Search results for: reverse logistics

Authors: Seyede Sanaz Seyedebrahimi, Shima Rahimi, Shohreh Fakhari, Ali Jalili

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Background: CXCR4, as a chemokine receptor, plays well-known roles in various types of cancers. Several studies have been conducted to overcome CXCR4 axis acts in multiple myeloma (MM) pathogenesis and progression. Dexamethasone, a standard treatment for multiple myeloma, has been shown to increase CXCR4 levels in multiple myeloma cell lines. Herein, we focused on the effects of metformin and dexamethasone on CXCR4 at the cellular level and the migration rate of cell lines after exposure to a combination compared to single-agent models. Materials and Method: Multiple myeloma cell lines (U266 and RPMI8226) were cultured with different metformin and dexamethasone concentrations in single-agent and combination models. The simultaneous combination doses were calculated by CompuSyn software. Cell surface and mRNA expression of CXCR4 were determined using flow cytometry and the quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay, respectively. The Transwell cell migration assay evaluated the migration ability. Results: In concurred with previous studies, our results showed a dexamethasone up-regulation effect on CXCR4 in a dose-dependent manner. Although, the metformin single-agent model could reduce CXCR4 expression of U266 and RPMI8226 in cell surface and mRNA expression level. Moreover, the administration of metformin and dexamethasone simultaneously exerted a higher suppression effect on CXCR4 expression than the metformin single-agent model. The migration rate through the combination model's matrigel membrane was remarkably lower than the metformin and dexamethasone single-agent model. Discussion: According to our findings, the combination of metformin and dexamethasone effectively inhibited dexamethasone-induced CXCR4 expression in multiple myeloma cell lines. As a result, metformin may be counted as an alternative medicine combined with other chemotherapies to combat multiple myeloma. However, more research is required.

Keywords: CXCR4, dexamethasone, metformin, migration, multiple myeloma

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Authors: Wen-Chyan Tsai, Syed S. H. Rizvi

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Organic solvent residues are always associated with liposomes produced by the traditional techniques like the thin film hydration and reverse phase evaporation methods, which limit the applications of these vesicles in the pharmaceutical, food and cosmetic industries. Our objective was to develop a novel and benign process of liposomal microencapsulation by using supercritical carbon dioxide (SC-CO2) as the sole phospholipid-dissolving medium and a green substitute for organic solvents. This process consists of supercritical fluid extraction followed by rapid expansion via a nozzle and automatic cargo suction. Lecithin and cholesterol mixed in 10:1 mass ratio were dissolved in SC-CO2 at 20 ± 0.5 MPa and 60 oC. After at least two hours of equilibrium, the lecithin/cholesterol-laden SC-CO2 was passed through a 1000-micron nozzle and immediately mixed with the cargo solution to form liposomes. Liposomal micro-encapsulation was conducted at three pressures (8.27, 12.41, 16.55 MPa), three temperatures (75, 83 and 90 oC) and two flow rates (0.25 ml/sec and 0.5 ml/sec). Liposome size, zeta potential and encapsulation efficiency were characterized as functions of the operating parameters. The average liposomal size varied from 400-500 nm to 1000-1200 nm when the pressure was increased from 8.27 to 16.55 MPa. At 12.41 MPa, 90 oC and 0.25 ml per second of 0.2 M glucose cargo loading rate, the highest encapsulation efficiency of 31.65 % was achieved. Under a confocal laser scanning microscope, large unilamellar vesicles and multivesicular vesicles were observed to make up a majority of the liposomal emulsion. This new approach is a rapid and continuous process for bulk production of liposomes using a green solvent. Based on the results to date, it is feasible to apply this technique to encapsulate hydrophilic compounds inside the aqueous core as well as lipophilic compounds in the phospholipid bilayers of the liposomes for controlled release, solubility improvement and targeted therapy of bioactive compounds.

Keywords: liposome, micro encapsulation, supercritical carbon dioxide, non-toxic process

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Authors: Xin Li, Yan-Rong Guo, Jin-Fang Lin, Yi Feng, Håkan Billig, Ruijin Shao

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Background: Young women with polycystic ovary syndrome (PCOS) have a high risk of developing endometrial carcinoma. There is a need for the development of new medical therapies that can reduce the need for surgical intervention so as to preserve the fertility of these patients. The aim of the study was to describe and discuss cases of PCOS and insulin resistance (IR) women with early endometrial carcinoma while being co-treated with Diane-35 and metformin. Methods: Five PCOS-IR women who were scheduled for diagnosis and therapy for early endometrial carcinoma were recruited. The hospital records and endometrial pathology reports were reviewed. All patients were co-treated with Diane-35 and metformin for 6 months to reverse the endometrial carcinoma and preserve their fertility. Before, during, and after treatment, endometrial biopsies and blood samples were obtained and oral glucose tolerance tests were performed. Endometrial pathology was evaluated. Body weight (BW), body mass index (BMI), follicle-stimulating hormone (FSH), luteinizing hormone (LH), total testosterone (TT), sex hormone-binding globulin (SHBG), free androgen index (FAI), insulin area under curve (IAUC), and homeostasis model assessment of insulin resistance (HOMA-IR) were determined. Results: Clinical stage 1a, low grade endometrial carcinoma was confirmed before treatment. After 6 months of co-treatment, all patients showed normal epithelia. No evidence of atypical hyperplasia or endometrial carcinoma was found. Co-treatment resulted in significant decreases in BW, BMI, TT, FAI, IAUC, and HOMA-IR in parallel with a significant increase in SHBG. There were no differences in the FSH and LH levels after co-treatment. Conclusions: Combined treatment with Diane-35 and metformin has the potential to revert the endometrial carcinoma into normal endometrial cells in PCOS-IR women. The cellular and molecular mechanisms behind this effect merit further investigation.

Keywords: PCOS, progesterone resistance, insulin resistance, steroid hormone receptors, endometrial carcinoma

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Authors: Paola Utreras, Yazmina Olmos, Loreto Sanhueza

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This work has the purpose of study and incorporate Problem Based Learning (PBL) for engineering students, through the analysis of several thermal images of dwellings located in different geographical points of the Region de los Ríos, Chile. The students analyze how heat is transferred in and out of the houses and how is the relation between heat transfer and climatic conditions that affect each zone. As a result of this activity students are able to acquire significant learning in the unit of heat and temperature, and manage to reverse previous conceptual errors related with energy, temperature and heat. In addition, student are able to generate prototype solutions to increase thermal efficiency using low cost materials. Students make public their results in a report using scientific writing standards and in a science fair open to the entire university community. The methodology used to measure previous Conceptual Errors has been applying diagnostic tests with everyday questions that involve concepts of heat, temperature, work and energy, before the unit. After the unit the same evaluation is done in order that themselves are able to evidence the evolution in the construction of knowledge. As a result, we found that in the initial test, 90% of the students showed deficiencies in the concepts previously mentioned, and in the subsequent test 47% showed deficiencies, these percent ages differ between students who carry out the course for the first time and those who have performed this course previously in a traditional way. The methodology used to measure Significant Learning has been by comparing results in subsequent courses of thermodynamics among students who have received problem based learning and those who have received traditional training. We have observe that learning becomes meaningful when applied to the daily lives of students promoting internalization of knowledge and understanding through critical thinking.

