model is used to calculate the distribution of fluid in the cranium. Known volumes and flows for normal conditions are used to determine reasonable parameters for the model, which can then be used to help understand pathological behaviour and suggest clinical interventions.

Keywords: cerebrovascular, compartmental model, CSF model, vascular network

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Authors: Aly Kassem, Eman Sapet, Eman Abdelbaset, Hosam Mahmoud

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Objective: Hypertension is an important public health challenge being one of the most common worldwide disease-affecting human. Our aim is to study the clinical characteristics, therapeutic regimens, treatment compliance, and risk factors in a sector of of hypertensive patients at Sohag City. Subject and Methods: A cross sectional study; conducted in Sohag city; it involved 520 patients; males (45.7 %) and females (54.3 %). Their ages ranged between 35-85 years. BP measurements, BMI, blood glucose, Serum creatinine, urine analysis, serum Lipids, blood picture and ECG were done all the studied patients. Results: Hypertension presented more between non-smokers (72.55%), females (54.3%), educated patients (50.99%) and patients with low SES (54.9%). CAD presented in (51.63%) of patients, while laboratory investigations showed hyperglycaemia in (28.7%), anemia in (18.3%), high serum creatinine level in (8.49%) and proteinuria in (10.45%) of patient. Adequate BP control was achieved in (49.67%); older patients had lower adequacy of BP control in spite of the extensive use of multiple-drug therapy. Most hypertensive patients had more than one coexistent CV risk factor. Aging, being a female (54.3%), DM (32.3%), family history of hypertension (28.7%), family history of CAD (25.4%), and obesity (10%) were the common contributing risk factors. ACE-inhibitors were prescribed in (58.16%), Beta-blockers in (34.64%) of the patients. Monotherapy was prescribed for (41.17%) of the patients. (75.81%) of patients had regular use of their drug regimens. (49.67%) only of patients had their condition under control, the number of drugs was inversely related to BP control. Conclusion: Hypertensive patients in Sohag city had a profile of high CV risks, and poor blood pressure control particularly in the elderly. A multidisciplinary approach for routine clinical check-up, follow-up, physicians and patients training, prescribing simple once-daily regimens and encouraging life style modifications are recommended. Anti hypertensives, hypertension, elderly patients, risk factors, treatment compliance.

Keywords: anti hypertensives, hypertension, elderly patients, risk factors, treatment compliance

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Authors: Nathalie Baeza-Kallee, Raphaël Bergès, Victoria Hein, Stéphanie Cabaret, Jeremy Garcia, Abigaëlle Gros, Emeline Tabouret, Aurélie Tchoghandjian, Carole Colin, Dominique Figarella-Branger

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Glioblastoma (GBM) contains cancer stem cells that are resistant to treatment. GBM cancer stem cell expresses glycolipids recognized by the A2B5 antibody. A2B5, induced by the enzyme ST8 alpha-N-acetyl-neuraminide alpha-2,8-sialyl transferase 3 (ST8Sia3), plays a crucial role in the proliferation, migration, clonogenicity, and tumorigenesis of GBM cancer stem cells. Our aim was to characterize the resulting effects of neuraminidase that remove A2B5 in order to target GBM cancer stem cells. To this end, we set up a GBM organotypic slice model; quantified A2B5 expression by flow cytometry in U87-MG, U87-ST8Sia3, and GBM cancer stem cell lines, treated or not by neuraminidase; performed RNAseq and DNA methylation profiling; and analyzed the ganglioside expression by liquid chromatography-mass spectrometry in these cell lines, treated or not with neuraminidase. Results demonstrated that neuraminidase decreased A2B5 expression, tumor size, and regrowth after surgical removal in the organotypic slice model but did not induce a distinct transcriptomic or epigenetic signature in GBM CSC lines. RNAseq analysis revealed that OLIG2, CHI3L1, TIMP3, TNFAIP2, and TNFAIP6 transcripts were significantly overexpressed in U87-ST8Sia3 compared to U87-MG. RT-qPCR confirmed these results and demonstrated that neuraminidase decreased gene expression in GBM cancer stem cell lines. Moreover, neuraminidase drastically reduced ganglioside expression in GBM cancer stem cell lines. Neuraminidase, by its pleiotropic action, is an attractive local treatment against GBM.

