Search results for: in vitro biofilm formation

Authors: Brajesh Tiwari, Yuvraj Dangi, Abhishek Jain, Ashok Jain

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The purpose of the investigation was to evaluate the potential of sphingosomes as nanoscale drug delivery units for site-specific delivery of anti-cancer agents. Doxorubicin Hydrochloride (DOX) was selected as a model anti-cancer agent. Sphingosomes were prepared and loaded with DOX and optimized for size and drug loading. The formulations were characterized by Malvern zeta-seizer and Transmission Electron Microscopy (TEM) studies. Sphingosomal formulations were further evaluated for in-vitro drug release study under various pH profiles. The in-vitro drug release study showed an initial rapid release of the drug followed by a slow controlled release. In vivo studies of optimized formulations and free drug were performed on albino rats for comparison of drug plasma concentration. The in- vivo study revealed that the prepared system enabled DOX to have had enhanced circulation time, longer half-life and lower elimination rate kinetics as compared to free drug. Further, it can be interpreted that the formulation would selectively enter highly porous mass of tumor cells and at the same time spare normal tissues. To summarize, the use of sphingosomes as carriers of anti-cancer drugs may prove to be a fascinating approach that would selectively localize in the tumor mass, increasing the therapeutic margin of safety while reducing the side effects associated with anti-cancer agents.

Keywords: sphingosomes, anti-cancer, doxorubicin, formulation

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Authors: Khaled M. Khleifat

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Pseudomonas aeruginosa was isolated from infected burn patients and characterized by standard biochemical tests. The in vitro copper uptake was compared between this isolated pathogenic strain and two non-pathogenic control strains of Gram-positive bacteria Bacillusthuringiensis strain Israelisas well as Gram-negative bacteria Enterobacter aerogenes. Maximum copper uptake of 470 ppm/g biomass was obtained by P. aeruginosa strain, while the control strains B. thuringiensis and Enterobacter aerogenes had copper uptake of 350 and 383 ppm/g biomass, respectively. However, the lowest copper uptake (60 ppm/g biomass) was observed with another control the saprophytic strain Pseudomonas (Shewanella) putrefaciens. A further investigation regarding the effect of copper toxicity on bacterial growth, gave an MIC score of 600 ppm for P. aeruginosa strain compared to 460 and 300 ppm for the two Gram positive and Gram negative control strains, respectively. In tandem with these in vitro findings, blood analysis on burn patients infected with P. aeruginosa has indicated a selective decrease of copper (hypocupremia) and ceruloplasmin plasma levels. The iron metabolism was also affected by this copper deprivation leading to a similar decrease in plasma levels of PCV, iron, total iron-binding capacity, and transferrin. All these hematological changes were significantly different (P < 0.05) from the matched group of non-infected burn patients. The observed hypocupremia in infected burn patients was attributed to demanding scavenger ability by P. aeruginosa strain for the copper of plasma.

Keywords: Pseudomonas aeruginosa, hypocupremia, correlation, PCV

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Authors: Adria Rae Abigail R. Eda, Arvin C. Diesmos, Vance T. Vredenburg, Merab A. Chan

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Mitigating strategies using symbiotic cutaneous bacteria is one of the major concerns in the conservation of amphibian population. Batrachochytrium dendrobatidis is the causative agent of chytridiomycosis associated with mass mortality and amphibian extinctions worldwide. In the Philippines, there is a lack of study on the cutaneous bacteria of Philippine amphibians that may have beneficial effects to ward off the deadly fungal infection. In this study, cutaneous bacteria from frogs were isolated and examined for anti-B. dendrobatidis activity. Eight species of frogs were collected at Mt. Palay-palay Mataas na Gulod National Park in Cavite, a site positive for the presence of B. dendrobatidis. Bacteria were isolated from the skin of frogs by swabbing the surfaces of the body and inoculated in Reasoner´s 2A (R2A) agar. Isolated bacteria were tested for potential inhibitory properties against B. dendrobatidis through zoospore inhibition assay. Results showed that frog cutaneous bacteria significantly inhibited the growth of B. dendrobatidis in vitro. By means of 16S rRNA gene primers, the anti-B. dendrobatidis bacteria were identified to be Enterobacter sp., Alcaligenes faecalis and Pseudomonas sp. Cutaneous bacteria namely Enterobacter sp. (isolates PLd33 and PCv4) and Pseudomonas (isolate PLd31) remarkably cleared the growth of B. dendrobatidis zoospore in 1% tryptone agar. Therefore, frog cutaneous bacteria inhibited B. dendrobatidis in vitro and could possibly contribute to the immunity and defense of frogs against the lethal chytridiomycosis.

Keywords: Batrachochytrium dendrobatidis, cutaneous bacteria, frogs, zoospore inhibition assay

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Authors: Ayesha Irum, Sadia Mumtaz, Abdul Rehman, Iffat Naz, Safia Ahmed

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The present study was conducted to evaluate the potential applicability of biological trickling filter system for the treatment of simulated textile wastewater containing reactive azo dyes with bacterial consortium under non-sterile conditions. The percentage decolorization for the treatment of wastewater containing structurally different dyes was found to be higher than 95% in all trials. The stable bacterial count of the biofilm on stone media of the trickling filter during the treatment confirmed the presence, proliferation, dominance and involvement of the added microbial consortium in the treatment of textile wastewater. Results of physicochemical parameters revealed the reduction in chemical oxygen demand (58.5-75.1%), sulphates (18.9-36.5%), and phosphates (63.6-73.0%). UV-Visible and FTIR spectroscopy confirmed decolorization of dye containing wastewater was the ultimate consequence of biodegradation. Toxicological studies revealed the nontoxic nature of degradative metabolites.

Keywords: biodegradation, textile dyes, waste water, trickling filters

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Authors: Ayda Khosravanihaghighi, Pramod Koshy, Bill Walsh, Vedran Lovric, Charles Christopher Sorrell

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There is an increasing demand for orthopedic implants owing to the increasing numbers of the aging population. Titanium alloy (Ti6Al4V) is a common material used for orthopedic implants owing to its advantageous properties in terms of good corrosion resistance, minimal elastic modulus mismatch with bone, bio-inertness, and high mechanical strength. However, it is important to improve the bioactivity and osseointegration of the titanium alloy and this can be achieved by coating the implant surface with suitable ceramic materials. In the present work, pure and doped-ceria (CeO₂) coatings were deposited by spin coating on the titanium alloy surface in order to enhance the biological interactions between the surface of the implant and the surrounding tissue. In order to examine the bone-binding ability of an implant, simulated body fluid (SBF) tests were conducted in order to assess the capability of apatite layer formation on the surface and thus predict in vivo bone bioactivity. Characterization was done using scanning electron microscopy (SEM) and X-ray diffraction (XRD) analyses to determine the extent of apatite formation. Preliminary tests showed that the CeO₂ coatings were biocompatible and that the extent of apatite formation and its characteristics can be enhanced by doping with suitable metal ions.