Keywords: engineering students, heat flow, problem-based learning, thermal images

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Authors: Cuiyang Feng, Xu Tang, Yi Jin

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Coal is the most important primary energy source in China, which exerts a significant influence on the rapid economic growth. However, it makes the water resources to be a constraint on coal industry development, on account of the reverse geographical distribution between coal and water. To ease the pressure on water shortage, the ‘3 Red Lines’ water policies were announced by the Chinese government, and then ‘water for coal’ plan was added to that policies in 2013. This study utilized a structural path analysis (SPA) based on the multi-regional input-output table to quantify the virtual water flows caused by coal consumption in different stages. Results showed that the direct water input (the first stage) was the highest amount in all stages of coal consumption, accounting for approximately 30% of total virtual water content. Regional analysis demonstrated that virtual water trade alleviated the pressure on water use for coal consumption in water shortage areas, but the import of virtual water was not from the areas which are rich in water. Sectoral analysis indicated that the direct inputs from the sectors of ‘production and distribution of electric power and heat power’ and ‘Smelting and pressing of metals’ took up the major virtual water flows, while the sectors of ‘chemical industry’ and ‘manufacture of non-metallic mineral products’ importantly but indirectly consumed the water. With the population and economic growth in China, the water demand-and-supply gap in coal consumption would be more remarkable. In additional to water efficiency improvement measures, the central government should adjust the strategies of the virtual water trade to address local water scarcity issues. Water resource as the main constraints should be highly considered in coal policy to promote the sustainable development of the coal industry.

Keywords: coal consumption, multi-regional input-output model, structural path analysis, virtual water

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Authors: Catarina Isabel Ramos Pereira, Jorge Machado, Begona Alonso Criado, Maria João Santos

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Introduction: Gait disorders are one of the symptoms that have severe implications on the quality of life in Parkinson's disease (PD). Currently, there is no therapy to reverse or treat this condition. None of the drugs used in conventional medical treatment is entirely efficient, and all have a high incidence of side effects. Acupuncture treatment is believed to improve motor ability, but there is still little scientific evidence in individuals with PD. Aim: The aim of the study is to investigate the acute effect of acupuncture on gait parameters in Parkinson's disease. Methods: This is a randomized and controlled crossover study. The same individual patient was part of both the experimental (real acupuncture) and control group (false acupuncture/sham), and the sequence was randomized. Gait parameters were measured at two different moments, before and after treatment, using four force platforms as well as the collection of 3D markers positions taken by 11 cameras. Images were quantitatively analyzed using Qualisys Track Manager software that let us extract data related to the quality of gait and balance. Seven patients with the diagnosis of Parkinson's disease were included in the study. Results: Statistically significant differences were found in gait speed (p = 0.016), gait cadence (p = 0.006), support base width (p = 0.0001), medio-lateral oscillation (p = 0.017), left-right step length (p = 0.0002), and stride length: right-right (p = 0.0000) and left-left (p = 0.0018), time of left support phase (p = 0.029), right support phase (p = 0.025) and double support phase (p = 0.015), between the initial and final moments for the experimental group. Differences in right-left stride length were found for both groups. Conclusion: Our results show that acupuncture could enhance gait in Parkinson's disease patients. Deep research involving a larger number of volunteers should be accomplished to validate these encouraging findings.

Keywords: acupuncture, traditional Chinese medicine, Parkinson's disease, gait

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Authors: J. Chakwizira, P. Bikam, T. A. Adeboyejo

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This paper seeks to illustrate the impact of rapid urbanization (in terms of both increase in people and vehicles) in the Gauteng region (which includes Johannesburg, Pretoria and Ekurhuleni). The impact that existing transport systems and options place on the capacity of residents from low income areas to travel and conduct various socio-economic activities is discussed. The findings are drawn from a 2013 analysis of a random transport household survey of 1550 households carried out in Gauteng province. 91.4% of the study respondents had access to public transport, while 8.6% had no access to public transport. Of the 91.4% who used public transport, the main reason used to explain this state of affairs was that it was affordable (54.3%), convenient (15.9%), Accessible (11.9%), lack of alternatives (6.4%) and reliable at 4.1%. Recommendations advanced revolve around the need to reverse land use and transportation effects of apartheid planning, growing and developing a sustainable critical mass of public transport interventions supported by appropriate transport systems that are environmentally sustainable through proper governance. 38.5% of the respondents indicated that developing compact, smart and integrated urban land spaces was key to reducing travel challenges in the study area. 23.4% indicated that the introduction and upgrading of BRT buses to cover all areas in the study area was a step in the right direction because it has great potential in shifting travel patterns to favor public modes of transport. 15.1% indicated that all open spaces should be developed so that fragmentation of land uses can be addressed. This would help to fight disconnected and fragmented space and trip making challenges in Gauteng. 13.4% indicated that improving the metro rail services was critical since this is a mass mover of commuters. 9.6% of the respondents highlighted that the bus subsidy policy has to be retained in the short to medium term since the spatial mismatches and challenges created by apartheid are yet to be fully reversed.

Keywords: urbanisation, population, public, transport systems, Gauteng

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Authors: Michael K. O. Ayomoh, Oyindamola A. Omotuyi

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Autonomous mobile robots deployed to explore or operate in a never explored or an abandoned sub-surface mine requires extreme effectiveness in coordination and communication. In a bid to transmit information from the depth of the mine to the external surface in real-time and amidst diverse physical, chemical and virtual impediments, the concept of unified cooperative robots is seen to be a proficient approach. This paper presents an effective [human → robot → task] coordination framework for effective exploration of an abandoned underground mine. The problem addressed in this research is basically the development of a globalized optimization model premised on time series differentiation and geometrical configurations for effective positioning of the two classes of robots in the cooperation namely the outermost stationary master (OSM) robots and the innermost dynamic task (IDT) robots for effective bi-directional signal transmission. In addition, the synchronization of a vision system and wireless communication system for both categories of robots, fiber optics system for the OSM robots in cases of highly sloppy or vertical mine channels and an autonomous battery recharging capability for the IDT robots further enhanced the proposed concept. The OSM robots are the master robots which are positioned at strategic locations starting from the mine open surface down to its base using a fiber-optic cable or a wireless communication medium all subject to the identified mine geometrical configuration. The OSM robots are usually stationary and function by coordinating the transmission of signals from the IDT robots at the base of the mine to the surface and in a reverse order based on human decisions at the surface control station. The proposed scheme also presents an optimized number of robots required to form the cooperation in a bid to reduce overall operational cost and system complexity.