Keywords: cancer stem cell, ganglioside, glioblastoma, targeted treatment

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Authors: Patience Manjengwa-Hungwe, Carmen White

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Background: Odyssean malaria refers to malaria acquired by infected mosquito bites from malaria endemic to non-endemic regions by mechanical modes of transport, such as airplanes, water vessels, trains and vehicles. Odyssean Malaria is rare and is characterised by absence of travel history to malaria endemic areas. As not anticipated in non-endemic areas, late diagnosis and treatment lead to a high case fatality rate. On 26 September 2014, the Outbreak Response Unit at the National Institute of Communicable Diseases was notified of a suspected death from Odyssean Malaria in Johannesburg, Gauteng Province, a non-endemic area. The main objective of this investigation was to identify the etiological agent's mode and source of transmission. Methods: Epidemiological surveys were conducted with the deceased’s family and clinical details were obtained from doctors who treated the victim in Southrand, Johannesburg. Blood samples were collected prior to death and sent to the National Health Laboratory Services, Johannesburg laboratory for a full blood count, urea electrolytes, creatinine, and C-reactive protein. Environmental assessments and entomological investigations, including collection of mosquito and larvae, were conducted at the deceased’s home and surrounding areas and sent to the laboratory for analysis. Results: Epidemiological surveys revealed no travel history, no mechanical transmission through blood transfusion and no previous possible exposure of the victim to malaria mosquitoes. Laboratory findings indicated that the platelet count was low. A further smear revealed that the malaria parasite was present and malaria antigen for P. falciparum was positive. Entomological findings revealed that none of the six adult or larval mosquitoes collected on site were malaria vectors. Dumping sites found at the back of the house were identified as possible sites where mosquitoes from endemic places could possibly breed. Conclusion: Given that there was no travel history or the possibility of mechanical transmission (blood transfusion or needle), the research team concluded that it is highly probable that the infection was acquired through an infective Anopheles mosquito inadvertently translocated from a Malaria endemic area by mechanical modes of transport. We recommend that clinicians in non-endemic malaria areas be aware of this type of malaria and test for malaria in patients showing malaria-like symptoms.

Keywords: Odyssean Malaria, vector Bourne, malaria, epidemiological surveys

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Authors: Mahbuba Rahman, Sabri Boughorbel, Scott Presnell, Charlie Quinn, Darawan Rinchai, Damien Chaussabel, Nico Marr

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Collections of large-scale datasets made available in public repositories can be used to identify and fill gaps in biomedical knowledge. But first, these data need to be made readily accessible to researchers for analysis and interpretation. Here a collection of transcriptome datasets was made available to investigate the functional programming of human hematopoietic cells in early life. Thirty two datasets were retrieved from the NCBI Gene Expression Omnibus (GEO) and loaded in a custom, interactive web application called the Gene Expression browser (GXB), designed for visualization and query of integrated large-scale data. Multiple sample groupings and gene rank lists were created based on the study design and variables in each dataset. Web links to customized graphical views can be generated by users and subsequently be used to graphically present data in manuscripts for publication. The GXB tool also enables browsing of a single gene across datasets, which can provide information on the role of a given molecule across biological systems. The dataset collection is available online. As a proof-of-principle, one of the datasets (GSE25087) was re-analyzed to identify genes that are differentially expressed by regulatory T cells in early life. Re-analysis of this dataset and a cross-study comparison using multiple other datasets in the above mentioned collection revealed that PMCH, a gene encoding a precursor of melanin-concentrating hormone (MCH), a cyclic neuropeptide, is highly expressed in a variety of other hematopoietic cell types, including neonatal erythroid cells as well as plasmacytoid dendritic cells upon viral infection. Our findings suggest an as yet unrecognized role of MCH in immune regulation, thereby highlighting the unique potential of the curated dataset collection and systems biology approach to generate new hypotheses which can be tested in future mechanistic studies.

Keywords: early-life, GEO datasets, PMCH, interactive query, systems biology

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Authors: R. A. Abdel Rahman, A. E. Abdel Wahab, E. A. Goghneimy, H. F. Mohamed, E. M. Salama

