Search results for: Paraptosis-like program cell death

Authors: Prabesh Karki, Shyam Thapa Chettri, Bajarang Prasad Sah, Manoj Bhattarai, Sudeep Mishra

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Introduction: Frontal sinus is frequently described as the most difficult sinus to access surgically due to its proximity to the cribriform plate, orbit, and anterior ethmoid artery. Frontal sinus surgery requires a detailed understanding of the cellular structure and FSDP unique to each patient, making high-resolution CT scans an indispensable tool to assess the difficulty of planned sinus surgery. International Frontal Sinus Anatomy Classification (IFAC) was developed to provide a more precise nomenclature for cells in the frontal recess, classifying cells based on their anatomic origin. Objectives: To assess the proportion of frontal cell variants defined by IFAC, variation with respect to age and gender. Methods: 54 cases were enrolled after a detailed clinical history, thorough general and physical examinations, and CT a report ordered in a film. Assessment and tabulation of the presence of frontal cells according to the IFAC analyzed. The prevalence of each cell type was calculated, and data were entered in MS Excel and analyzed using Statistical Package for the Social Sciences (SPSS). Descriptive statistics and frequencies were defined for categorical and numerical variables. Frequency, percentage, the mean and standard deviation were calculated. Result: Among 54 patients, 30 (55.6%) were male and 24 (44.4%) were female. The patient enrolled ranged from 18 to 78 years. Majority33.3% (n=18) were in age group of >50 years.According to IFAC, Agger nasi cells (92.6%) were most common, whereas supraorbital ethmoidal cells were least common 16 (29.6%). Prevalence of other frontoethmoidal cells was SAC- 57.4%, SAFC- 38.9%, SBC- 74.1%, SBFC- 33.3%, FSC- 38.9% of 54 cases. Conclusion: IFAC is an international consensus document that describes an anatomically precise nomenclature for classifying frontoethmoidal cells' anatomy. This study has defined the prevalence, symmetry and reliability of frontoethmoidal cells as established by the IFAC system as in other parts of the world.

Keywords: frontal sinus, frontoethmoidal cells, international frontal sinus anatomy classification

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Authors: M. Towhidur Rahman, Nasim Zaman Piyas, M. Sadiqul Baree, Shahnewaz Ahmed

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The design of high-speed craft has recently become one of the most active areas of naval architecture. Speed increase makes these vehicles more efficient and useful for military, economic or leisure purpose. The planing hull is designed specifically to achieve relatively high speed on the surface of the water. Speed on the water surface is closely related to the size of the vessel and the installed power. The Savitsky method was first presented in 1964 for application to non-monohedric hulls and for application to stepped hulls. This method is well known as a reliable comparative to CFD analysis of hull resistance. A computer program based on Savitsky’s method has been developed using MATLAB. The power of high-speed vessels has been computed in this research. At first, the program reads some principal parameters such as displacement, LCG, Speed, Deadrise angle, inclination of thrust line with respect to keel line etc. and calculates the resistance of the hull using empirical planning equations of Savitsky. However, some functions used in the empirical equations are available only in the graphical form, which is not suitable for the automatic computation. We use digital plotting system to extract data from nomogram. As a result, value of wetted length-beam ratio and trim angle can be determined directly from the input of initial variables, which makes the power calculation automated without manually plotting of secondary variables such as p/b and other coefficients and the regression equations of those functions are derived by using data from different charts. Finally, the trim angle, mean wetted length-beam ratio, frictional coefficient, resistance, and power are computed and compared with the results of Savitsky and good agreement has been observed.

Keywords: nomogram, planing hull, principal parameters, regression

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Authors: Yuri Bizzoni, Marianne Reboul

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Our program compares French and Italian translations of Homer’s Odyssey, from the XVIth to the XXth century. We focus on the third point, showing how distributional semantics systems can be used both to improve alignment between different French translations as well as between the Greek text and a French translation. Although we focus on French examples, the techniques we display are completely language independent.

Keywords: classical receptions, computational linguistics, distributional semantics, Homeric poems, machine translation, translation studies, text alignment

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Authors: Juan Heredia, Naci Dilekli

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The Ecuadorian Rainforest has been polluted for almost 60 years with little to no regard to oversight, law, or regulations. The consequences have been vast environmental damage such as pollution and deforestation, as well as sickness and the death of many people and animals. The aim of this paper is to quantify and localize the polluted zones, which something that has not been conducted and is the first step for remediation. To approach this problem, multi-spectral Remote Sensing imagery was utilized using a novel algorithm developed for this study, based on four normalized indices available in the literature. The algorithm classifies the pixels in polluted or healthy ones. The results of this study include a new algorithm for pixel classification and quantification of the polluted area in the selected image. Those results were finally validated by ground control points found in the literature. The main conclusion of this work is that using hyperspectral images, it is possible to identify polluted vegetation. The future work is environmental remediation, in-situ tests, and more extensive results that would inform new policymaking.

Keywords: remote sensing, oil pollution quatification, amazon forest, hyperspectral remote sensing

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Authors: Umar Saeed

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Peptidyl-prolyl cis/trans isomerases Par14 and Par17 in humans play crucial roles in diverse cellular processes, including protein folding, chromatin remodeling, DNA binding, ribosome biogenesis, and cell cycle progression. However, the effects of Par14 and Par17 on viral replication have been explored to a limited extent. We first time discovered their influential roles in promoting Hepatitis B Virus replication. In this study, we observed that in the presence of HBx, either Par14 or Par17 could upregulate HBV replication. However, in the absence of HBx, neither Par14 nor Par17 had any effect on replication. Their mechanism of action involves binding to specific motifs within HBc and HBx proteins. Notably, they target the conserved 133Arg-Pro134 (RP) motif of HBc and the 19RP20-28RP29 motifs of HBx. This interaction is fundamental for the stability of HBx, core particles, and HBc. Par14 and Par17 exhibit versatility by binding both outside and inside core particles, thereby facilitating core particle assembly through their participation in HBc dimer-dimer interactions. NAGE and immunoblotting analyses unveiled the binding of Par14/Par17 to core particles. Co-immunoprecipitation experiments further demonstrated the interaction of Par14/Par17 with core particle assembly-defective and dimer-positive HBc-Y132A. It's essential to emphasize that R133 is the key residue in the HBc RP motif that governs their interaction with Par14/Par17. Chromatin immunoprecipitation conducted on HBV-infected cells elucidated the participation of residues S19 and E46/D74 in Par14 and S44 and E71/D99 in Par17 in the recruitment of 133RP134 motif-containing HBc into cccDNA. Depleting PIN4 in liver cell lines results in a significant reduction in cccDNA levels, pgRNA, sgRNAs, HBc, core particle assembly, and HBV DNA synthesis. Notably, parvulin inhibitors like juglone and PiB have proven to be effective in substantially reducing HBV replication. These inhibitors weaken the interaction between HBV core particles and Par14/Par17, underscoring the dynamic nature of this interaction. It's also worth noting that specific Par14/Par17 inhibitors hold promise as potential therapeutic options for chronic hepatitis B.