Keywords: sub-surface mine, wireless communication, outermost stationary master robots, inner-most dynamic robots, fiber optic

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Authors: Lubomir Vecera, Tomas Gabrhelik, Benjamin Tolmaci, Josef Srovnal, Emil Berta, Petr Prasil, Petr Stourac

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Cancer is the second leading cause of death worldwide and colon cancer is the second most common type of cancer. Currently, there are only a few studies evaluating the effect of postoperative analgesia on the prognosis of patients undergoing radical colon cancer surgery. Postoperative analgesia in patients undergoing colon cancer surgery is usually managed in two ways, either with strong opioids (morphine, piritramide) or epidural analgesia. In our prospective study, we evaluated the effect of postoperative analgesia on the presence of circulating tumor cells or minimal residual disease after colon cancer surgery. A total of 60 patients who underwent radical colon cancer surgery were enrolled in this prospective, randomized, two-center study. Patients were randomized into three groups, namely piritramide, morphine and postoperative epidural analgesia. We evaluated the presence of carcinoembryonic antigen (CEA) and cytokeratin 20 (CK-20) mRNA positive circulating tumor cells in peripheral blood before surgery, immediately after surgery, on postoperative day two and one month after surgery. The presence of circulating tumor cells was assessed by quantitative real-time reverse transcriptase-polymerase chain reaction (qRT-PCR). In the priritramide postoperative analgesia group, the presence of CEA mRNA positive cells was significantly lower on a postoperative day two compared to the other groups (p=0.04). The value of CK-20 mRNA positive cells was the same in all groups on all days. In all groups, both types of circulating tumor cells returned to normal levels one month after surgery. Demographic and baseline clinical characteristics were similar in all groups. Compared with morphine and epidural analgesia, piritramide significantly reduces the amount of CEA mRNA positive circulating tumor cells after radical colon cancer surgery.

Keywords: cancer progression, colon cancer, minimal residual disease, perioperative analgesia.

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Authors: Hongping Lu, Kelbinur Tursun, Yaru Li, Yu Zhang, Shunai Liu, Ming Han

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Objective: To investigate whether NS5ATP9 is involved in IL-6 mediated autophagy and the relationship between IL-6 and NS5ATP9 in liver cancer cells. Methods: 1. Detect the mRNA and protein levels of Beclin 1 after HepG2 cells were treated with or without recombinant human IL-6 protein. 2. Measure and compare of the changes of autophagy-related genes with their respective control, after IL-6 was silenced or neutralized with monoclonal antibody against human IL-6. 3. HepG2 cells were incubated with 50 ng/ml of IL-6 in the presence or absence of PDTC. The expression of NS5ATP9 was analyzed by Western blot after 48 h. 4. After NS5ATP9-silenced HepG2 cells had been treated with 50 ng/ml recombinant IL-6 protein, we detected the Beclin 1 and LC3B (LC3Ⅱ/Ⅰ) expression. 5. HepG2 cells were transfected with pNS5ATP9, si-NS5ATP9, and their respective control. Total RNA was isolated from cells and analyzed for IL-6. 6. Silence or neutralization of IL-6 in HepG2 cells which has been transfected with NS5ATP9. Beclin 1 and LC3 protein levels were analyzed by Western blot. Result: 1. After HepG2 were treated with recombinant human IL-6 protein, the expression of endogenous Beclin 1 was up-regulated at mRNA and protein level, and the conversion of endogenous LC3-I to LC3-II was also increased. These results indicated that IL-6 could induce autophagy. 2. When HepG2 cells were treated with IL-6 siRNA or monoclonal antibody against human IL-6, the expression of autophagy-related genes were decreased. 3. Exogenous human IL-6 recombinant protein up-regulated NS5ATP9 via NF-κB activation. 4. The expression of Beclin 1 and LC3B was down-regulated after IL-6 treated NS5ATP9-silenced HepG2 cells. 5. NS5ATP9 could reverse regulates IL-6 expression in HepG2 cells. 6. Silence or neutralization of IL-6 attenuates NS5ATP9-induced autophagy slightly. Conclusion: Our results implied that in HCC patients, maybe the higher level of IL-6 in the serum promoted the expression of NS5ATP9 and induced autophagy in cancer cells. And the over-expression of NS5ATP9 which induced by IL-6, in turn, increased IL-6 expression, further, promotes the IL-6/NS5ATP9-mediated autophagy and affects the progression of tumor. Therefore, NS5ATP9 silence might be a potential target for HCC therapy.

Keywords: autophagy, Hepatocellular carcinoma, IL-6, microenvironment, NS5ATP9

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Authors: Vladimir Klepikov

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Countries of Central and Southern Europe have a typical feature: oil consumption in the region exceeds own commodity production capacity by far. So crude oil import prevails in the region’s crude oil consumption structure. Transportation using marine and pipeline transport is a common method of the imported oil delivery in the region. For certain refineries, in addition to possible transportation by oil pipelines from seaports, oil is delivered from Russian oil fields. With the view to these specific features and geographic location of the region’s refineries, three ways of imported oil delivery can be singled out: oil delivery by tankers to the port and subsequent transportation by pipeline transport of the port and the refinery; oil delivery by tanker fleet to the port and subsequent transportation by oil trunk pipeline transport; oil delivery from the fields by oil trunk pipelines to refineries. Oil is also delivered by road, internal water, and rail transport. However, the volumes transported this way are negligible in comparison to the three above transportation means. Multimodal oil transportation to refineries using the pipeline and marine transport is one of the biggest cargo flows worldwide. However, in scientific publications this problem is considered mainly for certain modes of transport. Therefore, this study is topical. To elaborate an efficient transportation policy of crude oil supply to Central and Southern Europe, in this paper the geographic concentration of oil refineries was determined and the capacities of the region’s refineries were assessed. The quantitative analysis method is used as a tool. The port infrastructure and the oil trunk pipeline system capacity were assessed in terms of delivery of raw materials to the refineries. The main groups of oil consuming countries were determined. The trends of crude oil production in the region were reviewed. The changes in production capacities and volumes at refineries in the last decade were shown. Based on the revealed refining trends, the scope of possible crude oil supplies to the refineries of the region under review was forecast. The existing transport infrastructure is able to handle the increased oil flow.

