Search results for: biological control agent

Authors: Rini Rahul, Manoj Kumar

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Cadmium (Cd) is a poisonous trace metal, which is responsible for excess reactive oxygen species generation (ROS) in plants, thereby adversely affecting their productivity and commercial potential. Ricinus communis (castor) is an industry-efficient non-edible bioenergy crop used for phytoremediation and re-vegetation. We have determined the total Cd content in castor genotypes and established a relationship between the Cd tolerance mechanism and physiological parameters like chlorophyll fluorescence, the total photosynthetic activity, chlorophyll and carotenoid content as well as ROS generation and malondialdehyde content. This study is an effort to comprehend the interrelation between Cd toxicity (control, 250 µM and 500 µM), proline, various ROS scavenging enzymes (anti-oxidative in nature), nicotianamine synthase (NAS) and Natural resistance-associated macrophage protein (NRAMP) gene. The antioxidant enzyme activity increased for WM hence conferring Cd toxicity in this genotype. RcNRAMP genes showed differential expression in GCH2 and WM genotypes; this can also be one of the reasons for Cd toxicity and sensitivity in WM and GCH2, respectively. The cause of pronounced Cd tolerance in WM leaves can be because of enhanced expression of RcNAS1, RcNAS2 and RcNAS3 genes. Our results demonstrate that there is an interrelation between Cd toxicity (control, 250 µM and 500 µM), proline, various ROS scavenging enzymes (anti-oxidative in nature), NAS and NRAMP gene.

Keywords: ricinus communis, cadmium, reactive oxygen species, nicotianamine synthase, NRAMP, malondialdehyde

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Authors: M. R. Mogarekar, Shraddha V. More, Pankaj Kumar

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Gastroenteritis, the third leading killer of children in India today is responsible for 13% of all deaths in children <5 years of age and kills an estimated 300,000 children in India each year. We decided to investigate parameters which can help in early disease detection and prompt treatment. Serum paraoxonase is calcium dependent esterase which is widely distributed among tissues such as liver, kidney, and intestine and is located in the chromosomal region 7q21.3 22.1. Studies show the presence of excessive reactive oxygen metabolites and antioxidant imbalance in the gastrointestinal tract leading to oxidative stress in gastroenteritis. To our knowledge, this is the first ever study done. The objective of present study is to investigate the role of paraoxonase 1 (PON 1) status i.e arylesterase and lactonase activities and Q192R polymorphism and conjugated dienes, in gastroenteritis of paediatric population. The study and control group consists of 40 paediatric patients with and without gastroenteritis. Paraoxonase arylesterase and lactonase activities were assessed and phenotyping was determined. Conjugated dienes were also assessed. PON 1 arylesterase activities in cases (61.494±13.220) and controls (70.942±15.385) and lactonase activities in cases (15.702±1.036) and controls (17.434±1.176) were significantly decreased (p<0.05). There is no significant difference of phenotypic distribution in cases and controls. Conjugated dienes were found significantly increased in patients (0.086±0.024) than the control group (0.064±0.019) (p<0.05). Paraoxonase 1 activities (arylesterase and lactonase) and conjugated dienes may be useful in risk assessment and management in gastroenteritis in paediatric population.

Keywords: paraoxonase 1 polymorphism, arylesterase, lactonase, conjugated dienes, p-nitrophenylacetate, DHC

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Authors: Suzanne Giasson, Alberto Guerron

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Stimuli-responsive polymer coatings can be used as functional elements in nanotechnologies, such as valves in microfluidic devices, as membranes in biomedical engineering, as substrates for the culture of biological tissues or in developing nanomaterials for targeted therapies in different diseases. However, such coatings usually suffer from major shortcomings, such as a lack of selectivity and poor environmental stability. The study will present multi-responsive hierarchical and hybrid polymer-based coatings aiming to overcome some of these limitations. Hierarchical polymer coatings, consisting of two-dimensional arrays of thermo-responsive cationic PNIPAM-based microgels and surface-functionalized with non-responsive or pH-responsive polymers, were covalently grafted to substrates to tune the surface chemistry and the elasticity of the surface independently using different stimuli. The characteristic dimensions (i.e., layer thickness) and surface properties (i.e., adhesion, friction) of the microgel coatings were assessed using the Surface Forces Apparatus. The ability to independently control the swelling and surface properties using temperature and pH as triggers were investigated for microgels in aqueous suspension and microgels immobilized on substrates. Polymer chain grafting did not impede the ability of cationic PNIPAM microgels to undergo a volume phase transition above the VPTT, either in suspension or immobilized on a substrate. Due to the presence of amino groups throughout the entirety of the microgel polymer network, the swelling behavior was also pH dependent. However, the thermo-responsive swelling was more significant than the pH-triggered one. The microgels functionalized with PEG exhibited the most promising behavior. Indeed, the thermo-triggered swelling of microgel-co-PEG did not give rise to changes in the microgel surface properties (i.e., surface potential and adhesion) within a wide range of pH values. It was possible for the immobilized microgel-co-PEG to undergo a volume transition (swelling/shrinking) with no change in adhesion, suggesting that the surface of the thermal-responsive microgels remains rather hydrophilic above the VPTT. This work confirms the possibility of tuning the swelling behavior of microgels without changing the adhesive properties. Responsive surfaces whose swelling properties can be reversibly and externally altered over space and time regardless of the surface chemistry are very innovative and will enable revolutionary advances in technologies, particularly in biomedical surface engineering and microfluidics, where advanced assembly of functional components is increasingly required.

Keywords: responsive materials, polymers, surfaces, cell culture

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Authors: Shasha Lv, Zhengcao Li

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Due to the CMOS compatibility, silicon-based field emission (FE) devices as potential electron sources have attracted much attention. The geometrical arrangement and dimensional features of aligned silicon nanowires (SiNWs) have a determining influence on the FE properties. We discuss a multistep template replication process of Ag-assisted chemical etching combined with polystyrene (PS) spheres to fabricate highly periodic and well-aligned silicon nanowires, then their diameter, aspect ratio and density were further controlled via dry oxidation and post chemical treatment. The FE properties related to proximity and aspect ratio were systematically studied. A remarkable improvement of FE propertiy was observed with the average nanowires tip interspace increasing from 80 to 820 nm. On the basis of adjusting SiNWs dimensions and morphology, addition of a secondary material whose properties complement the SiNWs could yield a combined characteristic. Three different nanoheterostructures were fabricated to control the FE performance, they are: NiSi/Si core-shell structures, ZnO/Si nanotrees, and Graphene/SiNWs. We successfully fabricated the high-quality NiSi/Si heterostructured nanowires with excellent conformality. First, nickle nanoparticles were deposited onto SiNWs, then rapid thermal annealing process were utilized to form NiSi shell. In addition, we demonstrate a new and simple method for creating 3D nanotree-like ZnO/Si nanocomposites with a spatially branched hierarchical structure. Compared with the as-prepared SiNRs and ZnO NWs, the high-density ZnO NWs on SiNRs have exhibited predominant FE characteristics, and the FE enhancement factors were attributed to band bending effect and geometrical morphology. The FE efficiency from flat sheet structure of graphene is low. We discussed an effective approach towards full control over the diameter of uniform SiNWs to adjust the protrusions of large-scale graphene sheet deposited on SiNWs. The FE performance regarding the uniformity and dimensional control of graphene protrusions supported on SiNWs was systematically clarified. Therefore, the hybrid SiNWs/graphene structures with protrusions provide a promising class of field emission cathodes.

