Search results for: reverse twin block

Authors: Ozan Ozkan, Hilal Turkoglu Sasmazel

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Natural polymers are widely used in tissue engineering applications, because of their biocompatibility, biodegradability and solubility in the physiological medium. On the other hand, synthetic polymers are also widely utilized in tissue engineering applications, because they carry no risk of infectious diseases and do not cause immune system reaction. However, the disadvantages of both polymer types block their individual usages as tissue scaffolds efficiently. Therefore, the idea of usage of natural and synthetic polymers together as a single 3D hybrid scaffold which has the advantages of both and the disadvantages of none has been entered to the literature. On the other hand, even though these hybrid structures support the cell adhesion and/or proliferation, various surface modification techniques applied to the surfaces of them to create topographical changes on the surfaces and to obtain reactive functional groups required for the immobilization of biomolecules, especially on the surfaces of synthetic polymers in order to improve cell adhesion and proliferation. In a study presented here, to improve the surface functionality and topography of the layer by layer electrospun 3D poly-epsilon-caprolactone/chitosan/poly-epsilon-caprolactone hybrid tissue scaffolds by using atmospheric pressure plasma method, thus to improve cell adhesion and proliferation of these tissue scaffolds were aimed. The formation/creation of the functional hydroxyl and amine groups and topographical changes on the surfaces of scaffolds were realized by using two different atmospheric pressure plasma systems (nozzle type and dielectric barrier discharge (DBD) type) carried out under different gas medium (air, Ar+O2, Ar+N2). The plasma modification time and distance for the nozzle type plasma system as well as the plasma modification time and the gas flow rate for DBD type plasma system were optimized with monitoring the changes in surface hydrophilicity by using contact angle measurements. The topographical and chemical characterizations of these modified biomaterials’ surfaces were carried out with SEM and ESCA, respectively. The results showed that the atmospheric pressure plasma modifications carried out with both nozzle type plasma and DBD plasma caused topographical and functionality changes on the surfaces of the layer by layer electrospun tissue scaffolds. However, the shelf life studies indicated that the hydrophilicity introduced to the surfaces was mainly because of the functionality changes. Therefore, according to the optimized results, samples treated with nozzle type air plasma modification applied for 9 minutes from a distance of 17 cm and Ar+O2 DBD plasma modification applied for 1 minute under 70 cm3/min O2 flow rate were found to have the highest hydrophilicity compared to pristine samples.

Keywords: biomaterial, chitosan, hybrid, plasma

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Authors: Dalal Alghawas, Jetty Lee, Kaisa Poutanen, Hani El-Nezami

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Oat β-glucan a water soluble non starch linear polysaccharide has been approved as a cholesterol lowering agent by various food safety administrations and is commonly used to reduce the risk of heart disease. The molecular weight of oat β-glucan can vary depending on the extraction and fractionation methods. It is not clear whether the molecular weight has a significant impact at reducing the acceleration of atherosclerosis. The aim of this study was to investigate three different oat β-glucan fractionations on the development of atherosclerosis in vivo. With special focus on plaque stability and the intestinal barrier function. To test this, ApoE-/- female mice were fed a high fat diet supplemented with oat bran, high molecular weight (HMW) oat β-glucan fractionate and low molecular weight (LMW) oat β-glucan fractionate for 16 weeks. Atherosclerosis risk markers were measured in the plasma, heart and aortic tree. Plaque size was measured in the aortic root and aortic tree. ICAM-1, VCAM-1, E-Selectin, P-Selectin, protein levels were assessed from the aortic tree to determine plaque stability at 16 weeks. The expression of p22phox at the aortic root was evaluated to study the NADPH oxidase complex involved in nitric oxide bioavailability and vascular elasticity. The tight junction proteins E-cadherin and beta-catenin from western blot analyses were analysed as an intestinal barrier function test. Plasma LPS, intestinal D-lactate levels and hepatic FMO gene expression were carried out to confirm whether the compromised intestinal barrier lead to endotoxemia. The oat bran and HMW oat β-glucan diet groups were more effective than the LMW β-glucan diet group at reducing the plaque size and showed marked improvements in plaque stability. The intestinal barrier was compromised for all the experimental groups however the endotoxemia levels were higher in the LMW β-glucan diet group. The oat bran and HMW oat β-glucan diet groups were more effective at attenuating the development of atherosclerosis. Reasons for this could be due to the LMW oat β-glucan diet group’s low viscosity in the gut and the inability to block the reabsorption of cholesterol. Furthermore the low viscosity may allow more bacterial endotoxin translocation through the impaired intestinal barrier. In future food technologists should carefully consider how to incorporate LMW oat β-glucan as a health promoting food.