Keywords: apatite layer, biocompatibility, ceria, orthopaedic implant, SBF, spin coater, Ti-implant

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Authors: Junior Nomani

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This paper presents a novel approach to modelling the metal cutting of duplex stainless steels, a two-phase alloy regarded as a difficult-to-machine material. Calculation and control of shear strain and stresses during cutting are essential to achievement of ideal cutting conditions. Too low or too high leads to higher required cutting force or excessive heat generation causing premature tool wear failure. A 2D finite element cutting model was created based on electron backscatter diffraction (EBSD) data imagery of duplex microstructure. A mesh was generated using ‘object-oriented’ software OOF2 version V2.1.11, converting microstructural images to quadrilateral elements. A virtual workpiece was created on ABAQUS modelling software where a rigid body toolpiece advanced towards workpiece simulating chip formation, generating serrated edge chip formation cutting. Model results found calculated stress strain contour plots correlated well with similar finite element models tied with austenite stainless steel alloys. Virtual chip form profile is also similar compared experimental frozen machining chip samples. The output model data provides new insight description of strain behavior of two phase material on how it transitions from workpiece into the chip.

Keywords: Duplex stainless steel, ABAQUS, OOF2, Chip formation

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Authors: Parnaz Boodagh, Danila Vella, Antonio Damore, Laura Modica De Mohac, Sang-Ho Ye, Garret Coyan, Gaetano Burriesci, William Wagner, Federica Cosentino

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The use of cardiac patches is among the main therapeutic solution for cardiovascular diseases, a leading mortality cause in the world with an increasing trend, responsible of 19 millions deaths in 2020. Several classes of biomaterials serve that purpose, both of synthetic origin and biological derivation, and many bioengineered treatment alternatives were proposed to satisfy two main requirements, providing structural support and promoting tissue remodeling. The objective of this paper is to compare the mechanical properties and the characterization of four cardiac patches: the Adeka, PhotoFix, CorPatch, and CardioCel patches. In vitro and in vivo tests included: biaxial, uniaxial, ball burst, suture retention for mechanical characterization; 2D surface topography, 3D volume and microstructure, and histology assessments for structure characterization; in vitro test to evaluate platelet deposition, calcium deposition, and macrophage polarization; rat right ventricular outflow tract (RVOT) models at 8- and 16-week time points to characterize the patch-host interaction. Lastly, the four patches were used to produce four stented aortic valve prosthesis, subjected to hydrodynamic assessment as well as durability testing to verify compliance with the standard ISO.

Keywords: cardiac patch, cardiovascular disease, cardiac repair, blood contact biomaterial

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Authors: Olha Oleshchuk

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Such companies as Apple, Coca-Cola, Microsoft, Samsung are well-known, but most people do not know about their economic and political influence on society at national and international levels. This ignorance is beneficial for transnational corporations because it reduces the risk of possible resistance of the society and creates favourable conditions for the formation of a new governmental political form. The aim of the research to analyse the opportunity of creating of a new governmental political form. The aim of the research is realized by implementation of such objectives: 1) to consider the possibility of the formation of new political forms of government; 2) to determine the influence of TNCs on the state, which is necessary for a mature, aware citizen; 3) to learn the roles of TNCs, which provide the awareness in the spheres of the world politics, economics, and everyday life. The relevance of the research is confirmed by the fact that new political forms of government are poorly studied.

Keywords: objective factors of losing leading position by state, transnational corporations, TNCs' economic influence, TNCs' political influence

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Authors: Armin Bodaghkhani, Bruce Colbourne, Yuri S. Muzychka

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The process of spray-cloud formation and flow kinematics produced from breaking wave impact on vertical and slant lab-scale bow-shaped models were experimentally investigated. Bubble Image Velocimetry (BIV) and Image Processing (IP) techniques were applied to study the various types of wave-model impacts. Different wave characteristics were generated in a tow tank to investigate the effects of wave characteristics, such as wave phase velocity, wave steepness on droplet velocities, and behavior of the process of spray cloud formation. The phase ensemble-averaged vertical velocity and turbulent intensity were computed. A high-speed camera and diffused LED backlights were utilized to capture images for further post processing. Various pressure sensors and capacitive wave probes were used to measure the wave impact pressure and the free surface profile at different locations of the model and wave-tank, respectively. Droplet sizes and velocities were measured using BIV and IP techniques to trace bubbles and droplets in order to measure their velocities and sizes by correlating the texture in these images. The impact pressure and droplet size distributions were compared to several previously experimental models, and satisfactory agreements were achieved. The distribution of droplets in front of both models are demonstrated. Due to the highly transient process of spray formation, the drag coefficient for several stages of this transient displacement for various droplet size ranges and different Reynolds number were calculated based on the ensemble average method. From the experimental results, the slant model produces less spray in comparison with the vertical model, and the droplet velocities generated from the wave impact with the slant model have a lower velocity as compared with the vertical model.

Keywords: spray charachteristics, droplet size and velocity, wave-body interactions, bubble image velocimetry, image processing

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Authors: Miroslava Rysová, Zdena Syrová, Tomáš Zajíc, Petr Exnar

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Currently, attention of tissue engineers has been attracted to novel nanofibrous materials having advanced properties and ability to mimic extracellular matrix (ECM) by structure which makes them interesting candidates for application in regenerative medicine as scaffolding and/or drug delivering material. Throughout the last decade, more than 200 synthetic and natural polymers have been successfully electrospun leading to the formation of nanofibres with a wide range of chemical, mechanical and degradation properties. In this family, inorganic nanofibres represent very specific group offering an opportunity to manufacture inert to body, well degradable and in properties tunable material. Aim of this work, was to reveal unique properties of silica (SiO2, CAS 7631-86-9) nanofibres and their potential in field of regenerative medicine. Silica nanofibres were prepared by sol-gel method from tetraethyl orthosilicate (TEOS, CAS 78-10-4) as a precursor and subsequently manufactured by needleless electrospinning on NanospiderTM device. Silica nanofibres thermally stabilized under 200°C were confirmed to be fully biodegradable and soluble in several simulated body fluids. In vitro cytotoxicity tests of eluate (ES ISO 10993-5:1999) and in direct contact (ES ISO 10993-5:2009) showed no toxicity - e.g. cell viabilities reached values exceeding 80%. Those results were obtained equally from two different cell lines (Vero, 3T3). Non-toxicity of silaca nanofibres´ eluate was additionally confirmed in real time by testing on xCelligence (ACEA Biosciences, Inc.) device. Both cell types also showed good adhesion to material. To conclude, all mentioned results lead to resumption that silica nanofibres have a potential as material for regenerative medicine which opens door to further research.

