Search results for: stem laboratory
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3016

Search results for: stem laboratory

2866 Study of Pseudomonas as Biofertiliser in Salt-Affected Soils of the Northwestern Algeria: Solubilisation of Calcium Phosphate and Growth Promoting of Broad Bean (Vcia faba)

Authors: A. Djoudi, R. Djibaou, H. A. Reguieg Yssaad

Abstract:

Our study focuses on the study of a bacteria belonging to Pseudomonas solubilizing tricalcium phosphate. They were isolated from rhizosphere of a variety of broad bean grown in salt-affected soils (electrical conductivity between 4 and 8 mmhos/cm) of the irrigated perimeter of Mina in northwestern Algeria. Isolates which have advantageous results in the calcium phosphate solubilization index test were subjected to identification using API20 then used to re-inoculate the same soil in pots experimentation to assess the effects of inoculation on the growth of the broad bean (Vicia faba). Based on the results obtained from the in-vitro tests, two isolates P5 and P8 showed a significant effect on the solubilization of tricalcium phosphate with an index I estimated at 314% and 283% sequentially. According to the results of in-vivo tests, the inoculation of the soil with P5 and P8 were significantly and positively influencing the growth in biometric parameters of the broad bean. Inoculation with strain P5 has promoted the growth of the broad bean in stem height, stem fresh weight and stem dry weight of 108.59%, 115.28%, 104.33%, respectively. Inoculation with strain P8 has fostered the growth of the broad bean stem fresh weight of 112.47%. The effect of Pseudomonas on the development of Vicia faba is considered as an interesting process by which PGPR can increase biological production and crop protection.

Keywords: Pseudomonas, Vicia faba, promoting of plant growth, solubilization tricalcium phosphate

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2865 A Strategy Therapy for Retinitis Pigmentosa Induced by Argon Laser in Rabbits by High Dose Adult Stem Cells

Authors: Hager E. Amer, Hany El Saftawy, Laila Rashed, Ahmed M. Ata, Fatma Metwally, Hesham Mettawei, Hossam E. Sayed, Tamer Adel, Kareem M. El Sawah

Abstract:

Aim: The purpose of this study is to regenerate the damaged photoreceptor cells as a result of argon laser as a model of Retinitis Pigmentosa in rabbits' retina by using adult stem cells from rabbits' bone marrow. Background: Retinitis pigmentosa (RP) is a group of inherited disorders that primarily affect photoreceptor and pigment epithelium function. RP leads to loss of the rod outer segment and shorten the photoreceptor layer and expose the photoreceptor cell body to high-pressure levels in oxygen (oxidative stress) leads to apoptosis to the rod and cone cells. In particular, there is no specific treatment for retinitis pigmentosa. Materials and Methods: Forty Two Giant (Rex) rabbits were used in this experiment divided into 3 groups: Group 1: Control (6 rabbits), Group 2: Argon laser radiated as a model of retinitis pigmentosa (12 rabbits), Group 3: Laser radiated and treated by 6 million stem cells (12 rabbits). The last two groups are divided each into two subgroups each subgroup contains 6 rabbits, the ophthalmological examination was performed on rabbits, blood samples and retina samples were taken after 25 days and after 36 days from the laser radiation (10 days and 21 days after stem cells insertion in group 3) to perform the biochemical analysis. Results: Compared to control Group, a decrease of ERG wave amplitude and antioxidant substances (Glutathione) in blood and retina in group 2, and an increase of oxidative stress substances (Nitric oxide, Malonaldehyde, and carponyl protein) and apoptotic substances (Advanced glycation end product and M-metalloproteinase) in blood and retina. Compared to group 2, mostly increases of antioxidant substances and ERG wave amplitude in group 3, and mostly decreases in oxidative stress substances and apoptotic substances. Conclusion: Insertion of 6 million stem cells intravitreous gives good results in regeneration of the damaged photoreceptor cells after 21 days.

Keywords: retinitis pigmentosa, stem cells, argon laser, oxidative stress, apoptosis

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2864 Co-Limitation of Iron Deficiency in Stem Allantoin and Amino-N Formation of Peanut Plants Intercropped with Cassava

Authors: Hong Li, Tingxian Li, Xudong Wang, Weibo Yang

Abstract:

Co-limitation of iron (Fe) deficiency in legume nitrogen fixation process is not well understood. Our objectives were to examine how peanut plants cope with Fe deficiency with the rhizobial inoculants and N-nutrient treatments. The study was conducted in the tropical Hainan Island during 2012-2013. The soil was strongly acidic (pH 4.6±0.7) and deficient in Fe (9.2±2.3 mg/kg). Peanut plants were intercropped with cassava. The inoculants and N treatments were arranged in a split-plot design with three blocks. Peanut root nodulation, stem allantoin, amino acids and plant N derived from fixation (P) reduced with declining soil Fe concentrations. The treatment interactions were significant on relative ureide % and peanut yields (P<0.05). Residual fixed N from peanut plants was beneficial to cassava plants. It was concluded that co-variance of Fe deficiency could influence peanut N fixation efficiency and rhizobia and N inputs could help improving peanut tolerance to Fe deficiency stress.

Keywords: amino acids, plant N derived from N fixation, root nodulation, soil Fe co-variance, stem ureide, peanuts, cassava

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2863 A Laboratory–Designed Activity in Ecology to Demonstrate the Allelopathic Property of the Philippine Chromolaena odorata L. (King and Robinson) Leaf Extracts

Authors: Lina T. Codilla

Abstract:

This study primarily designed a laboratory activity in ecology to demonstrate the allelopathic property of the Philippine Chromolaena odorata L. (hagonoy) leaf extracts to Lycopersicum esculentum (M), commonly known as tomatoes. Ethanol extracts of C. odorata leaves were tested on seed germination and seedling growth of L. esculentum in 7-day and 14-day observation periods. Analysis of variance and Tukey’s HSD post hoc test was utilized to determine differences among treatments while Pre–test – Post–test experimental design was utilized in the determination of the effectiveness of the designed laboratory activity. Results showed that the 0.5% concentration level of ethanol leaf extracts significantly inhibited germination and seedling growth of L. esculentum in both observation periods. These results were used as the basis in the development of instructional material in ecology. The laboratory activity underwent face validation by five (5) experts in various fields of specialization, namely, Biological Sciences, Chemistry and Science Education. The readability of the designed laboratory activity was determined using a Cloze Test. Pilot testing was conducted and showed that the laboratory activity developed is found to be a very effective tool in supplementing learning about allelopathy in ecology class. Thus, it is recommended for use among ecology classes but modification will be made in a small – scale basis to minimize time consumption.

