Search results for: car sequencing and operator allocation

Authors: Olanrewaju Aladeitan, Obiageli Phina Anaghara-Uzor

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Despite the abundance of natural gas deposits in Nigeria and the immense potential, this presents both for the domestic and export oriented revenue, there exists an imbalance in the preference for export as against the development and optimal utilization of natural gas for the domestic industry. Considerable amounts of gas are still being wasted by flaring in the country to this day. Although the government has set in place initiatives to harness gas at the flare and thereby reduce volumes flared, the gas producers would rather direct the gas produced to the export market whereas gas apportioned to the domestic market is often marred by the low domestic gas price which is often discouraging to the gas producers. The exported fraction of gas production no doubt yields healthy revenues for the government and an encouraging return on investment for the gas producers and for this reason export sales remain enticing and preferable to the domestic sale of gas. This export pull impacts negatively if left unchecked, on the domestic market which is in no position to match the price at the international markets. The issue of gas price remains critical to the optimal development of the domestic gas industry, in that it comprises the basis for investment decisions of the producers on the allocation of their scarce resources and to what project to channel their output in order to maximize profit. In order then to rebalance the domestic industry and streamline the market for gas, the Gas Aggregation Company of Nigeria, also known as the Strategic Aggregator was proposed under the Nigerian Gas Master Plan of 2008 and then established pursuant to the National Gas Supply and Pricing Regulations of 2008 to implement the domestic gas supply obligation which focuses on ramping-up gas volumes for domestic utilization by mandatorily requiring each gas producer to dedicate a portion of its gas production for domestic utilization before having recourse to the export market. The 2008 Regulations further stipulate penalties in the event of non-compliance. This study, in the main, assesses the adequacy of the legal framework for the Nigerian Gas Industry, given that the operational laws are structured more for oil than its gas counterpart; examine the legal basis for the Strategic Aggregator in the light of the Domestic Gas Supply and Pricing Policy 2008 and the National Domestic Gas Supply and Pricing Regulations 2008 and makes a case for a review of the pivotal role of the Aggregator in the Nigerian Gas market. In undertaking this assessment, the doctrinal research methodology was adopted. Findings from research conducted reveal the reawakening of the Federal Government to the immense potential of its gas industry as a critical sector of its economy and the need for a sustainable domestic natural gas market. A case for the review of the ownership structure of the Aggregator to comprise a balanced mix of the Federal Government, gas producers and other key stakeholders in order to ensure the effective implementation of the domestic supply obligations becomes all the more imperative.

Keywords: domestic supply obligations, natural gas, Nigerian gas sector, strategic gas aggregator

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Authors: Danylo Brindak, Viktor Liashko, Olexander Chepurniy

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Despite considerable experience in implementation of the best international approaches and services within response to epidemy of multi-drug resistant tuberculosis, the results of situation analysis indicate the presence of faults in this area. In 2014, Ukraine (for the first time) was included in the world’s five countries with the highest level of drug-resistant tuberculosis. The effectiveness of its treatment constitutes only 35% in the country. In this context, the increase in allocation of funds to control the epidemic of multidrug-resistant tuberculosis does not produce perceptible positive results. During 2001-2016, only the Global Fund to fight AIDS, Tuberculosis, and Malaria allocated to Ukraine more than USD 521,3 million for programs of tuberculosis and HIV/AIDS control. However, current conditions in post-Semashko system create little motivation for rational use of resources or cost control at inpatient TB facilities. There is no motivation to reduce overdue hospitalization and to target resources to priority sectors of modern tuberculosis control, including a model of care focused on the patient. In the presence of a line-item budget at medical institutions, based on the input factors as the ratios of beds and staff, there is a passive disposal of budgetary funds by health care institutions and their employees who have no motivation to improve quality and efficiency of service provision. Outpatient treatment of tuberculosis is being implemented in Ukraine since 2011 and has many risks, namely creation of parallel systems, low consistency through dependence on funding for the project, reduced the role of the family doctor, the fragmentation of financing, etc. In terms of reforming approaches to health system financing, which began in Ukraine in late 2016, NGO Infection Control in Ukraine conducted piloting of a new, motivating method of remuneration of employees in primary health care. The innovative aspect of this funding mechanism is cost according to results of treatment. The existing method of payment on the basis of the standard per inhabitant (per capita ratio) was added with motivating costs according to results of work. The effectiveness of such treatment of TB patients at the outpatient stage is 90%, while in whole on the basis of a current system the effectiveness of treatment of newly diagnosed pulmonary TB with positive swab is around 60% in the country. Even though Ukraine has 5.24 TB beds per 10 000 citizens. Implemented pilot model of ambulatory treatment will be used for the creation of costs system according to results of activities, the integration of TB and primary health and social services and their focus on achieving results, the reduction of inpatient treatment of tuberculosis.

Keywords: health care reform, multi-drug resistant tuberculosis, outpatient treatment efficiency, tuberculosis

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Authors: Nicole Vlasman, Karamjeet Dhillon

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Over the past four years, the Healthy Relationships Program has been delivered in community organizations and schools across Canada. More than 240 groups have been facilitated in collaboration with 33 organizations. As a result, 2157 youth have been engaged in the programming. The purpose and scope of the Healthy Relationships Program are to offer sustainable, evidence-based skills through small group implementation to prevent violence and promote positive, healthy relationships in youth. The program development has included extensive networking at regional and national levels. The Healthy Relationships Program is currently being implemented, adapted, and researched within the Resilience and Inclusion through Strengthening and Enhancing Relationships (RISE-R) project. Alongside the project’s research objectives, the RISE-R team has worked to virtually share the ongoing findings of the project through a slow ontology approach. Slow ontology is a practice integrated into project systems and structures whereby slowing the pace and volume of outputs offers creative opportunities. Creative production reveals different layers of success and complements the project, the building blocks for sustainability. As a result of integrating a slow ontology approach, the RISE-R team has developed a Geographic Information System (GIS) that documents local landscapes through a Story Map feature, and more specifically, video installations. Video installations capture the cartography of space and place within the context of singular diverse community spaces (case studies). By documenting spaces via human connections, the project captures narratives, which further enhance the voices and faces of the community within the larger project scope. This GIS project aims to create a visual and interactive flow of information that complements the project's mixed-method research approach. Conclusively, creative project development in the form of a geographic information system can provide learning and engagement opportunities at many levels (i.e., within community organizations and educational spaces or with the general public). In each of these disconnected spaces, fragmented stories are connected through a visual display of project outputs. A slow ontology practice within the context of the RISE-R project documents activities on the fringes and within internal structures; primarily through documenting project successes as further contributions to the Centre for School Mental Health framework (philosophy, recruitment techniques, allocation of resources and time, and a shared commitment to evidence-based products).

Keywords: community programming, geographic information system, project development, project management, qualitative, slow ontology

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Authors: Elena Collino, Dario A. Ronzio, Goffredo Decimi, Maurizio Riva

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The rapid increase of renewable energy in Italy is led by wind and solar installations. The 2017 Italian energy strategy foresees a further development of these sustainable technologies, especially solar. This fact has resulted in new opportunities, challenges, and different problems to deal with. The growth of renewables allows to meet the European requirements regarding energy and environmental policy, but these types of sources are difficult to manage because they are intermittent and non-programmable. Operationally, these characteristics can lead to instability on the voltage profile and increasing uncertainty on energy reserve scheduling. The increasing renewable production must be considered with more and more attention especially by the Transmission System Operator (TSO). The TSO, in fact, every day provides orders on energy dispatch, once the market outcome has been determined, on extended areas, defined mainly on the basis of power transmission limitations. In Italy, six market zone are defined: Northern-Italy, Central-Northern Italy, Central-Southern Italy, Southern Italy, Sardinia, and Sicily. An accurate hourly renewable power forecasting for the day-ahead on these extended areas brings an improvement both in terms of dispatching and reserve management. In this study, an operational forecasting tool of the hourly solar output for the six Italian market zones is presented, and the performance is analysed. The implementation is carried out by means of a numerical weather prediction model, coupled with a statistical post-processing in order to derive the power forecast on the basis of the meteorological projection. The weather forecast is obtained from the limited area model RAMS on the Italian territory, initialized with IFS-ECMWF boundary conditions. The post-processing calculates the solar power production with the Analog Ensemble technique (AN). This statistical approach forecasts the production using a probability distribution of the measured production registered in the past when the weather scenario looked very similar to the forecasted one. The similarity is evaluated for the components of the solar radiation: global (GHI), diffuse (DIF) and direct normal (DNI) irradiation, together with the corresponding azimuth and zenith solar angles. These are, in fact, the main factors that affect the solar production. Considering that the AN performance is strictly related to the length and quality of the historical data a training period of more than one year has been used. The training set is made by historical Numerical Weather Prediction (NWP) forecasts at 12 UTC for the GHI, DIF and DNI variables over the Italian territory together with corresponding hourly measured production for each of the six zones. The AN technique makes it possible to estimate the aggregate solar production in the area, without information about the technologic characteristics of the all solar parks present in each area. Besides, this information is often only partially available. Every day, the hourly solar power forecast for the six Italian market zones is made publicly available through a website.

