Search results for: plant tissues

Authors: Adrian Reynaldo Sudirman, Siti Farida, Aisyah Aminy Maulidina, Caren Andika Surbakti

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Sedentary lifestyle and imbalance consumption pattern has made metabolic syndrome as the global time bomb phenomenon in the world. The increasing tendency of people in consuming high amount of fructose and cholesterol food has worsened this issue in the society. Pancreas steatosis become one of the most comorbid when early diagnosis and prompt treatment has not been applied on hyperglycemic and hyperlipidemic condition in metabolic syndrome patient. Gemfibrozil become the drug of choice to prevent this issue, yet the efficacy of this regiment was still questionable. Acalypha indica Linn. is the herb that has protective effect on hyperlipidemic and hyperglycemic condition. This study was aimed to compare therapeutic effect of gemfibrozil (G) and Acalypha indica Linn. (AI) on high fructose and cholesterol diet-induced pancreas steatosis in Sprague-Dawley mice. The post induction mice were divided into four groups: control, gemfibrozil, AI extract, and G+AI combination regiment. Each group received four weeks intervention. The result of statistical analysis using the One-Way ANOVA test and Tukey Post Hoc test showed significant decrease in pancreatic steatosis between the control group and administered Acalypha indica group (p = 0.004, 95% CI: 0.170-0.959) and the group administered with a combination of Gemfibrozil-Acalypha indica (p = 0.023, 95% CI: 0.537-0.813). The protective effect of Acalypha indica Linn. shows that this plant has the potential as therapeutic option in overcoming the condition of pancreas steatosis in metabolic syndrome.

Keywords: Acalypha Indica Linn., Cholesterol, Fructose, Gemfibrozil, Pancreas Steatosis

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Authors: Daniel Martin, Roberto Rodriguez, Derek Woller, Chris Reuter, Lonnie Black, Mohamed Latheef

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The grasshoppers comprised of heterogeneous assemblages of Acrididae (Family: Orthoptera) species periodically reach outbreak levels by their gregarious behavior and voracious feeding habits, devouring stems and leaves of food crops and rangeland pasture. Cattle consume about 1.5-2.5% of their body weight in forage per day, so pound for pound, a grasshopper will eat 12-20 times as much plant material as a steer and cause serious economic damage to the cattle industry, especially during a drought when forage is already scarce. Grasshoppers annually consume more than 20% of rangeland forages in the western United States at an estimated loss of $1.25 billion per year in forage. A remotely piloted aerial application system with both a spreader and spray application system was used to apply granular insect bait and a liquid formulation of Carbaryl for control of grasshopper infestations on rangeland in New Mexico, United States. Pattern testing and calibration of both the granular and liquid application systems were conducted to determine proper application rate set up and distribution pattern. From these tests, an effective swath was calculated. Results showed that 14 days after application, granular baits were only effective on those grasshopper species that accepted the baits. The liquid formulation at 16 ounces per acre was highly successful at controlling all grasshopper species. Results of this study indicated that a remotely piloted aerial application system can be used to effectively deliver grasshopper control products in both granular and liquid form. However, the spray application treatment proved to be most effective and efficient for all grasshopper species present.

Keywords: Carbaryl, Grasshopper, Insecticidal Efficacy, Remotely Piloted Aerial Application System

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Authors: Nneka Augustina Akwu, Yougasphree Naidoo

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Molecular advancement in technology has created a means whereby the atoms and molecules (solid forms) of certain materials such as plants, can now be reduced to a range of 1-100 nanometres. Green synthesis of silver nanoparticles (AgNPs) was carried out at room temperature (RT) 25 ± 2°C and 80°C, using the metabolites in the aqueous extracts of the leaves and stem bark of Grewia lasiocarpa as reductants and stabilizing agents. The biosynthesized AgNPs were characterized by UV-Vis spectrophotometry, attenuated total reflectance - Fourier transforms infrared (ATR-FTIR) spectroscopy, nanoparticle tracking analysis (NTA), Energy Dispersive X-ray fluorescence scanning electron microscope (SEM-EDXRF) and high-resolution transmission electron microscopy (HRTEM). The AgNPs were biologically evaluated for antioxidant, antibacterial and cytotoxicity activities. The phytochemical and FTIR analyses revealed the presence of metabolites that act as reducing and capping agents, while the UV-Vis spectroscopy of the biosynthesized NPs showed absorption between 380-460 nm, confirming AgNP synthesis. The Zeta potential values were between -9.1 and -20.6 mV with a hydrodynamics diameter ranging from 38.3 to 46.7 nm. SEM and HRTEM analyses revealed that AgNPs were predominately spherical with an average particle size of 2- 31 nm for the leaves and 5-27 nm for the stem bark. The cytotoxicity IC50 values of the AgNPs against HeLa, Caco-2 and MCF-7 were >1 mg/mL. The AgNPs were sensitive to all strains of bacteria used, with methicillin-resistant Staphylococcus aureus (MRSA), Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922) being more sensitive to the AgNPs. Our findings propose that antibacterial and anticancer agents could be derived from these AgNPs of G. lasiocarpa, and warrant their further investigation.

Keywords: antioxidant, cytotoxicity, Grewia lasiocarpa, silver nanoparticles, Zeta potentials

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Authors: Shyh-Ming Chern, Te-Hsiu Tang

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Hemicellulose is one of the major constituents of all plant cell walls, making up 15-25% of dry wood. It is a biopolymer from many different sugar monomers, including pentoses, like xylose, and hexoses, like mannose. In an effort to gasify real biomass in subcritical and supercritical water in a single process, it is necessary to understand the gasification of hemicellulose, in addition to cellulose and lignin, in subcritical and supercritical water. In the present study, xylose is chosen as the model compound for hemicellulose, since it has the largest amount in most hardwoods. Xylose is gasified in subcritical and supercritical water for the production of higher-valued gaseous products. Experiments were conducted with a 16-ml autoclave batch-type reactor. Hydrogen peroxide is adopted as the oxidant in an attempt to promote the gasification yield. The major operating parameters for the gasification include reaction temperature (400 - 600°C), reaction pressure (5 - 25 MPa), the concentration of xylose (0.05 and 0.30 M), and level of oxidant added (0 and 0.25 chemical oxygen demand). 102 experimental runs were completed out of 46 different set of experimental conditions. Product gases were analyzed with a GC-TCD and determined to be mainly composed of H₂ (10 – 74 mol. %), CO (1 – 56 mol. %), CH₄ (1 – 27 mol. %), CO₂ (10 – 50 mol. %), and C₂H₆ (0 – 8 mol. %). It has been found that the gas yield (amount of gas produced per gram of xylose gasified), higher heating value (HHV) of the dry product gas, and energy yield (energy stored in the product gas divided by the energy stored in xylose) all increase significantly with rising temperature and moderately with reducing pressure. The overall best operating condition occurred at 873 K and 10 MPa, with a gas yield of 54 mmol/g of xylose, a gas HHV of 440 kJ/mol, and an energy yield of 1.3. A seemingly unreasonably energy yield of greater than unity resulted from the external heating employed in the experiments to drive the gasification process. It is concluded that xylose can be completely gasified in subcritical and supercritical water under proper operating conditions. The addition of oxidant does not promote the gasification of xylose.