Keywords: Par14Par17, HBx, HBc, cccDNA, HBV

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Authors: Soledad Carinelli, Iñigo Fernández, José Luis González-Mora, Pedro A. Salazar-Carballo

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Mastitis is the most relevant inflammatory disease in cattle, affecting the animal health and causing important economic losses on dairy farms. This disease takes place in the mammary gland or udder when some opportunistic microorganisms, such as Staphylococcus aureus, Streptococcus agalactiae, Corynebacterium bovis, etc., invade the teat canal. According to the severity of the inflammation, mastitis can be classified as sub-clinical, clinical and chronic. Standard methods for mastitis detection include counts of somatic cells, cell culture, electrical conductivity of the milk, and California test (evaluation of “gel-like” matrix consistency after cell lysed with detergents). However, these assays present some limitations for accurate detection of subclinical mastitis. Currently, haptoglobin, an acute phase protein, has been proposed as novel and effective biomarker for mastitis detection. In this work, an electrochemical biosensor based on polydopamine-modified magnetic nanoparticles (MNPs@pDA) for haptoglobin detection is reported. Thus, MNPs@pDA has been synthesized by our group and functionalized with hemoglobin due to its high affinity to haptoglobin protein. The protein was labeled with specific antibodies modified with alkaline phosphatase enzyme for its electrochemical detection using an electroactive substrate (1-naphthyl phosphate) by differential pulse voltammetry. After the optimization of assay parameters, the haptoglobin determination was evaluated in milk. The strategy presented in this work shows a wide range of detection, achieving a limit of detection of 43 ng/mL. The accuracy of the strategy was determined by recovery assays, being of 84 and 94.5% for two Hp levels around the cut off value. Milk real samples were tested and the prediction capacity of the electrochemical biosensor was compared with a Haptoglobin commercial ELISA kit. The performance of the assay has demonstrated this strategy is an excellent and real alternative as screen method for sub-clinical bovine mastitis detection.

Keywords: bovine mastitis, haptoglobin, electrochemistry, magnetic nanoparticles, polydopamine

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Authors: Roger G. Pagente Jr.

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This study seeks to investigate the language learning experiences of the thirty-nine AB-English majors who were selected through fish-bowl technique from the 157 students enrolled in the AB-English program. Findings taken from the diary, questionnaire and unstructured interview revealed that motivation, learners’ belief, self-monitoring, language anxiety, activities and strategies were the prevailing factors that influenced the learning of English of the participants.

Keywords: diary, English language learning experiences, self-monitoring, language anxiety

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Authors: Abdela Zeinu Yasin

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The current research aimed to assess the status of IMC in providing treatment for malnourished children and programs in water, sanitation, and hygiene (WASH), food and livelihood security, and comprehensive healthcare through Mobile health and nutrition programs during the last 5 years period. We have conducted 60 in-depth interviews with women during the period from conception to a child’s birthday, health facility staff, and female community health volunteers (FCHVs), as well as 12 focus group discussions with health facility staff and other household decision-makers. We employed thematic analysis using framework matrices and analytical memorandums. The study revealed that 78% of the respondents, of whom 97% were women, have benefited from the selected vulnerable areas. The use of the clear water and sanitization program has reached the 81% of selected households. The use of a modern baby delivery system among the respondent has been 68% of the women and health facilities among the decision-makers/focal person. More than 8 in 10 participants (84%) could read and understand the health facility instructions, and the majority (82%) of women, health facility staff, and male decision-makers can also read and write bulletins and instructions. We found that decision-maker women preferred participative education, whereas health facilities and the IMC desired educational and motivational bulletins. A Mobile Health and Nutrition program intervention by the IMC is acceptable in the conditions of the Ethiopian community and has the potential to improve community health and nutrition service utilization, particularly by providing clean water and sanitization; women’s birth control, and health improvement in the vulnerable regions of the country. The current research findings shall contribute to text IMC Mobile Health and Nutritional intervention design in under-resourced settings.

Keywords: clean water, health and nutrition services, hygiene, IMC, mobile health, sanitation

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Authors: Belal Bakheet, John Beardall, Xiwang Zhang, David McCarthy

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The potential risks associated with toxic cyanobacteria have raised growing environmental and public health concerns leading to an increasing effort into researching ways to bring about their removal from water, together with destruction of their associated cyanotoxins. A variety of toxins are synthesized by cyanobacteria and include hepatotoxins, neurotoxins, and cytotoxins which can cause a range of symptoms in humans from skin irritation to serious liver and nerve damage. Therefore drinking water treatment processes should ensure the consumers’ safety by removing both cyanobacterial cells, and cyanotoxins from the water. Cyanobacterial cells and cyanotoxins presented challenges to the conventional water treatment systems; their accumulation within drinking water treatment plants has been reported leading to plants shut down. Thus, innovative and effective water purification systems to tackle cyanobacterial pollution are required. In recent years there has been increasing attention to the electrochemical oxidation process as a feasible alternative disinfection method which is able to generate in situ a variety of oxidants that would achieve synergistic effects in the water disinfection process and toxin degradation. By utilizing only electric current, the electrochemical process through electrolysis can produce reactive oxygen species such as hydroxyl radicals from the water, or other oxidants such as chlorine from chloride ions present in the water. From extensive physiological and morphological investigation of cyanobacterial cells during electrolysis, our results show that these oxidants have significant impact on cell inactivation, simultaneously with cyanotoxins removal without the need for chemicals addition. Our research aimed to optimize existing electrochemical oxidation systems and develop new systems to treat water containing toxic cyanobacteria and cyanotoxins. The research covers detailed mechanism study on oxidants production and cell inactivation in the treatment under environmental conditions. Overall, our study suggests that the electrochemical treatment process e is an effective method for removal of toxic cyanobacteria and cyanotoxins.

Keywords: toxic cyanobacteria, cyanotoxins, electrochemical process, oxidants

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Authors: Ashwini Mohan, Haris Batley

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An orthodontic index is used to rate or categorise an individual’s occlusion using a numeric or alphanumeric score. Indexing of malocclusions and their correction is important in epidemiology, diagnosis, communication between clinicians as well as their patients and assessing treatment outcomes. Many useful indices have been put forward, but to the author’s best knowledge, no one method to this day appears to be equally suitable for the use of epidemiologists, public health program planners and clinicians. This article describes the common clinical orthodontic indices and classifications used in United Kingdom.