Keywords: field emission, silicon nanowires, heterostructures, controllable synthesis

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Authors: Humberto Miranda, Haroldo G. Santana, Gabriel A. Paz, Vicente P. Lima, Jeffrey M. Willardson

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Foam rolling is a practice that has increased in popularity before and after strength training. The purpose of this study was to compare the acute effects of different foam rolling periods for the lower body muscles on subsequent performance (total repetitions and training volume), myoelectric activity and rating of perceived exertion in trained men. Fourteen trained men (26.2 ± 3.2 years, 178 ± 0.04 cm height, 82.2 ± 10 kg weight and body mass index 25.9 ± 3.3kg/m2) volunteered for this study. Four repetition maximum (4-RM) loads were determined for hexagonal bar deadlift and 45º angled leg press during test and retest sessions over two nonconsecutive days. Five experimental protocols were applied in a randomized design, which included: a traditional protocol (control)—a resistance training session without prior foam rolling; or resistance training sessions performed following one (P1), two (P2), three (P3), or four (P4) sets of 30 sec. foam rolling for the lower extremity musculature. Subjects were asked to roll over the medial and lateral aspects of each muscle group with as much pressure as possible. All foam rolling was completed at a cadence of 50 bpm. These procedures were performed on both sides unilaterally as described below. Quadriceps: between the apex of the patella and the ASIS; Hamstring: between the gluteal fold and popliteal fossa; Triceps surae: between popliteal fossa and calcaneus tendon. The resistance training consisted of five sets with 4-RM loads and two-minute rest intervals between sets, and a four-minute rest interval between the hexagonal bar deadlift and the 45º angled leg press. The number of repetitions completed, the myoelectric activity of vastus lateralis (VL), vastus medialis oblique (VMO), semitendinosus (SM) and medial gastrocnemius (GM) were recorded, as well as the rating of perceived exertion for each protocol. There were no differences between the protocols in the total repetitions for the hexagonal bar deadlift (Control - 16.2 ± 5.9; P1 - 16.9 ± 5.5; P2 - 19.2 ± 5.7; P3 - 19.4 ± 5.2; P4 - 17.2 ± 8.2) (p > 0.05) and 45º angled leg press (Control - 23.3 ± 9.7; P1 - 25.9 ± 9.5; P2 - 29.1 ± 13.8; P3 - 28.0 ± 11.7; P4 - 30.2 ± 11.2) exercises. Similar results between protocols were also noted for myoelectric activity (p > 0.05) and rating of perceived exertion (p > 0.05). Therefore, the results of the present study indicated no deleterious effects on performance, myoelectric activity and rating of perceived exertion responses during lower body resistance training.

Keywords: self myofascial release, foam rolling, electromyography, resistance training

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Authors: Dinebari Akpee, Friday Aabe Gaage, Florence Fred Nwaigwu

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Natural disasters like flood affect many parts of the world including developing countries like Nigeria. As a result, many human lives are lost, properties damaged and so much money is lost in infrastructure damages. These hazards and losses can be mitigated and reduced by providing reliable spatial information to the generality of the people through about flood risks through flood inundation maps. Flood inundation maps are very crucial for emergency action plans, urban planning, ecological studies and insurance rates. Nigeria experience her worst flood in her entire history this year. Many cities were submerged and completely under water due to torrential rainfall. Poor city planning, lack of effective development control among others contributes to the problem too. Geographic information system (GIS) can be used to visualize the extent of flooding, analyze flood maps to produce flood damaged estimation maps and flood risk maps. In this research, the under listed steps were taken in preparation of flood risk maps for the study area: (1) Digitization of topographic data and preparation of digital elevation model using ArcGIS (2) Flood simulation using hydraulic model and integration and (3) Integration of the first two steps to produce flood risk maps. The results shows that GIS can play crucial role in Flood disaster control and mitigation.

Keywords: flood disaster, risk maps, geographic information system, hazards

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Authors: Olusola Akintunde Adediran

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This study examined differential effect of Gender difference in social interaction skill of pupils with autism using token economy and video modeling as intervention strategies. A pretest, posttest, control group, quasi-experimental research design was adopted in the study. 17 participants (11 males and 6 females) were selected purposively from 5 centres in Ibadan and randomized into three groups (token economy, video modeling and control groups). Two instruments were used in the study; Autism Spectrum Rating Scale (ASRS) for 299.00 Autistic Disorder (r = 0.82) and Children’s Self-report Social Skill Scale (CS4) (r= 0.93). A descriptive statistics was used to analyse the participants social interaction data based on intervention and gender, while inferential statistics of analysis of covariance (ANCOVA) and scheffe post-hoc measure was used to anlayse three null hypotheses tested at 0.05 level of significance. The results obtained indicated that there was a significant main effect of treatment on social interaction of participants, but there was no significant of main effect of gender on the social interaction of participants, hence, (F(2,14) = .741; p > .05, eta = .050). Lastly, there was no significant interaction effect of treatment and gender of the participants, hence (F(2,10) = 2.177; p > .05, eta 2 = 202). The study has contributed to the frontiers of knowledge by establishing that social interaction of autism is attainable when token economy and video modelling are used as treatment intervention, hence, they should be adopted by the teachers, curriculum planners and other stakeholders.