Keywords: European region, infrastructure, oil terminal capacity, pipeline capacity, refinery capacity, tanker draft

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Authors: ZhenlinJu, Christopher P. Vellano, RehanAkbani, Yiling Lu, Gordon B. Mills

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The epithelial–mesenchymal transition (EMT) is a process by which epithelial cells acquire mesenchymal characteristics, such as profound disruption of cell-cell junctions, loss of apical-basolateral polarity, and extensive reorganization of the actin cytoskeleton to induce cell motility and invasion. A hallmark of EMT is its capacity to promote metastasis, which is due in part to activation of several transcription factors and subsequent downregulation of E-cadherin. Unfortunately, current approaches have yet to uncover robust protein marker sets that can classify tumors as possessing strong EMT signatures. In this study, we utilize reverse phase protein array (RPPA) data and consensus clustering methods to successfully classify a subset of cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC) tumors into an EMT protein signaling group (EMT group). The overall survival (OS) of patients in the EMT group is significantly worse than those in the other Hormone and PI3K/AKT signaling groups. In addition to a shrinkage and selection method for linear regression (LASSO), we applied training/test set and Monte Carlo resampling approaches to identify a set of protein markers that predicts the EMT status of CESC tumors. We fit a logistic model to these protein markers and developed a classifier, which was fixed in the training set and validated in the testing set. The classifier robustly predicted the EMT status of the testing set with an area under the curve (AUC) of 0.975 by Receiver Operating Characteristic (ROC) analysis. This method not only identifies a core set of proteins underlying an EMT signature in cervical cancer patients, but also provides a tool to examine protein predictors that drive molecular subtypes in other diseases.

Keywords: consensus clustering, TCGA CESC, Silhouette, Monte Carlo LASSO

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Authors: Ka Ho Yau, Jan Frederick Engels, Kwok Kei Lai, Reinhard Renneberg

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Lateral flow test is a prevalent technology in various sectors such as food, pharmacology and biomedical sciences. Colloidal gold (CG) is widely used as the signalling molecule because of the ease of synthesis, bimolecular conjugation and its red colour due to intrinsic SPRE. However, the production of colloidal gold is costly and requires vigorous conditions. The stability of colloidal gold are easily affected by environmental factors such as pH, high salt content etc. Silica nanoparticles are well known for its ease of production and stability over a wide range of solvents. Using reverse micro-emulsion (w/o), silica nanoparticles with different sizes can be produced precisely by controlling the amount of water. By incorporating different water-soluble dyes, a rainbow colour of the silica nanoparticles could be produced. Conjugation with biomolecules such as antibodies can be achieved after surface modification of the silica nanoparticles with organosilane. The optimum amount of the antibodies to be labelled was determined by Bradford Assay. In this work, we have demonstrated the ability of the dye-doped silica nanoparticles as a signalling molecule in lateral flow test, which showed a semi-quantitative measurement of the analyte. The image was further analysed for the LOD=10 ng of the analyte. The working range and the linear range of the test were from 0 to 2.15μg/mL and from 0 to 1.07 μg/mL (R2=0.988) respectively. The performance of the tests was comparable to those using colloidal gold with the advantages of lower cost, enhanced stability and having a wide spectrum of colours. The positives lines can be imaged by naked eye or by using a mobile phone camera for a better quantification. Further research has been carried out in multicolour detection of different biomarkers simultaneously. The preliminary results were promising as there was little cross-reactivity being observed for an optimized system. This approach provides a platform for multicolour detection for a set of biomarkers that enhances the accuracy of diseases diagnostics.

Keywords: colorimetric detection, immunosensor, paper-based biosensor, silica

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Authors: Priya Varma, Denis John McKeown

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Psychological time, unlike physical time, is believed to be ‘compressive’ in the sense that the mental representations of a series of events may be internally arranged with ever decreasing inter-event spacing (looking back from the most recently encoded event). If this is true, the record within immediate memory of recent events is severely temporally distorted. Although this notion of temporal distortion of the memory record is captured within some theoretical accounts of human forgetting, notably temporal distinctiveness accounts, the way in which the fundamental nature of the distortion underpins memory and forgetting broadly is barely recognised or at least directly investigated. Our intention here was to manipulate the spacing of items for recall in order to ‘reverse’ this supposed natural compression within the encoding of the items. In Experiment 1 three schedules of presentation (expanding, contracting and fixed irregular temporal spacing) were created using logarithmic spacing of the words for both free and serial recall conditions. The results of recall of lists of 7 words showed statistically significant benefits of temporal isolation, and more excitingly the contracting word series (which we may think of as reversing the natural compression within the mental representation of the word list) showed best performance. Experiment 2 tested for effects of active verbal rehearsal in the recall task; this reduced but did not remove the benefits of our temporal scheduling manipulation. Finally, a third experiment used the same design but with Chinese characters as memoranda, in a further attempt to subvert possible verbal maintenance of items. One change to the design here was to introduce a probe item following the sequence of items and record response times to this probe. Together the outcomes of the experiments broadly support the notion of temporal compression within immediate memory.

Keywords: memory, serial position curves, temporal isolation, temporal schedules

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Authors: F. Christie, M. Staber

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Malnutrition is a predictor of poor clinical outcome in the elderly. Up to 60% of hip fracture patients are clinically malnourished on admission. This study assessed the perioperative nutritional state of patients admitted with a proximal femoral fracture and examined if adequate nutritional support was achieved. Methods: Prospective, the observational audit of 30 patients, admitted with a proximal femoral fracture, over a one-month period. We recorded: patient demographics; surgical delay; nutritional state on admission; documentation of Malnutrition Universal Screening Tool (MUST) score; dietician input and daily calorie intake through food charts. The nutritional state was re-assessed weekly and at discharge. The outcome was measured by the length of hospital stay and thirty-day mortality. Results: Mean age 87, M:F 1:2 and all patients were ASA three or four. Five patients (17%) had a prolonged ( >24 hours) fasting period. All patients had a MUST score completed on admission, 27% were underweight and 30% were high risk for malnutrition. Twenty-six patients (87%) were appropriately assessed for dietician referral. Thirteen patients had food charts; on average, hospital meals provided 1500kcal daily. No patient achieved > 75% of the provided calories with 69% of patients achieving 50% or less. Only three patients were started on nutritional supplements. Twenty-three patients (77%) lost weight, averaging 6% weight loss during admission. Mean length of stay (LOS) was 23 days and 30-day mortality 9%. Four patients (13%) gained weight, their mean LOS was 17 days and 30-day mortality 0%. Discussion: Malnutrition in the elderly originates in the community. Following major trauma it’s difficult to reverse nutritional deficits in hospitals. It’s therefore concerning that no high-risk patient achieved their recommended calorie intake. Perioperative optimisation needs to include early nutritional intervention, early anaesthetic review and adjusted anaesthetic techniques to support feeding.