Keywords: cytotoxicity, electrospinning, nanofibres, silica, tissue engineering

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Authors: Zhang Yu

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Shanxi Province is a province with a long history in China. The historical characteristics of Jinzhong Prefecture in Shaanxi Province are very prominent. This research has done a lot of field research and analysis, and has analyzed a large number of documents. The formation and characteristics of villages in Jinzhong Prefecture are summarized. But the remains of many areas have not been systematically discovered and analyzed. This study found that the reasons for the formation of villages are natural, cultural, traffic and economic reasons. It mainly includes water, mountain, and developed business culture during the Ming and Qing Dynasties. By analyzing the evolution characteristics of each period, the characteristics and remains of the existing villages are explained in detail. These types of relics mainly include courtyards, fortresses, and Exchange shops. This study can provide systematic guidance on the protection of future village remains.

Keywords: Jinzhong Prefecture, village, features, remains

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Authors: Abdul Karim Zulkarnain, Marchaban, Subagus Wahyono, Ratna Asmah Susidarti

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Mahkota Dewa contains phalerin which has activity as sun screen. In this study, 13 formulations of cream oil in water (o/w) were prepared and tested for their physical characteristics. The physical characteristics were then used for determining the optimum formula. This study aimed to explore the physical stability of optimized formulation of cream, its sun protecting factor (SPF) values using in vitro and in vivo tests. The optimum formula of o/w cream were prepared based on Simplex Lattice Design (LSD) method using software Design Expert®. The formulation of o/w cream were varied based on the proportion of cetyl alcohol, mineral oil and tween 80. The difference of physical characteristic of optimum and predicted formula was tested using t-test with significant level of 95%. The optimum formula of o/w cream was the formula which consists of cetyl alcohol 9.71%, mineral oil, 29%, and tween 80 3.29. Based on t-test, there was no significant difference of physical characteristics of optimum and predicted formulation. Viscosity, spread power, adhesive power, and separation volume ratio of o/w at week 0-4 were relatively stable. The o/w creams were relatively stable at extreme temperature. The o/w creams from mahkota dewa, phalerin, and benzophenone have SPF values of 21.32, 33.12, and 42.49, respectively. The formulas did not irritate the skin based on in vivo test.

Keywords: cream, stability, In vitro, In vivo

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Authors: Katarzyna Drela, Miroslaw Wielgos, Mikolaj Wrobel, Barbara Lukomska

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Mesenchymal stem cells (MSCs) have a great potential in regenerative medicine. Since the initial number of isolated MSCs is limited, in vitro propagation is often required to reach sufficient numbers of cells for therapeutic applications. During long-term culture MSCs may undergo genetic or epigenetic alterations that subsequently increase the probability of spontaneous malignant transformation. Thus, factors that influence genomic stability of MSCs following long-term expansions need to be clarified before cultured MSCs are employed for clinical application. The aim of our study was to investigate the potential for spontaneous transformation of human neonatal cord blood (HUCB-MSCs) and adult bone marrow (BM-MSCs) derived MSCs. Materials and Methods: HUCB-MSCs and BM-MSCs were isolated by standard Ficoll gradient centrifugations method. Isolated cells were initially plated in high density 106 cells per cm2. After 48 h medium were changed and non-adherent cells were removed. The malignant transformation of MSCs in vitro was evaluated by morphological changes, proliferation rate, ability to enter cell senescence, the telomerase expression and chromosomal abnormality. Proliferation of MSCs was analyzed with WST-1 reduction method and population doubling time (PDT) was calculated at different culture stages. Then the expression pattern of genes characteristic for mesenchymal or epithelial cells, as well as transcriptions factors were examined by RT-PCR. Concomitantly, immunocytochemical analysis of gene-related proteins was employed. Results: Our studies showed that MSCs from all bone marrow isolations ultimately entered senescence and did not undergo spontaneous malignant transformation. However, HUCB-MSCs from one of the 15 donors displayed an increased proliferation rate, failed to enter senescence, and exhibited an altered cell morphology. In this sample we observed two different cell phenotypes: one mesenchymal-like exhibited spindle shaped morphology and express specific mesenchymal surface markers (CD73, CD90, CD105, CD166) with low proliferation rate, and the second one with round, densely package epithelial-like cells with significantly increased proliferation rate. The PDT of epithelial-like populations was around 1day and 100% of cells were positive for proliferation marker Ki-67. Moreover, HUCB-MSCs showed a positive expression of human telomerase reverse transcriptase (hTERT), cMYC and exhibit increased number of CFU during the long-term culture in vitro. Furthermore, karyotype analysis revealed chromosomal abnormalities including duplications. Conclusions: Our studies demonstrate that HUCB-MSCs are susceptible to spontaneous malignant transformation during long-term culture. Spontaneous malignant transformation process following in vitro culture has enormous effect on the biosafety issues of future cell-based therapies and regenerative medicine regimens.

Keywords: mesenchymal stem cells, spontaneous, transformation, long-term culture

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Authors: Chenhao Zhu

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Quantitative mapping is playing a growing role in guiding urban planning, such as using a heat map created by CFX, CFD2000, or Envi-met, to adjust the master plan. However, there is no effective quantitative link between the mappings and planning formation. So, in many cases, the decision-making is still based on the planner's subjective interpretation and understanding of these mappings, which limits the improvement of scientific and accuracy brought by the quantitative mapping. Therefore, in this paper, an effort has been made to give a methodology of building a parametric link between the mapping and planning formation. A parametric planning process based on radiant mapping has been proposed for creating an urban green system. In the first step, a script is written in Grasshopper to build a road network and form the block, while the Ladybug Plug-in is used to conduct a radiant analysis in the form of mapping. Then, the research creatively transforms the radiant mapping from a polygon into a data point matrix, because polygon is hard to engage in the design formation. Next, another script is created to select the main green spaces from the road network based on the criteria of radiant intensity and connect the green spaces' central points to generate a green corridor. After that, a control parameter is introduced to adjust the corridor's form based on the radiant intensity. Finally, a green system containing greenspace and green corridor is generated under the quantitative control of the data matrix. The designer only needs to modify the control parameter according to the relevant research results and actual conditions to realize the optimization of the green system. This method can also be applied to much other mapping-based analysis, such as wind environment analysis, thermal environment analysis, and even environmental sensitivity analysis. The parameterized link between the mapping and planning will bring about a more accurate, objective, and scientific planning.