Keywords: social interaction, token economy, video modelling, autism, gender

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Authors: C. M. Choi, H. C. Tsang, W. K. Fong, Y. K. Cheng, T. K. Chui, L. Y. Chan, K. W. Lee, C. K. Yuen, P. W. Lau, Y. L. To, K. C. Chow

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Six-minute walk test (6MWT) is a sub-maximal exercise test to assess aerobic capacity and exercise tolerance of patients with chronic respiratory disease and heart failure. This has been proven to be a reliable and valid tool and commonly used in clinical situations. Traditional 6MWT is labour-intensive and time-consuming especially for patients who require assistance in ambulation and oxygen use. When performing the test with these patients, one staff will assist the patient in walking (with or without aids) while another staff will need to manually record patient’s oxygen saturation, heart rate and walking distance at every minute and/or carry oxygen cylinder at the same time. Physiotherapist will then have to document the test results in bed notes in details. With the use of electronic 6MWT (E-6MWT), patients wear a wireless oximeter that transfers data to a tablet PC via Bluetooth. Real-time recording of oxygen saturation, heart rate, and distance are displayed. No manual work on recording is needed. The tablet will generate a comprehensive report which can be directly attached to the patient’s bed notes for documentation. Data can also be saved for later patient follow up. This study was carried out in North District Hospital. Patients who followed commands and required 6MWT assessment were included. Patients were assigned to study or control groups. In the study group, patients adopted the E-6MWT while those in control group adopted the traditional 6MWT. Manpower and time consumed were recorded. Physiotherapists also completed a questionnaire about the use of E-6MWT. Total 12 subjects (Study=6; Control=6) were recruited during 11-12/2017. An average number of staff required and time consumed in traditional 6MWT were 1.67 and 949.33 seconds respectively; while in E-6MWT, the figures were 1.00 and 630.00 seconds respectively. Compared to traditional 6MWT, E-6MWT required 67.00% less manpower and 50.10% less in time spent. Physiotherapists (n=7) found E-6MWT is convenient to use (mean=5.14; satisfied to very satisfied), requires less manpower and time to complete the test (mean=4.71; rather satisfied to satisfied), has better data management (mean=5.86; satisfied to very satisfied) and is recommended to be used clinically (mean=5.29; satisfied to very satisfied). It is proven that E-6MWT requires less manpower input with higher efficiency and better data management. It is welcomed by the clinical frontline staff.

Keywords: electronic, physiotherapy, six-minute walk test, 6MWT

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Authors: Jenny Ngo, Harry De Boer, Jurgen Enders

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Indonesian higher education has experienced significant changes over the last decade. In 1999, the government published an overall strategy for decentralization and enhancement of local autonomy in many sectors, including (higher) education. Indonesian higher education reforms have forced universities to restructure their internal university governance to become more entrepreneurial. These new types of internal university governance are likely to affect the institutions’ leadership and management. This paper discusses the approach and findings of a study on the managerial leadership styles of deans in Indonesian universities. The study aims to get a better understanding of styles exhibited by deans manifested in their behaviors. Using the theories of reasoned action and planned behavior, in combination with the competing values framework, a large-scale survey was conducted to gather information on the deans’ behaviors, attitudes, subjective norms, and perceived behavioral control. Based on the responses of a sample of 218 deans, the study identifies a number of leadership styles: the Master, the Competitive Consultant, the Consensual Goal-Setter, the Focused Team Captain, and the Informed Trust-Builder style. The study demonstrates that attitudes are the primary determinant of the styles that were found. Perceived behavioral control is a factor that explains some managerial leadership styles. By understanding the attitudes of deans in Indonesian universities, and their leadership styles, universities can strengthen their management and governance, and thus improve their effectiveness.

Keywords: deans, Indonesian higher education, leadership and management styles, decentralization

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Authors: Amita Jain, Shantanu Prakash, Suruchi Shukla

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Introduction: Acute encephalitis syndrome (AES) is a complex multi etiology syndrome posing a great public health problem in the northern part of India. Japanese encephalitis (JE) virus is an established etiology of AES in this region. Recently, Scrub typhus (ST) is being recognized as an emerging aetiology of AES in JE endemic belt. This study was conducted to establish the direct evidence of Central nervous system invasion by Orientia tsutsugamushi leading to AES. Methodology: A total of 849 cases with clinical diagnosis of AES were enrolled from six districts (Deoria and its adjoining area) of the traditional north Indian Japanese encephalitis (JE) belt. Serum and Cerebrospinal fluid samples were collected and tested for major agent causing acute encephalitis. AES cases either positive for anti-ST IgM antibodies or negative for all tested etiologies were investigated for ST-DNA by real-time PCR. Results: Of these 505 cases, 250 patients were laboratory confirmed for O. tsutsugamushi infection either by anti-ST IgM antibodies positivity (n=206) on serum sample or by ST-DNA detection by real-time PCR assay on CSF sample (n=2) or by both (n=42).Total 29 isolate could be sequenced for 56KDa gene. Conclusion: All the strains were found to cluster with Gilliam strains. The majority of the isolates showed a 97–99% sequence similarity with Thailand and Cambodian strains. Gilliam strain of O.tsusugamushi is an emerging as one of the major aetiologies leading to AES in northern part of India.

Keywords: acute encephalitis syndrome, O. tsutsugamushi, Gilliam strain, North India, cerebrospinal fluid

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Authors: Suchismita Dattagupta

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Social rules and guidelines specify how a body should be clothed and how it should look. The social rules have made the body a space for expression, oppression and sexual 'commodification'. Being a malleable aspect of the human body, hair has always been worn in a number of ways and this characteristic of hair has made it an essential vehicle for conveying symbolic meaning. Hair, particularly women’s hair has always been considered to be associated with richness and beauty, apart from being associated with sexual power. Society has always had a preoccupation with hair bordering on obsession and has projected its moral and political supremacy by controlling and influencing how an individual wears their hair. Irrespective of the gender of the individual, society has tried to control an individual’s hair to express its control. However, with time, there has been a marked change in the way hair has been used by the individual. Hair has always been the focus of scholarly studies; not just aesthetically, but also in the cultural and social context. The fascination with hair rises from the fact that it is the only part of the human body that is always on display. Fetishization of hair is common in literature and goes ahead to reveal the character’s social and moral status. Modern authors for children and young adults have turned this concept on its head to point out how characters are breaking away from the mould and establishing their personal, moral and social boundaries. This paper will trace the change in hair symbolism in literature for children and young adults to understand how it has changed over the course of the time and what light it throws on the changing pattern of women’s position in society.