Keywords: classification, indices, orthodontics, validity

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Authors: Anita Conti, Riccardo Ossanna, Lindsey A. Quintero, Giamaica Conti, Andrea Sbarbati

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Ischemia/reperfusion (IR) injury induces muscle fiber atrophy and skeletal muscle fiber death with subsequently functionality loss. The heterogeneous pool of cells, especially mesenchymal stem cells, contained in the stromal vascular fraction (SVF) of adipose tissue could promote muscle fiber regeneration. To prevent SVF dispersion, it has been proposed the use of injectable biopolymers that work as cells carrier. A significant element of the extracellular matrix is hyaluronic acid (HA), which has been widely used in regenerative medicine as a cell scaffold given its biocompatibility, degradability, and the possibility of chemical functionalization. Connective tissue micro-fragments enriched with SVF obtained from mechanical disaggregation of adipose tissue were evaluated for IR muscle injury regeneration using low molecular weight HA as a scaffold. IR induction. Hindlimb ischemia was induced in 9 athymic nude mice through the clamping of the right quadriceps using a plastic band. Reperfusion was induced by cutting the plastic band after 3 hours of ischemic period. Contralateral (left) muscular tissue was used as healthy control. Treatment. Twenty-four hours after the IR induction, animals (n=3) were intramuscularly injected with 100 µl of SVF mixed with HA (SVF-HA). Animals treated with 100 µl of HA (n=3) and 100 µl saline solution (n=3) were used as control. Treatment monitoring. All animals were in vivo monitored by magnetic resonance imaging (MRI) at 5, 7, 14 and 18 days post-injury (dpi). High-resolution morphological T2 weighed, quantitative T2 map and Dynamic Contrast-Enhanced (DCE) images were acquired in order to assess the regenerative potential of SVF-HA treatment. Ex vivo evaluation. After 18 days from IR induction, animals were sacrificed, and the muscles were harvested for histological examination. At 5 dpi T2 high-resolution MR images clearly reveal the presence of an extensive edematous area due to IR damage for all groups identifiable as an increase of signal intensity (SI) of muscular and surrounding tissue. At 7 dpi, animals of the SVF-HA group showed a reduction of SI, and the T2relaxation time of muscle tissue of the HA-SVF group was 29±0.5ms, comparable with the T2relaxation time of contralateral muscular tissue (30±0.7ms). These suggest a reduction of edematous overflow and swelling. The T2relaxation time at 7dpi of HA and saline groups were 84±2ms and 90±5ms, respectively, which remained elevated during the rest of the study. The evaluation of vascular regeneration showed similar results. Indeed, DCE-MRI analysis revealed a complete recovery of muscular tissue perfusion after 14 dpi for the SVF-HA group, while for the saline and HA group, controls remained in a damaged state. Finally, the histological examination of SVF-HA treated animals exhibited well-defined and organized fibers morphology with a lateralized nucleus, similar to contralateral healthy muscular tissue. On the contrary, HA and saline-treated animals presented inflammatory infiltrates, with HA slightly improving the diameter of the fibers and less degenerated tissue. Our findings show that connective tissue micro-fragments enriched with SVF induce higher muscle homeostasis and perfusion restoration in contrast to control groups.

Keywords: ischemia/reperfusion injury, regenerative medicine, resonance imaging, stromal vascular fraction

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Authors: Justin P. Pool, Haruyo Yoshida

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This research focuses on the effects of creating an English as a foreign language curriculum that combines online learning and classroom teaching in a complementary manner. Through pre- and post-test results, teacher observation, and learner reflection, it will be shown that learners can benefit from online programs focusing on receptive skills if combined with a communicative classroom environment that encourages learners to develop their productive skills. Much research has lamented the fact that many modern mobile assisted language learning apps do not take advantage of the affordances of modern technology by focusing only on receptive skills rather than inviting learners to interact with one another and develop communities of practice. This research takes into account the realities of the state of such apps and focuses on how to best create a curriculum that complements apps which focus on receptive skills. The research involved 15 adult learners working for a business in Japan simultaneously engaging in 1) a commercial online English language learning application that focused on reading, listening, grammar, and vocabulary and 2) a 15-week class focused on communicative language teaching, presentation skills, and mitigation of error aversion tendencies. Participants of the study experienced large gains on a standardized test, increased motivation and willingness to communicate, and asserted that they felt more confident regarding English communication. Moreover, learners continued to study independently at higher rates after the study than they had before the onset of the program. This paper will include the details of the program, reveal the improvement in test scores, share learner reflections, and critically view current evaluation models for mobile assisted language learning applications.

Keywords: adult learners, communicative language teaching, mobile assisted language learning, motivation

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Authors: Debabrata Adhikari, Mikhail Matveev, Louise Brown, Jan Kočí, Andy Long

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Resin transfer molding (RTM) is increasingly used for manufacturing high-quality composite structures due to its additional advantages over prepregs of low-cost out-of-autoclave processing. However, to retain the advantages, it is critical to reduce the void content during the injection. Reinforcements commonly used in RTM, such as woven fabrics, have dual-scale porosity with mesoscale pores between the yarns and the micro-scale pores within the yarns. Due to the fabric geometry and the nature of the dual-scale flow, the flow front during injection creates a complicated fingering formation which leads to void formation. Analytical modeling of void formation for woven fabrics has been widely studied elsewhere. However, there is scope for improvement to the reduction in void formation in 3D fabrics wherein the in-plane yarn layers are confined by additional through-thickness binder yarns. In the present study, the structural morphology of the tortuous pore spaces in the 3D fabric has been studied and implemented using open-source software TexGen. An analytical model for the void and the fingering formation has been implemented based on an idealized unit cell model of the 3D fabric. Since the pore spaces between the yarns are free domains, the region is treated as flow-through connected channels, whereas intra-yarn flow has been modeled using Darcy’s law with an additional term to account for capillary pressure. Later the void fraction has been characterised using the criterion of void formation by comparing the fill time for inter and intra yarn flow. Moreover, the dual-scale two-phase flow of resin with air has been simulated in the commercial CFD solver OpenFOAM/ANSYS to predict the probable location of voids and validate the analytical model. The use of an idealised unit cell model will give the insight to optimise the mesoscale geometry of the reinforcement and injection parameters to minimise the void content during the LCM process.

Keywords: 3D fiber, void formation, RTM, process modelling

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Authors: Bogale Tadesse, Krutik Parikh, Ndagha Mkandawire, Boris Albijanic, Nimal Subasinghe

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It is well established that the floatability and selectivity of mineral particles are highly dependent on the particle size. Generally, a particle size of 10 micron is considered as the critical size below which both flotation selectivity and recovery decline sharply. It is widely accepted that the majority of ultrafine particles, including highly liberated valuable minerals, will be lost in tailings during a conventional flotation process. This is highly undesirable particularly in the processing of finely disseminated complex and refractory ores where there is a requirement for fine grinding in order to liberate the valuable minerals. In addition, the continuing decline in ore grade worldwide necessitates intensive processing of low grade mineral deposits. Recent advances in comminution allow the economic grinding of particles down to 10 micron sizes to enhance the probability of liberating locked minerals from low grade ores. Thus, it is timely that the flotation of fine and ultrafine particles is improved in order to reduce the amount of valuable minerals lost as slimes. It is believed that the use of fine bubbles in flotation increases the bubble-particle collision efficiency and hence the flotation performance. Electroflotation, where bubbles are generated by the electrolytic breakdown of water to produce oxygen and hydrogen gases, leads to the formation of extremely finely dispersed gas bubbles with dimensions varying from 5 to 95 micron. The sizes of bubbles generated by this method are significantly smaller than those found in conventional flotation (> 600 micron). In this study, microbubbles generated by electrolysis of water were injected into a bench top flotation cell to assess the performance electroflotation in enhancing the flotation of fine and ultrafine pyrite particles of sizes ranging from 5 to 53 micron. The design of the cell and the results from optimization of the process variables such as current density, pH, percent solid and particle size will be presented at this conference.