Keywords: allelopathy, chromolaena odorata l. (hagonoy), designed-laboratory activity, organic herbicide students’ performance

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2862 The Importance of Development in Laboratory Diagnosis at the Intersection

Authors: Agus Sahri, Cahya Putra Dinata, Faishal Andhi Rokhman

Abstract:

Intersection is a critical area on a highway which is a place of conflict points and congestion due to the meeting of two or more roads. Conflicts that occur at the intersection include diverging, merging, weaving, and crossing. To deal with these conflicts, a crossing control system is needed, at a plot of intersection there are two control systems namely signal intersections and non-signalized intersections. The control system at a plot of intersection can affect the intersection performance. In Indonesia there are still many intersections with poor intersection performance. In analyzing the parameters to measure the performance of a plot of intersection in Indonesia, it is guided by the 1997 Indonesian Road Capacity Manual. For this reason, this study aims to develop laboratory diagnostics at plot intersections to analyze parameters that can affect the performance of an intersection. The research method used is research and development. The laboratory diagnosis includes anamnesis, differential diagnosis, inspection, diagnosis, prognosis, specimens, analysis and sample data analysts. It is expected that this research can encourage the development and application of laboratory diagnostics at a plot of intersection in Indonesia so that intersections can function optimally.

Keywords: intersection, the laboratory diagnostic, control systems, Indonesia

Procedia PDF Downloads 154
2861 Rejuvenation of Premature Ovarian Failure with Stem Cells/IVA Technique

Authors: Elham Vojoudi, Marzieh Mehrafza, Ahmad Hosseini, Azadeh Raofi, Maryam Najafi

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Premature ovarian failure (POF) has become one of the main causes of infertility in women of childbearing age and the incidence of this disorder is increasing year by year. In these patients, poor ovarian response (POR) to gonadotropins reflects a diminished ovarian reserve (DOR) that gives place to few follicles despite aggressive stimulation. Up to now, egg donation is the only way to resolve infertility problems in POF patients. Therefore, some novel aspects such as activating (Akt signaling pathway) and inhibiting (Hippo-signaling) elements have been identified as IVA procedure that promotes primordial follicle activation. In this study, we used the newly developed technique (combination of in vitro activation of dormant follicles (IVA) and stem cell therapy) to promote ovarian follicle growth much more efficiently than the natural, in vivo process for women with POF. Transplantation of Warton Jelly-MSCs to the ovaries of POF patients rescued overall ovarian function. Participants (10 patients) were followed up monthly for a period of six months by hormonal (AMH, FSH, LH and E2), clinical (resuming menstruation), and US (folliculometry) outcomes after a laparoscopic operation. In summary, IVA/WJ-MSC transplantation may provide an effective treatment for POF.

Keywords: POF, in vitro activation, stem cell therapy, infertility

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2860 Two Taxa of Paradiacheopsis Genera Recordings of the Myxomycetes from Turkey

Authors: Dursun Yağız, Ahmet Afyon

Abstract:

The study materials were collected from Isparta province in 2008. These materials were moved to the laboratory. The 'Most Chamber Techniques' were applied to the materials in the laboratory. Materials were examined with a stereo microscope. As a result of investigations carried out on the samples of sporophores which were developed in the laboratory, Paradiacheopsis erythropodia (Ing) Nann.-Bremek. and Paradiacheopsis longipes Hooff & Nann.-Bremek. species were identified. As a result of the literature research, it is determined that these taxa were new recordings in Turkey. The identified taxa have been added to Turkey's myxomycota. These two taxa’ microscopic features, photos, localities and substrate information were given.

Keywords: myxomycete, paradiacheopsis, Turkey, slime mould

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2859 A Machine Learning Model for Dynamic Prediction of Chronic Kidney Disease Risk Using Laboratory Data, Non-Laboratory Data, and Metabolic Indices

Authors: Amadou Wurry Jallow, Adama N. S. Bah, Karamo Bah, Shih-Ye Wang, Kuo-Chung Chu, Chien-Yeh Hsu

Abstract:

Chronic kidney disease (CKD) is a major public health challenge with high prevalence, rising incidence, and serious adverse consequences. Developing effective risk prediction models is a cost-effective approach to predicting and preventing complications of chronic kidney disease (CKD). This study aimed to develop an accurate machine learning model that can dynamically identify individuals at risk of CKD using various kinds of diagnostic data, with or without laboratory data, at different follow-up points. Creatinine is a key component used to predict CKD. These models will enable affordable and effective screening for CKD even with incomplete patient data, such as the absence of creatinine testing. This retrospective cohort study included data on 19,429 adults provided by a private research institute and screening laboratory in Taiwan, gathered between 2001 and 2015. Univariate Cox proportional hazard regression analyses were performed to determine the variables with high prognostic values for predicting CKD. We then identified interacting variables and grouped them according to diagnostic data categories. Our models used three types of data gathered at three points in time: non-laboratory, laboratory, and metabolic indices data. Next, we used subgroups of variables within each category to train two machine learning models (Random Forest and XGBoost). Our machine learning models can dynamically discriminate individuals at risk for developing CKD. All the models performed well using all three kinds of data, with or without laboratory data. Using only non-laboratory-based data (such as age, sex, body mass index (BMI), and waist circumference), both models predict chronic kidney disease as accurately as models using laboratory and metabolic indices data. Our machine learning models have demonstrated the use of different categories of diagnostic data for CKD prediction, with or without laboratory data. The machine learning models are simple to use and flexible because they work even with incomplete data and can be applied in any clinical setting, including settings where laboratory data is difficult to obtain.

Keywords: chronic kidney disease, glomerular filtration rate, creatinine, novel metabolic indices, machine learning, risk prediction

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2858 Pre-Analytical Laboratory Performance Evaluation Utilizing Quality Indicators between Private and Government-Owned Hospitals Affiliated to University of Santo Tomas

Authors: A. J. Francisco, K. C. Gallosa, R. J. Gasacao, J. R. Ros, B. J. Viado

Abstract:

The study focuses on the use of quality indicators (QI)s based on the standards made by the (IFCC), that could effectively identify and minimize errors occurring throughout the total testing process (TTP), in order to improve patient safety. The study was conducted through a survey questionnaire that was given to a random sample of 19 respondents (eight privately-owned and eleven government-owned hospitals), mainly CMTs, MTs, and Supervisors from UST-affiliated hospitals. The pre-analytical laboratory errors, which include misidentification errors, transcription errors, sample collection errors and sample handling and transportation errors, were considered as variables according to the IFCC WG-LEPS. Data gathered were analyzed using the Mann-Whitney U test, Percentile, Linear Regression, Percentage, and Frequency. The laboratory performance of both hospitals is High level. There is no significant difference between the laboratory performance between the two stated variables. Moreover, among the four QIs, sample handling and transportation errors contributed most to the difference between the two variables. Outcomes indicate satisfactory performance between both variables. However, in order to ensure high-quality and efficient laboratory operation, constant vigilance and improvements in pre-analytical QI are still needed. Expanding the coverage of the study, the inclusion of other phases, utilization of parametric tests are recommended.