Keywords: gasification, subcritical water, supercritical water, xylose

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Authors: Dina Atmani-Kilani, Farah Yous, Djebbar Atmani

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The etiology of peptic ulcer is closely related to stress, excessive consumption of nonsteroidal anti-inflammatory drugs, or ethanol. Clematis flammula (Ranunculaceae) is a medicinal plant widely used by rural populations to treat inflammatory disorders. This study was designed to assess the gastroprotective potential of C. flammula extracts. Gastric ulcer was induced by stress, indomethacin, HCl / ethanol, and absolute ethanol on NMRI-type mice. The antioxidant potency of the ethanolic extract of Clematis flammula (EECF) was evaluated on catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) activities. Glutathione (GSH) and malonaldehyde (MDA) levels were also quantified. The anti-inflammatory potential was evaluated through the effect of EECF on myeloperoxidase activity (MPO) and vascular permeability. Complementary tests concerning the quantification of mucus levels, gastric motility, inhibition of ATPase H+/K+activity, as well as a histopathological study were also undertaken to explore the mechanism of action of the EECF. The EECF exhibited a significant (p <0.001) and optimal (100 mg/kg) gastroprotective effect by elevating SOD, CAT, and GSH levels, thereby minimizing the production of MDA and lowering the activity of MPO and vascular permeability. EECF also increased the rate of mucus production, decreased gastric motility, and completely suppressed the H+/K+ ATPase activity. Histopathological study confirmed the effectiveness of the extract in the prevention of peptic ulcer. The results obtained in this study demonstrated the gastro-protective effect of EECF via acidic antioxidant, anti-inflammatory, cytoprotective and anti-secretory mechanisms, which may justify its use as a substitute in peptic ulcer treatment.

Keywords: clematis flammula, superoxide dismutase, myeloperoxidase, ATPase, pump

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Authors: Hisham Mousa Mohammed Ahmed, Ahmed Wali Mohamed Salad, Yousif Hamed Dldom Gomaa

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A drip or Trickle irrigation system is designed to apply a precise amount of water near the plant with a certain degree of uniformity. This study was conducted at the Experimental Farm of the Faculty of Agricultural Sciences, University of Gezira, in March 2018. The study aimed to design and evaluate the hydraulic performance of three drip emitter types using: average discharge (Qavg), discharge variation (Qvar %), coefficient of uniformity (CU %), coefficient of manufacturer variation (CV %), distribution uniformity (DU %), statistical uniformity (Us %), clogging (%) wetted diameter (cm) and wetted depth (cm). The emitter types used are regular gauges (RG), high compensating pressure (HCP) and low compensating pressure (LCP). The treatments were laid out in a randomized complete block design (RCBD) with four replications. Results showed that there were significant differences (P≤0.05) in all tested parameters except clogging, wetted diameter and wetted depth. Discharge variation (Qvar %) values were 12.71, 15.57 and 19.17 for RG, LCP, and HCP, respectively. The variation is quite good and within the acceptable range. Results of coefficient of manufacture variation (CV %) were 10.9, 27.8 and 52.7 for RG, LCP and HCP, respectively. It is considered within the unacceptable range except for RG type, which is excellent. Statistical uniformity (Us %) values were 89.1, 72.2 and 45.7 for RG, LCP and HCP, respectively. It is considered good, acceptable and unacceptable, respectively. Results of the coefficient of uniformity (CU %) were 91.3, 77.7 and 56.7 for RG, LCP and HCP, respectively. It is considered excellent, fair and unacceptable, respectively. Distribution uniformity (DU %) was 90.2, 67.9 and 36.5 for RG, LCP and HCP, respectively. It is considered excellent, poor and poor, respectively. The study recommended regular gauges (RG) type emitters under the heavy clay soil conditions of the Gezira State, Sudan.

Keywords: drip irrigation, uniformity, clogging, coefficient, performance

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Authors: Levylee Bautista, Dawn Grace Santos, Aishwarya Veluchamy, Jesusa Santos, Ghafoor, Jr. I Haque, Rodolfo Rafael

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The booming interest in using natural compounds as an alternative to conventional medications has paved way to focus the attention on plants that provide rich sources of bioactive phytochemicals. For regulatory purposes, evaluation of the genotoxic effects of such alternatives is therefore empirical as part of the plant’s hazard assessment. Sesbania grandiflora is among the plants used as a traditional remedy in folk medicine and a subject of research for its medicinal benefits. This study aimed to evaluate the genotoxic potential induced by S. grandiflora flower methanol extract (SGFME) in terms of the frequency of micronucleus (MN) in polychromatic erythrocyte (PCE) (MNPCE) and PCE ratio employing the micronucleus assay. The frequency of MN was examined in bone marrow cells (BMCs) obtained from male Swiss albino mice exposed in vivo to four different concentrations (11.25, 22.5, 40, and 90 mg/kg) of SGFME and MMC (70 mg/kg; positive control) and sacrificed 24 hours post-intraperitoneal injection. Results showed a significant (p < 0.01) rate of MNPCEs for 11.25 and 22.5 tested concentrations of SGFME and is comparable with the MMC-treated mice. Although PCE ratio values in all doses of SGFME-treated mice were over 0.20, it is worth noting that 40 and 90 tested concentrations of SGFME-treated mice exhibited the lowest value, i.e., 0.22 and 0.28, respectively. The present study has demonstrated that S. grandiflora possesses genotoxic potential for murine BMCs. Such activity could be ascribed from the bioactive compounds present in S. grandiflora that require further isolation and characterization of the active molecules. Likewise, findings of this study warrant a caution of the use of S. grandiflora insomuch as further investigations do not demonstrate their safety.

Keywords: genotoxicity, micronucleus, phytochemicals, Sesbania grandiflora

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Authors: Nidhi Jindal, Ashok Chaudhury, Manisha Mangal

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Andrographis paniculata, (Burm f.) Wallich ex. Nees (Family Acanthaceae) popularly known as King of Bitters, is an important medicinal herb. It has an astonishingly wide range of medicinal properties such as anti-inflammatory,antidiarrhoeal, antiviral, antimalarial, hepatoprotective, cardiovascular, anticancer, and immunostimulatory activities. It is widely cultivated in southern Asia. Though propagation of this herb generally occurs through seeds, it has many germination problems which intrigued scientists to work out on the alternative techniques for its mass production. The potential of tissue culture techniques as an alternative tool for AP multiplication was found to be promising. However, the high mortality rate of explants caused by phenolic browning of explants is one of the difficulties reported. Low multiplication rates were reported in the proliferation phase, as well as cultures decline characterized by leaf fall and loss of overall vigor. In view of above problems, a study was undertaken to overcome seed dormancy to improve germination potential and to investigate further on the possible means for successful proliferation of cultures via preventive approaches to overcome failures caused by phenolic browning. Experiments were conducted to improve germination potential and among all the chemical and mechanical trials, scarification of seeds with sand paper proved to be the best method to enhance the germination potential (82.44%) within 7 days. Similarly, several pretreatments and media combinations were tried to overcome browning of explants leading to the conclusion that addition of 0.1% citric acid and 0.2% of ascorbic acid in the media followed by rapid sub culturing of explants controlled browning and decline of explants by 67.45%.