Keywords: electroflotation, fine bubbles, pyrite, ultrafine particles

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Authors: Shane-Anthony Smith

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Teachers across grade levels and content areas face a myriad of challenges in the classroom. From inconsistent attendance to disruptive behaviors, these challenges can have a dire impact on the educational space, untimely leading to a loss of instructional time and student disenfranchisement from learning. While these challenges are not new to the educational landscape, the post-COVID classroom has, in many instances, been more severely impacted by behaviors that are not conducive to learning. Despite the mounting challenges, the role of the teacher remains unchanged - that is, to create and maintain a safe environment that is conducive to learning and promotes successful learning outcomes. Accomplishing this feat is no easy task. Yet, there are steps teachers can - indeed, must - take to better set themselves and their students up for success. The key to achieving this success is effective classroom transitions. This paper presents a four-step approach for teachers to engage in successful classroom transitions to promote meaningful student engagement and active positive learning outcomes. The transition strategy I will explore is called PREP (Pause, Reset, Establish Expectations, and Proceed). I developed this strategy in my work as a Residency Director for my university’s teacher residency program. In this role, I am tasked with coaching emerging teachers and their in-service teaching mentors in the field, as well as providing mentorship to special education resident teachers pursuing teaching degrees in the program. As a teacher educator, being in Middle and High school classrooms provides an intricate and critical understanding of the challenges, opportunities, and possibilities in the classroom. For this paper, I will explore how teachers can optimize the opportunities PREP provides to keep students engaged and, thus, improve student achievement. I will describe the approach, explain its use, and provide case-study examples of its classroom application.

Keywords: classroom management, teaching strategies, student engagement, classroom transition

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Authors: Zinah Jabbar Mohammed Alrubaye

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Objective: The purpose of this study was to compare the efficacy of prophylactic and rescue cervical cerclages for pregnant patients with an incompetent cervix, and to assess the neonatal outcomes of both clinical conditions. Methods: This was a retrospective observational study of all women who had an elective or rescue cerclage between January 2008 and December 2016 in our hospital .Prophylactic cerclage was defined as a cerclage before 16 weeks of gestation, while rescue cerclages were performed between 16 and 23 weeks of gestation. Results: In total, we analyzed the outcomes of 212 cervical interventions; 71% of the recruited patients experienced prophylactic cerclage, while 29% underwent rescue cerclage. Most of the patients delivered vaginally (70%) and were able to leave the hospital with a healthy newborn (78%). The mean pregnancy prolongation time after cerclage in the prophylactic and rescue groups were 21 weeks and 10 weeks, respectively. Conclusion: Prophylactic cerclage interventions are most likely to be associated with a reduction of fetal demise because of the correlation between fetal prognosis and the gestational age at which cerclage is performed. Once the diagnosis of cervical insufficiency is confirmed, cerclage should be recommended as this will help to prolong the pregnancy.

Keywords: cervical, neonate, cerclage, Cervix

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Authors: Alireza Barari, Ahmad Abdi

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Regular body trainings cause adaption in various system in body. One of the important effect of body training is its effect on immune system. It seems that cytokines usually release after long period exercises or some exercises which cause skeletal muscular damages. If some of the cytokines which cause responses such as inflammation of cells in skeletal muscles, with manipulating of training program, it can be avoided or limited from those exercises which induct cytokines release. Ginger plant is a kind of medicinal plants which is known as a anti inflammation plant. This plant is as most precedence medicinal plants in medicine science especially in inflammation cure. The aim of the present study was the effect of selected resistance training and consumption of ginger extract on IL-1α and TNFα untrained young women. The population includes young women interested in participating in the study with the average of 30±2 years old from Abbas Abad city among which 32 participants were chosen randomly and divided into 4 four groups, resistance training (R), resistance training and ginger consumption(RG), Ginger consumption(G)and Control group(C). The training groups performed circuit resistance training at the intensity of 65-75% one repeat maximum, 3 days a week for 6 weeks. Besides resistance training, subjects were given either ginseng (5 mg/kg per day) or placebo. Prior to and 48 hours after interventions body composition was measured and blood samples were taken in order to assess serum levels of IL-1α and TNFα. Plasma levels of cytokines were measured with commercially available ELISA Kits.IL-1α kit and TNFα kit were used in this research. To demonstrate the effectiveness of the independent variable and the comparison between groups, t-test and ANOVA were used. To determine differences between the groups, the Scheffe test was used that showed significant changes in any of the variables. we observed that circuit resistance training in R and RG groups can significant decreased in weight and body mass index in untrained females (p<0.05). The results showed a significant decreased in the mean level of IL-1α levels before and after the training period in G group (p=0.046) and RG group (p=0.022). Comparison between groups also showed there was significant difference between groups R-RG and RG-C. Intergroup comparison results showed that the mean levels of TNFα before and after the training in group G (p=0.044) and RG (p=0.037), significantly decreased. Comparison between groups also showed there was significant difference between groups R–RG , R-G ,RG-C and G-C. The research shows that circuit resistance training with reducing overload method results in systemic inflammation had significant effect on IL-1α levels and TNFα. Of course, Ginger can counteract the negative effects of resistance training exercise on immune function and stability of the mast cell membrane. Considerable evidence supported the anti-inflammatory properties of ginger for several constituents, especially gingerols, shogaols, paradols, and zingerones, through decreased cytokine gene TNF α and IL-1Α expression and inhibition of cyclooxygenase 1 and 2. These established biological actions suggest that ingested ginger could block the increase in IL-1α.