Keywords: pre-analytical phase, quality indicators, laboratory performance, pre-analytical error

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2857 A Low Cost and Reconfigurable Experimental Platform for Engineering Lab Education

Authors: S. S. Kenny Lee, C. C. Kong, S. K. Ting

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Teaching engineering lab provides opportunity for students to practice theories learned through physical experiment in the laboratory. However, building laboratories to accommodate increased number of students are expensive, making it impossible for an educational institution to afford the high expenses. In this paper, we develop a low cost and remote platform to aid teaching undergraduate students. The platform is constructed where the real experiment setting up in laboratory can be reconfigure and accessed remotely, the aim is to increase student’s desire to learn at which they can interact with the physical experiment using network enabled devices at anywhere in the campus. The platform is constructed with Raspberry Pi as a main control board that provides communication between computer interfaces to the actual experiment preset in the laboratory. The interface allows real-time remote viewing and triggering the physical experiment in the laboratory and also provides instructions and learning guide about the experimental.

Keywords: engineering lab, low cost, network, remote platform, reconfigure, real-time

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2856 A Development of Science Instructional Model Based on Stem Education Approach to Enhance Scientific Mind and Problem Solving Skills for Primary Students

Authors: Prasita Sooksamran, Wareerat Kaewurai

Abstract:

STEM is an integrated teaching approach promoted by the Ministry of Education in Thailand. STEM Education is an integrated approach to teaching Science, Technology, Engineering, and Mathematics. It has been questioned by Thai teachers on the grounds of how to integrate STEM into the classroom. Therefore, the main objective of this study is to develop a science instructional model based on the STEM approach to enhance scientific mind and problem-solving skills for primary students. This study is participatory action research, and follows the following steps: 1) develop a model 2) seek the advice of experts regarding the teaching model. Developing the instructional model began with the collection and synthesis of information from relevant documents, related research and other sources in order to create prototype instructional model. 2) The examination of the validity and relevance of instructional model by a panel of nine experts. The findings were as follows: 1. The developed instructional model comprised of principles, objective, content, operational procedures and learning evaluation. There were 4 principles: 1) Learning based on the natural curiosity of primary school level children leading to knowledge inquiry, understanding and knowledge construction, 2) Learning based on the interrelation between people and environment, 3) Learning that is based on concrete learning experiences, exploration and the seeking of knowledge, 4) Learning based on the self-construction of knowledge, creativity, innovation and 5) relating their findings to real life and the solving of real-life problems. The objective of this construction model is to enhance scientific mind and problem-solving skills. Children will be evaluated according to their achievements. Lesson content is based on science as a core subject which is integrated with technology and mathematics at grade 6 level according to The Basic Education Core Curriculum 2008 guidelines. The operational procedures consisted of 6 steps: 1) Curiosity 2) Collection of data 3) Collaborative planning 4) Creativity and Innovation 5) Criticism and 6) Communication and Service. The learning evaluation is an authentic assessment based on continuous evaluation of all the material taught. 2. The experts agreed that the Science Instructional Model based on the STEM Education Approach had an excellent level of validity and relevance (4.67 S.D. 0.50).

Keywords: instructional model, STEM education, scientific mind, problem solving

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2855 Experimental Determination of Water Productivity of Improved Cassava Varieties Propagation under Rain-Fed Condition in Tropical Environment

Authors: Temitayo Abayomi Ewemoje, Isaac Olugbemiga Afolayan, Badmus Alao Tayo

Abstract:

Researchers in developing countries have worked on improving cassava resistance to diseases and pests, high yielding and early maturity However, water management has received little or no attention as cassava cultivation in Sub-Saharan Africa depended on available precipitation (rain-fed condition). Therefore the need for water management in Agricultural crop production cannot be overemphasized. As other sectors compete with agricultural sector for fresh water (which is not readily available), there is need to increase water productivity in agricultural production. Experimentation was conducted to examine water use, growth and yield of improved cassava varieties under rain fed condition using Latin- square design with four replications. Four improved disease free stem cassava varieties TMS (30572, 980505, 920326 and 090581) were planted and growth parameters of the varieties were monitored for 90 and 120 days after planting (DAP). Effective rainfall useful for the plant growth was calculated using CROPWAT8 for Windows. Results indicated TMS090581 was having the highest tuber yield and plant height while TMS30572 had highest number of nodes. Tuber stem and leaf water productivities at 90 and 120 DAP of TMS (30572, 980505, 920326 and 090581) are (1.27 and 3.58, 1.44 and 2.35, 0.89 and 1.86, 1.64 and 3.77) kg/m3 (1.56 and 2.59, 1.95 and 2.02, 1.98 and 2.05, 1.95 and 2.18) kg/m3, and (1.34 and 2.32, 1.94 and 2.16, 1.57 and 1.40, 1.27 and 1.80) kg/m3 respectively. Based on tuber water productivity TMS090581 are recommended while TMS30572 are recommended based on leaf and stem productivity in water scarce regions.Experimentation was conducted to examine water use, growth and yield of improved cassava varieties under rain fed condition using Latin- square design with four replications. Four improved disease free stem cassava varieties TMS (30572, 980505, 920326 and 090581) were planted and growth parameters of the varieties were monitored for 90 and 120 days after planting (DAP). Effective rainfall useful for the plant growth was calculated using CROPWAT8 for Windows. Results indicated TMS090581 was having the highest tuber yield and plant height while TMS30572 had the highest number of nodes. Tuber, stem and leaf water productivities at 90 and 120 DAP of TMS (30572, 980505, 920326 and 090581) are (1.27 and 3.58, 1.44 and 2.35, 0.89 and 1.86, 1.64 and 3.77) kg/m3 (1.56 and 2.59, 1.95 and 2.02, 1.98 and 2.05, 1.95 and 2.18) kg/m3, and (1.34 and 2.32, 1.94 and 2.16, 1.57 and 1.40, 1.27 and 1.80) kg/m3 respectively. Based on tuber water productivity TMS090581 are recommended while TMS30572 are recommended based on leaf and stem productivity in water scarce regions

Keywords: improved TMS varieties, leaf productivity, rain-fed cassava production, stem productivity, tuber productivity

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2854 The Effects of Science, Technology, Engineering and Math Problem-Based Learning on Native Hawaiians and Other Underrepresented, Low-Income, Potential First-Generation High School Students