Keywords: gender, hair, social symbols, society, women's role

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Authors: Patrick J. Kerpan, Ryan C. Koop, Margaret M. Walker, Chris P. Swan

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The User Datagram Protocol (UDP) multicast is a vital part of data center networking that is being left out of major cloud computing providers' network infrastructure. Enterprise users rely on multicast, and particularly UDP multicast to create and connect vital business operations. For example, UPD makes a variety of business functions possible from simultaneous content media updates, High-Performance Computing (HPC) grids, and video call routing for massive open online courses (MOOCs). Essentially, UDP multicast's technological slight is causing a huge effect on whether companies choose to use (or not to use) public cloud infrastructure as a service (IaaS). Allowing the ‘chatty’ UDP multicast protocol inside a cloud network could have a serious impact on the performance of the cloud as a whole. Cloud IaaS providers solve the issue by disallowing all UDP multicast. But what about enterprise use cases for multicast applications in organizations that want to move to the cloud? To re-allow multicast traffic, enterprises can build a layer 3 - 7 network over the top of a data center, private cloud, or public cloud. An overlay network simply creates a private, sealed network on top of the existing network. Overlays give complete control of the network back to enterprise cloud users the freedom to manage their network beyond the control of the cloud provider’s firewall conditions. The same logic applies if for users who wish to use IPsec or BGP network protocols inside or connected into an overlay network in cloud IaaS.

Keywords: cloud computing, protocols, UDP multicast, virtualization

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Authors: Anum Atiq, Haya Fatimah

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Lucid dream is a dream where one is aware that one is dreaming, and they also might be able to influence their dreaming states. Logically, since lucidity cues towards high awareness, it should be negatively associated with proneness to psychosis. However, this association is scarcely studied. Furthermore, although gender differences and similarities in psychopathology have been thoroughly studied, there is room for research in the influence of masculinity and femininity, regardless of one’s sex, on proneness to psychosis. The aim of this study is twofold: 1) We investigated if dream lucidity was negatively associated with psychosis proneness; and 2) We explored the influence of masculinity and femininity on psychosis proneness, over and above the sex. Data were collected by convenience sampling from the undergraduate students enrolled at the University of Management and Technology, Lahore. The sample consisted of 53 students among the age range of 18-26 (men=24, women=29). Masculinity and femininity were measured using the masculinity and femininity subscales of the Personality Attributes Questionnaire. Dream lucidity was measured with The Lucidity and Consciousness in Dreams Scale; and the reality testing sub scale of The Inventory of Personality Organization was used to measure proneness to psychosis. Pearson correlation analysis revealed that psychosis proneness was significantly and negatively correlated with dream lucidity-insight and negative emotion in dreams, but not with other aspects of dream lucidity. Furthermore, masculinity, in both men and women, was positively related with lucid dreaming, and negatively with psychosis proneness. Following this, linear regression analysis showed that psychosis proneness was negatively predicted by masculinity even after controlling for gender. Lucid dreamer and masculinity both have characteristic of independence, emotional control and internal locus of control. Therefore, masculinity makes lucid dreaming less risk of psychosis in both genders.

Keywords: lucid dreaming, psychosis, gender, masculinity and femininity

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Authors: Subhasree Majumdar, Sovan Dey, Sayari Mukherjee, Sourav Dutta, Dalia Dasgupta Mandal

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Bacterial isolates from Planococcus genus are known for the production of yellowish orange pigment that belongs to the carotenoid family. These pigments are of immense pharmacological importance as antioxidant, anticancer, eye and liver protective agent, etc. The production of this pigment in a cost effective manner is a challenging task. The present study explored paper mill sludge (PMS), a solid lignocellulosic waste generated in large quantities from pulp and paper mill industry as a substrate for carotenoid pigment production by Planococcus sp. TRC1. PMS was compared in terms of efficacy with sugarcane bagasse, which is a highly explored substrate for valuable product generation via solid state fermentation. The results showed that both the biomasses yielded the highest carotenoid during 48 hours of incubation, 31.6 mg/gm and 42.1 mg/gm for PMS and bagasse respectively. Compositional alterations of both the biomasses showed reduction in lignin, hemicellulose and cellulose content by 41%, 15%, 1% for PMS and 38%, 25% and 6% for sugarcane bagasse after 72 hours of incubation. Structural changes in the biomasses were examined by FT-IR, FESEM, and XRD which further confirmed modification of solid biomasses by bacterial isolate. This study revealed the potential of PMS to act as cheap substrate for carotenoid pigment production by Planococcus sp. TRC1, as it showed a significant production in comparison to sugarcane bagasse which gave only 1.3 fold higher production than PMS. Delignification of PMS by TRC1 during pigment production is another important finding for the reuse of this waste from the paper industry.

Keywords: carotenoid, lignocellulosic, paper mill sludge, Planococcus sp. TRC1, solid state fermentation, sugarcane bagasse

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Authors: Vani Chintapally

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The expansion of modest and minimized Global Positioning System (GPS) beneficiaries has prompted most Automatic Vehicle Location (AVL) frameworks today depending solely on satellite-based finding frameworks, as GPS is the most stable usage of these. This paper shows the attributes of a proposed framework for following and dissecting open transport in a run of the mill medium-sized city and complexities the qualities of such a framework to those of broadly useful AVL frameworks. Particular properties of the courses broke down by the AVL framework utilized for the examination of open transport in our study incorporate cyclic vehicle courses, the requirement for particular execution reports, and so forth. This paper particularly manages vehicle movement forecasts and the estimation of station landing time, combined with consequently produced reports on timetable conformance and other execution measures. Another side of the watched issue is proficient exchange of information from the vehicles to the control focus. The pervasiveness of GSM bundle information exchange advancements combined with decreased information exchange expenses have brought on today's AVL frameworks to depend predominantly on parcel information exchange administrations from portable administrators as the correspondences channel in the middle of vehicles and the control focus. This methodology brings numerous security issues up in this conceivably touchy application field.

Keywords: automatic vehicle location (AVL), expectation of landing times, AVL security, data administrations, wise transport frameworks (ITS), guide coordinating

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Authors: M. Knezevic, V. Marecic, M. Ozanic, I. Kelava, M. Mihelcic, M. Santic

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Francisella tularensis is a highly infectious, gram-negative intracellular bacterium and the causative agent of tularemia. The bacterium has been isolated from more than 250 wild species, including protozoa cells. Since Francisella is very virulent and persists in the environment for years, the aim of this study was to investigate whether Acanthamoeba castellanii-grown F. novicida exhibits an alteration in the resistance to disinfectants. It has been shown by other intracellular pathogens, including Legionella pneumophila that bacteria grown in amoeba exhibit more resistance to disinfectants. However, there is no data showing Francisella viability behaviour after intracellular life cycle in A. castellani. In this study, the bacterial suspensions of A. castellanii-grown or in vitro-grown Francisella were treated with three different disinfectants, and the bacterial viability after disinfection treatment was determined by a colony-forming unit (CFU) counting method, transmission electron microscopy (TEM), fluorescence microscopy as well as the leakage of intracellular fluid. Our results have shown that didecyldimethylammonium chloride (DDAC) combined with isopropyl alcohol was the most effective in bacterial killing; all in vitro-grown and A. castellanii-grown F. novicida were killed after only 10s. Surprisingly, in comparison to in vitro-grown bacteria, A. castellanii-grown F. novicida was more sensitive to decontamination by the benzalkonium chloride combined with DDAC and formic acid and the polyhexamethylene biguanide (PHMB). We can conclude that the tested disinfectants exhibit antimicrobial activity by causing a loss of structural organization and integrity of the Francisella cell wall and membrane and the subsequent leakage of the intracellular contents. Finally, the results of this study clearly demonstrate that Francisella grown in A. castellanii had become more susceptible to many disinfectants.