Keywords: parametric link, mapping, urban green system, radiant intensity, planning strategy, grasshopper

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Authors: Esra Turker, Umit Hakan Yildiz, Ahu Arslan Yildiz

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The key of regenerative medicine is mimicking natural three dimensional (3D) microenvironment of tissues by utilizing appropriate biomaterials. In this study, a synthetic biodegradable polymer; poly (L-lactide-co-ε-caprolactone) (PLLCL) and a natural polymer; collagen was used to mimic the biochemical structure of the natural extracellular matrix (ECM), and by means of electrospinning technique the real physical structure of ECM has mimicked. PLLCL/Collagen biocomposite scaffolds enables cell attachment, proliferation and nutrient transport through fabrication of micro to nanometer scale nanofibers. Biocomposite materials are commonly preferred due to limitations of physical and biocompatible properties of natural and synthetic materials. Combination of both materials improves the strength, degradation and biocompatibility of scaffold. Literature studies have shown that collagen is mostly solved with heavy chemicals, which is not suitable for cell culturing. To overcome this problem, a new approach has been developed in this study where polyvinylpyrrolidone (PVP) is used as co-electrospinning agent. PVP is preferred due to its water solubility, so PLLCL/collagen biocomposite scaffold can be easily and rapidly produced. Hydrolytic and enzymatic biodegradation as well as mechanical strength of scaffolds were examined in vitro. Cell adhesion, proliferation and cell morphology characterization studies have been performed as well. Further, on-chip drug screening analysis has been performed over 3D tumor models. Overall, the developed biocomposite scaffold was used for 3D tumor model formation and obtained results confirmed that developed model could be used for drug screening studies to predict clinical efficacy of a drug.

Keywords: biomaterials, 3D cell culture, drug screening, electrospinning, lab-on-a-chip, tissue engineering

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Authors: Sourav Ray, Christoph Stein, Marcus Weber

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A new class of drug candidates, initially derived from mathematical modeling of ligand-receptor interactions, activate the μ-opioid receptor (MOR) preferentially at acidic extracellular pH-levels, as present in injured tissues. This is of commercial interest because it may preclude the adverse effects of conventional MOR agonists like fentanyl, which include but are not limited to addiction, constipation, sedation, and apnea. Animal studies indicate the importance of taking the pH value of the chemical environment of MOR into account when designing new drugs. Hydrogen bonds (HBs) play a crucial role in stabilizing protein secondary structure and molecular interaction, such as ligand-protein interaction. These bonds may depend on the pH value of the chemical environment. For the MOR, antagonist naloxone and agonist [D-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin (DAMGO) form HBs with ionizable residue HIS 297 at physiological pH to modulate signaling. However, such interactions were markedly reduced at acidic pH. Although fentanyl-induced signaling is also diminished at acidic pH, HBs with HIS 297 residue are not observed at either acidic or physiological pH for this strong agonist of the MOR. Molecular dynamics (MD) simulations can provide greater insight into the interaction between the ligand of interest and the HIS 297 residue. Amino acid protonation states are adjusted to the model difference in system acidity. Unbiased and unrestrained MD simulations were performed, with the ligand in the proximity of the HIS 297 residue. Ligand-receptor complexes were embedded in 1-palmitoyl-2-oleoyl-sn glycero-3-phosphatidylcholine (POPC) bilayer to mimic the membrane environment. The occurrence of HBs between the different ligands and the HIS 297 residue of MOR at acidic and physiological pH values were tracked across the various simulation trajectories. No HB formation was observed between fentanyl and HIS 297 residue at either acidic or physiological pH. Naloxone formed some HBs with HIS 297 at pH 5, but no such HBs were noted at pH 7. Interestingly, DAMGO displayed an opposite yet more pronounced HB formation trend compared to naloxone. Whereas a marginal number of HBs could be observed at even pH 5, HBs with HIS 297 were more stable and widely present at pH 7. The HB formation plays no and marginal role in the interaction of fentanyl and naloxone, respectively, with the HIS 297 residue of MOR. However, HBs play a significant role in the DAMGO and HIS 297 interaction. Post DAMGO administration, these HBs might be crucial for the remediation of opioid tolerance and restoration of opioid sensitivity. Although experimental studies concur with our observations regarding the influence of HB formation on the fentanyl and DAMGO interaction with HIS 297, the same could not be conclusively stated for naloxone. Therefore, some other supplementary interactions might be responsible for the modulation of the MOR activity by naloxone binding at pH 7 but not at pH 5. Further elucidation of the mechanism of naloxone action on the MOR could assist in the formulation of cost-effective naloxone-based treatment of opioid overdose or opioid-induced side effects.

Keywords: effect of system acidity, hydrogen bond formation, opioid action, receptor activation

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Authors: Debraj Gangopadhyay, Moumita Das, Ranjan K. Singh, Poonam Tandon

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Creatinine is a toxic chemical waste generated from muscle metabolism. Abnormally high levels of creatinine in the body fluid indicate possible malfunction or failure of the kidneys. This leads to a condition termed as creatinine induced nephrotoxicity. N-acetylcysteine is an antioxidant drug which is capable of preventing creatinine induced nephrotoxicity and is helpful to treat renal failure in its early stages. Taurine is another antioxidant drug which serves similar purpose. The kidneys have a natural power that whenever reactive oxygen species radicals increase in the human body, the kidneys make an antioxidant shell so that these radicals cannot harm the kidney function. Taurine plays a vital role in increasing the power of that shell such that the glomerular filtration rate can remain in its normal level. Thus taurine protects the kidneys against several diseases. However, taurine also has some negative effects on the body as its chloramine derivative is a weak oxidant by nature. N-acetylcysteine is capable of inhibiting the residual oxidative property of taurine chloramine. Therefore, N-acetylcysteine is given to a patient along with taurine and this combination is capable of suppressing the negative effect of taurine. Both N-acetylcysteine and taurine being affordable, safe, and widely available medicines, knowledge of the mechanism of their combined effect on creatinine, the favored route of administration, and the proper dose may be highly useful in their use for treating renal patients. Raman spectroscopy is a precise technique to observe minor structural changes taking place when two or more molecules interact. The possibility of formation of a complex between a drug molecule and an analyte molecule in solution can be explored by analyzing the changes in the Raman spectra. The formation of a stable complex of creatinine with N-acetylcysteinein vitroin aqueous solution has been observed with the help of Raman spectroscopic technique. From the Raman spectra of the mixtures of aqueous solutions of creatinine and N-acetylcysteinein different molar ratios, it is observed that the most stable complex is formed at 1:1 ratio of creatinine andN-acetylcysteine. Upon drying, the complex obtained is gel-like in appearance and reddish yellow in color. The complex is hygroscopic and has much better water solubility compared to creatinine. This highlights that N-acetylcysteineplays an effective role in reducing the toxic effect of creatinine by forming this water soluble complex which can be removed through urine. Since the drug taurine is also known to be useful in reducing nephrotoxicity caused by creatinine, the aqueous solution of taurine with those of creatinine and N-acetylcysteinewere mixed in different molar ratios and were investigated by Raman spectroscopic technique. It is understood that taurine itself does not undergo complexation with creatinine as no additional changes are observed in the Raman spectra of creatinine when it is mixed with taurine. However, when creatinine, N-acetylcysteine and taurine are mixed in aqueous solution in molar ratio 1:1:3, several changes occurring in the Raman spectra of creatinine suggest the diminishing toxic effect of creatinine in the presence ofantioxidant drugs N-acetylcysteine and taurine.