Authors: Nahid Nariman

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The prosperity of any nation depends on its ability to use human potential, in particular, to offer an education that builds learners' competencies to become effective workforce participants and true citizens of the world. Ever since the Second World War, the United States has been a dominant player in the world politically, economically, socially, and culturally. The rapid rise of technological advancement and consumer technologies have made it clear that science, technology, engineering, and math (STEM) play a crucial role in today’s world economy. Exploring the top qualities demanded from new hires in the industry—i.e., problem-solving skills, teamwork, dependability, adaptability, technical and communication skills— sheds light on the kind of path that is needed for a successful educational system to effectively support STEM. The focus of 21st century education has been to build student competencies by preparing them to acquire and apply knowledge, to think critically and creatively, to competently use information, be able to work in teams, to demonstrate intellectual and moral values as well as cultural awareness, and to be able to communicate. Many educational reforms pinpoint various 'ideal' pathways toward STEM that educators, policy makers, and business leaders have identified for educating the workforce of tomorrow. This study will explore how problem-based learning (PBL), an instructional strategy developed in the medical field and adopted with many successful results in K-12 through higher education, is the proper approach to stimulate underrepresented high school students' interest in pursuing STEM careers. In the current study, the effect of a problem-based STEM model on students' attitudes and career interests was investigated using qualitative and quantitative methods. The participants were 71 low-income, native Hawaiian high school students who would be first-generation college students. They were attending a summer STEM camp developed as the result of a collaboration between the University of Hawaii and the Upward Bound Program. The project, funded by the National Science Foundation's Innovative Technology Experiences for Students and Teachers (ITEST) program, used PBL as an approach in challenging students to engage in solving hands-on, real-world problems in their communities. Pre-surveys were used before camp and post-surveys on the last day of the program to learn about the implementation of the PBL STEM model. A Career Interest Questionnaire provided a way to investigate students’ career interests. After the summer camp, a representative selection of students participated in focus group interviews to discuss their opinions about the PBL STEM camp. The findings revealed a significantly positive increase in students' attitudes towards STEM disciplines and STEM careers. The students' interview results also revealed that students identified PBL to be an effective form of instruction in their learning and in the development of their 21st-century skills. PBL was acknowledged for making the class more enjoyable and for raising students' interest in STEM careers, while also helping them develop teamwork and communication skills in addition to scientific knowledge. As a result, the integration of PBL and a STEM learning experience was shown to positively affect students’ interest in STEM careers.

Keywords: problem-based learning, science education, STEM, underrepresented students

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2853 Antioxidant and Acute Toxicity of Stem Extracts of the Ficus Iteophylla

Authors: Muhammad Mukhtar

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The aim of this study is to evaluate the antioxidant activity and acute toxicity of the extracts of Ficus iteophylla by reactions with 1, 1-diphenyl-2-picryhydrazyl radical (DPPH) and method developed by Lork 1983, respectively. Stem bark of Ficus iteophylla was collected, air dried, pulverized to fine powdered and sequentially extracted using acetone, methanol and water in order of increasing polarity. The result shows strong radical scavenging activity against DPPH for all the extracts when compared with ascorbic acid. The LD50 of 316 mg/kg was calculated for all the three extras, and the values were found to be within the practically toxic range, and therefore, care should be taken when using the plants in traditional medicine.

Keywords: antioxidant, acute toxicity, Ficus iteophylla

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2852 Laboratory Findings as Predictors of St2 and NT-Probnp Elevations in Heart Failure Clinic, National Cardiovascular Centre Harapan Kita, Indonesia

Authors: B. B. Siswanto, A. Halimi, K. M. H. J. Tandayu, C. Abdillah, F. Nanda , E. Chandra

Abstract:

Nowadays, modern cardiac biomarkers, such as ST2 and NT-proBNP, have important roles in predicting morbidity and mortality in heart failure patients. Abnormalities of serum electrolytes, sepsis or infection, and deteriorating renal function will worsen the conditions of patients with heart failure. It is intriguing to know whether cardiac biomarkers elevations are affected by laboratory findings in heart failure patients. We recruited 65 patients from the heart failure clinic in NCVC Harapan Kita in 2014-2015. All of them have consented for laboratory examination, including cardiac biomarkers. The findings were recorded in our Research and Development Centre and analyzed using linear regression to find whether there is a relationship between laboratory findings (sodium, potassium, creatinine, and leukocytes) and ST2 or NT-proBNP. From 65 patients, 26.9% of them are female, and 73.1% are male, 69.4% patients classified as NYHA I-II and 31.6% as NYHA III-IV. The mean age is 55.7+11.4 years old; mean sodium level is 136.1+6.5 mmol/l; mean potassium level is 4.7+1.9 mmol/l; mean leukocyte count is 9184.7+3622.4 /ul; mean creatinine level is 1.2+0.5 mg/dl. From linear regression logistics, the relationship between NT-proBNP and sodium level (p<0.001), as well as leukocyte count (p=0.002) are significant, while NT-proBNP and potassium level (p=0.05), as well as creatinine level (p=0.534) are not significant. The relationship between ST2 and sodium level (p=0.501), potassium level (p=0.76), leukocyte level (p=0.897), and creatinine level (p=0.817) are not significant. To conclude, laboratory findings are more sensitive in predicting NT-proBNP elevation than ST2 elevation. Larger studies are needed to prove that NT-proBNP correlation with laboratory findings is more superior than ST2.

Keywords: heart failure, laboratory, NT-proBNP, ST2

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2851 Antimicrobial and Phytochemical Screening of Stem Bark Extracts of Lovoa trichiliodes (Harm) and Trichilia heudelotii Planc (Harm)

Authors: Benjamin O. Opawale, Anthony K. Onifade, Ayodele O. Ogundare

Abstract:

The phytochemical and antimicrobial activities of stem bark extracts (cold water, ethanol and acetone) of Lovoa trichiliodes and Trichilia heudelotii were investigated using standard methods. The percentage yield of the extracts ranged from 3.90 to 6.53% and 9.63 to 10.20% respectively for the plant materials. Phytochemical screening of the plant materials revealed the presence of alkaloids, saponins, tannins, phlobatanins, phenols, anthraquinones and glycosides. Terpenes, cardenolides and flavonoids were absent in the two plants. All the extracts remarkably inhibited the growth of Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, Klebsiella pneumoniae, Salmonella typhii, Aspergillus flavus, Candida albicans and Candida glabrata. The mean diameter of the zone of inhibition exhibited by the extracts was between 8.00 and 22.33mm while the minimum inhibitory concentration (MIC) was between 2.5 and 200mg/ml. However, the cold water extracts of L. trichiliodes stem bark exhibited no inhibitory activity against the organisms. The results of this investigation confirmed the folkloric uses of these plants for the treatment of various infectious diseases.