Keywords: Acanthamoeba, disinfectant, Francisella, sensitivity

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Authors: Anita, Sari Septiani Tangke, Rusdina Bte Ladju, Nasrum Massi

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New diagnostic tools are urgently needed to interrupt the transmission of tuberculosis and multidrug-resistant tuberculosis. The microscopic-observation drug-susceptibility (MODS) assay is a rapid, accurate and simple liquid culture method to detect multidrug-resistant tuberculosis (MDR-TB). MODS were evaluated to determine a lower and same concentration of isoniazid and rifampin for detection of MDR-TB. Direct drug-susceptibility testing was performed with the use of the MODS assay. Drug-sensitive control strains were tested daily. The drug concentrations that used for both isoniazid and rifampin were at the same concentration: 0.16, 0.08 and 0.04μg per milliliter. We tested 56 M. tuberculosis clinical isolates and the control strains M. tuberculosis H37RV. All concentration showed same result. Of 53 M. tuberculosis clinical isolates, 14 were MDR-TB, 38 were susceptible with isoniazid and rifampin, 1 was resistant with isoniazid only. Drug-susceptibility testing was performed with the use of the proportion method using Mycobacteria Growth Indicator Tube (MGIT) system as reference. The result of MODS assay using lower concentration was significance (P<0.001) compare with the reference methods. A lower and same concentration of isoniazid and rifampin can be used to detect MDR-TB. Operational cost and application can be more efficient and easier in resource-limited environments. However, additional studies evaluating the MODS using lower and same concentration of isoniazid and rifampin must be conducted with a larger number of clinical isolates.

Keywords: isoniazid, MODS assay, MDR-TB, rifampin

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Authors: J. Hash, J. Converse, L. Gibson

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Plant medicines are of growing interest for addressing mental health concerns. Ayahuasca, a traditional plant-based medicine, has established itself as a powerful way of processing trauma and precipitating healing and mood stabilization. Eye Movement Desensitization and Reprocessing (EMDR) is another treatment modality that aids in the rapid processing and resolution of trauma. We investigated group EMDR therapy, G-TEP, as a preparatory practice before Ayahuasca ceremonies to determine if the combination of these modalities supports participants in their journeys of letting go of past experiences negatively impacting mental health, thereby accentuating the healing of the plant medicine. We surveyed 96 participants (51 experimental G-TEP, 45 control grounding prior to their ceremony; age M=38.6, SD=9.1; F=57, M=34; white=39, Hispanic/Latinx=23, multiracial=11, Asian/Pacific Islander=10, other=7) in a pre-post, mixed methods design. Participants were surveyed for demographic characteristics, symptoms of PTSD and cPTSD (International Trauma Questionnaire (ITQ), depression (Beck Depression Inventory, BDI), and stress (Perceived Stress Scale, PSS) before the ceremony and at the end of the ceremony weekend. Open-ended questions also inquired about their expectations of the ceremony and results at the end. No baseline differences existed between the control and experimental participants. Overall, participants reported a decrease in meeting the threshold for PTSD symptoms (p<0.01); surprisingly, the control group reported significantly fewer thresholds met for symptoms of affective dysregulation, 2(1)=6.776, p<.01, negative self-concept, 2 (1)=7.122, p<.01, and disturbance in relationships, 2 (1)=9.804, p<.01, on subscales of the ITQ as compared to the experimental group. All participants also experienced a significant decrease in scores on the BDI, t(94)=8.995, p<.001, and PSS, t(91)=6.892, p<.001. Similar to patterns of PTSD symptoms, the control group reported significantly lower scores on the BDI, t(65.115)=-2.587, p<.01, and a trend toward lower PSS, t(90)=-1.775, p=.079 (this was significant with a one-sided test at p<.05), compared to the experimental group following the ceremony. Qualitative interviews among participants revealed a potential explanation for these relatively higher levels of depression and stress in the experimental group following the ceremony. Many participants reported needing more time to process their experience to gain an understanding of the effects of the Ayahuasca medicine. Others reported a sense of hopefulness and understanding of the sources of their trauma and the necessary steps to heal moving forward. This suggests increased introspection and openness to processing trauma, therefore making them more receptive to their emotions. The integration process of an Ayahuasca ceremony is a week- to months-long process that was not accessible in this stage of research, yet it is an integral process to understanding the full effects of the Ayahuasca medicine following the closure of a ceremony. Our future research aims to assess participants weeks into their integration process to determine the effectiveness of EMDR, and if the higher levels of depression and stress indicate the initial reaction to greater awareness of trauma and receptivity to healing.

Keywords: ayahuasca, EMDR, PTSD, mental health

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Authors: Lukumba Phiri, Simon Tembo, Kumbuso Joshua Nyoni

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In Zambia recent initiatives by various power operators like ZESCO, CEC, and consumers like the mines to upgrade power systems into smart grids target an even tighter integration with information technologies to enable the integration of renewable energy sources, local and bulk generation, and demand response. Thus, for the reliable operation of smart grids, its information infrastructure must be secure and reliable in the face of both failures and cyberattacks. Due to the nature of the systems, ICS/SCADA cybersecurity and governance face additional challenges compared to the corporate networks, and critical systems may be left exposed. There exist control frameworks internationally such as the NIST framework, however, there are generic and do not meet the domain-specific needs of the SCADA systems. Zambia is also lagging in cybersecurity awareness and adoption, therefore there is a concern about securing ICS controlling key infrastructure critical to the Zambian economy as there are few known facts about the true posture. In this paper, we introduce a stochastic Edged-based Anomaly Detection for SCADA systems (SEADS) framework for threat modeling and risk assessment. SEADS enables the calculation of steady-steady probabilities that are further applied to establish metrics like system availability, maintainability, and reliability.