Keywords: resistance training, ginger, IL-1α , TNFα

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Authors: Dipendra Kumar Yadav, Nabaraj Paudel, Anjana Yadav

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Background: Low Birth weight (LBW) is defined as the weight at birth less than 2500 grams, irrespective of the period of their gestation. LBW is an important indicator of general health status of population and is considered as the single most important predictors of infant mortality especially of deaths within the first month of life that is birth weight determines the chances of newborn survival. Objective of this study was to identify the maternal risk factors associated with low birth weight neonates. Materials and Methods: A hospital based case-control study was conducted in maternity ward of Manipal Teaching Hospital, Pokhara, Nepal from 23 September 2014 to 12 November 2014. During study period 59 cases were obtained and twice number of control group were selected with frequency matching of the mother`s age with ± 3 years and total controls were 118. Interview schedule was used for data collection along with record review. Data were entered in Epi-data program and analysis was done with help of SPSS software program. Results: From bivariate logistic regression analysis, eighteen variables were found significantly associated with LBW and these were place of residence, family monthly income, education, previous still birth, previous LBW, history of STD, history of vaginal bleeding, anemia, ANC visits, less than four ANC visits, de-worming status, counseling during pregnancy, CVD, physical workload, stress, extra meal during pregnancy, smoking and alcohol consumption status. However after adjusting confounding variables, only six variables were found significantly associated with LBW. Mothers who had family monthly income up to ten thousand rupees were 4.83 times more likely to deliver LBW with CI (1.5-40.645) and p value 0.014 compared to mothers whose family income NRs.20,001-60,000. Mothers who had previous still birth were 2.01 times more likely to deliver LBW with CI (0.69-5.87) and p value 0.02 compared to mothers who did not has previous still birth. Mothers who had previous LBW were 5.472 times more likely to deliver LBW with CI (1.2-24.93) and p value 0.028 compared to mothers who did not has previous LBW. Mothers who had anemia during pregnancy were 3.36 times more likely to deliver LBW with CI (0.77-14.57) and p value 0.014 compared to mothers who did not has anemia. Mothers who delivered female newborn were 2.96 times more likely to have LBW with 95% CI (1.27-7.28) and p value 0.01 compared to mothers who deliver male newborn. Mothers who did not get extra meal during pregnancy were 6.04 times more likely to deliver LBW with CI (1.11-32.7) and p value 0.037 compared to mothers who getting the extra meal during pregnancy. Mothers who consumed alcohol during pregnancy were 4.83 times more likely to deliver LBW with CI (1.57-14.83) and p value 0.006 compared to mothers who did not consumed alcohol during pregnancy. Conclusions: To reduce low birth weight baby through economic empowerment of family and individual women. Prevention and control of anemia during pregnancy is one of the another strategy to control the LBW baby and mothers should take full dose of iron supplements with screening of haemoglobin level. Extra nutritional food should be provided to women during pregnancy. Health promotion program will be focused on avoidance of alcohol and strengthen of health services that leads increasing use of maternity services.

Keywords: low birth weight, case-control, risk factors, hospital based study

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Authors: Zia Ul Haq, Saleem Muhammad, Naeem Ullah, Abbas Shah, Abdullah Shah

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Pakistan being the developing country is facing double burden of communicable and non-communicable disease. The aspect of secondary prevention of ischemic heart disease in developing countries is the dire need for public health specialists, clinicians and policy makers. There is some evidence that psychotherapeutic measures, including psychotherapy, recreation, exercise and stress management training have positive impact on secondary prevention of cardiovascular diseases but there are some contradictory findings as well. Cardiac rehabilitation program (CRP) has not yet fully implemented in Pakistan. Psychological, physical and specific health-related quality of life (HRQoL) outcomes needs assessment with respect to its practicality, effectiveness, and success. Objectives: To determine the effect of cardiac rehabilitation program (CRP) on the health-related quality of life (HRQoL) measures of post MI patients compared to the usual care. Hypothesis: Post MI patients who receive the interventions (CRP) will have better HRQoL as compared to those who receive the usual cares. Methods: The randomized control trial was conducted at a Cardiac Rehabilitation Unit of Lady Reading Hospital (LRH), Peshawar. LRH is the biggest hospital of the Province Khyber Pakhtunkhwa (KP). A total 206 participants who had recent first myocardial infarction were inducted in the study. Participants were randomly allocated into two group i.e. usual care group (UCG) and cardiac rehabilitation group (CRG) by permuted-block randomization (PBR) method. CRP was conducted in CRG in two phases. Three HRQoL outcomes i.e. general health questionnaire (GHQ), self-rated health (SRH) and MacNew quality of life after myocardial infarction (MacNew QLMI) were assessed at baseline and follow-up visits among both groups. Data were entered and analyzed by appropriate statistical test in STATA version 12. Results: A total of 195 participants were assessed at the follow-up period due to lost-to-follow-up. The mean age of the participants was 53.66 + 8.3 years. Males were dominant in both groups i.e. 150 (76.92%). Regarding educational status, majority of the participants were illiterate in both groups i.e. 128 (65.64%). Surprisingly, there were 139 (71.28%) who were non-smoker on the whole. The comorbid status was positive in 120 (61.54%) among all the patients. The SRH at follow-up among UCG and CRG was 4.06 (95% CI: 3.93, 4.19) and 2.36 (95% CI: 2.2, 2.52) respectively (p<0.001). GHQ at the follow-up of UCG and CRG was 20.91 (95% CI: 18.83, 21.97) and 7.43 (95% CI: 6.59, 8.27) respectively (p<0.001). The MacNew QLMI at follow-up of UCG and CRG was 3.82 (95% CI: 3.7, 3.94) and 5.62 (95% CI: 5.5, 5.74) respectively (p<0.001). All the HRQoL measures showed strongly significant improvement in the CRG at follow-up period. Conclusion: HRQOL improved in post MI patients after comprehensive CRP. Education of the patients and their supervision is needed when they are involved in their rehabilitation activities. It is concluded that establishing CRP in cardiac units, recruiting post-discharged MI patients and offering them CRP does not impose high costs and can result in significant improvement in HRQoL measures. Trial registration no: ACTRN12617000832370

Keywords: cardiovascular diseases, cardiac rehabilitation, health-related quality of life, HRQoL, myocardial infarction, quality of life, QoL, rehabilitation, randomized control trial

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Authors: Nurul Aliaa Idris, David A. Collings

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Phi thickenings are specialized secondary cell wall thickenings that are found in the cortex of the roots in a wide range of plant species, including orchids. The role of phi thickenings in the root is still under debate through research have linked environmental conditions, particularly abiotic stresses such as water stress, heavy metal stress and salinity to their induction in the roots. It has also been suggested that phi thickenings may act as a barrier to regulate solute uptake, act as a physical barrier against fungal hyphal penetration due to its resemblance to the Casparian strip and play a mechanical role to support cortical cells. We have investigated phi thickening function in epiphytic orchids of the genus Miltoniopsis through induction experiment against factors such as soil compaction and water stress. The permeability of the phi thickenings in Miltoniopsis was tested through uptake experiments using the fluorescent tracer dyes Calcofluor white, Lucifer yellow and Propidium iodide then viewed with wide-field or confocal microscopy. To test whether phi thickening may prevent fungal colonization in the root cell, fungal re-infection experiment was conducted by inoculating isolated symbiotic fungus to sterile in vitro Miltoniopsis explants. As the movement of fluorescent tracers through the apoplast was not blocked by phi thickenings, and as phi thickenings developed in the roots of sterile cultures in the absence of fungus and did not prevent fungal colonization of cortical cells, the phi thickenings in Miltoniopsis do not function as a barrier. Phi thickenings were found to be absent in roots grown on agar and remained absent when plants were transplanted to moist soil. However, phi thickenings were induced when plants were transplanted to well-drained media, and by the application of water stress in all soils tested. It is likely that phi thickenings stabilize the root cortex during dehydration. Nevertheless, the varied induction responses present in different plant species suggest that the phi thickenings may play several adaptive roles, instead of just one, depending on species.