Keywords: antimicrobial, infectious diseases, phytochemical, T. heudelotii

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2850 Contour Defects of Face with Hyperpigmentation

Authors: Afzaal Bashir, Sunaina Afzaal

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Background: Facial contour deformities associated with pigmentary changes are of major concern for plastic surgeons, both being important and difficult in treating such issues. No definite ideal treatment option is available to simultaneously address both the contour defect as well as related pigmentation. Objectives: The aim of the current study is to compare the long-term effects of conventional adipose tissue grafting and ex-vivo expanded Mesenchymal Stem Cells enriched adipose tissue grafting for the treatment of contour deformities with pigmentary changes on the face. Material and Methods: In this study, eighty (80) patients with contour deformities of the face with hyperpigmentation were recruited after informed consent. Two techniques i.e., conventional fat grafting (C-FG) and fat grafts enriched with expanded adipose stem cells (FG-ASCs), were used to address the pigmentation. Both techniques were explained to patients, and enrolled patients were divided into two groups i.e., C-FG and FG-ASCS, per patients’ choice and satisfaction. Patients of the FG-ASCs group were treated with fat grafts enriched with expanded adipose stem cells, while patients of the C-FGs group were treated with conventional fat grafting. Patients were followed for 12 months, and improvement in face pigmentation was assessed clinically as well as measured objectively. Patient satisfaction was also documented as highly satisfied, satisfied, and unsatisfied. Results: Mean age of patients was 24.42(±4.49), and 66 patients were females. The forehead was involved in 61.20% of cases, the cheek in 21.20% of cases, the chin in 11.20% of cases, and the nose in 6.20% of cases. In the GF-ASCs group, the integrated color density (ICD) was decreased (1.08×10⁶ ±4.64×10⁵) as compared to the C-FG group (2.80×10⁵±1.69×10⁵). Patients treated with fat grafts enriched with expanded adipose stem cells were significantly more satisfied as compared to patients treated with conventional fat grafting only. Conclusion: Mesenchymal stem cell-enriched autologous fat grafting is a preferred option for improving the contour deformities related to increased pigmentation of face skin.

Keywords: hyperpigmentation, color density, enriched adipose tissue graft, fat grafting, contour deformities, Image J

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2849 Promoting Stem Education and a Cosmic Perspective by Using 21st Century Science of Learning

Authors: Rohan Roberts

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The purpose of this project was to collaborate with a group of high-functioning, more-able students (aged 15-18) to promote STEM Education and a love for science by bringing a cosmic perspective into the classroom and high school environment. This was done using 21st century science of learning, a focus on the latest research on Neuroeducation, and modern pedagogical methods based on Howard Gardner's theory of Multiple Intelligences, Bill Lucas’ theory of New Smarts, and Sir Ken Robinson’s recommendations on encouraging creativity. The result was an increased sense of passion, excitement, and wonder about science in general, and about the marvels of space and the universe in particular. In addition to numerous unique and innovative science-based initiatives, clubs, workshops, and science trips, this project also saw a marked rise in student-teacher collaboration in science learning and in student engagement with the general public through the press, social media, and community-based initiatives. This paper also outlines the practical impact that bringing a cosmic perspective into the classroom has had on the lives, interests, and future career prospects of the students involved in this endeavour.

Keywords: cosmic perspective, gifted and talented, neuro-education, STEM education

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2848 Disparities Versus Similarities; WHO Good Practices for Pharmaceutical Quality Control Laboratories and ISO/IEC 17025:2017: International Standards for Quality Management Systems in Pharmaceutical Laboratories

Authors: Mercy Okezue, Kari Clase, Stephen Byrn, Paddy Shivanand

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Medicines regulatory authorities expect pharmaceutical companies and contract research organizations to seek ways to certify that their laboratory control measurements are reliable. Establishing and maintaining laboratory quality standards are essential in ensuring the accuracy of test results. ‘ISO/IEC 17025:2017’ and ‘WHO Good Practices for Pharmaceutical Quality Control Laboratories (GPPQCL)’ are two quality standards commonly employed in developing laboratory quality systems. A review was conducted on the two standards to elaborate on areas on convergence and divergence. The goal was to understand how differences in each standard's requirements may influence laboratories' choices as to which document is easier to adopt for quality systems. A qualitative review method compared similar items in the two standards while mapping out areas where there were specific differences in the requirements of the two documents. The review also provided a detailed description of the clauses and parts covering management and technical requirements in these laboratory standards. The review showed that both documents share requirements for over ten critical areas covering objectives, infrastructure, management systems, and laboratory processes. There were, however, differences in standard expectations where GPPQCL emphasizes system procedures for planning and future budgets that will ensure continuity. Conversely, ISO 17025 was more focused on the risk management approach to establish laboratory quality systems. Elements in the two documents form common standard requirements to assure the validity of laboratory test results that promote mutual recognition. The ISO standard currently has more global patronage than GPPQCL.

Keywords: ISO/IEC 17025:2017, laboratory standards, quality control, WHO GPPQCL

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2847 Environmental Restoration Science in New York Harbor - Community Based Restoration Science Hubs, or “STEM Hubs”

Authors: Lauren B. Birney

Abstract:

The project utilizes the Billion Oyster Project (BOP-CCERS) place-based “restoration through education” model to promote computational thinking in NYC high school teachers and their students. Key learning standards such as Next Generation Science Standards and the NYC CS4All Equity and Excellence initiative are used to develop a computer science curriculum that connects students to their Harbor through hands-on activities based on BOP field science and educational programming. Project curriculum development is grounded in BOP-CCERS restoration science activities and data collection, which are enacted by students and educators at two Restoration Science STEM Hubs or conveyed through virtual materials. New York City Public School teachers with relevant experience are recruited as consultants to provide curriculum assessment and design feedback. The completed curriculum units are then conveyed to NYC high school teachers through professional learning events held at the Pace University campus and led by BOP educators. In addition, Pace University educators execute the Summer STEM Institute, an intensive two-week computational thinking camp centered on applying data analysis tools and methods to BOP-CCERS data. Both qualitative and quantitative analyses were performed throughout the five-year study. STEM+C – Community Based Restoration STEM Hubs. STEM Hubs are active scientific restoration sites capable of hosting school and community groups of all grade levels and professional scientists and researchers conducting long-term restoration ecology research. The STEM Hubs program has grown to include 14 STEM Hubs across all five boroughs of New York City and focuses on bringing in-field monitoring experience as well as coastal classroom experience to students. Restoration Science STEM Hubs activities resulted in: the recruitment of 11 public schools, 6 community groups, 12 teachers, and over 120 students receiving exposure to BOP activities. Field science protocols were designed exclusively around the use of the Oyster Restoration Station (ORS), a small-scale in situ experimental platforms which are suspended from a dock or pier. The ORS is intended to be used and “owned” by an individual school, teacher, class, or group of students, whereas the STEM Hub is explicitly designed as a collaborative space for large-scale community-driven restoration work and in-situ experiments. The ORS is also an essential tool in gathering Harbor data from disparate locations and instilling ownership of the research process amongst students. As such, it will continue to be used in that way. New and previously participating students will continue to deploy and monitor their own ORS, uploading data to the digital platform and conducting analysis of their own harbor-wide datasets. Programming the STEM Hub will necessitate establishing working relationships between schools and local research institutions. NYHF will provide introductions and the facilitation of initial workshops in school classrooms. However, once a particular STEM Hub has been established as a space for collaboration, each partner group, school, university, or CBO will schedule its own events at the site using the digital platform’s scheduling and registration tool. Monitoring of research collaborations will be accomplished through the platform’s research publication tool and has thus far provided valuable information on the projects’ trajectory, strategic plan, and pathway.