Keywords: plant tissue culture, empirical measure, germination, tissue culture

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Authors: Włodzimierz Przewodowski, Agnieszka Przewodowska

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Colloidal metal nanoparticles have drawn increasing attention in the field of phytopathology because of their unique properties and possibilities of applications. Their antibacterial activity, no induction of the development of pathogen resistance and the ability to penetrate most of biological barriers make them potentially useful in the fighting against dangerous pathogens. These properties are very important in the case of protection of strategic crops in the world, like potato - fourth crop in the world - which is host to numerous pathogenic microorganisms causing serious diseases, significantly affecting yield and causing the economic losses. One of the most important and difficult to reduce pathogen of potato plant is quarantine bacterium Clavibacter michiganensis ssp. sepedonicus (Cms) responsible for ring rot disease. Control and detection of these pathogens is very complicated. Application of healthy, certified seed material as well as hygiene in potato production and storage are the most efficient ways of preventing of ring rot disease. Currently used disinfectants and pesticides, have many disadvantages, such as toxicity, low efficiency, selectivity, corrosiveness, and the inability to eliminate the pathogens in potato tissue. In this situation, it becomes important to search for new formulations based on components harmful to health, yet efficient, stable during prolonged period of time and a with wide range of biocide activity. Such capabilities are offered by the latest generation of biocidal nanoparticles such as colloidal metals. Therefore the aim of the presented research was to develop newly antibacterial preparation based on colloidal metal nanoparticles and checking their influence on the Cms bacteria. Our preliminary results confirmed high efficacy of the nano-colloids in controlling the this selected pathogen.

Keywords: clavibacter michiganensis ssp. sepedonicus, colloidal metal nanoparticles, phytopathology, bacteria

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Authors: Anumol Jose, Sajana Nazar, M. R. Vishnu, M. Anilkumar

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Water yam is an underutilized tropical tuber crop and a rich source of polyphenol compounds and acylated anthocyanins. There is an inverse relationship between the risk of chronic human diseases and the consumption of polyphenolic rich diet. Dioscorea alata is a plant species with several undocumented landraces. In this study, several landraces of water yam with distinct morphological features were collected from all over kerala. Distinct variation in morphological feature among landraces was tuber colour and only those landraces which expressed consistent morphological characters for constitutively two growing seasons were included in the study. Plants were categorized according to the L*a*b* colour attributes of tuber extracts. There were five categories, red, pink, orange, yellow and white. Total phenol, flavanoid and anthocyanin content of the tuber extracts were measured spectroscopically and correlated with antioxidant properties determined by 2,2-diphenyl-1-picryl-hydrazyl-hydrate free radical method and ferric reducing antioxidant power assay. Landraces showed statistically significant difference in all the parameters studied and strong correlation were observed between total phenol and antioxidant activity. Out of the five categories orange coloured tubers showed relatively high phenol and flavanoid content.Colour variations of tuber extracts correlated with anthocyanin quantity and polymeric nature of anthocyanins. This study helps to identify and categorize landraces of D.alata with potential health benefits and commercial applications. Distinct colour characteristics of tuber could be useful in the field of natural colorants. This study also aimed to document and preserve landraces of water yams for further study and research in this area.

Keywords: the antioxidant property, anthocyanins, polyphenols, water yam

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Authors: Du Kim, Nelson Baro

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According to a report of International Atomic Energy Agency (IAEA), there are approximately 437 nuclear power stations are in operation in the present around the world in order to meet increasing energy demands. Indeed, nearly, a third of the world’s energy demands are met through nuclear power because it is one of the most efficient and long-lasting sources of energy. However, there are also consequences when a major event takes place at a nuclear power station. Over the past years, a few major nuclear accidents have occurred around the world. According to a report of International Nuclear and Radiological Event Scale (INES), there are six nuclear accidents that are considered to be high level (risk) of the events: Fukushima Dai-chi (Level 7), Chernobyl (Level 7), Three Mile Island (Level 5), Windscale (Level 5), Kyshtym (Level 6) and Chalk River (Level 5). Today, many people still have doubt about using nuclear power. There is growing number of people who are against nuclear power after the serious accident occurred at the Fukushima Dai-chi nuclear power plant in Japan. In other words, there are public concerns about radiation impacts which emphasize Linear-No-Threshold (LNT) Issues, Radiation Health Effects, Radiation Protection and Social Impacts. This paper will address those keywords by looking back at the history of these major nuclear accidents worldwide, based on INES. This paper concludes that all major mistake from nuclear accidents are preventable due to the fact that most of them are caused by human error. In other words, the human factor has played a huge role in the malfunction and occurrence of most of those events. The correct handle of a crisis is determined, by having a good radiation protection program in place, it’s what has a big impact on society and determines how acceptable people are of nuclear.

Keywords: linear-no-threshold (LNT) issues, radiation health effects, radiation protection, social impacts

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Authors: E. Locatelli, I. Monaco, R. C. Martin, Y. Li, R. Pini, M. Chiariello, M. Comes Franchini

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Despite the many advantages of application of nanomaterials in the field of nanomedicine, increasing concerns have been expressed on their potential adverse effects on human health. There is urgency for novel green strategies toward novel materials with enhanced biocompatibility using safe reagents. Photothermal ablation therapy, which exploits localized heat increase of a few degrees to kill cancer cells, has appeared recently as a non-invasive and highly efficient therapy against various cancer types; anyway new agents able to generate hyperthermia when irradiated are needed and must have precise biocompatibility in order to avoid damage to healthy tissues and prevent toxicity. Recently, there has been increasing interest in magnesium as a biomaterial: it is the fourth most abundant cation in the human body, and it is essential for human metabolism. However magnesium nanoparticles (Mg NPs) have had limited diffusion due to the high reduction potential of magnesium cations, which makes NPs synthesis challenging. Herein, we report the synthesis of Mg NPs and their surface functionalization for the obtainment of a stable and biocompatible nanomaterial suitable for photothermal ablation therapy against cancer. We synthesized the Mg crystals by reducing MgCl2 with metallic lithium and exploiting naphthalene as an electron carrier: the lithium–naphthalene complex acts as the real reducing agent. Firstly, the nanocrystal particles were coated with the ligand 12-ethoxy ester dodecanehydroxamic acid, and then entrapped into water-dispersible polymeric micelles (PMs) made of the FDA-approved PLGA-b-PEG-COOH copolymer using the oil-in-water emulsion technique. Lately, we developed a more straightforward methodology by introducing chitosan, a highly biocompatible natural product, at the beginning of the process, simultaneously using lithium–naphthalene complex, thus having a one-pot procedure for the formation and surface modification of MgNPs. The obtained MgNPs were purified and fully characterized, showing diameters in the range of 50-300 nm. Notably, when coated with chitosan the particles remained stable as dry powder for more than 10 months. We proved the possibility of generating a temperature rise of a few to several degrees once MgNPs were illuminated using a 810 nm diode laser operating in continuous wave mode: the temperature rise resulted significant (0-15 °C) and concentration dependent. We then investigated potential cytotoxicity of the MgNPs: we used HN13 epithelial cells, derived from a head and neck squamous cell carcinoma and the hepa1-6 cell line, derived from hepatocellular carcinoma and very low toxicity was observed for both nanosystems. Finally, in vivo photothermal therapy was performed on xenograft hepa1-6 tumor bearing mice: the animals were treated with MgNPs coated with chitosan and showed no sign of suffering after the injection. After 12 hours the tumor was exposed to near-infrared laser light. The results clearly showed an extensive damage to tumor tissue after only 2 minutes of laser irradiation at 3Wcm-1, while no damage was reported when the tumor was treated with the laser and saline alone in control group. Despite the lower photothermal efficiency of Mg with respect to Au NPs, we consider MgNPs a promising, safe and green candidate for future clinical translations.