Keywords: anomaly, availability, detection, edge, maintainability, reliability, stochastic

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Authors: Frederick Monyepao

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Crack cocaine first appeared on the scene in the form of cocaine freebasing in the late 1970s. Stockbrokers, investment bankers, rock stars, Hollywood elites, and a few pro athletes were regular users of the substance. As criminogenic factors associated with substance abuse began to surface, congress passed new legislation. The laws led to the increase of health coverage insurances and the expansion of hospitals. By the mid-1980s, crack use spread into America's inner cities among impoverished African Americans and Latinos. While substance abuse increased among minority communities, legislation pertaining to substance abuse evolved. The prison industry also expanded the number of cells available. A qualitative approach was taken, drawing from a range secondary sources for contextual analysis. This paper traces out the continued marginalisation and racist undertones towards minorities as perpetuated by certain drug policies. It was discovered that the new legislation on crack was instrumental in the largest incarcerations the United States ever faced. Drug offenders increased in prisons eightfold from 1986 to 2000. The paper concludes that American drug control policies are consistently irrational and ineffective when measured by levels of substance use and abuse. On the contrary, these policies have been successful as agents of social control in maintaining the stratification patterns of racial/ethnic minorities and women. To move beyond prohibition, radical law and policy reform may require a change in narratives on substance use.

Keywords: crack, drug policy, minorities, racism, substance abuse

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Authors: Susmita Goswami, Joyeeta Mitra, Indu Gaur, Neha Bhadauria, Shilpi Shilpi, Prabir K. Paul

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'Malbhog’, an indigenous banana variety, much prized for its flavour and delicacy suffers production losses due to Fusarium oxysporum f.sp. cubense. The pathogen enters young plants through feeder roots causing wilting of plants ultimately leading to death of plants. The pathogen spreads rapidly to other plants in the field. In eastern part of India, this variety escapes the onslaught of the pathogen when either co-cultivated or rotated with Amorphophallus campanulatus (yam). The present study provides an insight into the physiological aspect of the biocontrol by yam. In vitro application of sterile aqueous extract of yam tuber (100gm/100ml distilled water and its 1:10 and 1:100 dilutions) were mixed with PDA media which was substantially inoculated with spores of Fusarium oxysporum f.sp. cubense. The extract could significantly reduce germination of pathogen spores. Banana variety susceptible to Fusarium sp was raised in soil rite under aseptic conditions. Spores of the pathogen (106 spores/ml) were inoculated into the soil rite. The plants were spread with aqueous extract of yam. The control plants were treated with sterilized distilled water. The activity of phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO) and peroxidase (POX) were estimated in leaves and roots at interval of 24 hours for 5 days after treatment. The incidence of wilt disease was recorded after two weeks. The results demonstrated that yam extract could induce significant activity of PAL, PPO and POX along with accumulation of phenols in both roots and leaves of banana plants. However, significantly high activity of enzymes and phenol accumulation was observed in roots. The disease incidence was significantly low in yam treated plants. The results clearly demonstrated the control of the pathogen due to induction of defense mechanism in the host by the extract. The observed control of the pathogen in the field could possibly be due to induction of such defense responses in host by exudates leached into the soil from yam tubers. Yam extract could be a potential source of environment-friendly biocide against Panama wilt of banana.

Keywords: Amorphophallus campanulatus, banana, Fusarium oxysporum f.sp. cubense, phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), peroxidase (POX)

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Authors: Shirin jalili, Hadi Shirzad, Mahasti Modarresi, Samaneh Nabavi, Somayeh Khanjani

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Identifying the source tissue of biological material found at crime scenes can be very informative in a number of cases. Despite their usefulness, current visual, catalytic, enzymatic, and immunologic tests for presumptive and confirmatory tissue identification are applicable only to a subset of samples, might suffer limitations such as low specificity, lack of sensitivity, and are substantially impacted by environmental insults. In addition their results are operator-dependent. Recently the possibility of discriminating body fluids using mRNA expression differences in tissues has been described but lack of long term stability of that Molecule and the need to normalize samples for each individual are limiting factors. The use of DNA should solve these issues because of its long term stability and specificity to each body fluid. Cells in the human body have a unique epigenome, which includes differences in DNA methylation in the promoter of genes. DNA methylation, which occurs at the 5′-position of the cytosine in CpG dinucleotides, has great potential for forensic identification of body fluids, because tissue-specific patterns of DNA methylation have been demonstrated, and DNA is less prone to degradation than proteins or RNA. Previous studies have reported several body fluid-specific DNA methylation markers.The presence or absence of a methyl group on the 5’ carbon of the cytosine pyridine ring in CpG dinucleotide regions called ‘CpG islands’ dictates whether the gene is expressed or silenced in the particular body fluid. Were described methylation patterns at tissue specific differentially methylated regions (tDMRs) to be stable and specific, making them excellent markers for tissue identification. The results demonstrate that methylation-based tissue identification is more than a proof-of-concept. The methodology holds promise as another viable forensic DNA analysis tool for characterization of biological materials.

Keywords: DNA methylation, forensic science, epigenome, tDMRs

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Authors: Alireza Javed, Keivan Jamshidi

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Reactive amyloidosis is a condition that complicates a long list of chronic inflammation, chronic infectious, malignant, and hereditary disorders. In the present study the potential effects of two amyloidogenic substances: ie. AgNO3 and Freund's Adjuvant (AF) with that of vitamin free casein, on spatio-temporal pattern of experimental amyloidosis in mice, were compared. For this purpose, a total of 40 male Swees mice, obtained from Pasteur Institute Tehran, after being weighted were randomly divided into 4 groups including 2 treatments, 1 control (vitamin free casein) and 1 positive control (normal saline). At the end of 3rd, 5th and 7th weeks of experiment 3 mice were randomly selected and euthnised. Spleen sample of each animal obtained and preserved in 10% neutral buffer formalin. Sample were then processed through different stages of dehydration, clearing and impregnation and finally embedded in paraffin blocks. Sections of 5µm thickness then cut and stained by alkaline Congo red techniques. Spleen weights and the data obtained from the microscopic quantitative analysis did show no significant differences between groups A and B, A and C, and B and C. However, significant differences were observed between groups A and D, B and D, and C and D respectively. It is concluded that two compounds ie; AgNO3 and Freund's Adjuvant have the same potential, as does vitamin free casein have, in spatio – temporal pattern of experimental amyloidosis in mice.