Keywords: environmental science, citizen science, STEM, technology

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2846 Brain Stem Posterior Reversible Encephalopathy Syndrome in Nephrotic Syndrome

Authors: S. H. Jang

Abstract:

Posterior reversible encephalopathy syndrome (PRES) is characterized by acute neurologic symptoms (visual loss, headache, altered mentality and seizures) and by typical imaging findings (bilateral subcortical and cortical edema with predominatly posterior distribution). Nephrotic syndrome is a syndrome comprising signs of proteinuria, hypoalbuminemia, and edema. It is well known that hypertension predispose patient with nephrotic syndrome to PRES. A 45-year old male was referred for suddenly developed vertigo, disequilibrium. He had previous history of nephrotic syndrome. His medical history included diabetes controlled with medication. He was hospitalized because of generalized edema a few days ago. His vital signs were stable. On neurologic examination, his mental state was alert. Horizontal nystagmus to right side on return to primary position was observed. He showed good grade motor weakness and ataxia in right upper and lower limbs without other sensory abnormality. Brain MRI showed increased signal intensity in FLAIR image, decreased signal intensity in T1 image and focal enhanced lesion in T1 contrast image at whole midbrain, pons and cerebellar peduncle symmetrically, which was compatible with vasogenic edema. Laboratory findings showed severe proteinuria and hypoalbuminemia. He was given intravenous dexamethasone and diuretics to reduce vasogenic edema and raise the intra-vascular osmotic pressure. Nystagmus, motor weakness and limb ataxia improved gradually over 2 weeks; He recovered without any neurologic symptom and sign. Follow-up MRI showed decreased vasogenic edema fairly. We report a case of brain stem PRES in normotensive, nephrotic syndrome patient.

Keywords: posterior reversible encephalopathy syndrome, MRI, nephrotic syndrome, vasogenic brain edema

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2845 In silico Repopulation Model of Various Tumour Cells during Treatment Breaks in Head and Neck Cancer Radiotherapy

Authors: Loredana G. Marcu, David Marcu, Sanda M. Filip

Abstract:

Advanced head and neck cancers are aggressive tumours, which require aggressive treatment. Treatment efficiency is often hindered by cancer cell repopulation during radiotherapy, which is due to various mechanisms triggered by the loss of tumour cells and involves both stem and differentiated cells. The aim of the current paper is to present in silico simulations of radiotherapy schedules on a virtual head and neck tumour grown with biologically realistic kinetic parameters. Using the linear quadratic formalism of cell survival after radiotherapy, altered fractionation schedules employing various treatment breaks for normal tissue recovery are simulated and repopulation mechanism implemented in order to evaluate the impact of various cancer cell contribution on tumour behaviour during irradiation. The model has shown that the timing of treatment breaks is an important factor influencing tumour control in rapidly proliferating tissues such as squamous cell carcinomas of the head and neck. Furthermore, not only stem cells but also differentiated cells, via the mechanism of abortive division, can contribute to malignant cell repopulation during treatment.

Keywords: radiation, tumour repopulation, squamous cell carcinoma, stem cell

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2844 Evaluation of Cytotoxic Effect of Mitoxantrone Conjugated Magnetite Nanoparticles and Graphene Oxide-Magnetite Nanocomposites on Mesenchymal Stem Cells

Authors: Abbas Jafarizad, Duygu Ekinci

Abstract:

In this work targeted drug delivery is proposed to decrease adverse effect of drugs with concomitant reduces in consumption and treatment outgoings. Nanoparticles (NPs) can be prepared from a variety of materials such as lipid, biodegradable polymer that prevent the drugs cytotoxicity in healthy cells, etc. One of the most important drugs used in chemotherapy is mitoxantrone (MTX) which prevents cell proliferation by inhibition of topoisomerase II and DNA repair; however, it is not selective and has some serious side effects. In this study, mentioned aim is achieved by using several nanocarriers like magnetite (Fe3O4) and their composites with magnetic graphene oxide (Fe3O4@GO). Also, cytotoxic potential of Fe3O4, Fe3O4-MTX, and Fe3O4@GO-MTX nanocomposite were evaluated on mesenchymal stem cells (MSCs). In this study, we reported the synthesis of monodisperse Fe3O4 NPs and Fe3O4@GO nanocomposite and their structures were investigated by using field emission scanning electron microscope (FESEM), Fourier transform infrared (FTIR) spectra, atomic force microscopy (AFM), Brauneur Emmet Teller (BET) isotherm and contact angle studies. Moreover, we used 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay to evaluate cytotoxic effects of MTX, Fe3O4 NPs, Fe3O4-MTX and Fe3O4@GO-MTX nanocomposite on MSCs. The in-vitro MTT results indicated that all concentrations of MTX and Fe3O4@GO-MTX nanocomposites showed cytotoxic effects while all concentrations of Fe3O4 NPs and Fe3O4-MTX NPs did not show any cytotoxic effect on stem cells. The results from this study indicated that using Fe3O4 NPs as anticancer drug delivery systems could be a trustworthy method for cancer treatment. But for reaching excellent and accurate results, further investigation is necessary.

Keywords: mitoxantrone, magnetite, magnetic graphene oxide, MTT assay, mesenchymal stem cells

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2843 Entrepreneurship in Pakistan: Opportunities and Challenges

Authors: Bushra Jamil, Nudrat Baqri, Muhammad Hassan Saeed

Abstract:

Entrepreneurship is creating or setting up a business not only for the purpose of generating profit but also for providing job opportunities. Entrepreneurs are problem solvers and product developers. They use their financial asset for hiring a professional team and combine the innovation, knowledge, and leadership leads to a successful startup or a business. To be a successful entrepreneur, one should be people-oriented and have perseverance. One must have the ability to take risk, believe in his/her potential, and have the courage to move forward in all circumstances. Most importantly, have the ability to take risk and can assess the risk. For STEM students, entrepreneurship is of specific importance and relevance as it helps them not just to be able to solve real life existing complications but to be able to recognize and identify emerging needs and glitches. It is becoming increasingly apparent that in today’s world, there is a need as well as a desire for STEM and entrepreneurship to work together. In Pakistan, entrepreneurship is slowly emerging, yet we are far behind. It is high time that we should introduce modern teaching methods and inculcate entrepreneurial initiative in students. A course on entrepreneurship can be included in the syllabus, and we must invite businessmen and policy makers to motivate young minds for entrepreneurship. This must be pitching competitions, opportunities to win seed funding, and facilities of incubation centers. In Pakistan, there are many good public sector research institutes, yet there is a void gap in the private sector. Only few research institute are meant for research and development. BJ Micro Lab is one of them. It is SECP registered company and is working in academia to promote and facilitate research in STEM. BJ Micro Lab is a women led initiative, and we are trying to promote research as a passion, not as an arduous burden. For this, we are continuously arranging training workshops and sessions. More than 100 students have been trained in ten different workshops arranged at BJ Micro Lab.