Keywords: creatinine, creatinine induced nephrotoxicity, N-acetylcysteine, taurine

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Authors: Tamer El-Messery, Teresa Sanchez-Moya, Ruben Lopez-Nicolas, Gaspar Ros, Esmat Aly

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Olive leaves (OLs), the main by-product of the olive oil industry, have a considerable amount of phenolic compounds. The exploitation of these compounds represents the current trend in food processing. In this study, OLs polyphenols were microencapsulated with polycaprolactone (PCL) and utilized in formulating novel functional yoghurt. PCL-microcapsules were characterized by scanning electron microscopy, and Fourier transform infrared spectrometry analysis. Their total phenolic (TPC), total flavonoid (TFC) contents, and antioxidant activities (DPPH, FRAP, ABTS), and polyphenols bioaccessibility were measured after oral, gastric, and intestinal steps of in vitro digestion. The four yoghurt formulations (containing 0, 25, 50, and 75 mg of PCL-microsphere/100g yoghurt) were evaluated for their pH, acidity, syneresis viscosity, and color during storage. In vitro digestion significantly affected the phenolic composition in non-encapsulated extract while had a lower impact on encapsulated phenolics. Higher protection was provided for encapsulated OLs extract, and their higher release was observed at the intestinal phase. Yoghurt with PCL-microsphere had lower viscosity, syneresis, and color parameters, as compared to control yoghurt. Thus, OLs represent a valuable and cheap source of polyphenols which can be successfully applied, in microencapsulated form, to formulate functional yoghurt.

Keywords: yoghurt quality attributes, olive leaves, phenolic and flavonoids compounds, antioxidant activity, polycaprolactone as microencapsulant

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Authors: Sambit Supriya Dash, Rahul Ravi R, Vikram Ramanan, Vinayak Malhotra

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Combustion in itself is a complex phenomenon that involves the interaction and interplay of multiple phenomena, the combined effect of which gives rise to the common flame that we see and use in our daily life applications from cooking to propelling our vehicles to space. The most important thing that goes unnoticed about these flames is the effect of the various phenomena from its surrounding environment that affects its behavior and properties. These phenomena cause a variety of energy interactions that lead to various types of energy transformations which in turn affect the flame behavior. This paper focuses on experimentally investigating the effect of one such phenomenon, which is the acoustics or sound energy on diffusion flames. The subject in itself is extensively studied upon as thermo-acoustics globally, whereas the current work focuses on studying its effect on soot formation on diffusion flames. The said effect is studied in this research work by the use of a butane as fuel, fitted with a nozzle that houses 3 arrays consisting of 4 holes each that are placed equidistant to each other and the resulting flame impinged with sound from two independent and similar sound sources that are placed equidistant from the centre of the flame. The entire process is systematically video graphed using a 60 fps regular CCD and analysed for variation in flame heights and flickering frequencies where the fuel mass flow rate is maintained constant and the configuration of entrainment holes and frequency of sound are varied, whilst maintaining constant ambient atmospheric conditions. The current work establishes significant outcomes on the effect of acoustics on soot formation; it is noteworthy that soot formation is the main cause of pollution and a major cause of inefficiency of current propulsion systems. This work is one of its kinds, and its outcomes are widely applicable to commercial and domestic appliances that utilize combustion for energy generation or propulsion and help us understand them better, so that we can increase their efficiency and decrease pollution.

Keywords: thermoacoustics, entrainment, propulsion system, efficiency, pollution

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Authors: Vahid Changizi, Aram Rostami, Akbar Mosavi

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Purpose of study: Critical macromolecules of cells such as DNA are in exposure to damage of free radicals that induced from interaction of ionizing radiation with biological systems. Selenium and vitamin-E are natural compound that has been shown to be a direct free radical scavenger. The aim of this study was to investigate the in vivo/in vitro radioprotective effect of selenium and vitamin-E separately and synergistically against genotoxicity induced by 6MV x-rays irradiation in cultured blood lymphocytes from 15 human volunteers. Methods: Fifteen volunteers were divided in three groups include A, B and C. These groups were given slenium(800 IU), vitamin-E(100 mg) and selenium(400 IU) + vitamin-E(50 mg), respectively. Peripheral blood samples were collected from each group before(0 hr) and 1, 2 and 3 hr after selenium and vitamin-E administration (separately and synergistically). Then the blood samples were irradiated to 200 cGy of 6 Mv x-rays. After that, lymphocyte samples were cultured with mitogenic stimulation to determine the chromosomal aberrations wih micronucleus assay in cytokinesis-blocked binucleated cells. Results: The lymphocytes in the blood samples collected at 1 hr after ingestion selenium and vitamin-E, exposed in vitro to x-rays exhibited a significant decrease in the incidence of micronuclei, compared with control group at 0 hr. The maximum protection and decrease in frequency of micronuclei(50%) was observed at 1 hr after administration of selenium and vitamin-E synergistically. Conclusion: The data suggest that ingestion of selenium and vitamin-E as a radioprotector substances before exposures may reduce genetic damage caused by x-rays irradiation.