Keywords: Atherosclerosis, beta glucan, endotoxemia, intestinal barrier function

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Authors: Misheck Musokwa, Paramu Mafongoya, Simon Lorentz

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Seasonal shortages of fodder during the dry season is a major constraint to smallholder livestock farmers in South Africa. To mitigate the shortage of fodder, legume trees can be intercropped with pastures which can diversify the sources of feed and increase the amount of protein for grazing animals. The objective was to evaluate dry matter yield of Panicum maximum and land productivity under different fodder production systems during 2016/17-2017/18 seasons at Empangeni (28.6391° S and 31.9400° E). A randomized complete block design, replicated three times was used, the treatments were sole Panicum maximum, Panicum maximum + Sesbania sesban, Panicum maximum + pigeonpea, sole Sesbania sesban, Sole pigeonpea. Three months S.sesbania seedlings were transplanted whilst pigeonpea was direct seeded at spacing of 1m x 1m. P. maximum seeds were drilled at a respective rate of 7.5 kg/ha having an inter-row spacing of 0.25 m apart. In between rows of trees P. maximum seeds were drilled. The dry matter yield harvesting times were separated by six months’ timeframe. A 0.25 m² quadrant randomly placed on 3 points on the plot was used as sampling area during harvesting P. maximum. There was significant difference P < 0.05 across 3 harvests and total dry matter. P. maximum had higher dry matter yield as compared to both intercrops at first harvest and total. The second and third harvest had no significant difference with pigeonpea intercrop. The results was in this order for all 3 harvest: P. maximum (541.2c, 1209.3b and 1557b) kg ha¹ ≥ P. maximum + pigeonpea (157.2b, 926.7b and 1129b) kg ha¹ > P. maximum + S. sesban (36.3a, 282a and 555a) kg ha¹. Total accumulation of dry matter yield of P. maximum (3307c kg ha¹) > P. maximum + pigeonpea (2212 kg ha¹) ≥ P. maximum + S. sesban (874 kg ha¹). There was a significant difference (P< 0.05) on seed yield for trees. Pigeonpea (1240.3 kg ha¹) ≥ Pigeonpea + P. maximum (862.7 kg ha¹) > S.sesbania (391.9 kg ha¹) ≥ S.sesbania + P. maximum. The Land Equivalent Ratio (LER) was in the following order P. maximum + pigeonpea (1.37) > P. maximum + S. sesban (0.84) > Pigeonpea (0.59) ≥ S. Sesbania (0.57) > P. maximum (0.26). Results indicates that it is beneficial to have P. maximum intercropped with pigeonpea because of higher land productivity. Planting grass with pigeonpea was more beneficial than S. sesban with grass or sole cropping in terms of saving the shortage of arable land. P. maximum + pigeonpea saves a substantial (37%) land which can be subsequently be used for other crop production. Pigeonpea is recommended as an intercrop with P. maximum due to its higher LER and combined production of livestock feed, human food, and firewood. Panicum grass is low in crude protein though high in carbohydrates, there is a need for intercropping it with legume trees. A farmer who buys concentrates can reduce costs by combining P. maximum with pigeonpea this will provide a balanced diet at low cost.