Keywords: entrepreneurship, STEM, challenges, oppurtunties

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2842 Nanowire Substrate to Control Differentiation of Mesenchymal Stem Cells

Authors: Ainur Sharip, Jose E. Perez, Nouf Alsharif, Aldo I. M. Bandeas, Enzo D. Fabrizio, Timothy Ravasi, Jasmeen S. Merzaban, Jürgen Kosel

Abstract:

Bone marrow-derived human mesenchymal stem cells (MSCs) are attractive candidates for tissue engineering and regenerative medicine, due to their ability to differentiate into osteoblasts, chondrocytes or adipocytes. Differentiation is influenced by biochemical and biophysical stimuli provided by the microenvironment of the cell. Thus, altering the mechanical characteristics of a cell culture scaffold can directly influence a cell’s microenvironment and lead to stem cell differentiation. Mesenchymal stem cells were cultured on densely packed, vertically aligned magnetic iron nanowires (NWs) and the effect of NWs on the cell cytoskeleton rearrangement and differentiation were studied. An electrochemical deposition method was employed to fabricate NWs into nanoporous alumina templates, followed by a partial release to reveal the NW array. This created a cell growth substrate with free-standing NWs. The Fe NWs possessed a length of 2-3 µm, with each NW having a diameter of 33 nm on average. Mechanical stimuli generated by the physical movement of these iron NWs, in response to a magnetic field, can stimulate osteogenic differentiation. Induction of osteogenesis was estimated using an osteogenic marker, osteopontin, and a reduction of stem cell markers, CD73 and CD105. MSCs were grown on the NWs, and fluorescent microscopy was employed to monitor the expression of markers. A magnetic field with an intensity of 250 mT and a frequency of 0.1 Hz was applied for 12 hours/day over a period of one week and two weeks. The magnetically activated substrate enhanced the osteogenic differentiation of the MSCs compared to the culture conditions without magnetic field. Quantification of the osteopontin signal revealed approximately a seven-fold increase in the expression of this protein after two weeks of culture. Immunostaining staining against CD73 and CD105 revealed the expression of antibodies at the earlier time point (two days) and a considerable reduction after one-week exposure to a magnetic field. Overall, these results demonstrate the application of a magnetic NW substrate in stimulating the osteogenic differentiation of MSCs. This method significantly decreases the time needed to induce osteogenic differentiation compared to commercial biochemical methods, such as osteogenic differentiation kits, that usually require more than two weeks. Contact-free stimulation of MSC differentiation using a magnetic field has potential uses in tissue engineering, regenerative medicine, and bone formation therapies.

Keywords: cell substrate, magnetic nanowire, mesenchymal stem cell, stem cell differentiation

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2841 Evaluation of Gene Expression after in Vitro Differentiation of Human Bone Marrow-Derived Stem Cells to Insulin-Producing Cells

Authors: Mahmoud M. Zakaria, Omnia F. Elmoursi, Mahmoud M. Gabr, Camelia A. AbdelMalak, Mohamed A. Ghoneim

Abstract:

Many protocols were publicized for differentiation of human mesenchymal stem cells (MSCS) into insulin-producing cells (IPCs) in order to excrete insulin hormone ingoing to treat diabetes disease. Our aim is to evaluate relative gene expression for each independent protocol. Human bone marrow cells were derived from three volunteers that suffer diabetes disease. After expansion of mesenchymal stem cells, differentiation of these cells was done by three different protocols (the one-step protocol was used conophylline protein, the two steps protocol was depending on trichostatin-A, and the three-step protocol was started by beta-mercaptoethanol). Evaluation of gene expression was carried out by real-time PCR: Pancreatic endocrine genes, transcription factors, glucose transporter, precursor markers, pancreatic enzymes, proteolytic cleavage, extracellular matrix and cell surface protein. Quantitation of insulin secretion was detected by immunofluorescence technique in 24-well plate. Most of the genes studied were up-regulated in the in vitro differentiated cells, and also insulin production was observed in the three independent protocols. There were some slight increases in expression of endocrine mRNA of two-step protocol and its insulin production. So, the two-step protocol was showed a more efficient in expressing of pancreatic endocrine genes and its insulin production than the other two protocols.

Keywords: mesenchymal stem cells, insulin producing cells, conophylline protein, trichostatin-A, beta-mercaptoethanol, gene expression, immunofluorescence technique

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2840 Effects of Reclamation on Seasonal Dynamic of Carbon, Nitrogen and Phosphorus Stoichiometry in Suaeda salsa

Authors: Yajun Qiao, Yaner Yan, Ning Li, Shuqing An

Abstract:

In order to relieve the pressure on a land resource from a huge population, reclamation has occurred in many coastal wetlands. Plants can maintain their elemental composition within normal limits despite the variations of external conditions. Reclamation may affect carbon (C), nitrogen (N) and phosphorus (P) stoichiometry in the plant to some extent by altering physical and chemical properties of soil in a coastal wetland. We reported the seasonal dynamic of C, N and P stoichiometry in root, stem and leaf of Suaeda salsa (L.) Pall. and in soil between reclamation plots and natural plots. Our results of three-way ANOVA indicated that sampling season always had significant effect on C, N, P concentrations and their ratios; organ had no significant effect on N, P concentration and N:P; plot type had no significant effect on N concentration and C:N. Sampling season explained the most variability of tissue N and P contents, C:N, C:P and N:P, while it’s organ for C using the restricted maximum likelihood (REML) method. By independent sample T-test, we found that reclamation affect more on C, N and P stoichiometry of stem than that of root or leaf on the whole. While there was no difference between reclamation plots and natural plots for soil in four seasons. For three organs, C concentration had peak values in autumn and minimum values in spring while N concentration had peak values in spring and minimum values in autumn. For P concentration, three organs all had peak values in spring; however, the root had minimum value in winter, the stem had that in autumn, and leaf had that in summer. The seasonal dynamic of C, N and P stoichiometry in a leaf of Suaeda salsa were much steadier than that in root or stem under the drive of reclamation.