Keywords: analog ensemble, electricity market, PV forecast, solar energy

Procedia PDF Downloads 137

Authors: Dejene Ermias Mekango, Mussie Alemayehu, Gebremedhin Berhe Gebregergs, Araya Abrha Medhanye, Gelila Goba

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Background: Maternal near miss (MNM) can be used as a proxy indicator of maternal mortality ratio. There is a huge gap in life time risk between Sub-Saharan Africa and developed countries. In Ethiopia, a significant number of women die each year from complications during pregnancy, childbirth and the post-partum period. Besides, a few studies have been performed on MNM, and little is known regarding determinant factors. This study aims to identify determinants of MNM among women in Tigray region, Northern Ethiopia. Methods: a case-control study in hospital found in Tigray region, Ethiopia was conducted from January 30 - March 30, 2016. The sample included 103 cases and 205 controls recruited from women seeking obstetric care at six public hospitals. Clients having a life-threatening obstetric complication including haemorrhage, hypertensive diseases of pregnancy, dystocia, infections, and anemia or clinical signs of severe anemia in women without haemorrhage were taken as cases and those with normal obstetric outcomes were considered as controls. Cases were selected based on proportional to size allocation while systematic sampling was employed for controls. Data were analyzed using SPSS version 20.0. Binary and multiple variable logistic regression (odds ratio) analyses were calculated with 95% CI. Results: The largest proportion of cases and controls was among the ages of20–29 years, accounting for37.9 %( 39) of cases and 31.7 %( 65) of controls. Roughly 90% of cases and controls were married. About two-thirds of controls and 45.6 %( 47) of cases had gestational age between 37-41 weeks. History of chronic medical conditions was reported in 55.3 %(57) of cases and 33.2%(68) of controls. Women with no formal education [AOR=3.2;95%CI:1.24, 8.12],being less than 16 years old at first pregnancy [AOR=2.5; 95%CI:1.12,5.63],induced labor[AOR=3; 95%CI:1.44, 6.17], history of Cesarean section (C-section) [AOR=4.6; 95%CI: 1.98, 7.61] or chronic medical disorder[AOR=3.5;95%CI:1.78, 6.93], and women who traveled more than 60 minutes before reaching their final place of care[AOR=2.8;95% CI: 1.19,6.35] all had higher odds of experiencing MNM. Conclusions: The Government of Ethiopia should continue its effort to address the lack of road and health facility access as well as education, which will help reduce MNM. Work should also be continued to educate women and providers about common predictors of MNM like the history of C-section, chronic illness, and teenage pregnancy. These efforts should be carried out at the facility, community, and individual levels. The targeted follow-up to women with a history of chronic disease and C-section could also be a practical way to reduce MNM.

Keywords: maternal near miss, severe obstetric hemorrhage, hypertensive disorder, c-section, Tigray, Ethiopia

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Authors: Reham Khalaf-Nazzal, Imad Dweikat, Paula Gimenez, Iker Oyenarte, Alfonso Martinez-Cruz, Domonik Muller

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Metal cation transport mediator (CNNM) gene family comprises 4 isoforms that are expressed in various human tissues. Structurally, CNNMs are complex proteins that contain an extracellular N-terminal domain preceding a DUF21 transmembrane domain, a ‘Bateman module’ and a C-terminal cNMP-binding domain. Mutations in CNNM2 cause familial dominant hypomagnesaemia. Growing evidence highlights the role of CNNM2 in neurodevelopment. Mutations in CNNM2 have been implicated in epilepsy, intellectual disability, schizophrenia, and others. In the present study, we aim to elucidate the function of CNNM2 in the developing brain. Thus, we present the genetic origin of symptoms in two family cohorts. In the first family, three siblings of a consanguineous Palestinian family in which parents are first cousins, and consanguinity ran over several generations, presented a varying degree of intellectual disability, cone-rod dystrophy, and autism spectrum disorder. Exome sequencing and segregation analysis revealed the presence of homozygous pathogenic mutation in the CNNM2 gene, the parents were heterozygous for that gene mutation. Magnesium blood levels were normal in the three children and their parents in several measurements. They had no symptoms of hypomagnesemia. The CNNM2 mutation in this family was found to locate in the CBS1 domain of the CNNM2 protein. The crystal structure of the mutated CNNM2 protein was not significantly different from the wild-type protein, and the binding of AMP or MgATP was not dramatically affected. This suggests that the CBS1 domain could be involved in pure neurodevelopmental functions independent of its magnesium-handling role, and this mutation could have affected a protein partner binding or other functions in this protein. In the second family, another autosomal dominant CNNM2 mutation was found to run in a large family with multiple individuals over three generations. All affected family members had hypomagnesemia and hypermagnesuria. Oral supplementation of magnesium did not increase the levels of magnesium in serum significantly. Some affected members of this family have defects in fine motor skills such as dyslexia and dyslalia. The detected mutation is located in the N-terminal part, which contains a signal peptide thought to be involved in the sorting and routing of the protein. In this project, we describe heterogenous clinical phenotypes related to CNNM2 mutations and protein functions. In the first family, and up to the authors’ knowledge, we report for the first time the involvement of CNNM2 in retinal photoreceptor development and function. In addition, we report the presence of a neurophenotype independent of magnesium status related to the CNNM2 protein mutation. Taking into account the different modes of inheritance and the different positions of the mutations within CNNM2 and its different structural and functional domains, it is likely that CNNM2 might be involved in a wide spectrum of neuropsychiatric comorbidities with considerable varying phenotypes.

Keywords: magnesium transport, autosomal recessive, autism, neurodevelopment, CBS domain

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Authors: Saiyed M Ishtiaque, Priyal Dixit

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Objectives: An approach is being suggested to design the filter media to maximize the filtration efficiency with minimum possible pressure drop of composite nonwoven by incorporating the layers of different packing densities induced by fibre of different deniers and punching parameters by using the concept of sequential punching technique in specific order in layered composite nonwoven structure. X-ray computed tomography technique is used to measure the packing density along the thickness of layered nonwoven structure composed by placing the layer of differently oriented fibres influenced by fibres of different deniers and punching parameters in various combinations to minimize the pressure drop at maximum possible filtration efficiency. Methodology Used: This work involves preparation of needle punched layered structure with batts 100g/m2 basis weight having fibre denier, punch density and needle penetration depth as variables to produce 300 g/m2 basis weight nonwoven composite. X-ray computed tomography technique is used to measure the packing density along the thickness of layered nonwoven structure composed by placing the layers of differently oriented fibres influenced by considered variables in various combinations. to minimize the pressure drop at maximum possible filtration efficiencyFor developing layered nonwoven fabrics, batts made of fibre of different deniers having 100g/m2 each basis weight were placed in various combinations. For second set of experiment, the composite nonwoven fabrics were prepared by using 3 denier circular cross section polyester fibre having 64 mm length on needle punched nonwoven machine by using the sequential punching technique to prepare the composite nonwoven fabrics. In this technique, three semi punched fabrics of 100 g/m2 each having either different punch densities or needle penetration depths were prepared for first phase of fabric preparation. These fabrics were later punched altogether to obtain the overall basis weight of 300 g/m2. The total punch density of the composite nonwoven fabric was kept at 200 punches/ cm2 with a needle penetration depth of 10 mm. The layered structures so formed were subcategorised into two groups- homogeneous layered structure in which all the three batts comprising the nonwoven fabric were made from same denier of fibre, punch density and needle penetration depth and were placed in different positions in respective fabric and heterogeneous layered structure in which batts were made from fibres of different deniers, punch densities and needle penetration depths and were placed in different positions. Contributions: The results concluded that reduction in pressure drop is not derived by the overall packing density of the layered nonwoven fabric rather sequencing of layers of specific packing density in layered structure decides the pressure drop. Accordingly, creation of inverse gradient of packing density in layered structure provided maximum filtration efficiency with least pressure drop. This study paves the way for the possibility of customising the composite nonwoven fabrics by the incorporation of differently oriented fibres in constituent layers induced by considered variablres for desired filtration properties.

Keywords: filtration efficiency, layered nonwoven structure, packing density, pressure drop

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Authors: Benjamin Glasner, Carlos Henriquez, Fernando Alfaro, Nicole Trefault, Santiago Andrade, Rodrigo De La Iglesia

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A central question in ecology refers to the relative importance that local-scale variables have over community composition, when compared with regional-scale variables. In coastal environments, strong seasonal abiotic influence dominates these systems, weakening the impact of other parameters like micronutrients. After the industrial revolution, micronutrients like trace metals have increased in ocean as pollutants, with strong effects upon biotic entities and biological processes in coastal regions. Coastal picoplankton communities had been characterized as a cyanobacterial dominated fraction, but in recent years the eukaryotic component of this size fraction has gained relevance due to their high influence in carbon cycle, although, diversity patterns and responses to disturbances are poorly understood. South Pacific upwelling coastal environments represent an excellent model to study seasonal changes due to a strong influence in the availability of macro- and micronutrients between seasons. In addition, some well constrained coastal bays of this region have been subjected to strong disturbances due to trace metal inputs. In this study, we aim to compare the influence of seasonality and trace metals concentrations, on the community structure of planktonic picoeukaryotes. To describe seasonal patterns in the study area, satellite data in a 6 years time series and in-situ measurements with a traditional oceanographic approach such as CTDO equipment were performed. In addition, trace metal concentrations were analyzed trough ICP-MS analysis, for the same region. For biological data collection, field campaigns were performed in 2011-2012 and the picoplankton community was described by flow cytometry and taxonomical characterization with next-generation sequencing of ribosomal genes. The relation between the abiotic and biotic components was finally determined by multivariate statistical analysis. Our data show strong seasonal fluctuations in abiotic parameters such as photosynthetic active radiation and superficial sea temperature, with a clear differentiation of seasons. However, trace metal analysis allows identifying strong differentiation within the study area, dividing it into two zones based on trace metals concentration. Biological data indicate that there are no major changes in diversity but a significant fluctuation in evenness and community structure. These changes are related mainly with regional parameters, like temperature, but by analyzing the metal influence in picoplankton community structure, we identify a differential response of some plankton taxa to metal pollution. We propose that some picoeukaryotic plankton groups respond differentially to metal inputs, by changing their nutritional status and/or requirements under disturbances as a derived outcome of toxic effects and tolerance.