Keywords: chitosan, magnesium nanoparticles, nanomedicine, photothermal therapy

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Authors: Muhammad Aown Sammar Raza, Salman Ahmad, Muhammad Farrukh Saleem, Muhammad Saqlain Zaheer, Rashid Iqbal, Imran Haider, Muhammad Usman Aslam, Muhammad Adnan Nazar

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One of the main negative effects of climate change is the increasing scarcity of water worldwide, especially for irrigation purpose. In order to ensure food security with less available water, there is a need to adopt easy and economic techniques. Two of the effective techniques are; use of ground covers and partial root zone drying (PRD). A field experiment was arranged to find out the most suitable mulch for PRD irrigation system in wheat. The experiment was comprised of two irrigation methods (I0 = irrigation on both sides of roots and I1= irrigation to only one side of the root as alternate irrigation) and four ground covers (M0= open ground without any cover, M1= black plastic cover, M2= wheat straw cover and M4= cotton sticks cover). More plant height, spike length, number of spikelets and number of grains were found in full irrigation treatment. While water use efficiency and grain nutrient (NPK) contents were more in PRD irrigation. All soil covers suppress the weeds and significantly influenced the yield attributes, final yield as well as the grain nutrient contents. However black plastic cover performed the best. It was concluded that joint use of both techniques was more effective for water conservation and increasing grain yield than their sole application and combination of PRD with black plastic mulch performed the best than other ground covers combination used in the experiment.

Keywords: ground covers, partial root zone drying, grain yield, quality traits, WUE, weed control efficiency

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Authors: Mhuji Kilonzo, Patrick Ndakidemi, Musa Chacha

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Objective: To evaluate antibacterial activity from four selected medicinal plants namely Mystroxylon aethiopicum, Lonchocarpus capassa, Albizia anthelmentica and Myrica salicifolia used for management of bacterial infection in Tanzania. Methods: Minimum Inhibitory Concentration (MIC) of plants extracts against the tested bacterial species was determined by using 96 wells microdilution method. In this method, 50 μL of nutrient broth were loaded in each well followed by 50 μL of extract (100 mg/mL) to make a final volume of 100 μL. Subsequently, 50 μL were transferred from first rows of each well to the second rows and the process was repeated down the columns to the last wells from which 50 μL were discarded. Thereafter, 50 μL of the selected bacterial suspension were added to each well thus making a final volume of 100 μL. The lowest concentration which showed no bacterial growth was considered as MIC. Results: It was revealed that L. capassa leaf ethyl acetate extract exhibited antibacterial activity against Salmonella kisarawe and Salmonella typhi with MIC values of 0.39 and 0.781 mg/mL respectively. Likewise, L. capassa root bark ethyl acetate extracts inhibited growth of S. typhi and E. coli with MIC values of 0.39 and 0.781 mg/mL respectively. The M. aethiopicum leaf and root bark chloroform extracts displayed antibacterial activity against S. kisarawe and S. typhi respectively with MIC value of 0.781 mg/mL. The M. salicifolia stem bark ethyl acetate exhibited antibacterial activity against P. aeruginosa with MIC value of 0.39 mg/mL whereas the methanolic stem and root bark of the same plant inhibited the growth of Proteus mirabilis and Klebsiella pneumoniae with MIC value of 0.781 mg/mL. Conclusion: It was concluded that M. aethiopicum, L. capassa, A. anthelmentica and M. salicifolia are potential source of antibacterial agents. Further studies to establish structures of antibacterial and evaluate active ingredients are recommended.

Keywords: Albizia anthelmentica, Lonchocarpus capassa, Mystroxylon aethiopicum, Myrica salicifolia

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Authors: Richard G. Boakye, Dara A Stanley, Mathavan Vickneswaran, Blanaid White

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The cultivation of cocoa (Theobroma cocoa), an important cash crop that contributes immensely towards the economic growth of several Western African countries, depends almost entirely on pesticide application owing to the plant’s vulnerability to pest and disease attacks. However, the extent to which pesticides inputted for cocoa cultivation impact bees and bee products has rarely received attention in research. Through this study, the effects of pesticides applied for cocoa cultivation on honey in Ghana were examined by evaluating honey samples from cocoa and forest ecosystems in Ghana. An analysis of five honey samples from each land use type confirmed pesticide contaminants from these land use types at measured concentrations for acetamiprid (0.051mg/kg); imidacloprid (0.004-0.02 mg/kg), thiamethoxam (0.013-0.017 mg/kg); indoxacarb (0.004-0.045 mg/kg) and sulfoxaflor (0.004-0.026 mg/kg). None of the observed pesticide concentrations exceeded EU maximum residue levels, indicating no compromise of the honey quality for human consumption. However, from the results, it could be inferred that toxic effects on bees may not be ruled out because observed concentrations largely exceeded the threshold of 0.001 mg/kg at which sublethal effects on bees have previously been reported. One of the most remarkable results to emerge from this study is the detection of imidacloprid in all honey samples analyzed, with sulfoxaflor and thiamethoxam also being detected in 93% and 73% of the honey samples, respectively. This suggests the probable prevalence of pesticide use in the landscape. However, the conclusions reached in this study should be interpreted within the scope of pesticide applications within Bia West District and not necessarily extended to other cocoa-producing districts in Ghana. Future studies should therefore include multiple cocoa-growing districts and other non-cocoa farming landscapes. Such an approach can give a broader outlook on pesticide residues in honey produced in Ghana.

Keywords: honey, cocoa, pesticides, bees, land use, landscape, residues, Ghana

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Authors: Asnate Kirse, Daina Karklina, Sandra Muizniece-Brasava, Ruta Galoburda

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Pulses are high in plant protein and dietary fiber, and contain slowly digestible starches. Innovative products from pulses could increase their consumption and benefit consumer health. This study was conducted to evaluate physicochemical stability of processed cowpea (Vigna unguiculata (L.) Walp. cv. Fradel) and maple pea (Pisum sativum var. arvense L. cv. Bruno) spreads at 5 °C temperature during 62-day storage. Physicochemical stability of pulse spreads was compared after sous vide treatment (80 °C/15 min) and high pressure processing (700 MPa/10 min/20 °C). Pulse spreads were made by homogenizing cooked pulses in a food processor together with salt, citric acid, oil, and bruschetta seasoning. A total of four different pulse spreads were studied: Cowpea spread without and with seasoning, maple pea spread without and with seasoning. Transparent PA/PE and light proof PET/ALU/PA/PP film pouches were used for packaging of pulse spreads under vacuum. The parameters investigated were pH, water activity and mass losses. Pulse spreads were tested on days 0, 15, 29, 42, 50, 57 and 62. The results showed that sous-vide treatment and high pressure processing had an insignificant influence on pH, water activity and mass losses after processing, irrespective of packaging material did not change (p>0.1). pH and water activity of sous-vide treated and high pressure processed pulse spreads in different packaging materials proved to be stable throughout the storage. Mass losses during storage accounted to 0.1% losses. Chosen sous-vide treatment and high pressure processing regimes and packaging materials are suitable to maintain consistent physicochemical quality of the new products during 62-day storage.