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Infectious diseases are a major cause of death worldwide. The treatment of infections continues to be problematic in modern time because of the severe side effects of some drugs and the growing resistance to antimicrobial agents. Hence, the search for newer, safer and more potent antimicrobials is a pressing need. Herbal medicines have received much attention as a source of new antibacterial drugs since they are considered time-tested and comparatively safe both for human use and the environment. In the present study, the antimicrobial activity of marjoram (Origanum majorana L.) essential oil on some gram positive and gram negative reference bacteria, as well as some hospital resistant microbes, was tested. Marjoram oil was extracted and the oil chemical constituents were identified using GC/MS analysis. Staphylococcus aureas ATCC 6923, Pseudomonus auregonosa ATCC 9027, Bacillus subtilis ATCC 6633, E. coli ATCC 8736 and two hospital resistant microbes isolates 16 and 21 were used. The two isolates were identified by biochemical tests and 16s rRNA as proteus spp. and Enterococcus facielus. The effect of different concentrations of essential oils on bacterial growth was tested using agar disk diffusion assay method to determine the minimum inhibitory concentrations and using micro dilution method to determine the minimum bactericidal concentrations. Marjoram oil was found to be effective against both reference and hospital resistance strains. Hospital strains were more resistant to marjoram oil than reference strains. P. auregonosa growth was completely inhibited at a low concentration of oil (4µl/ml). The other reference strains showed sensitivity to marjoram oil at concentrations ranged from 5 to 7µl/ml. The two hospital strains showed sensitivity at media containing 10 and 15µl/ml oil. The major components of oil were terpineol, cis-beta (23.5%), 1,6 – octadien –3-ol,3,7-dimethyl, 2 aminobenzoate (10.9%), alpha terpieol (8.6%) and linalool (6.3%). Scanning electron microscope (SEM) and transmission electron microscope (TEM) analysis were used to determine the difference between treated and untreated hospital strains. SEM results showed that treated cells were smaller in size than control cells. TEM data showed that cell lysis has occurred to treated cells. Treated cells have ruptured cell wall and appeared empty of cytoplasm compared to control cells which shown to be intact with normal volume of cytoplasm. The results indicated that marjoram oil has a positive antimicrobial effect on hospital resistance microbes. Natural crude extracts can be perfect resources for new antimicrobial drugs.

Keywords: antimicrobial activity, essential oil, hospital resistance microbes, marjoram

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Authors: Gloria Pinilla, Jose Manuel Baena, Patricia Gálvez-Martín, Juan Antonio Marchad

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Cartilage is a dense connective tissue with limited self-repair properties. Currently, the therapeutic use of autologous or allogenic chondrocytes makes up an alternative therapy to the pharmacological treatment. The design of a bioprinted 3D cartilage with chondrocytes and biodegradable biomaterials offers a new therapeutic alternative able of bridging the limitations of current therapies in the field. We have developed an enhanced printing processes-Injection Volume Filling (IVF) to increase the viability and survival of the cells when working with high-temperature thermoplastics without the limitation of the scaffold geometry in contact with cells. We have demonstrated the viability of the printing process using chondrocytes for cartilage regeneration. This development will accelerate the clinical uptake of the technology and overcomes the current limitation when using thermoplastics as scaffolds. An alginate-based hydrogel combined with human chondrocytes (isolated from osteoarthritis patients) was formulated as bioink-A and the polylactic acid as bioink-B. The bioprinting process was carried out with the REGEMAT V1 bioprinter (Regemat 3D, Granada-Spain) through a IVF. The printing capacity of the bioprinting plus the viability and cell proliferation of bioprinted chondrociytes was evaluated after five weeks by confocal microscopy and Alamar Blue Assay (Biorad). Results showed that the IVF process does not decrease the cell viability of the chondrocytes during the printing process as the cells do not have contact with the thermoplastic at elevated temperatures. The viability and cellular proliferation of the bioprinted artificial 3D cartilage increased after 5 weeks. In conclusion, this study demonstrates the potential use of Regemat V1 for 3D bioprinting of cartilage and the viability of bioprinted chondrocytes in the scaffolds for application in regenerative medicine.

Keywords: cartilage regeneration, bioprinting, bioink, scaffold, chondrocyte

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Authors: Wei-Chia Huang, Jane Wang

Abstract:

Damage to nerves is considered one of the most irreversible injuries. The regeneration of nerves has always been an important topic in regenerative medicine. In general, damage to human tissue will naturally repair overtime. However, when the nerves are damaged, healed flesh wound cannot guarantee full restoration to its original function, as truncated nerves are often irreversible. Therefore, the development of treatment methods to successfully guide and accelerate the regeneration of nerves has been highly sought after. In order to induce nerve tissue growth, nerve conduits are commonly used to help reconnect broken nerve bundles to provide protection to the location of the fracture while guiding the growth of the nerve bundles. To prevent the protected tissue from becoming necrotic and to ensure the growth rate, the conduits used are often modified with microstructures or blended with neuron growth factors that may facilitate nerve regeneration. Electrical stimulation is another attempted treatment for medical rehabilitation. With appropriate range of voltages and stimulation frequencies, it has been demonstrated to promote cell proliferation and migration. Biodegradability are critical for medical devices like nerve conduits, while conductive polymers pose great potential toward the differentiation and growth of nerve cells. In this work, biodegradability and conductivity were combined into a novel biodegradable, photocurable, conductive polymer composite materials by embedding conductive nanoparticles in poly(glycerol sebacate) acrylate (PGSA) and 3D-printed into nerve conduits. Rat pheochromocytoma cells and rat neuronal Schwann cells were chosen for the in vitro tests of the conduits and had demonstrate selective growth upon culture in the conductive conduits with built-in microchannels and electrical stimulation.

Keywords: biodegradable polymer, 3d printing, neural regeneration, electrical stimulation

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