Keywords: Picoeukaryotes, plankton communities, trace metals, seasonal patterns

Procedia PDF Downloads 156

Authors: K. S. Vimal, D. Bodhini, K. Ramya, N. Lakshmipriya, R. M. Anjana, V. Sudha, J. A. Lovegrove, V. Mohan, V. Radha

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Gene-lifestyle interaction studies have been carried out in various populations. However, to date there are no studies in an Asian Indian population. Hence, we examined whether lifestyle factors such as diet and physical activity modify the association between fat mass and obesity–associated (FTO) gene variants and obesity and type 2 diabetes (T2D) in an Asian Indian population. We studied 734 unrelated T2D and 884 normal glucose-tolerant (NGT) participants randomly selected from the Chennai Urban Rural Epidemiology Study (CURES) in Southern India. Obesity was defined according to the World Health Organization Asia Pacific Guidelines (non-obese, BMI < 25 kg/m2; obese, BMI ≥ 25 kg/m2). Six single nucleotide polymorphisms (SNPs) in the FTO gene (rs9940128, rs7193144, rs8050136, rs918031, rs1588413 and rs11076023) identified from recent genome-wide association studies for T2D were genotyped by polymerase chain reaction-restriction fragment length polymorphism and direct sequencing. Dietary assessment was carried out using a validated food frequency questionnaire and physical activity was based upon the self-report. Interaction analyses were performed by including the interaction terms in the model. A joint likelihood ratio test of the main SNP effects and the SNP-diet/physical activity interaction effects was used in the linear regression analyses to maximize statistical power. Statistical analyses were performed using STATA version 13. There was a significant interaction between FTO SNP rs8050136 and carbohydrate energy percentage (Pinteraction=0.04) on obesity, where the ‘A’ allele carriers of the SNP rs8050136 had 2.46 times higher risk of obesity than those with ‘CC’ genotype (P=3.0x10-5) among individuals in the highest tertile of carbohydrate energy percentage. Furthermore, among those who had lower levels of physical activity, the ‘A’ allele carriers of the SNP rs8050136 had 1.89 times higher risk of obesity than those with ‘CC’ genotype (P=4.0x10-5). We also found a borderline interaction between SNP rs11076023 and carbohydrate energy percentage (Pinteraction=0.08) on T2D, where the ‘A’ allele carriers in the highest tertile of carbohydrate energy percentage, had 1.57 times higher risk of T2D than those with ‘TT’ genotype (P=0.002). There was also a significant interaction between SNP rs11076023 and physical activity (Pinteraction=0.03) on T2D. No further significant interactions between SNPs and macronutrient intake or physical activity on obesity and T2D were observed. In conclusion, this is the first study to provide evidence for a gene-diet and gene-physical activity interaction on obesity and T2D in an Asian Indian population. These findings suggest that the association between FTO gene variants and obesity and T2D is influenced by carbohydrate intake and physical activity levels. Greater understanding of how FTO gene influences obesity and T2D through dietary and exercise interventions will advance the development of behavioral intervention and personalised lifestyle strategies predicted to reduce the development of metabolic diseases in ‘A’ allele carriers of both SNPs in this Asian Indian population.

Keywords: dietary intake, FTO, obesity, physical activity, type 2 diabetes, Asian Indian.

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Authors: Ngabirano Levis

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One of Africa’s major environmental challenges remains air pollution. In 2017, UNICEF estimated over 400,000 children in Africa died as a result of indoor pollution, while 350 million children remain exposed to the risks of indoor pollution due to the use of biomass and burning of wood for cooking. Over time, indeed, the major causes of mortality across Africa are shifting from the unsafe water, poor sanitation, and malnutrition to the ambient and household indoor pollution, and greenhouse gas (GHG) emissions remain a key factor in this. In addition, studies by the OECD estimated that the economic cost of premature deaths due to Ambient Particulate Matter Pollution (APMP) and Household Air Pollution across Africa in 2013 was about 215 Billion US Dollars and US 232 Billion US Dollars, respectively. This is not only a huge cost for a continent where over 41% of the Sub-Saharan population lives on less than 1.9 US Dollars a day but also makes the people extremely vulnerable to the negative climate change and environmental degradation effects. Such impacts have led to extended droughts, flooding, health complications, and reduced crop yields hence food insecurity. Climate change, therefore, poses a threat to global targets like poverty reduction, health, and famine. Despite efforts towards mitigation, air contributors like carbon dioxide emissions are on a generally upward trajectory across Africa. In Egypt, for instance, emission levels had increased by over 141% in 2010 from the 1990 baseline. Efforts like the climate change adaptation and mitigation financing have also hit obstacles on the continent. The International Community and developed nations stress that Africa still faces challenges of limited human, institutional and financial systems capable of attracting climate funding from these developed economies. By using the qualitative multi-case study method supplemented by interviews of key actors and comprehensive textual analysis of relevant literature, this paper dissects the key emissions and air pollutant sources, their impact on the well-being of the African people, and puts forward suggestions as well as a remedial mechanism to these challenges. The findings reveal that whereas climate change mitigation plans appear comprehensive and good on paper for many African countries like Uganda; the lingering political interference, limited research guided planning, lack of population engagement, irrational resource allocation, and limited system and personnel capacity has largely impeded the realization of the set targets. Recommendations have been put forward to address the above climate change impacts that threaten the food security, health, and livelihoods of the people on the continent.

Keywords: Africa, air pollution, climate change, mitigation, emissions, effective planning, institutional strengthening

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Authors: Wei Xiao, Gangzhi Cai, Xingliang Qin, Hongyan Ren, Zaidong Hua, Zhe Zhu, Hongwei Xiao, Ximin Zheng, Jie Yao, Yanzhen Bi

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Myostatin (MSTN) is the master regulator of double muscling phenotype with overgrown muscle and decreased fatness in animals, but its action mode to regulate fat deposition remains to be elucidated. In this study a swin-specific pathway through which MSTN acts to regulate the fat deposition was deciphered. Deep sequenincing of the mRNA and miRNA of fat tissues of MSTN knockout (KO) and wildtype (WT) pigs discovered the positive correlation of myocyte enhancer factor 2C (MEF2C) and fat-inhibiting miR222 expression, and the inverse correlation of miR222 and stearoyl-CoA desaturase 5 (SCD5) expression. SCD5 is rodent-absent and expressed only in pig, sheep and cattle. Fatty acid spectrum of fat tissues revealed a lower percentage of oleoyl-CoA (18:1) and palmitoleyl CoA (16:1) in MSTN KO pigs, which are the catalyzing products of SCD5-mediated desaturation of steroyl CoA (18:0) and palmitoyl CoA (16:0). Blood metrics demonstrated a 45% decline of triglyceride (TG) content in MSTN KO pigs. In light of these observations we hypothesized that MSTN might act through MEF2C/miR222/SCD5 pathway to regulate desaturation of fatty acid as well as triglyceride synthesis in pigs. To this end, real-time PCR and Western blotting were carried out to detect the expression of the three genes stated above. These experiments showed that MEF2C expression was up-regulated by nearly 2-fold, miR222 up-regulated by nearly 3-fold and SCD5 down-regulated by nearly 50% in MSTN KO pigs. These data were consistent with the expression change in deep sequencing analysis. Dual luciferase reporter was then used to confirm the regulation of MEF2C upon the promoter of miR222. Ecotopic expression of MEF2C in preadipocyte cells enhanced miR222 expression by 3.48-fold. CHIP-PCR identified a putative binding site of MEF2C on -2077 to -2066 region of miR222 promoter. Electrophoretic mobility shift assay (EMSA) demonstrated the interaction of MEF2C and miR222 promoter in vitro. These data indicated that MEF2C transcriptionally regulates the expression of miR222. Next, the regulation of miR222 on SCD5 mRNA as well as its physiological consequences were examined. Dual luciferase reporter testing revealed the translational inhibition of miR222 upon the 3´ UTR (untranslated region) of SCD5 in preadipocyte cells. Transfection of miR222 mimics and inhibitors resulted in the down-regulation and up-regulation of SCD5 in preadipocyte cells respectively, consistent with the results from reporter testing. RNA interference of SCD5 in preadipocyte cells caused 26.2% reduction of TG, in agreement with the results of TG content in MSTN KO pigs. In summary, the results above supported the existence of a molecular pathway that MSTN signals through MEF2C/miR222/SCD5 to regulate the fat deposition in pigs. This swine-specific pathway offers potential molecular markers for the development and breeding of a new pig line with optimised fatty acid composition. This would benefit human health by decreasing the takeup of saturated fatty acid.