Keywords: cowpea, flexible packaging, maple pea, water activity

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Authors: Hosein I. Hosein, Ragab Azzam, Ahmed M. S. Menshawy, Sherin Rouby, Khaled Hendy, Ayman Mahrous, Hany Hussien

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Brucellosis is a highly contagious bacterial zoonotic disease of a worldwide spread and has different names; Infectious or enzootic abortion and Bang's disease in animals; and Mediterranean or Malta fever, Undulant Fever and Rock fever in humans. It is caused by the different species of genus Brucella which is a Gram-negative, aerobic, non-spore forming, facultative intracellular bacterium. Brucella affects a wide range of mammals including bovines, small ruminants, pigs, equines, rodents, marine mammals as well as human resulting in serious economic losses in animal populations. In human, Brucella causes a severe illness representing a great public health problem. The disease was reported in Egypt for the first time in 1939; since then the disease remained endemic at high levels among cattle, buffalo, sheep and goat and is still representing a public health hazard. The annual economic losses due to brucellosis were estimated to be about 60 million Egyptian pounds yearly, but actual estimates are still missing despite almost 30 years of implementation of the Egyptian control programme. Despite being the gold standard, bacterial isolation has been reported to show poor sensitivity for samples with low-level of Brucella and is impractical for regular screening of large populations. Thus, serological tests still remain the corner stone for routine diagnosis of brucellosis, especially in developing countries. In the present study, a total of 1533 cows (256 from Beni-Suef Governorate, 445 from Al-Fayoum Governorate and 832 from Damietta Governorate), were employed for estimation of relative sensitivity, relative specificity, positive predictive value and negative predictive value of buffered acidified plate antigen test (BPAT), rose bengal test (RBT) and complement fixation test (CFT). The overall seroprevalence of brucellosis revealed (19.63%). Relative sensitivity, relative specificity, positive predictive value and negative predictive value of BPAT,RBT and CFT were estimated as, (96.27 %, 96.76 %, 87.65 % and 99.10 %), (93.42 %, 96.27 %, 90.16 % and 98.35%) and (89.30 %, 98.60 %, 94.35 %and 97.24 %) respectively. BPAT showed the highest sensitivity among the three employed serological tests. RBT was less specific than BPAT. CFT showed the least sensitivity 89.30 % among the three employed serological tests but showed the highest specificity. Different tissues specimens of 22 seropositive cows (spleen, retropharyngeal udder, and supra-mammary lymph nodes) were subjected for bacteriological studies for isolation and identification of Brucella organisms. Brucella melitensis biovar 3 could be recovered from 12 (54.55%) cows. Bacteriological examinations failed to classify 10 cases (45.45%) and were culture negative. Bruce-ladder PCR was carried out for molecular identification of the 12 Brucella isolates at the species level. Three fragments of 587 bp, 1071 bp and 1682 bp sizes were amplified indicating Brucella melitensis. The results indicated the importance of using several procedures to overcome the problem of escaping of some infected animals from diagnosis.Bruce-ladder PCR is an important tool for diagnosis and epidemiologic studies, providing relevant information for identification of Brucella spp.

Keywords: brucellosis, relative sensitivity, relative specificity, Bruce-ladder, Egypt

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Authors: Ying-Ming Su, Mei-Shu Huang

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To mitigate the urban heat island effect has become a global issue facing the challenge of climate change. Through literature reviews, plant photosynthesis can reduce the carbon dioxide and mitigate the urban heat island effect relatively. Because there are not enough open space and park, green roof has become an important policy in Taiwan. We selected elementary school buildings in northern New Taipei City as research subjects since elementary school is asked priority to build green roof and important educational place to promote green roof concept. Testo 175-H1 recording device was used to record the temperature and humidity difference between roof surface and interior space below roof with and without green roof for the long-term. We also use questionnaire to investigate the awareness of comfort level of green roof and sensation of teachers and students of the elementary school. The results indicated the temperature of roof without greening was higher than that with greening about 2°C. But sometimes during noontime, the temperature of green roof was higher than that of non-green roof related to the character of the accumulation and dissipation of heat of greening probably. The temperature of interior space below green roof was normally lower than that without green roof about 1°C showed that green roof could lower the temperature. The humidity of the green roof was higher than the one without greening also indicated that green roof retained water better. Teachers liked to combine green roof concept in the curriculum, students wished all classes can take turns to maintain the green roof. Teachers and students that school had integrated green roof concept in the curriculum were more willing to participate in the maintenance work of green roof. Teachers and students who may access and touch the green roof can be more aware of the green roof benefit. We suggest architect to increase the accessibility and visibility of green roof, such as a part of the activity space. This idea can be a reference of the green roof curriculum design.

Keywords: comfort level, elementary school, green roof, heat island effect

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Authors: Piotr Wadolowski, Grzegorz Krzesinski, Piotr Gutowski

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The aim of the presented work is to recognize the impact of mini-plate application approach on the stress and displacement within the stabilization devices and surrounding bones. The mini-plate osteosynthesis technique is widely used by craniofacial surgeons as an improved replacement of wire connection approach. Many different types of metal plates and screws are used to the physical connection of fractured bones. Below investigation is based on a clinical observation of patient hospitalized with mini-plate stabilization system. Analysis was conducted on a solid mandible geometry, which was modeled basis on the computed tomography scan of the hospitalized patient. In order to achieve most realistic connected system behavior, the cortical and cancellous bone layers were assumed. The temporomandibular joint was simplified to the elastic element to allow physiological movement of loaded bone. The muscles of mastication system were reduced to three pairs, modeled as shell structures. Finite element grid was created by the ANSYS software, where hexahedral and tetrahedral variants of SOLID185 element were used. A set of nonlinear contact conditions were applied on connecting devices and bone common surfaces. Properties of particular contact pair depend on screw - mini-plate connection type and possible gaps between fractured bone around osteosynthesis region. Some of the investigated cases contain prestress introduced to the mini-plate during the application, what responds the initial bending of the connecting device to fit the retromolar fossa region. Assumed bone fracture occurs within the mandible angle zone. Due to the significant deformation of the connecting plate in some of the assembly cases the elastic-plastic model of titanium alloy was assumed. The bone tissues were covered by the orthotropic material. As a loading were used the gauge force of magnitude of 100N applied in three different locations. Conducted analysis shows significant impact of mini-plate application methodology on the stress distribution within the miniplate. Prestress effect introduces additional loading, which leads to locally exceed the titanium alloy yield limit. Stress in surrounding bone increases rapidly around the screws application region, exceeding assumed bone yield limit, what indicate the local bone destruction. Approach with the doubled mini-plate shows increased stress within the connector due to the too rigid connection, where the main path of loading leads through the mini-plates instead of plates and connected bones. Clinical observations confirm more frequent plate destruction of stiffer connections. Some of them could be an effect of decreased low cyclic fatigue capability caused by the overloading. The executed analysis prove that the mini-plate system provides sufficient support to mandible fracture treatment, however, many applicable solutions shifts the entire system to the allowable material limits. The results show that connector application with the initial loading needs to be carefully established due to the small material capability tolerances. Comparison to the clinical observations allows optimizing entire connection to prevent future incidents.