Keywords: x-rays, selenium, vitamin-e, lymphocyte, micronuclei

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Authors: Hamdi Omar Omar, Hela Fakhfakh, Chokri Yaich

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This work reflects the integration of different techniques, such as field sampling and observations, magnetic susceptibility measurement, cyclostratigaraphy and sequence stratigraphy. The combination of these results allows us to reconstruct the environmental evolution of the Sidi Khalif Formation in the North-South Axis (NOSA), aged of Upper Tithonian, Berriasian and Lower Valanginian. Six sedimentary facies were identified and are primarily influenced by open marine sedimentation receiving increasing terrigenous influx. Spectral analysis, based on MS variation (for the outcropped section) and wireline logging gamma ray (GR) variation (for the sub-area section) show a pervasive dominance of 405-kyr eccentricity cycles with the expression of 100-kyr eccentricity, obliquity and precession. This study provides (for the first time) a precise duration of 2.4 myr for the outcropped Sidi Khalif Formation with a sedimentation rate of 5.4 cm/kyr and the sub-area section to 3.24 myr with a sedimentation rate of 7.64 cm/kyr. We outlined 27 5th-order depositional sequences, 8 Milankovitch depositional sequences and 2 major 3rd-order cycles for the outcropping section, controlled by the long eccentricity (405 kyr) cycles and the precession index cycles. This study has demonstrated the potential of MS and GR to be used as proxies to develop an astronomically calibrated time-scale for the Mesozoic era.

Keywords: Berriasian, magnetic susceptibility, orbital tuning, Sidi Khalif Formation

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Authors: A. Aulinger, A. M. Backes, J. Bieser, V. Matthias, M. Quante

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High concentrations of particles pose a threat to human health. Thus, legal maximum concentrations of PM10 and PM2.5 in ambient air have been steadily decreased over the years. In central Europe, the inorganic species ammonium sulphate and ammonium nitrate make up a large fraction of fine particles. Many studies investigate the influence of emission reductions of sulfur- and nitrogen oxides on aerosol concentration. Here, we focus on the influence of ammonia (NH3) emissions. While emissions of sulphate and nitrogen oxides are quite well known, ammonia emissions are subject to high uncertainty. This is due to the uncertainty of location, amount, time of fertilizer application in agriculture, and the storage and treatment of manure from animal husbandry. For this study, we implemented a crop growth model into the SMOKE emission model. Depending on temperature, local legislation, and crop type individual temporal profiles for fertilizer and manure application are calculated for each model grid cell. Additionally, the diffusion from soils and plants and the direct release from open and closed barns are determined. The emission data was used as input for the Community Multiscale Air Quality (CMAQ) model. Comparisons to observations from the EMEP measurement network indicate that the new ammonia emission module leads to a better agreement of model and observation (for both ammonia and ammonium). Finally, the ammonia emission model was used to create emission scenarios. This includes emissions based on future European legislation, as well as a dynamic evaluation of the influence of different agricultural sectors on particle formation. It was found that a reduction of ammonia emissions by 50% lead to a 24% reduction of total PM2.5 concentrations during winter time in the model domain. The observed reduction was mainly driven by reduced formation of ammonium nitrate. Moreover, emission reductions during winter had a larger impact than during the rest of the year.

Keywords: ammonia, ammonia abatement strategies, ctm, seasonal impact, secondary aerosol formation

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Authors: Faraj M. Elkhatri, Hana Ali Allafi

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The spatial and temporal distribution of diagenetic alterations related impact on the reservoir quality of the Sarir Formation. (present day burial depth of about 9000 feet) Depositional facies and diagenetic alterations are the main controls on reservoir quality of Sarir Formation Sirt Ba-sin Libya; these based on lithology and grain size as well as authigenic clay mineral types and their distributions. However, petrology investigation obtained on study area with five sandstone wells concentrated on main rock components and the parameters that may have impacts on reservoirs. the main authigenic clay minerals are kaolinite and dickite, these investigations have confirmed by X.R.D analysis and clay fraction. mainly Kaolinite and Dickite were extensively presented on all of wells with high amounts. As well as trace of detrital smectite and less amounts of illitized mud-matrix are possibly find by SEM image. Thin layers of clay presented as clay-grain coatings in local depth interpreted as remains of dissolved clay matrix is partly transformed into kaolinite adjacent and towards pore throat. This also may have impacts on most of the pore throats of this sandstone which are open and relatively clean with some of fine martial have been formed on occluded pores. This material is identified by EDS analysis to be collections of not only kaolinite booklets, but also small, disaggregated kaolinite platelets derived from the dis-aggregation of larger kaolinite booklets. These patches of kaolinite not only fill this pore, but also coat some of the sur-rounding framework grains. Quartz grains often enlarged by authigenic quartz overgrowths partially occlude and re-duce porosity. Scanning Electron Microscopy with Energy Dispersive Spectroscopy (SEM) was conducted on the post-test samples to examine any mud filtrate particles that may be in the pore throats. Semi-qualitative elemental data on select-ed minerals observed during the SEM study were obtained using an Energy Dispersive Spectroscopy (EDS) unit. The samples showed mostly clean open pore throats, with limited occlusion by kaolinite.

Keywords: por throat, formation damage, porosity lose, solids plugging

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Authors: Akbar Esmaeili, Zahra Salarieh

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Cerium oxide nanoparticles (nano-series) are used as catalysts in industrial applications due to their free radical scavenging properties. Given that free radicals play an essential role in the pathology of many neurological diseases, we investigated the use of nanocrystals as a potential therapeutic agent for oxidative damage. This project synthesized nano-series from a new and environmentally friendly bio-pathway. Investigation of cerium nitrate in culture medium containing inoculated Lactobacillus acidophilus strain before incubation produces nano-series. Loaded with glatiramer acetate (GA) was formed by coating carboxymethylcellulose (CMC) and CeO2. FE-SEM analysis showed nano-series in the 9-11 nm range, spherical shape, and uniform particle size distribution. Cubic nanoparticles containing anti-multiple sclerosis (anti-Ms) treatment called GA were used. Glycerol monostearate (GMS) was used as a fat base, and evening primrose extract was used as an anti-inflammatory in cubosomes. Design-Expert® software was used to study the effects of different formulation factors on the properties of GAloaded cubic dispersions. Thirty GA-labeled cubic dispersions were prepared with GA-labeled carboxymethylcellulose and evaluated in vitro. The results showed an average nano-series size of 89.02 and a zeta potential of -49.9. Cubosomes containing GA-CMC/CeO2 showed a stable release profile for 180 min. The results showed that cubosomes containing GA-CMC/CeO2 could be a promising drug carrier with normal release behavior.