Keywords: fat deposition, MEF2C, miR222, myostatin, SCD5, pig

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Authors: Md. M. Hossain, Regan Roat, Jenica Christopherson, Colette Free, Zhiguang Guo

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Long noncoding RNA (lncRNA) mediated post-transcriptional gene regulation, and their epigenetic landscapes have been shown to be involved in many human diseases. However, their regulation in diabetes through governing islet’s β-cell function and survival needs to be elucidated. Due to the technical and ethical constraints, it is difficult to study their role in β-cell function and survival in human under in vivo condition. In this study, humanized mice have been developed through transplanting human pancreatic islet under the kidney capsule of NOD.SCID mice and induced β-cell death leading to diabetes condition to study lncRNA mediated regulation. For this, human islets from 3 donors (3000 IEQ, purity > 80%) were transplanted under the kidney capsule of STZ induced diabetic NOD.scid mice. After at least 2 weeks of normoglycecemia, lymphocytes from diabetic NOD mice were adoptively transferred and islet grafts were collected once blood glucose reached > 200 mg/dl. RNA from human donor islets, islet grafts from humanized mice with either adoptive lymphocyte transfer (ALT) or PBS control (CTL) were ribodepleted; barcoded fragment libraries were constructed and sequenced on the Ion Proton sequencer. lncRNA expression in isolated human islets, islet grafts from humanized mice with and without induced β-cell death and their regulation in human islets function in vitro under glucose challenge, cytokine mediated inflammation and induced apoptotic condition were investigated. Out of 3155 detected lncRNAs, 299 that highly expressed in islets were found to be significantly downregulated and 224 upregulated in ALT compared to CTL. Most of these are found to be collocated within 5 kb upstream and 1 kb downstream of 788 up- and 624 down-regulated mRNAs. Genomic Regions Enrichment of Annotations Analysis revealed deregulated and collocated genes are related to pancreas endocrine development; insulin synthesis, processing, and secretion; pancreatitis and diabetes. Many of them, that found to be located within enhancer domains for islet specific gene activity, are associated to the deregulation of known islet/βcell specific transcription factors and genes that are important for β-cell differentiation, identity, and function. RNA sequencing analysis revealed aberrant lncRNA expression which is associated to the deregulated mRNAs in β-cell function as well as in molecular pathways related to diabetes. A distinct set of candidate lncRNA isoforms were identified as highly enriched and specific to human islets, which are deregulated in human islets from donors with different BMIs and with type 2 diabetes. These RNAs show an interesting regulation in cultured human islets under glucose stimulation and with induced β-cell death by cytokines. Aberrant expression of these lncRNAs was detected in the exosomes from the media of islets cultured with cytokines. Results of this study suggest that the islet specific lncRNAs are deregulated in human islet with β-cell death, hence important in diabetes. These lncRNAs might be important for human β-cell function and survival thus could be used as biomarkers and novel therapeutic targets for diabetes.

Keywords: β-cell, humanized mouse, pancreatic islet, LncRNAs

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Authors: Oleg Reva, Ilya Korotetskiy, Marina Lankina, Murat Kulmanov, Aleksandr Ilin

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Molecular docking approaches are widely used for design of new antibiotics and modeling of antibacterial activities of numerous ligands which bind specifically to active centers of indispensable enzymes and/or key signaling proteins of pathogens. Widespread drug resistance among pathogenic microorganisms calls for development of new antibiotics specifically targeting important metabolic and information pathways. A generally recognized problem is that almost all molecular targets have been identified already and it is getting more and more difficult to design innovative antibacterial compounds to combat the drug resistance. A promising way to overcome the drug resistance problem is an induction of reversion of drug resistance by supplementary medicines to improve the efficacy of the conventional antibiotics. In contrast to well established computer-based drug design, modeling of drug resistance reversion still is in its infancy. In this work, we proposed an approach to identification of compensatory genetic variants reducing the fitness cost associated with the acquisition of drug resistance by pathogenic bacteria. The approach was based on an analysis of the population genetic of Mycobacterium tuberculosis and on results of experimental modeling of the drug resistance reversion induced by a new anti-tuberculosis drug FS-1. The latter drug is an iodine-containing nanomolecular complex that passed clinical trials and was admitted as a new medicine against MDR-TB in Kazakhstan. Isolates of M. tuberculosis obtained on different stages of the clinical trials and also from laboratory animals infected with MDR-TB strain were characterized by antibiotic resistance, and their genomes were sequenced by the paired-end Illumina HiSeq 2000 technology. A steady increase in sensitivity to conventional anti-tuberculosis antibiotics in series of isolated treated with FS-1 was registered despite the fact that the canonical drug resistance mutations identified in the genomes of these isolates remained intact. It was hypothesized that the drug resistance phenotype in M. tuberculosis requires an adjustment of activities of many genes to compensate the fitness cost of the drug resistance mutations. FS-1 cased an aggravation of the fitness cost and removal of the drug-resistant variants of M. tuberculosis from the population. This process caused a significant increase in genetic heterogeneity of the Mtb population that was not observed in the positive and negative controls (infected laboratory animals left untreated and treated solely with the antibiotics). A large-scale search for linkage disequilibrium associations between the drug resistance mutations and genetic variants in other genomic loci allowed identification of target proteins, which could be influenced by supplementary drugs to increase the fitness cost of the drug resistance and deprive the drug-resistant bacterial variants of their competitiveness in the population. The approach will be used to improve the efficacy of FS-1 and also for computer-based design of new drugs to combat drug-resistant infections.

Keywords: complete genome sequencing, computational modeling, drug resistance reversion, Mycobacterium tuberculosis

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Authors: M. Kazou, A. Gavriil, O. Kalagkatsi, T. Paschos, E. Tsakalidou

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Kopanisti cheese is a Greek soft Protected Designation of Origin (PDO) cheese made of raw cow, sheep or goat milk, or mixtures of them, with similar organoleptic characteristics to that of Roquefort cheese. Traditional manufacturing of Kopanisti cheese is limited in small-scale dairies, without the addition of starter cultures. Instead, an amount of over-mature Kopanisti cheese, called Mana Kopanisti, is used to initiate ripening. Therefore, the selection of proper starter cultures and the understanding of the contribution of various microbial groups to its overall quality is crucial for the production of a high-quality final product with standardized organoleptic and physicochemical characteristics. Taking the above into account, the aim of the present study was the investigation of Kopanisti cheese microbiota and its role in cheese quality. For this purpose, four different types of Kopanisti were produced in triplicates, all with pasteurized cow milk, with the addition of (A) the typical mesophilic species Lactococcus lactis and Lactobacillus paracasei used as starters in the production of soft spread cheeses, (B) strains of Lactobacillus acidipiscis and Lactobacillus rennini previously isolated from Kopanisti and Mana Kopanisti, (C) all the species from (A) and (B) as inoculum, and finally (D) the species from (A) and Mana Kopanisti. Physicochemical and microbiological analysis was performed for milk and cheese samples during ripening. Enumeration was performed for major groups of lactic acid bacteria (LAB), total mesophilic bacteria, yeasts as well as hygiene indicator microorganisms. Bacterial isolates from all the different LAB groups, apart from enterococci, alongside yeasts isolates, were initially grouped using repetitive sequence-based polymerase chain reaction (rep-PCR) and then identified at the species level using 16S rRNA gene and internal transcribed spacer (ITS) DNA region sequencing, respectively. Sensory evaluation was also performed for final cheese samples at the end of the ripening period (35 days). Based on the results of the classical microbiological analysis, the average counts of the total mesophilic bacteria and LAB, apart from enterococci, ranged between 7 and 10 log colony forming unit (CFU) g⁻¹, phychrotrophic bacteria, and yeast extract glucose chloramphenicol (YGC) isolates between 4 and 8 log CFU g⁻¹, while coliforms and enterococci up to 2 log CFU g⁻¹ throughout ripening in cheese samples A, C and D. In contrast, in cheese sample B, the average counts of the total mesophilic bacteria and LAB, apart from enterococci, phychrotrophic bacteria, and YGC isolates ranged between 0 and 10 log CFU g⁻¹ and coliforms and enterococci up to 2 log CFU g⁻¹. Although the microbial counts were not that different among samples, identification of the bacterial and yeasts isolates revealed the complex microbial community structure present in each cheese sample. Differences in the physicochemical characteristics among the cheese samples were also observed, with pH ranging from 4.3 to 5.3 and moisture from 49.6 to 58.0 % in the final cheese products. Interestingly, the sensory evaluation also revealed differences among samples, with cheese sample B ranking first based on the total score. Overall, the combination of these analyses highlighted the impact of different starter cultures on the Kopanisti microbiota as well as on the physicochemical and sensory characteristics of the final product.

Keywords: Kopanisti cheese, microbiota, classical microbiological analysis, physicochemical analysis

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Authors: Alvarez M., Chapela M. J., Balboa E., Rubianes D., Sinde E., Fernandez de Ana C., Rodríguez-Blanco A.

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The gut microbiota plays an important role as gut inflammation could contribute to colorectal cancer development; however, this role is still not fully understood, and tools able to prevent this progression are yet to be developed. The main objective of this study was to monitor the effects of a mushroom extracts formulation in gut microbial community composition of an Azoxymethane (AOM)/Dextran sodium sulfate (DSS) mice model of inflammation-linked carcinogenesis. For the in vivo study, 41 adult male mice of the C57BL / 6 strain were obtained. 36 of them have been induced in a state of colon carcinogenesis by a single intraperitoneal administration of AOM at a dose of 12.5 mg/kg; the control group animals received instead of the same volume of 0.9% saline. DSS is an extremely toxic polysaccharide sulfate that causes chronic inflammation of the colon mucosa, favoring the appearance of severe colitis and the production of tumors induced by AOM. Induction by AOM/DSS is an interesting platform for chemopreventive intervention studies. This time the model was used to monitor gut microbiota changes as a result of supplementation with a specific mushroom extracts formulation previously shown to have prebiotic activity. The animals have been divided into three groups: (i) Cancer + mushroom extracts formulation experimental group: to which the MicoDigest2.0 mushroom extracts formulation developed by Hifas da Terra S.L has been administered dissolved in drinking water at an estimated concentration of 100 mg / ml. (ii) Control group of animals with Cancer: to which normal water has been administered without any type of treatment. (iii) Control group of healthy animals: these are the animals that have not been induced cancer or have not received any treatment in drinking water. This treatment has been maintained for a period of 3 months, after which the animals were sacrificed to obtain tissues that were subsequently analyzed to verify the effects of the mushroom extract formulation. A microbiological analysis has been carried out to compare the microbial communities present in the intestines of the mice belonging to each of the study groups. For this, the methodology of massive sequencing by molecular analysis of the 16S gene has been used (Ion Torrent technology). Initially, DNA extraction and metagenomics libraries were prepared using the 16S Metagenomics kit, always following the manufacturer's instructions. This kit amplifies 7 of the 9 hypervariable regions of the 16S gene that will then be sequenced. Finally, the data obtained will be compared with a database that makes it possible to determine the degree of similarity of the sequences obtained with a wide range of bacterial genomes. Results obtained showed that, similarly to certain natural compounds preventing colorectal tumorigenesis, a mushroom formulation enriched the Firmicutes and Proteobacteria phyla and depleted Bacteroidetes. Therefore, it was demonstrated that the consumption of the mushroom extracts’ formulation developed could promote the recovery of the microbial balance that is disrupted in the mice model of carcinogenesis. More preclinical and clinical studies are needed to validate this promising approach.