Keywords: mandible fracture, mini-plate connection, numerical analysis, osteosynthesis

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Authors: Mohammed A. Alshehri

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Salvia officinalis is an aromatic plant member of the mint (Labiatae) family. It is popular kitchen herb. Not surprise to find that the name of this herb related to cure, in Latin language Salvia means to cure where officinalis means medicinal which answer why the sage has a top place in the list of medicinal plants. The aim of the present study was to assess the genetic damage and cytological changes caused by exposure of the test organism (Rattusrattus) to Salvia officinals. For this purpose, adult female rats, weighing 200–250 g, were used as donors. A total of 36 adult Wister male rats were randomly assigned to five groups: the experimental groups (rats were intraperitonealy injected with Salvia officinalis pure extract at (0.1, 0.2, 0.5, 0.1mg/kg body weight, the same dose was administered once a day. Control group (rats were injected intraperitonealy physiological saline. And positive control were injected with Cyclophosphamide. On the 21st days following Salvia officinalis pure extract exposure, rats were sacrificed, and samples of bone marrow were collected. Following that, we performed a micronuclei (MN) test using MNNCE (Micro-nucleated normocromatic erythrocytes) and MNPCE (Micronucleated polychromatic erythrocytes), NDI (Nuclear division index), and cytological parameters using NDCI (nuclear division cytotoxicity index), necrotic, and apoptotic cells in rat's bone marrow samples. Results showed that there was a no significant increase in the frequency of micro-nucleatedas well as in cytological parameters in bone marrow cells. In light of these results, if Salvia officinalis pure extract may considered to be safe from the stand point of genotoxicity and cytotoxicity effects.

Keywords: Salvia officinalis, micronucleus, NDI, NDCI, toxicity, chromosomal aberrations

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Authors: Muhammad Iqbal, Hafiz Muhammad Tauqeer, Hamza Rafaqat, Muhammad Naveed, Muhammad Awais Irshad

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Soil pollution with chromium (Cr) has become one of the most important concerns due to its toxicity for humans. To date, various remediation approaches have been employed for the remediation and management of Cr contaminated soils. Phytoextraction is an eco-friendly and emerging remediation approach which has gained attention due to several advantages over conventional remediation approach. The use of energy crops for phytoremediation is an emerging trend worldwide. These energy crops have high tolerance against various environmental stresses, the potential to grow in diverse ecosystems and high biomass production make them a suitable candidate for phytoremediation of contaminated soils. The removal efficiency of plants in phytoextraction depends upon several soil and plant factors including solubility, bioavailability and metal speciation in soils. A pot scale experiment was conducted to evaluate the phytoextraction potential of Arundo donax L. with the application of acetic acid (A.A) in Cr contaminated soils. Plants were grown in pots filled with 5 kg soils for 90 days. After 30 days plants acclimatization in pot conditions, plants were treated with various levels of Cr (2.5 mM, 5 mM, 7.5 mM, 10 mM) and A.A (Cr 2.5 mM + A.A 2.5 mM, Cr 5 mM + A.A 2.5 mM, Cr 7.5 mM + A.A 2.5 mM, Cr 10 mM + A.A 2.5 mM). The application of A.A significantly increased metal uptake and in roots and shoots of A. donax. This increase was observed at Cr 7.5 mM + A.A 2.5 mM but at high concentrations, visual symptoms of Cr toxicity were observed on leaves. Similarly, A.A applications also affect the activities of key enzymes including catalase (CAT), superoxidase dismutase (SOD), and ascorbate peroxidase (APX) in leaves of A. donax. Based on results it is concluded that the applications of A.A acid for phytoextraction is an alternative approach for the management of Cr affected soils and synthetic chelators should be replaced with organic acids.

Keywords: acetic acid, A. donax, chromium, energy crop, phytoextraction

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Authors: Yakubu Suleiman, S. A. Musa

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The research was conducted to determine the physical characteristics of banana fruits that influenced consumer preferences for the fruit in Northern Nigeria. Socio-economic characteristics of the respondents were also identified. Simple descriptive statistics and Hedonic prices model were used to analyze the data collected for socio-economic and consumer preference respectively with the aid of 1000 structured questionnaires. The result revealed the value of R2 to be 0.633, meaning that, 63.3% of the variation in the banana price was brought about by the explanatory variables included in the model and the variables are: colour, size, degree of ripeness, softness, surface blemish, cleanliness of the fruits, weight, length, and cluster size of fruits. However, the remaining 36.7% could be attributed to the error term or random disturbance in the model. It could also be seen from the calculated result that the intercept was 1886.5 and was statistically significant (P < 0.01), meaning that about N1886.5 worth of banana fruits could be bought by consumers without considering the variables of banana included in the model. Moreover, consumers showed that they have significant preference for colours, size, degree of ripeness, softness, weight, length and cluster size of banana fruits and they were tested to be significant at either P < 0.01, P < 0.05, and P < 0.1 . Moreover, the result also shows that consumers did not show significance preferences to surface blemish, cleanliness and variety of the banana fruit as all of them showed non-significance level with negative signs. Based on the findings of the research, it is hereby recommended that plant breeders and research institutes should concentrate on the production of banana fruits that have those physical characteristics that were found to be statistically significance like cluster size, degree of ripeness,’ softness, length, size, and skin colour.

Keywords: analysis, consumers, preference, variables

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Authors: N. Hachemi, I. Benmehdi, O. Hasnaoui

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The air like water is an essential element for living beings. Each day, a man breathes about 20m3 of air. It originally consists of a set of gas whose presence and concentrations correspond to the needs of life. This study focuses on air pollution by smoke and dust emitted from the chimney of the cement works of Beni Saf, pathological and their impact on the environment. Dust of the cement plant are harmless to permissible levels for living organisms, but the two combined phenomena namely the release of dust and aridity of the climate, which severely marked area of Beni Saf; have contributed adverse effects in on human health and the degradation of vegetation cover and species especially weakened by environmental stress. The most visible impact is certainly the deposition of dust on the surrounding areas of the cement factory, and seriously affecting the aesthetics of the landscape. Health problems are more important inside and outside the factory. Among the diseases notable caused by the cement works are: deafness, heart disease, asthma and mental. The dust of the cement works is mainly composed of fine particles of limestone, clay, free lime, silicates and also loaded of the gases such as carbon dioxide gas CO2. The accumulation of this gas in the atmosphere is directly involved in the phenomenon of increasing of greenhouse effect. Some gases, for example, are directly toxic. They can change the climate, changing precipitation types and become a greater source of stress by drought, etc. The environment also suffers from air pollution indirectly; it is more precisely the acid rain. They are produced by the combustion of non-metals in air. Acid rain has consequences for contaminating the soil, weakening the flora, fauna and acidifies lakes. Finally, the pollution problems are multiple and specific dust. It can worsen and change, it has reached epidemic proportions quantitatively and qualitatively disturbing and unpredictable.