Keywords: trauma, nutrition, neck of femur fracture

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Authors: M. Gomaa Abdoelatef, Robert M. Field, Lee, Yong-Kwan

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Growing human populations have placed increased demands on water supplies and a heightened interest in desalination infrastructure. Key elements of the economics of desalination projects are thermal and electrical inputs. With growing concerns over the use of fossil fuels to (indirectly) supply these inputs, coupling of desalination with nuclear power production represents a significant opportunity. Individually, nuclear and desalination technologies have a long history and are relatively mature. For desalination, Reverse Osmosis (RO) has the lowest energy inputs. However, the economically driven output quality of the water produced using RO, which uses only electrical inputs, is lower than the output water quality from thermal desalination plants. Therefore, modern desalination projects consider that RO should be coupled with thermal desalination technologies (MSF, MED, or MED-TVC) with attendant steam inputs to permit blending to produce various qualities of water. A large nuclear facility is well positioned to dispatch large quantities of both electrical and thermal power. This paper considers the supply of thermal energy to a large desalination facility to examine heat balance impact on the nuclear steam cycle. The APR1400 nuclear plant is selected as prototypical from both a capacity and turbine cycle heat balance perspective to examine steam supply and the impact on electrical output. Extraction points and quantities of steam are considered parametrically along with various types of thermal desalination technologies to form the basis for further evaluations of economically optimal approaches to the interface of nuclear power production with desalination projects. In our study, the thermodynamic evaluation will be executed by DE-TOP which is the IAEA desalination program, it is approved to be capable of analyzing power generation systems coupled to desalination systems through various steam extraction positions, taking into consideration the isolation loop between the APR-1400 and the thermal desalination plant for safety concern.

Keywords: APR-1400, desalination, DE-TOP, IAEA, MSF, MED, MED-TVC, RO

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Authors: Mohammed Sghir Taleb

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Introduction The date palm (Phoenix dactylifera L.) represents an essential element of the oasis ecosystem for Saharan and pre-Saharan regions of Morocco. It plays an important role, not only due to its economic importance, but also its ecological adaptation to, firstly, to ensure necessary protection for crops against underlying warm and dry sales, and secondly to contribute to the fight against desertification. This is one of the oldest cultivated plant species best adapted to difficult climatic conditions of the Saharan and pre-Saharan regions, because of its ecological requirements and economically most suitable for investing in oasis agriculture. Methodology The methodology is mainly based on a literature review of principal theses and projects for the conservation of flora and vegetation. Results The date palm has multiple uses. Indeed, it produces fruits rich in nutrients, provides a multitude of secondary products and generates needed revenue for the survival of oasis populations. In Morocco, the development and modernization of the date palm sector face, both upstream and downstream of the industry, several major constraints. In addition to climate constraints (prolonged drought), in its environment (lack of water resources), to the incessant invasion of disease Bayoud, Moroccan palm ecosystem suffers from a low level of technical and traditional practices prevail and traditional, from the choice of variety and site preparation up to harvesting and recycling of products. Conclusion The date palm plays an important role in the socioeconomic development of local and national level. However, this ecosystem however, is subject to numerous degradation factors caused by anthropogenic action and climate change. to reverse the trends, several programs have been developed by Morocco for the restoration of degraded areas and the development of the Phoenix dactylifera ecosystem to meet the needs of local populations and the development of the national economy.

Keywords: efforts, flora, ecosystem, forest, conservation, Morocco

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Authors: S. Mohajeri

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Electrocodeposition of Ni-TiO2 nanocomposite single layers and Ni-TiO2/TiO2 multilayers from Watts bath containing TiO2 sol was carried out on copper substrate. Pulse plating and pulse reverse plating techniques were applied to facilitate higher incorporations of TiO2 nanoparticles in Ni-TiO2 nanocomposite single layers, and the results revealed that by prolongation of the current-off durations and the anodic cycles, deposits containing 11.58 wt.% and 13.16 wt.% TiO2 were produced, respectively. Multilayer coatings which consisted of Ni-TiO2 and TiO2-rich layers were deposited by pulse potential deposition through limiting the nickel deposition by diffusion control mechanism. The TiO2-rich layers thickness and accordingly, the content of TiO2 reinforcement reached 104 nm and 18.47 wt.%, respectively in the optimum condition. The phase structure and surface morphology of the nanocomposite coatings were characterized by X-ray diffraction (XRD) and scanning electron microscopy (SEM). The cross sectional morphology and line scans of the layers were studied by field emission scanning electron microscopy (FESEM). It was confirmed that the preferred orientations and the crystallite sizes of nickel matrix were influenced by the deposition technique parameters, and higher contents of codeposited TiO2 nanoparticles refined the microstructure. The corrosion behavior of the coatings in 1M NaCl and 0.5M H2SO4 electrolytes were compared by means of potentiodynamic polarization and electrochemical impedance spectroscopy (EIS) techniques. Increase of corrosion resistance and the passivation tendency were favored by TiO2 incorporation, while the degree of passivation declined as embedded particles disturbed the continuity of passive layer. The role of TiO2 incorporation on the improvement of mechanical properties including hardness, elasticity, scratch resistance and friction coefficient was investigated by the means of atomic force microscopy (AFM). Hydrophilicity and wettability of the composite coatings were investigated under UV illumination, and the water contact angle of the multilayer was reduced to 7.23° after 1 hour of UV irradiation.

Keywords: electrodeposition, hydrophilicity, multilayer, pulse-plating

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Authors: M. H. Rajikin, S. M. M. Syairah, A. R. Sharaniza

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Effects of nicotine on pre-partum body weight and preimplantation embryonic development has been reported previously. Present study was conducted to determine the effects of annatto (Bixa orellana)-derived delta-tocotrienol (TCT) (with presence of 10% gamma-TCT isomer) on the nicotine-induced reduction in body weight and 8-cell embryonic growth in mice. Twenty four 6-8 weeks old (23-25g) female balb/c mice were randomly divided into four groups (G1-G4; n=6). Those groups were subjected to the following treatments for 7 consecutive days: G1 (control) were gavaged with 0.1 ml tocopherol stripped corn oil, G2 was subcutaneously (s.c.) injected with 3 mg/kg/day of nicotine, G3 received concurrent treatment of nicotine (3 mg/kg/day) and 60 mg/kg/day of δ-TCT mixture (contains 90% delta & 10% gamma isomers) and G4 was given 60 mg/kg/day of δ-TCT mixture alone. Body weights were recorded daily during the treatment. On Day 8, females were superovulated with 5 IU Pregnant Mare’s Serum Gonadotropin (PMSG) for 48 hours followed with 5 IU human Chorionic Gonadotropin (hCG) before mated with males at the ratio of 1:1. Females were sacrificed by cervical dislocation for embryo collection 48 hours post-coitum. Collected embryos were cultured in vitro. Results showed that throughout Day 1 to Day 7, the body weight of nicotine treated group (G2) was significantly lower (p<0.05) than that of G1, G3 and G4. Intervention with δ-TCT mixture (G3) managed to increase the body weight close to the control group. This is also observed in the group treated with δ-TCT mixture alone (G4). The development of 8-cell embryos following in vitro culture (IVC) was totally inhibited in G2. Intervention with δ-TCT mixture (G3) resulted in the production of 8-cell embryos, although it was not up to that of the control group. Treatment with δ-TCT mixture alone (G4) caused significant increase in the average number of produced 8-cell embryo compared to G1. Present data indicated that δ-TCT mixture was able to reverse the body weight loss in nicotine treated mice and the development of 8-cell embryos was also improved.