Keywords: carcinogenesis, microbiota, mushroom extracts, inflammation

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Authors: Julio Jesus Salazar, Manuel Andres Jesus De Lama

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There are very few models that serve to analyze the use of water in the socio-economic process. On the supply side, the joint use of groundwater has been considered in addition to the simple limits on the availability of surface water. In addition, we have worked on waterlogging and the effects on water quality (mainly salinity). In this paper, a 'complex' water economy is examined; one in which demands grow differentially not only within but also between sectors, and one in which there are limited opportunities to increase consumptive use. In particular, high-value growth, the growth of the production of irrigated crops of high value within the basins of the case study, together with the rapidly growing urban areas, provides a rich context to examine the general problem of water management at the basin level. At the same time, the long-term aridity of nature has made the eco-environment in the basins located on the coast of Peru very vulnerable, and the exploitation and immediate use of water resources have further deteriorated the situation. The presented methodology is the optimization with embedded simulation. The wide basin simulation of flow and water balances and crop growth are embedded with the optimization of water allocation, reservoir operation, and irrigation scheduling. The modeling framework is developed from a network of river basins that includes multiple nodes of origin (reservoirs, aquifers, water courses, etc.) and multiple demand sites along the river, including places of consumptive use for agricultural, municipal and industrial, and uses of running water on the coast of Peru. The economic benefits associated with water use are evaluated for different demand management instruments, including water rights, based on the production and benefit functions of water use in the urban agricultural and industrial sectors. This work represents a new effort to analyze the use of water at the regional level and to evaluate the modernization of the integrated management of water resources and socio-economic territorial development in Peru. It will also allow the establishment of policies to improve the process of implementation of the integrated management and development of water resources. The input-output analysis is essential to present a theory about the production process, which is based on a particular type of production function. Also, this work presents the Computable General Equilibrium (CGE) version of the economic model for water resource policy analysis, which was specifically designed for analyzing large-scale water management. As to the platform for CGE simulation, GEMPACK, a flexible system for solving CGE models, is used for formulating and solving CGE model through the percentage-change approach. GEMPACK automates the process of translating the model specification into a model solution program.

Keywords: water economy, simulation, modeling, integration

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Authors: C. Villamizar, M. Serrato

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In the pursuit of athletic performance, the role of physical training which is determined by a number of charges or taxes on physiological stress and musculoskeletal systems of the human body generated by the intensity and duration is fundamental. Given the physical demands of these activities both training and competitive must take into account the optimal relationship with a straining process recovery post favoring the process of overcompensation which aims to facilitate the return and rising energy potential and protein synthesis also of different tissues. Allowing muscle function returns to baseline or pre-exercise states. If this recovery process is not performed or is not allowed in a proper way, will result in an increased state of fatigue. Active recovery, is one of the strategies implemented in the sport for a return to pre-exercise physiological states. However, there are some adverse assumptions regarding the negative effects, as is the possibility of increasing the degradation of muscle glycogen and thus delaying the synthesis thereof. For them, it is necessary to investigate what would be the effects generated application made at different times after the effort. The aim of this study was to determine the effects of active recovery post effort made at three different times: immediately, at 12 and 24 hours on biochemical markers creatine kinase in youth soccer player’s categories. A randomized controlled trial with allocation to three groups was performed: A. active recovery immediately after the effort; B. active recovery performed at 12 hours after the effort; C. active recovery made at 24 hours after the effort. This study included 27 subjects belonging to a Colombian soccer team of the second division. Vital signs, weight, height, BMI, the percentage of muscle mass, fat mass percentage, personal medical history, and family were valued. The velocity, explosive force and Creatin Kinase (CK) in blood were tested before and after interventions. SAFT 90 protocol (Soccer Field specific Aerobic Test) was applied to participants for generating fatigue. CK samples were taken one hour before the application of the fatigue test, one hour after the fatigue protocol and 48 of the initial CK sample. Mean age was 18.5 ± 1.1 years old. Improvements in jumping and speed recovery the 3 groups (p < 0.05), but no statistically significant differences between groups was observed after recuperation. In all participants, there was a significant increment of CK when applied SAFT 90 in all the groups (median 103.1-111.1). The CK measurement after 48 hours reflects a recovery in all groups, however the group C, a decline below baseline levels of -55.5 (-96.3 /-20.4) which is a significant find. Other research has shown that CK does not return quickly to their baseline, but our study shows that active recovery favors the clearance of CK and also to perform recovery 24 hours after the effort generates higher clearance of this biomarker.

Keywords: active recuperation, creatine phosphokinase, post training, young soccer players

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Authors: Anna Solovyeva, Ivan Levakin, Nickolai Galaktionov, Olga Podgornaya

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Animals that reproduce asexually were thought to have the same genotypes within generations for a long time. However, some refuting examples were found, and mobile genetic elements (MGEs) or transposons are considered to be the most probable source of genetic instability. Dispersed nature and the ability to change their genomic localization enables MGEs to be efficient mutators. Hence the study of MGEs genomic impact requires an appropriate object which comprehends both representative amounts of various MGEs and options to evaluate the genomic influence of MGEs. Animals that reproduce asexually seem to be a decent model to study MGEs impact in genomic variability. We found a small marine trematode Himasthla elongata (Himasthlidae) to be a good model for such investigation as it has a small genome size, diverse MGEs and parthenogenetic stages in the lifecycle. In the current work, clonal diversity of cercaria was traced with an AFLP (Amplified fragment length polymorphism) method, diverse zones from electrophoretic patterns were cloned, and the nature of the fragments explored. Polymorphic patterns of individual cercariae AFLP-based fingerprints are enriched with retrotransposons of different families. The bulk of those sequences are represented by open reading frames of non-Long Terminal Repeats containing elements(non-LTR) yet Long-Terminal Repeats containing elements (LTR), to a lesser extent in variable figments of AFLP array. The CR1 elements expose both in polymorphic and conservative patterns are remarkably more frequent than the other non-LTR retrotransposons. This data was confirmed with shotgun sequencing-based on Illumina HiSeq 2500 platform. Individual cercaria of the same clone (i.e., originated from a single miracidium and inhabiting one host) has a various distribution of MGE families detected in sequenced AFLP patterns. The most numerous are CR1 and RTE-Bov retrotransposons, typical for trematode genomes. Also, we identified LTR-retrotransposons of Pao and Gypsy families among DNA transposons of CMC-EnSpm, Tc1/Mariner, MuLE-MuDR and Merlin families. We detected many of them in H. elongata transcriptome. Such uneven MGEs distribution in AFLP sequences’ sets reflects the different patterns of transposons spreading in cercarial genomes as transposons affect the genome in many ways (ectopic recombination, gene structure interruption, epigenetic silencing). It is considered that they play a key role in the origins of trematode clonal polymorphism. The authors greatly appreciate the help received at the Kartesh White Sea Biological Station of the Russian Academy of Sciences Zoological Institute. This work is funded with RSF 19-74-20102 and RFBR 17-04-02161 grants and the research program of the Zoological Institute of the Russian Academy of Sciences (project number AAAA-A19-119020690109-2).

Keywords: AFLP, clonal polymorphism, Himasthla elongata, mobile genetic elements, NGS

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Authors: Oliver Fuo

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One of the central features of post-apartheid constitutional reform was the establishment of local government as a distinct sphere of government in the Constitution of the Republic of South Africa, 1996. Local government, constituted by about 279 wall-to-wall municipalities, have legislative and executive powers vested in democratically elected municipal councils to govern areas within their jurisdiction subject only to limits imposed by the Constitution. In addition, unlike the past where municipalities merely played a service delivery role, they are now mandated to realise an expanded developmental mandate – pursue social justice and sustainable development; contribute, together with national and provincial government, to the realisation of socio-economic rights entrenched in the Bill of Rights; and facilitate public participation in local governance. In order to finance their developmental programmes, municipalities receive equitable allocations from national government and have legal powers to generate additional finances by charging rates on property and imposing surcharges on services provided. In addition to its general obligation to foster public participation in local governance, the law requires municipalities to facilitate public participation in their budgeting processes. This requirement is generally consistent with recent trends in local government democratic reforms which call for inclusive budget planning and implementation whereby citizens, civil society and NGOs participate in the allocation of resources. This trend is best captured in the concept of participatory budgeting. This paper specifically analyses the legal and policy framework for participatory budgeting at the local government level in South Africa. Using Borbet South Africa (Pty) Ltd and Others v Nelson Mandela Bay Municipality 2014 (5) SA 256 (ECP) as an example, this paper argues that the legal framework for participatory budgeting creates an illusory right for citizens to participate in municipal budgeting processes. This challenge is further compounded by the barrenness of the jurisprudence of courts that interpret the obligation of municipalities in this regard. It is submitted that the wording of s 27(4) of the Municipal Finance Management Act (MFMA) 53 of 2003 - which expressly stipulates that non-compliance by a municipality with a provision relating to the budget process or a provision in any legislation relating to the approval of a budget-related policy, does not affect the validity of an annual or adjustments budget – is problematic as it seems to trivialise the obligation to facilitate public participation in budgeting processes. It is submitted that where this provision is abused by municipal officials, this could lead to the sidelining of the real interests of communities in local budgets. This research is based on a critical and integrated review of primary and secondary sources of law.