Keywords: atmospheric pollution, cement, dust, environment

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Authors: Vibeke Sorensen, Nagaraju Thummanapalli, J. Stephen Lansing

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Inspired by the indigenous people of Borneo, Indonesia and their traditional bark cloth, artist and professor Vibeke Sorensen executed a “digital unwrapping” of several trees in Southeast Asia using a digital panorama camera and digitally “stitched” them together for printing onto sustainable silk and fashioning into the “Tree Dress”. This dress is a symbolic “un-wrapping” and “re-wrapping” of the tree’s bark onto a person as a second skin. The “digital bark” is directly responsive to the real tree through embedded and networked electronics that connect in real-time to sensors at the physical site of the living tree. LEDs and circuits inserted into the dress display the continuous measurement of the O2 / CO2, temperature, humidity, and light conditions at the tree. It is an “Internet of Living Things” (IOLT) textile that can be worn to track and interact with it. The computer system connecting the dress and the tree converts the gas emission data at the site of the real tree into sound and music as sonification. This communicates not only the scientific data but also translates it into a poetic representation. The wearer of the garment can symbolically identify with the tree, or “become one” with it by adorning its “skin.” In this way, the wearer also becomes a human agent for the tree, bringing its actual condition to direct perception of the wearer and others who may engage it. This project is an attempt to bring greater awareness to issues of deforestation by providing a direct access to living things separated by physical distance, and hopefully, to increase empathy for them by providing a way to sense individual trees and their daily existential condition through remote monitoring of data. Further extensions to this project and related issues of sustainability include the use of recycled and alternative plant materials such as bamboo and air plants, among others.

Keywords: IOLT, sonification, sustainability, tree, wearable technology

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Authors: Sunday Olufemi Adesogan

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The supply of potable water at least is a very important index in national development. Water tariffs depend on the treatment cost which carries the highest percentage of the total operation cost in any water supply scheme. In order to keep water tariffs as low as possible, treatment costs have to be minimized. The University of Ibadan, Nigeria, water supply scheme consists of a treatment plant with three distribution stations (Amina way, Kurumi and Lander) and two raw water supply sources (Awba dam and Eleyele dam). An operational study of the scheme was carried out to ascertain the efficiency of the supply of potable water on the campus to justify the need for water supply schemes in tertiary institutions. The study involved regular collection, processing and analysis of periodic operational data. Data collected include supply reading (water production on daily basis) and consumers metered reading for a period of 22 months (October 2013 - July 2015), and also collected, were the operating hours of both plants and human beings. Applying the required mathematical equations, total loss was determined for the distribution system, which was translated into monetary terms. Adequacies of the operational functions were also determined. The study revealed that water supply scheme is justified in tertiary institutions. It was also found that approximately 10.7 million Nigerian naira (N) is lost to leakages during the 22-month study period; the system’s storage capacity is no longer adequate, especially for peak water production. The capacity of the system as a whole is insufficient for the present university population and that the existing water supply system is not being operated in an optimal manner especially due to personnel, power and system ageing constraints.

Keywords: development, panacea, supply, water

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Authors: Larry V. Aceres, Jesryl B. Paulite, Emelie M. Pelicano, J. B. Anciano, J. A. Esteban

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Enhancing the population of beneficial arthropods under less diverse agroecosystem is the most sought by many researchers and plant growers. This strategy was done through the establishment of pintoi peanut, Arachis pintoi as live mulch or cover crop in pummelo orchard of the University of Southeastern Philippines (USeP), Mabini, Compostela Valley Province, Philippines. This study was conducted to compare and compute population dynamics and diversity of beneficial arthropods in pummelo in with and without Arachis pintoi cover crop. Data collections were done for the 12-month period (from June 2013 to May 2014) at the pummelo orchard of USeP Mabini Campus, COMVAL Province, Philippines and data were analyzed using the Independent Samples T-Test to compare the effect of the presence and absence of Arachis pintoi on beneficial arthropods incidence in pummelo orchard. Moreover, diversity and family richness analyses were computed using the Margalef’s diversity index for family richness; the Shannon index of general diversity and the evenness index; and the Simpson index of dominance. Results revealed numerically and statistically higher density of important beneficial arthropods such as microhymenopterans, macrohymenopterans, spiders, tachinid flies and ground beetles were recorded in pummelo orchard with Arachis pintoi than from without Arachis pintoi cover crop for the 12-month observation period. Further, the result of the study revealed the high family richness and diversity index with more or less even distribution of individuals within the family and low dominance index were documented in pummelo with Arachis pintoi cover crop than from pummelo without Arachis pintoi cover crop. The study revealed that planting A. pintoi in pummelo orchard could enhance natural enemy populations.

Keywords: Arachis pintoi, cover crop, beneficial arthropods, pummelo

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Authors: Kabange Kasumbwe, Viresh Mohanlall, Bharti Odhav, Venu Narayanaswamy

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Coumarins are naturally occurring plant metabolites known for their pharmacological properties such as anticoagulant, antimicrobial, anticancer, antioxidant, anti-inflammatory and antiviral properties. The pharmacological and biochemical properties and curative applications of coumarins depend on the substitution around the coumarin core structure. In the present study, seven halogenated coumarins CMRN1-CMRN7 were synthesized and evaluated for their anticancer activity. The cytotoxicity potential of the test compounds was evaluated against UACC62 (Melanoma), MCF-7 (Breast cancer) and PBM (Peripheral Blood Mononuclear) cell lines using MTT assay keeping doxorubicin as standard drug. The apoptotic potential of the coumarin compounds was evaluated against UACC62 (Melanoma) cell by assessing their morphological changes, membrane change, mitochondria membrane potential; pro-apoptotic changes were investigated using the AnnexinV-PI staining, JC-1, caspase-3 enzyme kits respectively on flow cytometer. The synthetic coumarin has strongly suppressed the cell proliferation of UACC-62 (Melanoma) and MCF-7 (Breast) Cancer cells, the higher toxicity of these compounds against UACC-62 (Melanoma) and MCF-7 (Breast) were CMRN3, CMRN4, CMRN5, CMRN6. However, compounds CMRN1, CMRN2, and CMRN7 had no significant inhibitory effect. Furthermore the active compounds CMRN3, CMRN4, CMRN5, CMRN6 exerted antiproliferative effects through apoptosis induction against UACC-62 (Melanoma), suggesting their potential could be considered as attractive lead molecules in the future for the development of potential anticancer agents since one of the important criteria in the development of therapeutic drugs for cancer treatment is to have high selectivity and less or no side-effects on normal cells and these compounds had no inhibitory effect against the PBMC cells.