Keywords: abiotic stress, Miltoniopsis, orchid, phi thickening

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Authors: S. A. Hashemvarzi, A. Heidarianpour, Z. Fallahmohammadi, M. Pourghasem, M. Kaviani

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Introduction: Parkinson’s disease (PD) is a progressive neurodegenerative in the central nervous system characterized by the loss of dopaminergic neurons in the substantia nigra resulting in loss of dopamine release in the striatum. Non-drug treatment options such as Stem cell transplantation and exercise have been considered for treatment of Parkinson's disease. Purpose: The purpose of this study was to evaluate the effect of aerobic exercise after bone marrow stem cells transplantation on recovery of dopaminergic neurons and promotion of angiogenesis markers in the striatum of parkinsonian rats. Materials and Methods: 42 male Wistar rats were divided randomly into six groups: Normal (N), Sham (S), Parkinson’s (P), Stem cells transplanted Parkinson’s (SP), Exercised Parkinson’s (EP) and Stem cells transplanted + Exercised Parkinson’s (SEP). To create a model of Parkinson's, the striatum was destroyed by injection of 6-hydroxy-dopamine into the striatum through stereotaxic apparatus. Stem cells were derived from the bone marrow of femur and tibia of male rats with 6-8 weeks old. After cultivation, approximately 5×105 cells in 5 microliter of medium were injected into the striatum of rats through the channel. Aerobic exercise was included 8 weeks of running on the treadmill with a speed of 15 meters per minute. At the end, all subjects were decapitated and striatum tissues were separately isolated for measurement of vascular endothelial growth factor (VEGF), dopamine (DA) and tyrosine hydroxylase (TH) levels. Results: VEGF, DA and TH levels in the striatum of parkinsonian rats significantly increased in treatment groups (SP, EP and SEP), especially in SEP group compared to P group after treatment (P<0.05). Conclusion: The findings implicate that the BMSCs transplantation in combination with exercise would have synergistic effects leading to functional recovery, dopaminergic neurons recovery and promotion of angiogenesis marker in the striatum of parkinsonian rats.

Keywords: stem cells, treadmill training, neurotrophic factors, Parkinson

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Authors: Sharmila Rathee

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Individuals with disability/ies often face negative attitudes, discrimination, exclusion, and inequality of treatment due to stigmatization and stigmatized treatment. While a significant number of studies in field of stigma suggest that group-identification has positive consequences for stigmatized individuals, ironically very miniscule empirical work in sight has attempted to investigate in-group identification as a coping measure against stigma, humiliation and related experiences among disability group. In view of death of empirical research on in-group identification among disability group, through present work, an attempt has been made to examine the experiences of stigma, humiliation, and in-group identification among disability group. Results of the study suggest that use of in-group identification as a coping strategy is not uniform across members of disability group and degree of in-group identification differs across different sub-groups of disability groups. Further, in-group identification among members of disability group depends on variables like degree and impact of disability, factors like onset of disability, nature, and visibility of disability, educational experiences and resources available to deal with disabling conditions.

Keywords: disability, stigma, in-group identification, social identity

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Authors: Sumeet Rai, Anuj Tyagi, B. T. Naveen Kumar, Shubhkaramjeet Kaur, Niraj K. Singh

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Since their discovery, indiscriminate use of antibiotics in human, veterinary and aquaculture systems has resulted in global emergence/spread of multidrug-resistant bacterial pathogens. Thus, the need for alternative approaches to control bacterial infections has become utmost important. High selectivity/specificity of bacteriophages (phages) permits the targeting of specific bacteria without affecting the desirable flora. In this study, a lytic phage (Ahp1) specific to Aeromonas hydrophila subsp. hydrophila was isolated from finfish aquaculture pond. The host range of Ahp1 range was tested against 10 isolates of A. hydrophila, 7 isolates of A. veronii, 25 Vibrio cholerae isolates, 4 V. parahaemolyticus isolates and one isolate each of V. harveyi and Salmonella enterica collected previously. Except the host A. hydrophila subsp. hydrophila strain, no lytic activity against any other bacterial was detected. During the adsorption rate and one-step growth curve analysis, 69.7% of phage particles were able to get adsorbed on host cell followed by the release of 93 ± 6 phage progenies per host cell after a latent period of ~30 min. Phage nucleic acid was extracted by column purification methods. After determining the nature of phage nucleic acid as dsDNA, phage genome was subjected to next-generation sequencing by generating paired-end (PE, 2 x 300bp) reads on Illumina MiSeq system. De novo assembly of sequencing reads generated circular phage genome of 42,439 bp with G+C content of 58.95%. During open read frame (ORF) prediction and annotation, 22 ORFs (out of 49 total predicted ORFs) were functionally annotated and rest encoded for hypothetical proteins. Proteins involved in major functions such as phage structure formation and packaging, DNA replication and repair, DNA transcription and host cell lysis were encoded by the phage genome. The complete genome sequence of Ahp1 along with gene annotation was submitted to NCBI GenBank (accession number MF683623). Stability of Ahp1 preparations at storage temperatures of 4 °C, 30 °C, and 40 °C was studied over a period of 9 months. At 40 °C storage, phage counts declined by 4 log units within one month; with a total loss of viability after 2 months. At 30 °C temperature, phage preparation was stable for < 5 months. On the other hand, phage counts decreased by only 2 log units over a period of 9 during storage at 4 °C. As some of the phages have also been reported as glycerol sensitive, the stability of Ahp1 preparations in (0%, 15%, 30% and 45%) glycerol stocks were also studied during storage at -80 °C over a period of 9 months. The phage counts decreased only by 2 log units during storage, and no significant difference in phage counts was observed at different concentrations of glycerol. The Ahp1 phage discovered in our study had a very narrow host range and it may be useful for phage typing applications. Moreover, the endolysin and holin genes in Ahp1 genome could be ideal candidates for recombinant cloning and expression of antimicrobial proteins.