Keywords: courts and jurisprudence, local government law, participatory budgeting, South Africa

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Authors: Shagufta Jabeen, Faez Jesse Firdaus Abdullah, Zunita Zakaria, Nurulfiza Mat Isa, Yung Chie Tan, Wai Yan Yee, Abdul Rahman Omar

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Pasteurella multocida is associated with acute, as well as, chronic infections in avian and bovine such as pasteurellosis and hemorrhagic septicemia (HS) in cattle and buffaloes. Iron is one of the most important nutrients for pathogenic bacteria including Pasteurella and acts as a cofactor or prosthetic group in several essential enzymes and is needed for amino acid, pyrimidine, and DNA biosynthesis. In our recent study, we showed that 2% of Pasteurella multocida serotype A strain PMTB2.1 encode for iron regulating genes (Accession number CP007205.1). Genome sequencing of other Pasteurella multocida serotypes namely PM70 and HB01 also indicated up to 2.5% of the respective genome encode for iron regulating genes, suggesting that Pasteurella multocida genome comprises of multiple systems for iron uptake. Since P. multocida PMTB2.1 has more than 40 CDs out of 2097 CDs (approximately 2%), encode for iron-regulated. The gene expression profiling of four iron-regulating genes namely fbpb, yfea, fece and fur were characterized under iron-restricted environment. The P. multocida strain PMTB2.1 was grown in broth with and without iron chelating agent and samples were collected at different time points. Relative mRNA expression profile of these genes was determined using Taqman probe based real-time PCR assay. The data analysis, normalization with two house-keeping genes and the quantification of fold changes were carried out using Bio-Rad CFX manager software version 3.1. Results of this study reflect that iron reduced environment has significant effect on expression profile of iron regulating genes (p < 0.05) when compared to control (normal broth) and all evaluated genes act differently with response to iron reduction in media. The highest relative fold change of fece gene was observed at early stage of treatment indicating that PMTB2.1 may utilize its periplasmic protein at early stage to acquire iron. Furthermore, down-regulation expression of fece with the elevated expression of other genes at later time points suggests that PMTB2.1 control their iron requirements in response to iron availability by down-regulating the expression of iron proteins. Moreover, significantly high relative fold change (p ≤ 0.05) of fbpb gene is probably associated with the ability of P. multocida to directly use host iron complex such as hem, hemoglobin. In addition, the significant increase (p ≤ 0.05) in fbpb and yfea expressions also reflects the utilization of multiple iron systems in P. multocida strain PMTB2.1. The findings of this study are very much important as relative scarcity of free iron within hosts creates a major barrier to microbial growth inside host and utilization of outer-membrane proteins system in iron acquisition probably occurred at early stage of infection with P. multocida. In conclusion, the presence and utilization of multiple iron system in P. multocida strain PMTB2.1 revealed the importance of iron in the survival of P. multocida.

Keywords: iron-related genes, real-time PCR, gene expression profiling, fold changes

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Authors: Seungsik Yu, Minho Kang

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As the threat of terrorism to nuclear facilities is increasing globally after the attacks of September 11, we are striving to recognize the physical protection system and strengthen the emergency response system. Since 2015, Korea has implemented physical protection security exercise for nuclear facilities. The exercise should be carried out with full cooperation between the operator and response forces. Performance testing of the physical protection system should include appropriate exercises, for example, force-on-force exercises, to determine if the response forces can provide an effective and timely response to prevent sabotage. Significant deficiencies and actions taken should be reported as stipulated by the competent authority. The IAEA(International Atomic Energy Agency) is also preparing force-on-force exercise program documents to support exercise of member states. Currently, ROK(Republic of Korea) is implementing exercise on the force-on-force exercise evaluation system which is developed by itself for the nuclear power plant, and it is necessary to establish the exercise procedure considering the use of the force-on-force exercise evaluation system. The purpose of this study is to establish the work procedures of the three major organizations related to the force-on-force exercise of nuclear power plants in ROK, which conduct exercise using force-on-force exercise evaluation system. The three major organizations are composed of licensee, KINAC (Korea Institute of Nuclear Nonproliferation and Control), and the NSSC(Nuclear Safety and Security Commission). Major activities are as follows. First, the licensee establishes and conducts an exercise plan, and when recommendations are derived from the result of the exercise, it prepares and carries out a force-on-force result report including a plan for implementation of the recommendations. Other detailed tasks include consultation with surrounding units for adversary, interviews with exercise participants, support for document evaluation, and self-training to improve the familiarity of the MILES (Multiple Integrated Laser Engagement System). Second, KINAC establishes a force-on-force exercise plan review report and reviews the force-on-force exercise plan report established by licensee. KINAC evaluate force-on-force exercise using exercise evaluation system and prepare training evaluation report. Other detailed tasks include MILES training, adversary consultation, management of exercise evaluation systems, and analysis of exercise evaluation results. Finally, the NSSC decides whether or not to approve the force-on-force exercise and makes a correction request to the nuclear facility based on the exercise results. The most important part of ROK's force-on-force exercise system is the analysis through the exercise evaluation system implemented by KINAC after the exercise. The analytical method proceeds in the order of collecting data from the exercise evaluation system and analyzing the collected data. The exercise application process of the exercise evaluation system introduced in ROK in 2016 will be concretely set up, and a system will be established to provide objective and consistent conclusions between exercise sessions. Based on the conclusions drawn up, the ultimate goal is to complement the physical protection system of licensee so that the system makes licensee respond effectively and timely against sabotage or unauthorized removal of nuclear materials.

Keywords: Force-on-Force exercise, nuclear power plant, physical protection, sabotage, unauthorized removal

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Authors: A. Odoom, A. Salama, H. Ibrahim

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Hydrogen is a clean source of energy for power production and transportation. The main source of hydrogen in this research is biodiesel. Glycerol also called glycerine is a by-product of biodiesel production by transesterification of vegetable oils and methanol. This is a reliable and environmentally-friendly source of hydrogen production than fossil fuels. A typical composition of crude glycerol comprises of glycerol, water, organic and inorganic salts, soap, methanol and small amounts of glycerides. Crude glycerol has limited industrial application due to its low purity thus, the usage of crude glycerol can significantly enhance the sustainability and production of biodiesel. Reforming techniques is an approach for hydrogen production mainly Steam Reforming (SR), Autothermal Reforming (ATR) and Partial Oxidation Reforming (POR). SR produces high hydrogen conversions and yield but is highly endothermic whereas POR is exothermic. On the downside, PO yields lower hydrogen as well as large amount of side reactions. ATR which is a fusion of partial oxidation reforming and steam reforming is thermally neutral because net reactor heat duty is zero. It has relatively high hydrogen yield, selectivity as well as limits coke formation. The complex chemical processes that take place during the production phases makes it relatively difficult to construct a reliable and robust numerical model. Numerical model is a tool to mimic reality and provide insight into the influence of the parameters. In this work, we introduce a finite volume numerical study for an 'in-house' lab-scale experiment of ATR. Previous numerical studies on this process have considered either using Comsol or nodal finite difference analysis. Since Comsol is a commercial package which is not readily available everywhere and lab-scale experiment can be considered well mixed in the radial direction. One spatial dimension suffices to capture the essential feature of ATR, in this work, we consider developing our own numerical approach using MATLAB. A continuum fixed bed reactor is modelled using MATLAB with both pseudo homogeneous and heterogeneous models. The drawback of nodal finite difference formulation is that it is not locally conservative which means that materials and momenta can be generated inside the domain as an artifact of the discretization. Control volume, on the other hand, is locally conservative and suites very well problems where materials are generated and consumed inside the domain. In this work, species mass balance, Darcy’s equation and energy equations are solved using operator splitting technique. Therefore, diffusion-like terms are discretized implicitly while advection-like terms are discretized explicitly. An upwind scheme is adapted for the advection term to ensure accuracy and positivity. Comparisons with the experimental data show very good agreements which build confidence in our modeling approach. The models obtained were validated and optimized for better results.