Keywords: δ-tocotrienol, body weight, nicotine, preimplantation embryonic development

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Authors: Mitiku Tefera

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Background: Meconium-stained amniotic fluid is one of the primary causes of birth asphyxia. Each year, over five million neonatal deaths occur worldwide due to meconium-stained amniotic fluid, with 90% of these deaths due to birth asphyxia. In Ethiopia meconium-stained amniotic fluid is under investigated, specifically in North Shoa Zone Amhara region Ethiopia. Objective: The aim of this study was to assess the magnitude of meconium-stained amniotic fluid and associated factors among women who gave birth in the North Shoa Zone Hospital’s Amhara Region, Ethiopia, in 2022. Methods: An institutional-based, cross-sectional study was conducted among 628 women who gave birth at North Shoa Zone Hospitals, Amhara, Ethiopia. The study was conducted from 08/June-08/August 2022. Two-stage cluster sampling was used to recruit study participants. The data was collected by using a structured interview-administered questionnaire and chart review. The collected data was entered into Epi-Data Version 4.6 and exported to SPSS Version 25. Logistics regression was employed, and a p-value <0.05 was considered significant. Result: The magnitude of meconium-stained amniotic fluid was 30.3%. Women presented with normal hematocrit level 83% less likely develop meconium-stained amniotic fluid. Women had mid-upper arm circumference value was less than 22.9cm(AOR=1.9; 95% CI;1.18-3.20), obstructed labor(AOR=3.6; 95% CI;1.48-8.83), prolonged labor ≥ 15hr (AOR=7.5; 95% CI ;7.68-13.3), the premature rapture of the membrane (AOR=1.7; 95% CI; 3.22-7.40), fetal tachycardia(AOR=6.2; 95% CI; 2.41-16.3) and Bradycardia (AOR=3.1; 95% CI;1.93-5.28) were significant association with meconium stained amniotic fluid. Conclusion: The magnitude of meconium-stained amniotic fluid, which was high. In this study, MUAC value <22.9 cm, obstructed and prolonged labor, PROM, bradycardia, and tachycardia were factors associated with meconium-stained amniotic fluid. A follow-up study and pooled similar articles will be mentioned for better evidence, enhancing intrapartum services and strengthening early detection of meconium-stained amniotic fluid for the health of the mother and baby.

Keywords: women, meconium-staned amniotic fluid, magnitude, Ethiopia

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Authors: Kedar Vishnu, Parmod Kumar

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India is becoming most favored retail destination in the world. The organized retail has presented many opportunities to farmers to increase income by shifting cropping pattern from food grains to commercial crops. Previous research revealed potential benefits for farmers by supplying fruits and vegetables to organized retail channels. However the supply of fruits and vegetables from small and marginal farmers remain low than expected. The main objective of this paper is to identify the factors determining market participation of smallholder farmers in modern organized retail chains. Attempt is also made to find out factors influencing the choice of participation in particular organized retail collection centers as compared to other organized retail. The paper was based on primary survey of 40 Beans and Tomato farmers who supply to organized retail collection centers from Karnataka, India. Multiple regression technique is used to identify the factors determining quantity sold at collection centers. The regression result, show that area under vegetables, yield, and price from modern collection center and having access to technical help were found significantly affecting quantity sold into modern organized retail channels. On the opposite, increased rejection rates and vegetable prices at APMC were found influencing farmers decision into the reverse side. Empirical result of the multinomial logit model show that Reliance fresh has tendency to prefer large farmers who can supply more quality and better quantity compared with TESCO and More collection centers. The negative sign of area, having access to technical help, transportation cost, and number of bore wells led to higher probability of farmers to participate in Reliance Fresh collection centers as compared with More and TESCO.

Keywords: fruits, vegetables, organized retail markets, multinomial logit model

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Authors: Lubna Azmi, Ila Shukla, Shyam Sundar Gupta, Padam Kant, Ch. V. Rao

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Gastric ulcer is a major global health threat, and it is the leading cause of stomach cancer death worldwide. Helicobacter pylori bacteriumis the most important etiologic factor for gastric ulcer. This infection is highly pervasive in South Asian developing countries, especially in India, Nepal, Srilanka etc. due to diversification in geographic area. Pathophysiology of gastric mucosal damage associated with non-invasive bacterium has not justified in detail, but it leads to change in histopathology, immunochemistry of the gastric and duodenal reason of host. The mechanism responsible for bacteria tissue tropism and mucosal damage in stomach variance during the disease is not clearly described and understood scientifically in treatment and control of pathogenic organisms. Polyphenols are secondary metabolites of plants and are generally involved in defense against aggression by pathogens. 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxychromen-4-one and 1-hydroxy-5,7-dimethoxy-2-naphthalene-carboxaldehyde are polyphenolic compound obtained from popular Indian medicinal plants ghavpatta (ArgeriaspeciosaLinn.f) andBael (Aeglemarmelos) have long been used in traditional Ayurvedic Indian medicine for various diseases. They have promising effects on ulcer, as detailed investigation has made in our laboratory. Therefore, the aim of present study is to explore membrane –dependent morphogenesis of H. pylori and associated apoptosis-mediated cell death. Based on this we analyzed immune gene expression in stomach of experimental animals with H. pylori, using quantitative reverse transcription polymerase chain reaction(q RT-PCR). This revealed rapid induction of prostaglandin, interferon I (INF-I), interferon II (INF-II) and INF-I associated genes in the infected animal. Ultrastructural changes associated with H. pylori will be taken for advanced studies. This investigation shows that the biomarkers eradicate H. pylori bacterium caused gastric ulcer which is a major risk factor for gastric cancer.

Keywords: gastric ulcer, Helicobacter pylori, immunochemistry, polyphenols

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Authors: Ballari Bagchi

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The dynamism ingrained in the process of urban-rural integration is manifested in the emergence of rurban settlements, referring to areas that combine the characteristics of agricultural activities found in rural zones with those of suburban living areas and industrialised zones. The concept of rurbanisation refers to the idea of introducing urban conveniences and opportunities, to rural areas in an attempt to stem rural urban migration. In the backdrop of the worldwide problem of disharmonised urban-rural dependence and the associated problems in urban and rural areas, the present study seeks to explore the potentialities of few settlements having a blend of rural and urban characteristics in the urban field of Koch Bihar. The prime concern of the present paper is three-fold: (i) to identify the rurban centres, (ii) to analyse the spatial integration of these identified centres with the rural areas situated in the urban periphery, and (iii) to suggest the necessities to be introduced in these settlements. The methodology applied here includes rurban index, gravity model, and functional classification of rurban centres, correlation and regression analysis and cartographic representation of data collected through primary and secondary sources. The investigation has identified a number of settlements potentially viable to be termed as rurban centres which may render services to the other less equipped rural areas in all aspects of life and thereby would lessen the burden on Koch Bihar urban centre. The levels of infrastructure of these settlements should be such that it might even attract the urban population in a reverse direction. The villages belonging to the lower rung of these service settlements would require metalled road connection with these intermediate settlements in addition to their connection with the core town. That is to say, a proper policy needs to be adopted in this regard to furnish these settlements with required infrastructures for serving their own population as well as the population of other villages. As a consequence of that, the idea of a well-coordinated settlement hierarchy may emerge in future.