Keywords: Aeromonas hydrophila, endolysin, phage, narrow host range

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Authors: R. Hasterok, A. Betekhtin, N. Borowska, A. Braszewska-Zalewska, E. Breda, K. Chwialkowska, R. Gorkiewicz, D. Idziak, J. Kwasniewska, M. Kwasniewski, D. Siwinska, A. Wiszynska, E. Wolny

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In contrast to animals, the organisation of plant genomes at the cytomolecular level is still relatively poorly studied and understood. However, the Brachypodium genus in general and B. distachyon in particular represent exceptionally good model systems for such study. This is due not only to their highly desirable ‘model’ biological features, such as small nuclear genome, low chromosome number and complex phylogenetic relations, but also to the rapidly and continuously growing repertoire of experimental tools, such as large collections of accessions, WGS information, large insert (BAC) libraries of genomic DNA, etc. Advanced cytomolecular techniques, such as fluorescence in situ hybridisation (FISH) with evermore sophisticated probes, empowered by cutting-edge microscope and digital image acquisition and processing systems, offer unprecedented insight into chromatin organisation at various phases of the cell cycle. A good example is chromosome painting which uses pools of chromosome-specific BAC clones, and enables the tracking of individual chromosomes not only during cell division but also during interphase. This presentation outlines the present status of molecular cytogenetic analyses of plant genome structure, dynamics and evolution using B. distachyon and some of its relatives. The current projects focus on important scientific questions, such as: What mechanisms shape the karyotypes? Is the distribution of individual chromosomes within an interphase nucleus determined? Are there hot spots of structural rearrangement in Brachypodium chromosomes? Which epigenetic processes play a crucial role in B. distachyon embryo development and selective silencing of rRNA genes in Brachypodium allopolyploids? The authors acknowledge financial support from the Polish National Science Centre (grants no. 2012/04/A/NZ3/00572 and 2011/01/B/NZ3/00177)

Keywords: Brachypodium, B. distachyon, chromosome, FISH, molecular cytogenetics, nucleus, plant genome organisation

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Authors: Lynn Louis, Bor Shin Chee, Marion McAfee, Michael Nugent

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This article aims to develop a sustained release formulation of microspheres containing the mTOR inhibitor Everolimus (EVR) using Polyvinyl alcohol (PVA) to enhance the bioavailability of the drug and to overcome poor solubility characteristics of Everolimus. This paper builds on recent work in the manufacture of microspheres using the sessile droplet technique by freezing the polymer-drug solution by suspending the droplets into pre-cooled ethanol vials immersed in liquid nitrogen. The spheres were subjected to 6 freezing cycles and 3 freezing cycles with thawing to obtain proper geometry, prevent aggregation, and achieve physical cross-linking. The prepared microspheres were characterised for surface morphology by SEM, where a 3-D porous structure was observed. The in vitro release studies showed a 62.17% release over 12.5 days, indicating a sustained release due to good encapsulation. This result is comparatively much more than the 49.06% release achieved within 4 hours from the solvent cast Everolimus film as a control with no freeze-thaw cycles performed. The solvent cast films were made in this work for comparison. A prolonged release of Everolimus using a polymer-based drug delivery system is essential to reach optimal therapeutic concentrations in treating SEGA tumours without systemic exposure. These results suggest that the combination of PVA and Everolimus via a rheological synergism enhanced the bioavailability of the hydrophobic drug Everolimus. Physical-chemical characterisation using DSC and FTIR analysis showed compatibility of the drug with the polymer, and the stability of the drug was maintained owing to the high molecular weight of the PVA. The obtained results indicate that the developed PVA/EVR microsphere is highly suitable as a potential drug delivery system with improved bioavailability in treating Subependymal Giant cell astrocytoma (SEGA).

Keywords: drug delivery system, everolimus, freeze-thaw cycles, polyvinyl alcohol

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Authors: Satoshi Furukawa, Satomu Morita, Katsuji Nishi, Masahito Hitosugi

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Coronary artery dissection leading to acute myocardial infarction after blunt chest trauma is extremely rare. A 67-year-old woman suffered blunt chest trauma following a traffic accident. The electrocardiogram revealed acute posterior ST-segment elevation and myocardial infarction and coronary angiography demonstrated acute right coronary artery dissection. Following the death of the victim an autopsy was performed after cardiopulmonary support had been carried out. In this case report, we describe the case of a woman with blunt chest trauma, who developed an acute myocardial infarction secondary to right coronary artery dissection. Although there was additional the blunt chest trauma due to chest compression, we confirmed the injury at autopsy and by histological findings.

Keywords: blunt chest trauma, right coronary artery dissection, coronary angiography, autopsy, histological examination

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Authors: Sapna Bhargav

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Contemporary fiction is an intricate exercise where novelist acquires the role of a philosopher, a sociologist and a psychologist. The dilemma of fragmented self that a man experiences, is a crucial subject of contemporary fiction. Paul Auster's fiction is exemplary of the merger of Existentialism and Postmodernism, and while both of these movements insist on isolation of the self from all aspects of social affiliation, Auster's unique blend of these concepts presents man in a state which is not just alienated, but stranded in a desolate abyss, rendering even the release of death as questionable. The conundrums of the self is a compulsory consequence of the existentialist alienation that postmodern man is subjected to, and is further accentuated by the fact that existentialist freedom dictates that not only are one's actions not dictated by any form of external entity, but also the onus of one's destiny lies on an individual's own deeds. This paper will analyse Auster's The Brooklyn Follies from an Existentialist perspective, and will attempt to trace the alienation and identity conflicts of the Auster’s characters along with some of the common Austerian themes. An emphasis will be laid on the characters’ endeavour to reconstruct their lost self.

Keywords: alienation, existentialism, identity, postmodernism, self

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Authors: Tagel Gebrehiwot, Carolina Castilla

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Food insecurity and child malnutrition are among the most critical issues in Ethiopia. Accordingly, different reform programs have been carried to improve household food security. The Food Security Program (FSP) (among others) was introduced to combat the persistent food insecurity problem in the country. The FSP combines a safety net component called the Productive Safety Net Program (PSNP) started in 2005. The goal of PSNP is to offer multi-annual transfers, such as food, cash or a combination of both to chronically food insecure households to break the cycle of food aid. Food or cash transfers are the main elements of PSNP. The case for cash transfers builds on the Sen’s analysis of ‘entitlement to food’, where he argues that restoring access to food by improving demand is a more effective and sustainable response to food insecurity than food aid. Cash-based schemes offer a greater choice of use of the transfer and can allow a greater diversity of food choice. It has been proven that dietary diversity is positively associated with the key pillars of food security. Thus, dietary diversity is considered as a measure of household’s capacity to access a variety of food groups. Studies of dietary diversity among Ethiopian rural households are somewhat rare and there is still a dearth of evidence on the impact of PSNP on household dietary diversity. In this paper, we examine the impact of the Ethiopia’s PSNP on household dietary diversity and child nutrition using panel household surveys. We employed different methodologies for identification. We exploit the exogenous increase in kebeles’ PSNP budget to identify the effect of the change in the amount of money households received in transfers between 2012 and 2014 on the change in dietary diversity. We use three different approaches to identify this effect: two-stage least squares, reduced form IV, and generalized propensity score matching using a continuous treatment. The results indicate the increase in PSNP transfers between 2012 and 2014 had no effect on household dietary diversity. Estimates for different household dietary indicators reveal that the effect of the change in the cash transfer received by the household is statistically and economically insignificant. This finding is robust to different identification strategies and the inclusion of control variables that determine eligibility to become a PSNP beneficiary. To identify the effect of PSNP participation on children height-for-age and stunting we use a difference-in-difference approach. We use children between 2 and 5 in 2012 as a baseline because by then they have achieved long-term failure to grow. The treatment group comprises children ages 2 to 5 in 2014 in PSNP participant households. While changes in height-for-age take time, two years of additional transfers among children who were not born or under the age of 2-3 in 2012 have the potential to make a considerable impact on reducing the prevalence of stunting. The results indicate that participation in PSNP had no effect on child nutrition measured as height-for-age or probability of beings stunted, suggesting that PSNP should be designed in a more nutrition-sensitive way.