Keywords: autothermal reforming, crude glycerol, hydrogen, numerical model

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Authors: Binder Hans

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Cancer is no longer seen as solely a genetic disease where genetic defects such as mutations and copy number variations affect gene regulation and eventually lead to aberrant cell functioning which can be monitored by transcriptome analysis. It has become obvious that epigenetic alterations represent a further important layer of (de-)regulation of gene activity. For example, aberrant DNA methylation is a hallmark of many cancer types, and methylation patterns were successfully used to subtype cancer heterogeneity. Hence, unraveling the interplay between different omics levels such as genome, transcriptome and epigenome is inevitable for a mechanistic understanding of molecular deregulation causing complex diseases such as cancer. This objective requires powerful downstream integrative bioinformatics methods as an essential prerequisite to discover the whole genome mutational, transcriptome and epigenome landscapes of cancer specimen and to discover cancer genesis, progression and heterogeneity. Basic challenges and tasks arise ‘beyond sequencing’ because of the big size of the data, their complexity, the need to search for hidden structures in the data, for knowledge mining to discover biological function and also systems biology conceptual models to deduce developmental interrelations between different cancer states. These tasks are tightly related to cancer biology as an (epi-)genetic disease giving rise to aberrant genomic regulation under micro-environmental control and clonal evolution which leads to heterogeneous cellular states. Machine learning algorithms such as self organizing maps (SOM) represent one interesting option to tackle these bioinformatics tasks. The SOMmethod enables recognizing complex patterns in large-scale data generated by highthroughput omics technologies. It portrays molecular phenotypes by generating individualized, easy to interpret images of the data landscape in combination with comprehensive analysis options. Our image-based, reductionist machine learning methods provide one interesting perspective how to deal with massive data in the discovery of complex diseases, gliomas, melanomas and colon cancer on molecular level. As an important new challenge, we address the combined portrayal of different omics data such as genome-wide genomic, transcriptomic and methylomic ones. The integrative-omics portrayal approach is based on the joint training of the data and it provides separate personalized data portraits for each patient and data type which can be analyzed by visual inspection as one option. The new method enables an integrative genome-wide view on the omics data types and the underlying regulatory modes. It is applied to high and low-grade gliomas and to melanomas where it disentangles transversal and longitudinal molecular heterogeneity in terms of distinct molecular subtypes and progression paths with prognostic impact.

Keywords: integrative bioinformatics, machine learning, molecular mechanisms of cancer, gliomas and melanomas

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Authors: Gideon J. Bonthuys, Marco van Dijk, Jay N. Bhagwan

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Small scale hydropower used to play a very important role in the provision of energy to urban and rural areas of South Africa. The national electricity grid, however, expanded and offered cheap, coal generated electricity and a large number of hydropower systems were decommissioned. Unfortunately, large numbers of households and communities will not be connected to the national electricity grid for the foreseeable future due to high cost of transmission and distribution systems to remote communities due to the relatively low electricity demand within rural communities and the allocation of current expenditure on upgrading and constructing of new coal fired power stations. This necessitates the development of feasible alternative power generation technologies. A feasibility and implementation model was developed to assist in designing and financially evaluating small-scale hydropower (SSHP) plants. Several sites were identified using the model. The SSHP plants were designed for the selected sites and the designs for the different selected sites were priced using pricing models (civil, mechanical and electrical aspects). Following feasibility studies done on the designed and priced SSHP plants, a feasibility analysis was done and a design chart developed for future similar potential SSHP plant projects. The methodology followed in conducting the feasibility analysis for other potential sites consisted of developing cost and income/saving formulae, developing net present value (NPV) formulae, Capital Cost Comparison Ratio (CCCR) and levelised cost formulae for SSHP projects for the different types of plant installations. It included setting up a model for the development of a design chart for a SSHP, calculating the NPV, CCCR and levelised cost for the different scenarios within the model by varying different parameters within the developed formulae, setting up the design chart for the different scenarios within the model and analyzing and interpreting results. From the interpretation of the develop design charts for feasible SSHP in can be seen that turbine and distribution line cost are the major influences on the cost and feasibility of SSHP. High head, short transmission line and islanded mini-grid SSHP installations are the most feasible and that the levelised cost of SSHP is high for low power generation sites. The main conclusion from the study is that the levelised cost of SSHP projects indicate that the cost of SSHP for low energy generation is high compared to the levelised cost of grid connected electricity supply; however, the remoteness of SSHP for rural electrification and the cost of infrastructure to connect remote rural communities to the local or national electricity grid provides a low CCCR and renders SSHP for rural electrification feasible on this basis.

Keywords: cost, feasibility, rural electrification, small-scale hydropower

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Authors: Naziema Begum Jappie

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The internationalization of higher education has become a focal point for academic institutions worldwide, including those in South Africa. This paper explores the multifaceted implications of internationalization on management and practice within the South African higher education landscape. Universities all over the world are increasingly recognizing the challenges of globalization and the pressures towards internationalization. Internationalization in higher education encompasses a range of activities, including academic exchange programs, research collaborations, joint degree programs, and the recruitment of international students and faculty. In South Africa, this process is driven by various factors, including the quest for global competitiveness, the pursuit of academic excellence, and the promotion of cultural diversity. However, while internationalization presents numerous opportunities, it also brings forth significant challenges that require careful consideration by management and practitioners in higher education institutions. Furthermore, the internationalization of higher education in South Africa has significant implications for teaching and learning practices. With an increasingly diverse student body, educators must employ innovative pedagogical approaches that cater to the needs and preferences of a multicultural cohort. This may involve the integration of global perspectives into the curriculum, the use of technology-enhanced learning platforms, and the promotion of intercultural competence among students and faculty. Additionally, the exchange of knowledge and ideas with international partners can enrich research activities and contribute to the advancement of knowledge in various fields. The internationalization of higher education in South Africa has profound implications for management and practice within academic institutions. While it offers opportunities for enhancing academic quality, promoting cultural exchange, and advancing research agendas, it also presents challenges that require strategic planning, resource allocation, and stakeholder engagement. By addressing these challenges proactively and leveraging the opportunities presented by internationalization, South African universities can position themselves as global leaders in higher education while contributing to the socio-economic development of the country and the continent at large. This paper draws together the international experience in South Africa to explore the emerging patterns of strategy and practice in internationalizing Higher Education and will highlight some critical notions of how the concepts of internationalization and globalization in the context of higher education are understood by those who lead universities and what new challenges are being created as universities seek to become more international. Institutions cannot simply have bullet points in the strategic plan for the recruitment of international students; there has to be a complete commitment to a national strategy of inclusivity. This paper will further examine the leadership styles that ensure transformation together with the goals set out for internationalization. Discussions around adding the international relations dimension to the curriculum. Addressing the issues relevant to cross-border delivery of higher education.

Keywords: challenges, higher education, internationalization, strategic focus

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Authors: Ombir Singh

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In spite of the religious and cultural pre-dominance of the river Ganga in the Indian ethos, fragmentation and degradation of the river continued down the ages. Recognizing the national concern on environmental degradation of the river and its basin, Ministry of Water Resources, River Development & Ganga Rejuvenation (MoWR,RD&GR), Government of India has initiated a number of pilot schemes for the rejuvenation of river Ganga under the ‘Namami Gange’ Programme. Considering the diversity, complexity, and intricacies of forest ecosystems and pivotal multiple functions performed by them and their inter-connectedness with highly dynamic river ecosystems, forestry interventions all along the river Ganga from its origin at Gaumukh, Uttarakhand to its mouth at Ganga Sagar, West Bengal has been planned by the ministry. For that Forest Research Institute (FRI) in collaboration with National Mission for Clean Ganga (NMCG) has prepared a Detailed Project Report (DPR) on Forestry Interventions for Ganga. The Institute has adopted an extensive consultative process at the national and state levels involving various stakeholders relevant in the context of river Ganga and employed a science-based methodology including use of remote sensing and GIS technologies for geo-spatial analysis, modeling and prioritization of sites for proposed forestation and conservation interventions. Four sets of field data formats were designed to obtain the field based information for forestry interventions, mainly plantations and conservation measures along the river course. In response, five stakeholder State Forest Departments had submitted more than 8,000 data sheets to the Institute. In order to analyze a voluminous field data received from five participating states, the Institute also developed a software to collate, analyze and generation of reports on proposed sites in Ganga basin. FRI has developed potential plantation and treatment models for the proposed forestry and other conservation measures in major three types of landscape components visualized in the Ganga riverscape. These are: (i) Natural, (ii) Agriculture, and (iii) Urban Landscapes. Suggested plantation models broadly varied for the Uttarakhand Himalayas and the Ganga Plains in five participating states. Besides extensive plantations in three type of landscapes within the riverscape, various conservation measures such as soil and water conservation, riparian wildlife management, wetland management, bioremediation and bio-filtration and supporting activities such as policy and law intervention, concurrent research, monitoring and evaluation, and mass awareness campaigns have been envisioned in the DPR. The DPR also incorporates the details of the implementation mechanism, budget provisioned for different components of the project besides allocation of budget state-wise to five implementing agencies, national partner organizations and the Nodal Ministry.

Keywords: conservation, Ganga, river, water, forestry interventions

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Authors: Bello Gonzalez T. D. J., Setten Van M., Essen Van A., Brouwer M., Veldman K. T.