Keywords: Hinterland, rurban, settlement hierarchy, urban-rural integration

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Authors: Shih-Wen Hsiao, Yi-Cheng Tsao

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In the field of reverse engineering and creative industries, applying 3D scanning process to obtain geometric forms of the objects is a mature and common technique. For instance, organic objects such as faces and nonorganic objects such as products could be scanned to acquire the geometric information for further application. However, although the data resolution of 3D scanning device is increasing and there are more and more abundant complementary applications, the penetration rate of 3D scanning for the public is still limited by the relative high price of the devices. On the other hand, Kinect, released by Microsoft, is known for its powerful functions, considerably low price, and complete technology and database support. Therefore, related studies can be done with the applying of Kinect under acceptable cost and data precision. Due to the fact that Kinect utilizes optical mechanism to extracting depth information, limitations are found due to the reason of the straight path of the light. Thus, various angles are required sequentially to obtain the complete 3D information of the object when applying a single Kinect for 3D scanning. The integration process which combines the 3D data from different angles by certain algorithms is also required. This sequential scanning process costs much time and the complex integration process often encounter some technical problems. Therefore, this paper aimed to apply multiple Kinects simultaneously on the field of developing a rapid 3D mannequin scan platform and proposed suggestions on the number and angles of Kinects. In the content, a method of establishing the coordination based on the relation between mannequin and the specifications of Kinect is proposed, and a suggestion of angles and number of Kinects is also described. An experiment of applying multiple Kinect on the scanning of 3D mannequin is constructed by Microsoft API, and the results show that the time required for scanning and technical threshold can be reduced in the industries of fashion and garment design.

Keywords: 3D scan, depth sensor, fashion and garment design, mannequin, multiple Kinect sensor

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Authors: Tariq Ahmad Dar, Moinuddin, M. Masroor A. Khan

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There is considerable need in enhancing the content and yield of active constituents of medicinal plants keeping in view their massive demand worldwide. Different strategies have been employed to enhance the active constituents of medicinal plants and the use of phytohormones has been proved effective in this regard. Gamma-irradiated Sodium alginate (ISA) is known to elicit an array of plant defense responses and biological activities in plants. Considering the medicinal importance, a pot experiment was conducted to explore the effect of ISA and phosphorus on growth, yield and quality of fenugreek (Trigonella foenum-graecum L.). ISA spray treatments (0, 40, 80 and 120 mg L-1) were applied alone and in combination with 40 kg P ha-1 (P40). Crop performance was assessed in terms of plant growth characteristics, physiological attributes, seed yield and the content of seed trigonelline. Of the ten-treatments, P40 + 80 mg L−1 of ISA proved the best. The results showed that foliar spray of ISA alone or in combination with P40 augmented the plant vegetative growth, enzymatic activities, trigonelline content, trigonelline yield and economic yield of fenugreek. Application of 80 mg L−1 of ISA applied with P40 gave the best results for almost all the parameters studied compared to control or to 80 mg L−1 of ISA applied alone. This treatment increased the total content of chlorophyll, carotenoids, leaf -N, -P and -K and trigonelline compared to the control by 24.85 and 27.40%, 15 and 23.52%, 18.70 and 16.84%, 15.88 and 18.92%, 12 and 14.44%, at 60 and 90 DAS respectively. The combined application of 80 mg L−1 of ISA along with P40 resulted in the maximum increase in seed yield, trigonelline content and trigonelline yield by146, 34 and 232.41%, respectively, over the control. Gel permeation chromatography revealed the formation of low molecular weight fractions in ISA samples, containing even less than 20,000 molecular weight oligomers, which might be responsible for plant growth promotion in this study. Trigonelline content was determined by reverse phase high performance liquid chromatography (HPLC) with C-18 column.

Keywords: gamma-irradiated sodium alginate, phosphorus, gel permeation chromatography, HPLC, trigonelline content, yield

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Authors: Babak Rezaei, Arash Zargar Shoushtari

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Kangan anticline is located in the Coastal Fars area, southwest of Nar and west of west Assaluyeh anticlines and north of Kangan harbor in Boushehr province. The Kangan anticline is nearly asymmetric and with 55Km long and 6Km wide base on structural map of Kangan Formation. The youngest and the oldest Formations on surface are Bakhtiyari (Pliocene) and Sarvak (Cenomanian) respectively. The highest dip angles of 30 and 40 degree were observed in north and south flanks of Kangan anticline respectively and two reverse faults cut these flanks parallel to structure strike. Existence of sweet gas in Kangan Fm. and Upper Dalan in this structure is confirmed with probable Silurian shales origin. Main facies belts in these formations include super tidal and intertidal flat, lagoon, oolitic-bioclastic shoals and open marine sub environments that expand in a homoclinal and shallow water carbonate ramp under the arid climates. Digenetic processes studies, indicates the influence of all digenetic environments (marine, meteoric, burial) in the reservoir succession. These processes sometimes has led to reservoir quality improvement (such as dolomitization and dissolution) but in many instances reservoir units has been destroyed (such as compaction, anhydrite and calcite cementation). In this study, petrophysical evaluation is made in Kangan and upper Dalan formations by using well log data of five selected wells. Probabilistic method is used for petrophysical evaluation by applying appropriate soft wares. According to this evaluation the lithology of Kangan and upper Dalan Formations mainly consist of limestone and dolomite with thin beds of Shale and evaporates. In these formations 11 Zones with different reservoir characteristic have been identified. Based on wire line data analyses, in some part of these formations, high porosity can be observed. The range of porosity (PHIE) and water saturation (Sw) are estimated around 10-20% and 20-30%, respectively.