Keywords: amyloidosis, mice, AgNO3, Freund's Adjuvant

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Authors: Chia-Pei Chen, Yu-Ju Chen, Yu-Juei Hsu

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The prevalence and incidence of end-stage renal disease in Taiwan ranks the highest in the world. According to the statistical survey of the Ministry of Health and Welfare in 2019, kidney disease is the ninth leading cause of death in Taiwan. It leads to autonomic dysfunction, inflammatory response and symptom distress, and further increases the damage to the structure and function of the kidneys, leading to increased demand for renal replacement therapy and risks of cardiovascular disease, which also has medical costs for the society. If we can intervene in a feasible manual to effectively regulate the autonomic nerve function of CKD patients, reduce the inflammatory response and symptom distress. To prolong the progression of the disease, it will be the main goal of caring for CKD patients. This study aims to test the effect of heart rate variability biofeedback (HRVBF) on improving autonomic nerve function (Heart Rate Variability, HRV), inflammatory response (Interleukin-6 [IL-6], C reaction protein [CRP] ), symptom distress (Piper fatigue scale, Pittsburgh Sleep Quality Index [PSQI], and Beck Depression Inventory-II [BDI-II] ) in patients with chronic kidney disease. This study was experimental research, with a convenience sampling. Participants were recruited from the nephrology clinic at a medical center in northern Taiwan. With signed informed consent, participants were randomly assigned to the HRVBF or control group by using the Excel BINOMDIST function. The HRVBF group received four weekly hospital-based HRVBF training, and 8 weeks of home-based self-practice was done with StressEraser. The control group received usual care. We followed all participants for 3 months, in which we repeatedly measured their autonomic nerve function (HRV), inflammatory response (IL-6, CRP), and symptom distress (Piper fatigue scale, PSQI, and BDI-II) on their first day of study participation (baselines), 1 month, and 3 months after the intervention to test the effects of HRVBF. The results were analyzed by SPSS version 23.0 statistical software. The data of demographics, HRV, IL-6, CRP, Piper fatigue scale, PSQI, and BDI-II were analyzed by descriptive statistics. To test for differences between and within groups in all outcome variables, it was used by paired sample t-test, independent sample t-test, Wilcoxon Signed-Rank test and Mann-Whitney U test. Results: Thirty-four patients with chronic kidney disease were enrolled, but three of them were lost to follow-up. The remaining 31 patients completed the study, including 15 in the HRVBF group and 16 in the control group. The characteristics of the two groups were not significantly different. The four-week hospital-based HRVBF training combined with eight-week home-based self-practice can effectively enhance the parasympathetic nerve performance for patients with chronic kidney disease, which may against the disease-related parasympathetic nerve inhibition. In the inflammatory response, IL-6 and CRP in the HRVBF group could not achieve significant improvement when compared with the control group. Self-reported fatigue and depression significantly decreased in the HRVBF group, but they still failed to achieve a significant difference between the two groups. HRVBF has no significant effect on improving the sleep quality for CKD patients.

Keywords: heart rate variability biofeedback, autonomic nerve function, inflammatory response, symptom distress, chronic kidney disease

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Authors: Majid Hejazian, Nam-Trung Nguyen

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Over the past few years, microfluidic devices have generated significant attention from industry and academia due to advantages such as small sample volume, low cost and high efficiency. Microfluidic devices have applications in chemical, biological and industry analysis and can facilitate assay of bio-materials and chemical reactions, separation, and sensing. Micromixers are one of the important microfluidic concepts. Micromixers can work as stand-alone devices or be integrated in a more complex microfluidic system such as a lab on a chip (LOC). Micromixers are categorized as passive and active types. Passive micromixers rely only on the arrangement of the phases to be mixed and contain no moving parts and require no energy. Active micromixers require external fields such as pressure, temperature, electric and acoustic fields. Rapid and efficient mixing is important for many applications such as biological, chemical and biochemical analysis. Achieving fast and homogenous mixing of multiple samples in the microfluidic devices has been studied and discussed in the literature recently. Improvement in mixing rely on effective mass transport in microscale, but are currently limited to molecular diffusion due to the predominant laminar flow in this size scale. Using magnetic field to elevate mass transport is an effective solution for mixing enhancement in microfluidics. The use of a non-uniform magnetic field to improve mass transfer performance in a microfluidic device is demonstrated in this work. The phenomenon of mixing ferrofluid and DI-water streams has been reported before, but mass transfer enhancement for other non-magnetic species through magnetic field have not been studied and evaluated extensively. In the present work, permanent magnets were used in a simple microfluidic device to create a non-uniform magnetic field. Two streams are introduced into the microchannel: one contains fluorescent dye mixed with diluted ferrofluid to induce enhanced mass transport of the dye, and the other one is a non-magnetic DI-water stream. Mass transport enhancement of fluorescent dye is evaluated using fluorescent measurement techniques. The concentration field is measured for different flow rates. Due to effect of magnetic field, a body force is exerted on the paramagnetic stream and expands the ferrofluid stream into non-magnetic DI-water flow. The experimental results demonstrate that without a magnetic field, both magnetic nanoparticles of the ferrofluid and the fluorescent dye solely rely on molecular diffusion to spread. The non-uniform magnetic field, created by the permanent magnets around the microchannel, and diluted ferrofluid can improve mass transport of non-magnetic solutes in a microfluidic device. The susceptibility mismatch between the fluids results in a magnetoconvective secondary flow towards the magnets and subsequently the mass transport of the non-magnetic fluorescent dye. A significant enhancement in mass transport of the fluorescent dye was observed. The platform presented here could be used as a microfluidics-based micromixer for chemical and biological applications.

Keywords: ferrofluid, mass transfer, micromixer, microfluidics, magnetic

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Authors: Stella Y. Erinosho

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In an attempt to impact on girls’ interest in science, an instructional package on ‘Learn Science by Doing (LSD)’ was developed to support science teachers in teaching integrated science at the junior secondary level in Nigeria. LSD provides an instructional framework aimed at actively engaging girls in beginners’ science through activities that are discovery-oriented and allow for experiential learning. The goal of this study was to show the impact of application of LSD on girls’ performance and interest in science. The major hypothesis that was tested in the study was that students would exhibit higher learning outcomes (achievement and attitude) in science as effect of exposure to LSD instructional package. A quasi-experimental design was adopted, incorporating four all-girls schools. Three of the schools (comprising six classes) were randomly designated as experimental and one as the control. The sample comprised 357 girls (275 experimental and 82 control) and nine science teachers drawn from the experimental schools. The questionnaire was designed to gather data on students’ background characteristics and their attitude toward science while the cognitive outcomes were measured using tests, both within a group and between groups, the girls who had exposure to LSD exhibited improved cognitive outcomes and more positive attitude towards science compared with those who had conventional teaching. The data are consistent with previous studies indicating that interactive learning activities increase student performance and interest.