Keywords: continuous treatment, dietary diversity, impact, nutrition security

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Authors: Sam Bartlett, Thomas Bacquart, Arul Murugan, Abigail Morris

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Fuel cell electric vehicles provide the potential to decarbonise road transport, create new economic opportunities, diversify national energy supply, and significantly reduce the environmental impacts of road transport. A potential issue, however, is that the catalyst used at the fuel cell cathode is susceptible to degradation by impurities, especially sulphur-containing compounds. A recent European Directive (2014/94/EU) stipulates that, from November 2017, all hydrogen provided to fuel cell vehicles in Europe must comply with the hydrogen purity specifications listed in ISO 14687-2; this includes reactive and toxic chemicals such as ammonia and total sulphur-containing compounds. This requirement poses great analytical challenges due to the instability of some of these compounds in calibration gas standards at relatively low amount fractions and the difficulty associated with undertaking measurements of groups of compounds rather than individual compounds. Without the available reference materials and analytical infrastructure, hydrogen refuelling stations will not be able to demonstrate compliance to the ISO 14687 specifications. The hydrogen purity laboratory at NPL provides world leading, accredited purity measurements to allow hydrogen refuelling stations to evidence compliance to ISO 14687. Utilising state-of-the-art methods that have been developed by NPL’s hydrogen purity laboratory, including a novel method for measuring total sulphur compounds at 4 nmol/mol and a hydrogen impurity enrichment device, we provide the capabilities necessary to achieve these goals. An overview of these capabilities will be given in this paper. As part of the EMPIR Hydrogen co-normative project ‘Metrology for sustainable hydrogen energy applications’, NPL are developing a validated analytical methodology for the measurement of speciated sulphur-containing compounds in hydrogen at low amount fractions pmol/mol to nmol/mol) to allow identification and measurement of individual sulphur-containing impurities in real samples of hydrogen (opposed to a ‘total sulphur’ measurement). This is achieved by producing a suite of stable gravimetrically-prepared primary reference gas standards containing low amount fractions of sulphur-containing compounds (hydrogen sulphide, carbonyl sulphide, carbon disulphide, 2-methyl-2-propanethiol and tetrahydrothiophene have been selected for use in this study) to be used in conjunction with novel dynamic dilution facilities to enable generation of pmol/mol to nmol/mol level gas mixtures (a dynamic method is required as compounds at these levels would be unstable in gas cylinder mixtures). Method development and optimisation are performed using gas chromatographic techniques assisted by cryo-trapping technologies and coupled with sulphur chemiluminescence detection to allow improved qualitative and quantitative analyses of sulphur-containing impurities in hydrogen. The paper will review the state-of-the art gas standard preparation techniques, including the use and testing of dynamic dilution technologies for reactive chemical components in hydrogen. Method development will also be presented highlighting the advances in the measurement of speciated sulphur compounds in hydrogen at low amount fractions.

Keywords: gas chromatography, hydrogen purity, ISO 14687, sulphur chemiluminescence detector

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Authors: Amelia J. McFarland, Andrew K. Davey, Shailendra Anoopkumar-Dukie

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Microglia are the resident macrophage population of the central nervous system (CNS), contributing to both innate and adaptive immune response, and brain homeostasis. Activation of microglia occurs in response to a multitude of pathogenic stimuli in their microenvironment; this induces morphological and functional changes, resulting in a state of acute neuroinflammation which facilitates injury resolution. Adequate microglial function is essential for the health of the neuroparenchyma, with microglial dysfunction implicated in numerous CNS pathologies. Given the critical role that these macrophage-derived cells play in CNS homeostasis, there is a high demand for microglial models suitable for use in neuroscience research. The isolation of primary human microglia, however, is both difficult and costly, with microglial activation an unwanted but inevitable result of the extraction process. Consequently, there is a need for the development of alternative experimental models which exhibit morphological, biochemical and functional characteristics of human microglia without the difficulties associated with primary cell lines. In this study, our aim was to evaluate whether THP-1 human peripheral blood monocytes would display microglial-like qualities following an induced differentiation, and, therefore, be suitable for use as surrogate microglia. To achieve this aim, THP-1 human peripheral blood monocytes from acute monocytic leukaemia were differentiated with a range of phorbol 12-myristate 13-acetate (PMA) concentrations (50-200 nM) using two different protocols: a 5-day continuous PMA exposure or a 3-day continuous PMA exposure followed by a 5-day rest in normal media. In each protocol and at each PMA concentration, microglial-like cell morphology was assessed through crystal violet staining and the presence of CD-14 microglial / macrophage cell surface marker. Lipopolysaccharide (LPS) from Escherichia coli (055: B5) was then added at a range of concentrations from 0-10 mcg/mL to activate the PMA-differentiated THP-1 cells. Functional microglial-like behavior was evaluated by quantifying the release of prostaglandin (PG)-E2 and pro-inflammatory cytokines interleukin (IL)-1β and tumour necrosis factor (TNF)-α using mediator-specific ELISAs. Furthermore, production of global reactive oxygen species (ROS) and nitric oxide (NO) were determined fluorometrically using dichlorodihydrofluorescein diacetate (DCFH-DA) and diaminofluorescein diacetate (DAF-2-DA) respectively. Following PMA-treatment, it was observed both differentiation protocols resulted in cells displaying distinct microglial morphology from 10 nM PMA. Activation of differentiated cells using LPS significantly augmented IL-1β, TNF-α and PGE2 release at all LPS concentrations under both differentiation protocols. Similarly, a significant increase in DCFH-DA and DAF-2-DA fluorescence was observed, indicative of increases in ROS and NO production. For all endpoints, the 5-day continuous PMA treatment protocol yielded significantly higher mediator levels than the 3-day treatment and 5-day rest protocol. Our data, therefore, suggests that the differentiation of THP-1 human monocyte cells with PMA yields a homogenous microglial-like population which, following stimulation with LPS, undergo activation to release a range of pro-inflammatory mediators associated with microglial activation. Thus, the use of PMA-differentiated THP-1 cells represents a suitable microglial model for in vitro research.

Keywords: differentiation, lipopolysaccharide, microglia, monocyte, neuroscience, THP-1

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