Keywords: microfacies, electrofacies, petrophysics, diagenese, gas fields

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Authors: Sadia H. Syeda, Zain H. Malik

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As systems continue to become more complex and the interdependencies of processes and sub-systems continue to grow and transform, the need for a comprehensive method of tracking and linking the lifecycle of the systems in a digital form becomes ever more critical. Digital Engineering (DE) provides an approach to managing an authoritative data source that links, tracks, and updates system data as it evolves and grows throughout the system development lifecycle. DE enables the developing, tracking, and sharing system data, models, and other related artifacts in a digital environment accessible to all necessary stakeholders. The DE environment provides an integrated electronic repository that enables traceability between design, engineering, and sustainment artifacts. The DE activities' primary objective is to develop a set of integrated, coherent, and consistent system models for the program. It is envisioned to provide a collaborative information-sharing environment for various stakeholders, including operational users, acquisition personnel, engineering personnel, and logistics and sustainment personnel. Examining the processes that DE can support in the systems engineering life cycle (SELC) is a primary step in the DE adoption and implementation journey. Through an analysis of the U.S Department of Defense’s (DoD) Office of the Secretary of Defense (OSD’s) Digital Engineering Strategy and their implementation, examples of DE implementation by the industry and technical organizations, this paper will provide descriptions of the current DE processes and best practices of implementing DE across an enterprise. This will help identify the capabilities, environment, and infrastructure needed to develop a potential roadmap for implementing DE practices consistent with its business strategy. A capability maturity matrix will be provided to assess the organization’s DE maturity emphasizing how all the SELC elements interlink to form a cohesive ecosystem. If implemented, DE can increase efficiency and improve the systems engineering processes' quality and outcomes.

Keywords: digital engineering, digital environment, digital maturity model, single source of truth, systems engineering life-cycle

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Authors: Olorunsanya E. O., Adedeji S. O., Anyanwu A. A.

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Food insecurity is a challenge to many nations Nigeria inclusive. It is increasingly becoming a major problem among farm households due to many factors chief of which is low labour productivity. This study therefore profiles the food security status of a representative randomly selected 90 farming households in Chanchaga area of Nigeria’s Guinea Savana using structured interview schedule Descriptive and inferential statistics were used as analytical tools for the study. The results of the descriptive statistics show that majority (35.56%) of the surveyed household heads fall within the age range of 40 – 49 years and (88.89%) are male while (78.89) are married. More than half of the respondents have formal education. About 43.3% of the household heads have farm experience of 11- 20 years and a modal household size class range of 7 – 12. The results further reveal that majority (68.8%) earned more than N12, 500 (22.73 US Dollar) per month. The result of households’ food expenditure pattern reveals that an average household spends about N3, 644.44 (6.63 US Dollar) on food and food items on a weekly basis. The result of the analysis of food diversity intake in the study area shows that 63.33% of the sampled households fell under the low household food diversity intake, while 33 households, representing 36.67% ranks high in term of household food diversity intake. The result for the food security status shows that the sampled population was food secure (58.89%) while 41.11% falls below the recommended threshold. The result for the logistics regression model shows that age, engagement in off farm employment and household size are significant in determining the food security status of farm household in the study area. The three variables were significant at 10%, 5% and 1% respectively. The study therefore recommends among others, that measures be put in place by stakeholders to make agriculture attractive for youth since age is a significant determinant of food security in the study area. Awareness should also be created by stakeholders on the needs for effective family planning methods to be adopted by farm household in the study area.

Keywords: Niger State, Guinea Savana, food diversity, logit regression model and food security

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Authors: Yun-Chin Chung, Nileema Divate, Gen-Hung Chen, Pei-Ru Huang, Rupesh Divate

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Many environmental factors, such as glucose concentration, ethanol, temperature, osmotic pressure and pH, decrease the production rate of ethanol using yeast as a starter. Fermentation starters with high tolerance to various stresses are always demanded for brewing industry. Trehalose, a storage carbohydrate in cell wall of yeast, plays an important role in tolerance of environmental stress by preserving integrity of plasma membrane and stabilizing proteins. Furan aldehydes are toxic to yeast and the growth rate of yeast is significantly reduced if furan aldehydes were present in the fermentation medium. In yeast, aldehyde reductase is involved in the detoxification of reactive aldehydes and consequently the growth of yeast is improved. The aims of this study were to construct a genetic recombinant Saccharomyces cerevisiae or Pichia pastoris with furfural and HMF degrading and high ethanol tolerance capacities. Yeast strains were engineered by genetic recombination for overexpression of trehalose-6-phosphate synthase gene (tps1) and aldehyde reductase gene (ari1). TPS1 gene was cloned from S. cerevisiae by reverse transcription-polymerase chain reaction (RT-PCR) and then ligated with pGAPZαC vector. The constructed vector, pGAPZC-tps1, was transformed to recombinant yeasts strain with overexpression of ari1. The transformants with pGAPZC-tps1-ari1 were generated called STA (S. cerevisiae) and PTA (P. pastoris) with overexpression of tps1, ari1. PCR with tps1-specific primers and western blot with his-tag confirmed the gene insertion and protein expression of tps1 in the transformants, respectively. The neutral trehalase gene (nth1) of STA was successfully deleted and the novel strain STAΔN will be used for further study, including the measurement of trehalose concentration and ethanol, furfural tolerance assay.

Keywords: genetic recombinant, yeast, ethanol tolerance, trehalase, aldehyde reductase

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Authors: Akram Abi, Clarissa Müller, Hans-Joachim Jördening

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Laminaribiose is a beta-1,3-glycoside which is used in the medical field for the treatment of dermatitis and also can be used as a building block for new pharmaceutics. The conventional process of laminaribiose production is the uneconomical process of hydrolysis of laminarin extracted from natural polysaccharides of plant origin. A more economical approach however is attainable by enzymatically synthesis of laminaribiose via a reverse phosphorylase reaction catalyzed by laminaribiose phosphorylase (LP) from Euglena gracilis. Different cultivation methods of Euglena gracilis and the effect on LP production have been investigated. Buffered/unbuffered heterotrophic and mixotrophic cultivations of Euglena gracilis has been carried out. Changes of biomass and LP production, glucose level and pH, cell count and shape has been monitored in the course of time. The results obtained from experiments each in three repetitions, show that in the heterotrophic cultivation of Euglena gracilis not only more biomass is produced compared to mixotrophic cultivation, but also higher specific protein concentration is achieved. Furthermore, the LP activity test showed that the protein extracted from heterotrophically cultured cells has a higher LP activity. It was also observed that the cells develop in a distinctive different shape between these two cultures and have different length to width ratios. Taking the heterotrophic culture as the more efficient cultivation method in LP production, another comparative experiment between buffered and unbuffered heterothrophic culture was carried out that showed the unbuffered culture has advantages over the other one in respect of both LP production and resulting activity. A hetrotrophic cultivation of Euglena gracilis in a 5L bioreactor with controlled operating conditions showed a distinctive improvement of all the aspects of culture compared to the shaking flask cultivations. Biomass production was improved from 5 to more than 8 g/l (dry weight) which resulted in a specific protein concentration of 45 g/l in the heterotrophic cultivation in the bioreactor. In further attempts to improve LP production, different purification methods were tested and each method was checks through an activity assay. A laminaribiose yield of 35% was achieved which was by far the highest amount amongst different methods tested.

Keywords: euglena gracilis, heterotrophic culture, laminaribiose production, mixotrophic culture

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