Keywords: active learning, school science, teaching and learning, Nigeria

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Authors: Isil Cakir, Mustafa Uluhan

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Elevated serum Cardiotrophin-1 (CT-1) and leptin levels are important risk factors for cardiovascular diseases (CVDs). Obstructive sleep apnea syndrome (OSAS) has been reported to increase the risk of CVDs, too. The aim of this study was to evaluate the concentrations of serum CT-1 and leptin in these patients and whether their possible association with the disease severity. Fifty newly diagnosed patients with OSAS and thirty nonapneic snoring subjects were participated in this study. The mean ages of patients and control groups were 47.40±13.30 and 43.23±10.50 years, respectively (P=0.128). Fasting serum triglyseride, total cholesterol, LDL and HDL cholesterol, also CT-1 and leptin levels were evaluated. A significant difference was found in the serum CT-1 and leptin levels between the patients and the controls:serum median CT-1 levels in patients and control groups, respectively, were 19.47 and 8.23 pg/mL (P < 0.001) and leptin levels were 2.07 and 1.29 ng/mL (P < 0.001). In severe patients group (n=39), serum median CT-1 level was found statistically significantly higher than the median level in mild/moderate patients (n=11) group. Patients CT-1 concentrations were not associated with lipoprotein levels and there was no correlation between patients’ leptin and lipid profile parameters. Two risk factors for CVDs, CT-1 and leptin, have significantly elevated and they were associated with OSAS. Furthermore, CT-1 was associated with the severity of disease. We recommend the use of increased serum CT-1 and leptin concentrations as markers of the presence and severity of OSAS.They can be used as early markers in male OSAS patients without known CVDs.

Keywords: obstructive sleep apnea syndrome, cardiotrophin-1, leptin, cardiovascular disease

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Authors: Ben Mansour Mouin, Elloumi Abdelkarim

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We will discuss the problem of routing a fleet of different vehicles from a central depot to different types of infrastructure-defaults with dynamic maintenance requests, modified time windows, and control of default maintained. For this reason, we propose a modified metaheuristicto to solve our mathematical model. SVRDP-CMTW is a variant VRP of an optimal vehicle plan that facilitates the maintenance task of different types of infrastructure-defaults. This task will be monitored after the maintenance, based on its priorities, the degree of danger associated with each default, and the neighborhood at the black-spots. We will present, in this paper, a multidimensional genetic algorithm “MGA” by detailing its characteristics, proposed mechanisms, and roles in our work. The coding of this algorithm represents the necessary parameters that characterize each infrastructure-default with the objective of minimizing a combination of cost, distance and maintenance times while satisfying the priority levels of the most urgent defaults. The developed algorithm will allow the dynamic integration of newly detected defaults at the execution time. This result will be displayed in our programmed interactive system at the routing time. This multidimensional genetic algorithm replaces N genetic algorithm to solve P different type problems of infrastructure defaults (instead of N algorithm for P problem we can solve in one multidimensional algorithm simultaneously who can solve all these problemsatonce).

Keywords: mathematical model, VRP, multidimensional genetic algorithm, metaheuristics

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Authors: M. G. Tanga, P. B. Telefo, D. N. Tarla

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Even though the WHO, the EPA and other regulatory bodies have recognized the effects of acute pesticide poisoning little data exists on health effects after long-term low-dose exposures especially in Africa and Cameroon. The aim of this study was to evaluate the impact of pesticides on the hepatic functions of market gardeners in the Western Region of Cameroon by studying some biochemical parameters. Sixty six male market gardeners in Foumbot, Massangam, and Bantoum were interviewed on their health status, habits and pesticide use in agriculture, including the spray frequency, application method, and pesticide dosage. Thirty men with no history of pesticide exposure were recruited as control group. Thereafter, their blood samples were collected for assessment of hepatic function biomarkers (ALT, AST, and albumin). The results showed that 56 pesticides containing 25 active ingredients were currently used by market gardeners enrolled in our study and most of their symptoms (headache, fatigue, skin rashes, eye irritation, and nausea) were related to the use of these chemicals. Compared to the control subjects market gardeners’ ALT levels (32.9 ± 7.19 UL-1 vs. 82.11 ± 35.40 UL-1; P < 0.001) and, AST levels (40.63 ± 6.52 UL-1 vs. 112.11 UL-1 ± 47.15 UL-1; P < 0.001) were significantly increased. These results suggest that liver function tests can be used as biomarkers to indicate toxicity before overt clinical signs occur. The market gardeners’ chronic exposure to pesticides due to poor application measures could lead to hepatic function impairment. Further research on larger scale is needed to confirm these findings and to establish a mechanism of toxicity.

Keywords: biomarkers, liver, pesticides, occupational exposure

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Authors: Lethukuthula Ngobese, Abin Gupthar, Patrick Govender

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The ability of yeast cell wall-derived mannoproteins (glycoproteins) to positively contribute to oenological properties has been a key factor that stimulates research initiatives into these industrially important glycoproteins. In addition, and from a fundamental research perspective, yeast cell wall glycoproteins are involved in a wide range of biological interactions. To date, and to the best of our knowledge, our understanding of the fine molecular structure of these mannoproteins is fairly limited. Generally, the amino acid sequences of their protein moieties have been established from structural and functional analysis of the genomic sequence of these yeasts whilst far less information is available on the glycosyl moieties of these mannoproteins. A novel strategy was devised in this study that entails the genetic engineering of yeast strains that over-express and release cell wall-associated glycoproteins into the liquid growth medium. To this end, the Flo1p mannoprotein was overexpressed in Saccharomyces cerevisiae laboratory strains bearing a specific deletion in KNR4 and GPI7 genes involved in cell wall biosynthesis that have been previously shown to extracellularly hyper-secrete cell wall-associated glycoproteins. A polymerase chain reaction (PCR) -based cloning strategy was employed to generate transgenic yeast strains in which the native cell wall FLO1 glycoprotein-encoding gene is brought under transcriptional control of the constitutive PGK1 promoter. The modified Helm’s flocculation assay was employed to assess flocculation intensities of a Flo1p over-expressing wild type and deletion mutant as an indirect measure of their abilities to release the desired mannoprotein. The flocculation intensities of the transformed strains were assessed and all the strains showed similar intensities (>98% flocculation). To assess if mannoproteins were released into the growth medium, the supernatant of each strain was subjected to the BCA protein assay and the transformed Δknr4 strain showed a considerable increase in protein levels. This study has the potential to produce mannoproteins in sufficient quantities that may be employed in future investigations to understand their molecular structures and mechanisms of interaction to the benefit of both fundamental and industrial applications.

Keywords: glycoproteins, genetic engineering, flocculation, over-expression

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