Keywords: nitrogen, phosphorus, reclamation, seasonal dynamic, Suaeda salsa

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2839 Effects of Planned Pre-laboratory Discussion on Physics Students’ Acquisition of Science Process Skills in Kontagora, Niger State

Authors: Akano Benedict Ubawuike

Abstract:

This study investigated the effects of pre-laboratory discussion on physics students’ acquisition of science process skills. The study design was quasi-experimental and purposive sampling technique was applied in selecting two schools in Kontagora Town for the research based on the availability of a good physics laboratory. Intact classes already grouped by the school for the sake of small laboratory space and equipment, comprising Thirty (30) students, 15 for experimental group in School A and 15 for control in school B were the subjects for the research. The instrument used for data collection was the lesson prepared for pre – practical discussion and researcher made Science Process Skill Test (SPST ) and two (2) research questions, and two (2) research hypotheses were developed to guide the study. The data collected were analyzed using means and t-Test statistics at 0.05 level of significance. The study revealed that pre-laboratory discussion was found to be more efficacious in enhancing students’ acquisition of science process skills. It also revealed that gender, had no significant effect on students’ acquisition of science process skills. Based on the findings, it was recommended among others that teachers should encourage students to develop interest in practical activities by engaging them in pre-laboratory discussion and providing instructional materials that will challenge them to be actively involved during practical lessons. It is also recommended that Ministries of Education and professional organizations like Science Teachers' Association of Nigeria (STAN) should organize workshops, seminars and conferences for physics teachers and Physics concepts should be taught with practical activity so that the students will do science instead of learning about science.

Keywords: physics, laboratory, discussion, students, acquisition, science process skills

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2838 Dynamic Changes in NT-proBNP Levels in Unrelated Donors during Hematopoietic Stem Cells Mobilization

Authors: Natalia V. Minaeva, Natalia A. Zorina, Marina N. Khorobrikh, Philipp S. Sherstnev, Tatiana V. Krivokorytova, Alexander S. Luchinin, Maksim S. Minaev, Igor V. Paramonov

Abstract:

Background. Over the last few decades, the Center for International Blood and Marrow Transplant Research (CIBMTR) and the World Marrow Donor Association (WMDA) have been actively working to ensure the safety of the hematopoietic stem cell (HSC) donation process. Registration of adverse events that may occur during the donation period and establishing a relationship between donation and side effects are included in the WMDA international standards. The level of blood serum N-terminal pro-brain natriuretic peptide (NT-proBNP) is an early marker of myocardial stress. Due to the high analytical sensitivity and specificity, laboratory assessment of NT-proBNP makes it possible to objectively diagnose myocardial dysfunction. It is well known that the main stimulus for proBNP synthesis and secretion from atrial and ventricular cardiac myocytes is myocyte stretch and increasement of myocardial extensibility and pressure in the heart chambers. Аim. The aim of the study was to assess the dynamic changes in the levels of blood serum N-terminal pro-brain natriuretic peptide of unrelated donors at various stages of hematopoietic stem cell mobilization. Materials. We have examined 133 unrelated donors, including 92 men and 41 women, that have been included into the study. The NT-proBNP levels were measured before the start of mobilization, then on the day of apheresis, and after the donation of allogeneic HSC. The relationship between NT-proBNP levels and body mass index (BMI), ferritin, hemoglobin, and white blood cells (WBC) levels was assessed on the day of apheresis. The median age of donors was 34 years. Mobilization of HSCs was managed with filgrastim administration at a dose of 10 μg/kg daily for 4-5 days. The first leukocytapheresis was performed on day 4 from the start of filgrastim administration. Quantitative values of the blood serum NT-proBNP level are presented as a median (Me), first and third quartiles (Q1-Q3). Comparative analysis was carried out using the t-test and correlation analysis as well by Spearman method. Results. The baseline blood serum NT-proBNP levels in all 133 donors were within the reference values (<125 pg/ml) and equaled 21,6 (10,0; 43,3) pg/ml. At the same time, the level of NT-proBNP in women was significantly higher than that of men. On the day of the HSC apheresis, a significant increase of blood serum NT-proBNP levels was detected and equald 131,2 (72,6; 165,3) pg/ml (p<0,001), with higher rates in female donors. A statistically significant weak inverse correleation was established between the level of NT-proBNP and the BMI of donors (-0.18, p = 0,03), as well as the level of hemoglobin (-0.33, p <0,001), and ferritin levels (-0.19, p = 0,03). No relationship has been established between the magnitude of WBC levels achieved as a result of the mobilization of HSC on the day of leukocytapheresis. A day after the apheresis, the blood serum NT-proBNP levels still exceeded the reference values, but there was a decreasing tendency. Conclusion. An increase of the blood serum NT-proBNP level in unrelated donors during the mobilization of HSC was established. Future studies should clarify the reason for this phenomenon, as well as its effects on donors' long-term health.

Keywords: unrelated donors, mobilization, hematopoietic stem cells, N-terminal pro-brain natriuretic peptide

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2837 A Serum- And Feeder-Free Culture System for the Robust Generation of Human Stem Cell-Derived CD19+ B Cells and Antibody-Secreting Cells

Authors: Kirsten Wilson, Patrick M. Brauer, Sandra Babic, Diana Golubeva, Jessica Van Eyk, Tinya Wang, Avanti Karkhanis, Tim A. Le Fevre, Andy I. Kokaji, Allen C. Eaves, Sharon A. Louis, , Nooshin Tabatabaei-Zavareh

Abstract:

Long-lived plasma cells are rare, non-proliferative B cells generated from antibody-secreting cells (ASCs) following an immune response to protect the host against pathogen re-exposure. Despite their therapeutic potential, the lack of in vitro protocols in the field makes it challenging to use B cells as a cellular therapeutic tool. As a result, there is a need to establish robust and reproducible methods for the generation of B cells. To address this, we have developed a culture system for generating B cells from hematopoietic stem and/or progenitor cells (HSPCs) derived from human umbilical cord blood (CB) or pluripotent stem cells (PSCs). HSPCs isolated from CB were cultured using the StemSpan™ B Cell Generation Kit and produced CD19+ B cells at a frequency of 23.2 ± 1.5% and 59.6 ± 2.3%, with a yield of 91 ± 11 and 196 ± 37 CD19+ cells per input CD34+ cell on culture days 28 and 35, respectively (n = 50 - 59). CD19+IgM+ cells were detected at a frequency of 31.2 ± 2.6% and were produced at a yield of 113 ± 26 cells per input CD34+ cell on culture day 35 (n = 50 - 59). The B cell receptor loci of CB-derived B cells were sequenced to confirm V(D)J gene rearrangement. ELISpot analysis revealed that ASCs were generated at a frequency of 570 ± 57 per 10,000 day 35 cells, with an average IgM+ ASC yield of 16 ± 2 cells per input CD34+ cell (n = 33 - 42). PSC-derived HSPCs were generated using the STEMdiff™ Hematopoietic - EB reagents and differentiated to CD10+CD19+ B cells with a frequency of 4 ± 0.8% after 28 days of culture (n = 37, 1 embryonic and 3 induced pluripotent stem cell lines tested). Subsequent culture of PSC-derived HSPCs increased CD19+ frequency and generated ASCs from 1 - 2 iPSC lines. This method is the first report of a serum- and feeder-free system for the generation of B cells from CB and PSCs, enabling further B lineage-specific research for potential future clinical applications.

Keywords: stem cells, B cells, immunology, hematopoiesis, PSC, differentiation

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