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Resistance to fluoroquinolones (FQ) has evolved increasingly over the years, posing a significant challenge for the treatment of human infections, particularly gastrointestinal tract infections caused by zoonotic bacteria transmitted through the food chain and environment. In broiler chickens, a relatively high proportion of FQ resistance has been observed in Escherichia coli indicator, Salmonella and Campylobacter isolates. We hypothesize that flumequine (Flu), used as a secondary choice for the treatment of poultry infections, could potentially be associated with a high proportion of FQ resistance. To evaluate this hypothesis, we used an in vitro fermentation chicken caeca model. Two continuous single-stage fermenters were used to simulate in real time the physiological conditions of the chicken caeca microbial content (temperature, pH, caecal content mixing, and anoxic environment). A pool of chicken caecal content containing FQ-resistant E. coli obtained from chickens at slaughter age was used as inoculum along with a spiked FQ-susceptible Campylobacter jejuni strain isolated from broilers. Flu was added to one of the fermenters (Flu-fermenter) every 24 hours for two days to evaluate the selection and maintenance of FQ resistance over time, while the other served as a control (C-Fermenter). The experiment duration was 5 days. Samples were collected at three different time points: before, during and after Flu administration. Serial dilutions were plated on Butzler culture media with and without Flu (8mg/L) and enrofloxacin (4mg/L) and on MacConkey culture media with and without Flu (4mg/L) and enrofloxacin (1mg/L) to determine the proportion of resistant strains over time. Positive cultures were identified by mass spectrometry and matrix-assisted laser desorption/ionization (MALDI). A subset of the obtained isolates were used for Whole Genome Sequencing analysis. Over time, E. coli exhibited positive growth in both fermenters, while C. jejuni growth was detected up to day 3. The proportion of Flu-resistant E. coli strains recovered remained consistent over time after antibiotic selective pressure, while in the C-fermenter, a decrease was observed at day 5; a similar pattern was observed in the enrofloxacin-resistant E. coli strains. This suggests that Flu might play a role in the selection and persistence of enrofloxacin resistance, compared to C-fermenter, where enrofloxacin-resistant E. coli strains appear at a later time. Furthermore, positive growth was detected from both fermenters only on Butzler plates without antibiotics. A subset of C. jejuni strains from the Flu-fermenter revealed that those strains were susceptible to ciprofloxacin (MIC < 0.12 μg/mL). A selection of E. coli strains from both fermenters revealed the presence of plasmid-mediated quinolone resistance (PMQR) (qnr-B19) in only one strain from the C-fermenter belonging to sequence type (ST) 48, and in all from Flu-fermenter belonged to ST189. Our results showed that Flu selective impact on PMQR-positive E. coli strains, while no effect was observed in C. jejuni. Maintenance of Flu-resistance was correlated with antibiotic selective pressure. Further studies into antibiotic resistance gene transfer among commensal and zoonotic bacteria in the chicken caeca content may help to elucidate the resistance spread mechanisms.

Keywords: fluoroquinolone-resistance, escherichia coli, campylobacter jejuni, in vitro model

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Authors: Dalia Ambrazaitienė, Monika Vilkienė, Danute Karcauskienė, Gintaras Siaudinis

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The soil is a complex ecosystem that is part of our biosphere. The ability of soil to provide ecosystem services is dependent on microbial diversity. T Tillage is one of the major factors that affect soil properties. The no-till systems or shallow ploughless tillage are opposite of traditional deep ploughing, no-tillage systems, for instance, increase soil organic matter by reducing mineralization rates and stimulating litter concentrations of the top soil layer, whereas deep ploughing increases the biological activity of arable soil layer and reduces the incidence of weeds. The role of soil organisms is central to soil processes. Although the number of microbial species in soil is still being debated, the metagenomic approach to estimate microbial diversity predicted about 2000 – 18 000 bacterial genomes in 1 g of soil. Despite the key role of bacteria in soil processes, there is still lack of information about the bacterial diversity of soils as affected by tillage practices. This study focused on metagenomic analysis of bacterial diversity in long-term experimental plots of Dystric Epihypogleyic Albeluvisols in western part of Lithuania. The experiment was set up in 2013 and had a split-plot design where the whole-plot treatments were laid out in a randomized design with three replicates. The whole-plot treatments consisted of two tillage methods - deep ploughing (22-25 cm) (DP), ploughless tillage (7-10 cm) (PT). Three subsamples (0-20 cm) were collected on October 22, 2015 for each of the three replicates. Subsamples from the DP and PT systems were pooled together wise to make two composition samples, one representing deep ploughing (DP) and the other ploughless tillage (PT). Genomic DNA from soil sample was extracted from approximately 200 mg field-moist soil by using the D6005 Fungal/Bacterial Miniprep set (Zymo Research®) following the manufacturer’s instructions. To determine bacterial diversity and community composition, we employed a culture – independent approach of high-throughput pyrosequencing of the 16S rRNA gene. Metagenomic sequencing was made with Illumina MiSeq platform in Base Clear Company. The microbial component of soil plays a crucial role in cycling of nutrients in biosphere. Our study was a preliminary attempt at observing bacterial diversity in soil under two common but contrasting tillage practices. The number of sequenced reads obtained for PT (161 917) was higher than DP (131 194). The 10 most abundant genus in soil sample were the same (Arthrobacter, Candidatus Saccharibacteria, Actinobacteria, Acidobacterium, Mycobacterium, Bacillus, Alphaproteobacteria, Longilinea, Gemmatimonas, Solirubrobacter), just the percent of community part was different. In DP the Arthrobacter and Acidobacterium consist respectively 8.4 % and 2.5%, meanwhile in PT just 5.8% and 2.1% of all community. The Nocardioides and Terrabacter were observed just in PT. This work was supported by the project VP1-3.1-ŠMM-01-V-03-001 NKPDOKT and National Science Program: The effect of long-term, different-intensity management of resources on the soils of different genesis and on other components of the agro-ecosystems [grant number SIT-9/2015] funded by the Research Council of Lithuania.

Keywords: deep ploughing, metagenomics, ploughless tillage, soil community analysis

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Authors: Maria C. Almario, Pam Remer, Jeff Resse, Kathy Moran, Linda Theander Adam

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The identification of victims of human trafficking and consequential service provision is characterized by a significant disconnection between the estimated prevalence of this issue and the number of cases identified. This poses as tremendous problem for human rights advocates as it prevents data collection, information sharing, allocation of resources and opportunities for international dialogues. The current paper introduces the Inclusive Human Trafficking Checklist (IHTC) as a measurement methodology with theoretical underpinnings derived from dialectic theory. The presence of human trafficking in a person’s life is conceptualized as a dynamic and dialectic interaction between vulnerability and exploitation. The current papers explores the operationalization of exploitation and vulnerability, evaluates the metric qualities of the instrument, evaluates whether there are differences in assessment based on the participant’s profession, level of knowledge, and training, and assesses if users of the instrument perceive it as useful. A total of 201 participants were asked to rate three vignettes predetermined by experts to qualify as a either human trafficking case or not. The participants were placed in three conditions: business as usual, utilization of the IHTC with and without training. The results revealed a statistically significant level of agreement between the expert’s diagnostic and the application of the IHTC with an improvement of 40% on identification when compared with the business as usual condition While there was an improvement in identification in the group with training, the difference was found to have a small effect size. Participants who utilized the IHTC showed an increased ability to identify elements of identity-based vulnerabilities as well as elements of fraud, which according to the results, are distinctive variables in cases of human trafficking. In terms of the perceived utility, the results revealed higher mean scores for the groups utilizing the IHTC when compared to the business as usual condition. These findings suggest that the IHTC improves appropriate identification of cases and that it is perceived as a useful instrument. The application of the IHTC as a multidisciplinary instrumentation that can be utilized in legal and human services settings is discussed as a pivotal piece of helping victims restore their sense of dignity, and advocate for legal, physical and psychological reparations. It is noteworthy that this study was conducted with a sample in the United States and later re-tested in Colombia. The implications of the instrument for treatment conceptualization and intervention in human trafficking cases are discussed as opportunities for enhancement of victim well-being, restoration engagement and activism. With the idea that what is personal is also political, we believe that the careful observation and data collection in specific cases can inform new areas of human rights activism.

Keywords: exploitation, human trafficking, measurement, vulnerability, screening

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Authors: N. Perini, M. C. Thaller, F. Mercuri, S. Orlanducci, A. Rubechini, L. Migliore

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The preservation of cultural heritage is one of the major challenges of today’s society, because of the fundamental right of future generations to inherit it as the continuity with their historical and cultural identity. Parchments, consisting of a semi-solid matrix of collagen produced from animal skin (i.e., sheep or goats), are a significant part of the cultural heritage, being used as writing material for many centuries. Due to their animal origin, parchments easily undergo biodeterioration. The most common biological damage is characterized by isolated or coalescent purple spots that often leads to the detachment of the superficial layer and the loss of the written historical content of the document. Although many parchments with the same biodegradative features were analyzed, no common causative agent has been found so far. Very recently, a study was performed on a purple-damaged parchment roll dated back 1244 A.D, the A.A. Arm. I-XVIII 3328, belonging to the oldest collection of the Vatican Secret Archive (Fondo 'Archivum Arcis'), by comparing uncolored undamaged and purple damaged areas of the same document. As a whole, the study gave interesting results to hypothesize a model of biodeterioration, consisting of a microbial succession acting in two main phases: the first one, common to all the damaged parchments, is characterized by halophilic and halotolerant bacteria fostered by the salty environment within the parchment maybe induced by bringing of the hides; the second one, changing with the individual history of each parchment, determines the identity of its colonizers. The design of this model was pivotal to this study, performed by different labs of the Tor Vergata University (Rome, Italy), in collaboration with the Vatican Secret Archive. Three documents, belonging to a collection of dramatically damaged parchments archived as 'Faldone Patrizi A 19' (dated back XVII century A.D.), were analyzed through a multidisciplinary approach, including three updated technologies: (i) Next Generation Sequencing (NGS, Illumina) to describe the microbial communities colonizing the damaged and undamaged areas, (ii) RAMAN spectroscopy to analyze the purple pigments, (iii) Light Transmitted Analysis (LTA) to evaluate the kind and entity of the damage to native collagen. The metagenomic analysis obtained from NGS revealed DNA sequences belonging to Halobacterium salinarum mainly in the undamaged areas. RAMAN spectroscopy detected pigments within the purple spots, mainly bacteriorhodopsine/rhodopsin-like pigments, a purple transmembrane protein containing retinal and present in Halobacteria. The LTA technique revealed extremely damaged collagen structures in both damaged and undamaged areas of the parchments. In the light of these data, the study represents a first confirmation of the microbial succession model described above. The demonstration of this model is pivotal to start any possible new restoration strategy to bring back historical parchments to their original beauty, but also to open opportunities for intervention on a huge amount of documents.

Keywords: biodeterioration, parchments, purple spots, ecological succession

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