Keywords: ciochemistry, biotechnology, molecular, biology

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Authors: Carlos Amnael Orozco-Diaz, Robert David Moorehead, Gwendolen Reilly, Fiona Gilchrist, Cheryl Ann Miller

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The current gold-standard for maxillofacial reconstruction surgery (MRS) utilizes auto-grafted cancellous bone as a filler. This study was aimed towards developing a polylactic-acid/hydroxyapatite (PLA-HA) composite suitable for fused-deposition 3D printing. Functionalization of the polymer through the addition of HA was directed to promoting bone-regeneration properties so that the material can rival the performance of cancellous bone grafts in terms of bone-lesion repair. This kind of composite enables the production of MRS implants based off 3D-reconstructions from image studies – namely computed tomography – for anatomically-correct fitting. The present study encompassed in-vitro degradation and in-vitro biocompatibility profiling for 3D-printed PLA and PLA-HA composites. PLA filament (Verbatim Co.) and Captal S hydroxyapatite micro-scale HA powder (Plasma Biotal Ltd) were used to produce PLA-HA composites at 5, 10, and 20%-by-weight HA concentration. These were extruded into 3D-printing filament, and processed in a BFB-3000 3D-Printer (3D Systems Co.) into tensile specimens, and were mechanically challenged as per ASTM D638-03. Furthermore, tensile specimens were subjected to accelerated degradation in phosphate-buffered saline solution at 70°C for 23 days, as per ISO-10993-13-2010. This included monitoring of mass loss (through dry-weighing), crystallinity (through thermogravimetric analysis/differential thermal analysis), molecular weight (through gel-permeation chromatography), and tensile strength. In-vitro biocompatibility analysis included cell-viability and extracellular matrix deposition, which were performed both on flat surfaces and on 3D-constructs – both produced through 3D-printing. Discs of 1 cm in diameter and cubic 3D-meshes of 1 cm3 were 3D printed in PLA and PLA-HA composites (n = 6). The samples were seeded with 5000 MG-63 osteosarcoma-like cells, with cell viability extrapolated throughout 21 days via resazurin reduction assays. As evidence of osteogenicity, collagen and calcium deposition were indirectly estimated through Sirius Red staining and Alizarin Red staining respectively. Results have shown that 3D printed PLA loses structural integrity as early as the first day of accelerated degradation, which was significantly faster than the literature suggests. This was reflected in the loss of tensile strength down to untestable brittleness. During degradation, mass loss, molecular weight, and crystallinity behaved similarly to results found in similar studies for PLA. All composite versions and pure PLA were found to perform equivalent to tissue-culture plastic (TCP) in supporting the seeded-cell population. Significant differences (p = 0.05) were found on collagen deposition for higher HA concentrations, with composite samples performing better than pure PLA and TCP. Additionally, per-cell-calcium deposition on the 3D-meshes was significantly lower when comparing 3D-meshes to discs of the same material (p = 0.05). These results support the idea that 3D-printable PLA-HA composites are a viable resorbable material for artificial grafts for bone-regeneration. Degradation data suggests that 3D-printing of these materials – as opposed to other manufacturing methods – might result in faster resorption than currently-used PLA implants.

Keywords: bone regeneration implants, 3D-printing, in vitro testing, biocompatibility, polymer degradation, polymer-ceramic composites

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Authors: Akshita Jindal, Renu Chadha, Maninder Karan

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Supramolecular chemistry has gained recent eminence in a flurry of research documents demonstrating the formation of new crystalline forms with potentially advantageous characteristics. Mesalamine (5-amino salicylic acid) belongs to anti-inflammatory class of drugs, is used to treat ulcerative colitis and Crohn’s disease. Unfortunately, mesalamine suffer from poor solubility and therefore very low bioavailability. This work is focused on preparation and characterization of cocrystal of mesalamine with nicotinamide (MNIC) a coformer of GRAS status. Cocrystallisation was achieved by solvent drop grinding in stoichiometric ratio of 1:1 using acetonitrile as solvent and was characterized by various techniques including DSC (Differential Scanning Calorimetry), PXRD (X-ray Powder Diffraction), and FTIR (Fourier Transform Infrared Spectrometer). The co-crystal depicted single endothermic transitions (254°C) which were different from the melting peaks of both drug (288°C) and coformer (128°C) indicating the formation of a new solid phase. Different XRPD patterns and FTIR spectrums for the co-crystals from those of individual components confirms the formation of new phase. Enhancement in apparent solubility study and intrinsic dissolution study showed effectiveness of this cocrystal. Further improvement in pharmacokinetic profile has also been observed with 2 folds increase in bioavailability. To conclude, our results show that application of nicotinamide as a coformer is a viable approach towards the preparation of cocrystals of potential drug molecule having limited solubility.

Keywords: cocrystal, mesalamine, nicotinamide, solvent drop grinding

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Authors: Mike Wei, Nebu Koshy, Koen van Besien, Giorgio Inghirami, Steven M. Horwitz

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T-cell prolymphocytic leukemia (T-PLL) is a rare hematologic disease characterized by a T-cell phenotype, rapid progression, and poor prognosis with median survival of less than a year. Alemtuzumab-based chemotherapy has increased the rate of complete remissions but these are often short-lived, and allogeneic transplant is considered the only curative therapy. In recent studies, JAK3 activating mutations have been identified in T-cell cancers, with T-PLL having the highest rate of JAK3 mutations (30 – 42%). As such, T-PLL is a model disease for evaluating the utility of JAK3 inhibitors. We present a case of a 64-year-old man with relapsed-refractory T-PLL. He was initially treated with alemtuzumab and obtained complete response and was consolidated with matched unrelated donor stem cell transplant. His disease stayed in remission for approximately 1.5 years before relapse, which was then treated with a clinical trial of romidepsin-lenalidomide (partial responses then progression at 6 months) and later alemtuzumab. Due to complications of myelosuppression and CMV reactivation, his treatment was interrupted leading to disease progression. The doubling time of lymphocyte count was approximately 20 days and over a span of 60 days the lymphocyte count rose from 8 x 109/L to 68 x 109/L. Exon sequencing showed a JAK3 mutation. The patient consented to and was treated with FDA-approved tofacitinib (initially 5 mg BID, increased to 10 mg BID after 15 days of treatment). An initial decrease in lymphocyte count was followed by progression. In vitro treatment of the patient’s cells showed modest effects of tofacitinib and ruxolitinib as single agents, in the range of doxorubicin, but synergy between the agents. After 40 days of treatment with tofacitinib and with a lymphocyte count of 150 x 109/L, ruxolitinib (5mg BID) was added. Over the 60 days since dual inhibition was started, the lymphocyte count has stabilized. The patient has remained completely asymptomatic during treatment with tofacitinib and ruxolitinib. Neutrophil count has remained normal. Platelet count and hemoglobin have however declined from ~50 x109/L to ~30 x109/L and from 11 g/dL to 8.1 g/dL respectively, since the introduction of ruxolitinib. The stabilization in lymphocyte count confirms the clinical activity of JAK inhibitors in T-PLL as suggested by the presence of JAK3 mutations and by in-vitro assays. It also suggests clinical synergy between ruxolitinib and tofacitinib in this setting. Prospective studies of JAK inhibitors in PLL patients with formal dose-finding studies are needed.