Keywords: coumarin, MTT, apoptosis, cytotoxicity

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Authors: Martyna Ekiert, Natalia Grzechnik, Joanna Karbowniczek, Urszula Stachewicz, Andrzej Mlyniec

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Tendon grafting is a common procedure performed to treat tendon rupture. Before the surgical procedure, tissues intended for grafts (i.e., Achilles tendon) are stored in ultra-low temperatures for a long time and also may be subjected to unfavorable conditions, such as repetitive freezing (F) and thawing (T). Such storage protocols may highly influence the graft mechanical properties, decrease its functionality and thus increase the risk of complications during the transplant procedure. The literature reports on the influence of multiple F/T cycles on internal structure and mechanical properties of tendons stay inconclusive, confirming and denying the negative influence of multiple F/T at the same time. An inconsistent research methodology and lack of clear limit of F/T cycles, which disqualifies tissue for surgical graft purposes, encouraged us to investigate the issue of multiple F/T cycles by the mean of biomechanical tensile tests supported with Scanning Electron Microscope (SEM) imaging. The study was conducted on male bovine Achilles tendon-derived from the local abattoir. Fresh tendons were cleaned of excessive membranes and then sectioned to obtained fascicle bundles. Collected samples were randomly assigned to 6 groups subjected to 1, 2, 4, 6, 8 and 12 cycles of freezing-thawing (F/T), respectively. Each F/T cycle included deep freezing at -80°C temperature, followed by thawing at room temperature. After final thawing, thin slices of the side part of samples subjected to 1, 4, 8 and 12 F/T cycles were collected for SEM imaging. Then, the width and thickness of all samples were measured to calculate the cross-sectional area. Biomechanical tests were performed using the universal testing machine (model Instron 8872, INSTRON®, Norwood, Massachusetts, USA) using a load cell with a maximum capacity of 250 kN and standard atmospheric conditions. Both ends of each fascicle bundle were manually clamped in grasping clamps using abrasive paper and wet cellulose wadding swabs to prevent tissue slipping while clamping and testing. Samples were subjected to the testing procedure including pre-loading, pre-cycling, loading, holding and unloading steps to obtain stress-strain curves for representing tendon stretching and relaxation. The stiffness of AT fascicles bundle samples was evaluated in terms of modulus of elasticity (Young’s modulus), calculated from the slope of the linear region of stress-strain curves. SEM imaging was preceded by chemical sample preparation including 24hr fixation in 3% glutaraldehyde buffered with 0.1 M phosphate buffer, washing with 0.1 M phosphate buffer solution and dehydration in a graded ethanol solution. SEM images (Merlin Gemini II microscope, ZEISS®) were taken using 30 000x mag, which allowed measuring a diameter of collagen fibrils. The results confirm a decrease in fascicle bundles Young’s modulus as well as a decrease in the diameter of collagen fibrils. These results confirm the negative influence of multiple F/T cycles on the mechanical properties of tendon tissue.

Keywords: biomechanics, collagen, fascicle bundles, soft tissue

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Authors: Ajay Kumar, Satwinderjeet Kaur

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Onosma bracteata Wall. (Boraginaceae), is known to be a medicinal plant, useful in the treatment of body swellings, abdominal pain and urinary calculi, etc. The present study focused on the radical scavenging and cancer growth inhibitory properties of isolates from O. bracteata. Obea fraction demonstrated noticeable free radical scavenging ability along with antiproliferative activity in human osteosarcoma MG-63, human neuroblastoma IMR-32, and human lung cancer A549 cell lines using MTT assay with GI50 values of 88.56, 101.61 and 112.7 μg/ml, respectively. The scanning electron and confocal microscopy studies showed morphological alterations including nuclear condensation and formation of apoptotic bodies in osteosarcoma MG-63 cells. Obea fraction in osteosarcoma MG-63 cells augmented the reactive oxygen species (ROS) level and decreased the mitochondrial membrane potential. Flow cytometry analysis revealed the Obea treated cells to be arrested in the G0/G1 phase in a dose dependent manner supported by the observed increase in the early apoptotic cell population. Western blotting analysis showed that the expression of p-NF-kB, COX-2, p-Akt, and Bcl-xL decreased whereas, the expression of GSK-3β, p53, caspase-3 and caspase-9 proteins increased. The downregulation of Bcl-2, Cyclin E, CDK2 and mortalin gene expression and upregulation of p53 genes was unfolded in RT-qPCR studies. The presence of catechin, kaempferol, Onosmin A and epicatechin, as revealed in high-performance liquid chromatography (HPLC) studies, contributes towards the chemopreventive potential of O. bracteata which can be tapped for chemotherapeutic use.

Keywords: apoptosis, confocal microscopy, HPLC, mitochondria membrane potential, reactive oxygen species

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Authors: Michael Josef Schwerer

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Background: Analyzing any aircraft accident is mandatory based on the regulations of the International Civil Aviation Organization and the respective country’s criminal prosecution authorities. Legal medicine investigations are unavoidable when fatalities involve the flight crew or when doubts arise concerning the pilot’s aeromedical health status before the event. As a result of frequently tremendous blunt and sharp force trauma along with the impact of the aircraft to the ground, consecutive blast or fire exposition of the occupants or putrefaction of the dead bodies in cases of delayed recovery, relevant findings can be masked or destroyed and therefor being inaccessible in standard pathology practice comprising just forensic autopsy and histopathology. Such cases are of considerable risk of remaining unsolved without legal consequences for those responsible. Further, no lessons can be drawn from these scenarios to improve flight safety and prevent future mishaps. Aims and Methods: To learn from previously unsolved aircraft accidents, re-evaluations of the investigation files and modern molecular pathology studies were performed. Genetic testing involved predominantly PCR-based analysis of gene regulation, studying DNA promotor methylations, RNA transcription and posttranscriptional regulation. In addition, the presence or absence of infective agents, particularly DNA- and RNA-viruses, was studied. Technical adjustments of molecular genetic procedures when working with archived sample material were necessary. Standards for the proper interpretation of the respective findings had to be settled. Results and Discussion: Additional molecular genetic testing significantly contributes to the quality of forensic pathology assessment in aviation mishaps. Previously undetected cardiotropic viruses potentially explain e.g., a pilot’s sudden incapacitation resulting from cardiac failure or myocardial arrhythmia. In contrast, negative results for infective agents participate in ruling out concerns about an accident pilot’s fitness to fly and the aeromedical examiner’s precedent decision to issue him or her an aeromedical certificate. Care must be taken in the interpretation of genetic testing for pre-existing diseases such as hypertrophic cardiomyopathy or ischemic heart disease. Molecular markers such as mRNAs or miRNAs, which can establish these diagnoses in clinical patients, might be misleading in-flight crew members because of adaptive changes in their tissues resulting from repeated mild hypoxia during flight, for instance. Military pilots especially demonstrate significant physiological adjustments to their somatic burdens in flight, such as cardiocirculatory stress and air combat maneuvers. Their non-pathogenic alterations in gene regulation and expression will likely be misinterpreted for genuine disease by inexperienced investigators. Conclusions: The growing influence of molecular pathology on legal medicine practice has found its way into aircraft accident investigation. As appropriate quality standards for laboratory work and data interpretation are provided, forensic genetic testing supports the medico-legal analysis of aviation mishaps and potentially reduces the number of unsolved events in the future.

Keywords: aviation medicine, aircraft accident investigation, forensic pathology, molecular pathology

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