Keywords: tofacitinib, ruxolitinib, T-cell prolymphocytic leukemia, JAK3

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Authors: Anton G. Kalmykov, Andrew Yu. Bychkov, Georgy A. Kalmykov

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Nowadays, most oil reserves in Russia and all over the world are hard to recover. That is the reason oil companies are searching for new sources for hydrocarbon production. One of the sources might be high-carbon formations with unconventional reservoirs. Bazhenov formation is a huge source rock formation located in West Siberia, which contains unconventional reservoirs on some of the areas. These reservoirs are formed by secondary processes with low predicting ratio. Only one of five wells is drilled through unconventional reservoirs, in others kerogen has low thermal maturity, and they are of low petroliferous. Therefore, there was a request for tertiary methods for in-situ cracking of kerogen and production of oil. Laboratory experiments of Bazhenov formation rock hydrous pyrolysis were used to investigate features of the oil generation process. Experiments on Bazhenov rocks with a different mineral composition (silica concentration from 15 to 90 wt.%, clays – 5-50 wt.%, carbonates – 0-30 wt.%, kerogen – 1-25 wt.%) and thermal maturity (from immature to late oil window kerogen) were performed in a retort under reservoir conditions. Rock samples of 50 g weight were placed in retort, covered with water and heated to the different temperature varied from 250 to 400°C with the durability of the experiments from several hours to one week. After the experiments, the retort was cooled to room temperature; generated hydrocarbons were extracted with hexane, then separated from the solvent and weighted. The molecular composition of this synthesized oil was then investigated via GC-MS chromatography Characteristics of rock samples after the heating was measured via the Rock-Eval method. It was found, that the amount of synthesized oil and its composition depending on the experimental conditions and composition of rocks. The highest amount of oil was produced at a temperature of 350°C after 12 hours of heating and was up to 12 wt.% of initial organic matter content in the rocks. At the higher temperatures and within longer heating time secondary cracking of generated hydrocarbons occurs, the mass of produced oil is lowering, and the composition contains more hydrocarbons that need to be recovered by catalytical processes. If the temperature is lower than 300°C, the amount of produced oil is too low for the process to be economically effective. It was also found that silica and clay minerals work as catalysts. Selection of heating conditions allows producing synthesized oil with specified composition. Kerogen investigations after heating have shown that thermal maturity increases, but the yield is only up to 35% of the maximum amount of synthetic oil. This yield is the result of gaseous hydrocarbons formation due to secondary cracking and aromatization and coaling of kerogen. Future investigations will allow the increase in the yield of synthetic oil. The results are in a good agreement with theoretical data on kerogen maturation during oil production. Evaluated trends could be tooled up for in-situ oil generation by shale rocks thermal action.

Keywords: Bazhenov formation, fossil fuels, hydropyrolysis, synthetic oil

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Authors: Masoud Sakhinia, Sajjad Ahmad

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Introduction: Though knowledge of the compositional makeup and structure of the limbal niche has progressed exponentially during the past decade, much is yet to be understood. Identifying the precise profile and role of the stromal makeup which spans the ocular surface may inform researchers of the most optimum conditions needed to effectively expand LESCs in vitro, whilst preserving their differentiation status and phenotype. Limbal fibroblasts, as opposed to corneal fibroblasts are thought to form an important component of the microenvironment where LESCs reside. Methods: The corneal stroma was tissue engineered in vitro using both limbal and corneal fibroblasts embedded within a tissue engineered 3D collagen matrix. The effect of these two different fibroblasts on LESCs and hTCEpi corneal epithelial cell line were then subsequently determined using phase contrast microscopy, histolological analysis and PCR for specific stem cell markers. The study aimed to develop an in vitro model which could be used to determine whether limbal, as opposed to corneal fibroblasts, maintained the stem cell phenotype of LESCs and hTCEpi cell line. Results: Tissue culture analysis was inconclusive and required further quantitative analysis for remarks on cell proliferation within the varying stroma. Histological analysis of the tissue-engineered cornea showed a comparable structure to that of the human cornea, though with limited epithelial stratification. PCR results for epithelial cell markers of cells cultured on limbal fibroblasts showed reduced expression of CK3, a negative marker for LESC’s, whilst also exhibiting a relatively low expression level of P63, a marker for undifferentiated LESCs. Conclusion: We have shown the potential for the construction of a tissue engineered human cornea using a 3D collagen matrix and described some preliminary results in the analysis of the effects of varying stroma consisting of limbal and corneal fibroblasts, respectively, on the proliferation of stem cell phenotype of primary LESCs and hTCEpi corneal epithelial cells. Although no definitive marker exists to conclusively illustrate the presence of LESCs, the combination of positive and negative stem cell markers in our study were inconclusive. Though it is less traslational to the human corneal model, the use of conditioned medium from that of limbal and corneal fibroblasts may provide a more simple avenue. Moreover, combinations of extracellular matrices could be used as a surrogate in these culture models.

Keywords: cornea, Limbal Stem Cells, tissue engineering, PCR

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Authors: Priyanka Sharma, Niti Puri

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Anemia develops when blood lacks enough healthy erythrocytes. Past studies indicated that anemia, inflammatory process, and oxidative stress are interconnected. Erythrocytes are continuously exposed to reactive oxygen species (ROS) during circulation, due to normal aerobic cellular metabolism and also pathology of inflammatory diseases. Systemic mastocytosis and genetic depletion of mast cells have been shown to affect anaemia. In the present study, we attempted to reveal whether mast cells have a direct role in clearance or erythrophagocytosis of normal or oxidatively damaged erythrocytes. Murine erythrocytes were treated with tert-butyl hydroperoxidase (t-BHP), an agent that induces oxidative damage and mimics in vivo oxidative stress. Normal and oxidatively damaged erythrocytes were labeled with carboxyfluorescein succinimidyl ester (CFSE) to track erythrophagocytosis. We show, for the first time, direct erythrophagocytosis of oxidatively damaged erythrocytes in vitro by RBL-2H3 mast cells as well as in vivo by murine peritoneal mast cells. Also, activated mast cells, as may be present in inflammatory conditions, showed a significant increase in the uptake of oxidatively damaged erythrocytes than resting mast cells. This suggests the involvement of mast cells in erythrocyte clearance during oxidative stress or inflammatory disorders. Partial inhibition of phagocytosis by various inhibitors indicated that this process may be controlled by several pathways. Hence, our study provides important evidence for involvement of mast cells in severe anemia due to inflammation and oxidative stress and might be helpful to circumvent the adverse anemic disorders.

Keywords: mast cells, anemia, erythrophagocytosis, oxidatively damaged erythrocytes

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