Keywords: fodder, livestock, productivity, smallholder farmers

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Authors: Iryna Biskub

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The interdisciplinary approach to modern linguistic studies is exemplified by the merge of various research methods, which sometimes causes complications related to the verification of the research results. This methodological confusion can be resolved by means of creating new techniques of linguistic analysis combining several scientific paradigms. Modern linguistics has developed really productive and efficient methods for the investigation of cognitive and communicative phenomena of which language is the central issue. In the field of discourse studies, one of the best examples of research methods is the method of Critical Discourse Analysis (CDA). CDA can be viewed both as a method of investigation, as well as a critical multidisciplinary perspective. In CDA the position of the scholar is crucial from the point of view exemplifying his or her social and political convictions. The generally accepted approach to obtaining scientifically reliable results is to use a special well-defined scientific method for researching special types of language phenomena: cognitive methods applied to the exploration of cognitive aspects of language, whereas communicative methods are thought to be relevant only for the investigation of communicative nature of language. In the recent decades discourse as a sociocultural phenomenon has been the focus of careful linguistic research. The very concept of discourse represents an integral unity of cognitive and communicative aspects of human verbal activity. Since a human being is never able to discriminate between cognitive and communicative planes of discourse communication, it doesn’t make much sense to apply cognitive and communicative methods of research taken in isolation. It is possible to modify the classical CDA procedure by means of mapping human cognitive procedures onto the strategic communicative planning of discourse communication. The analysis of the electronic petition 'Block Donald J Trump from UK entry. The signatories believe Donald J Trump should be banned from UK entry' (584, 459 signatures) and the parliamentary debates on it has demonstrated the ability to map cognitive and communicative levels in the following way: the strategy of discourse modeling (communicative level) overlaps with the extraction of semantic macrostructures (cognitive level); the strategy of discourse management overlaps with the analysis of local meanings in discourse communication; the strategy of cognitive monitoring of the discourse overlaps with the formation of attitudes and ideologies at the cognitive level. Thus, the experimental data have shown that it is possible to develop a new complex methodology of discourse analysis, where cognition would meet communication, both metaphorically and literally. The same approach may appear to be productive for the creation of computational models of human-computer interaction, where the automatic generation of a particular type of a discourse could be based on the rules of strategic planning involving cognitive models of CDA.

Keywords: cognition, communication, discourse, strategy

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Authors: Ahmad Kayvani Fard, Yehia Manawi

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Qatar’s primary source of fresh water is through seawater desalination. Amongst the major processes that are commercially available on the market, the most common large scale techniques are Multi-Stage Flash distillation (MSF), Multi Effect distillation (MED), and Reverse Osmosis (RO). Although commonly used, these three processes are highly expensive down to high energy input requirements and high operating costs allied with maintenance and stress induced on the systems in harsh alkaline media. Beside that cost, environmental footprint of these desalination techniques are significant; from damaging marine eco-system, to huge land use, to discharge of tons of GHG and huge carbon footprint. Other less energy consuming techniques based on membrane separation are being sought to reduce both the carbon footprint and operating costs is membrane distillation (MD). Emerged in 1960s, MD is an alternative technology for water desalination attracting more attention since 1980s. MD process involves the evaporation of a hot feed, typically below boiling point of brine at standard conditions, by creating a water vapor pressure difference across the porous, hydrophobic membrane. Main advantages of MD compared to other commercially available technologies (MSF and MED) and specially RO are reduction of membrane and module stress due to absence of trans-membrane pressure, less impact of contaminant fouling on distillate due to transfer of only water vapor, utilization of low grade or waste heat from oil and gas industries to heat up the feed up to required temperature difference across the membrane, superior water quality, and relatively lower capital and operating cost. To achieve the objective of this study, state of the art flat-sheet cross-flow DCMD bench scale unit was designed, commissioned, and tested. The objective of this study is to analyze the characteristics and morphology of the membrane suitable for DCMD through SEM imaging and contact angle measurement and to study the water quality of distillate produced by DCMD bench scale unit. Comparison with available literature data is undertaken where appropriate and laboratory data is used to compare a DCMD distillate quality with that of other desalination techniques and standards. Membrane SEM analysis showed that the PTFE membrane used for the study has contact angle of 127º with highly porous surface supported with less porous and bigger pore size PP membrane. Study on the effect of feed solution (salinity) and temperature on water quality of distillate produced from ICP and IC analysis showed that with any salinity and different feed temperature (up to 70ºC) the electric conductivity of distillate is less than 5 μS/cm with 99.99% salt rejection and proved to be feasible and effective process capable of consistently producing high quality distillate from very high feed salinity solution (i.e. 100000 mg/L TDS) even with substantial quality difference compared to other desalination methods such as RO and MSF.

Keywords: membrane distillation, waste heat, seawater desalination, membrane, freshwater, direct contact membrane distillation

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Authors: Tadesse Kebede, Orkun Baris Kovanci

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Tomatoes leaf minor (Tutaabasoluta) is one of the most economically important insect pest in tomatoes production. The use of biological control such as entomopathogen fungi isolates would be a long-term and cost-effective solution to control insects pest. Therefore, identifying the most virulent and pathogenic entomopathogen fungi is one of the basic requirements for effective management options to combat Tomatoes leaf minor (Tutaabasoluta). Furthermore, the pathogenicity and virulence difference among entomopathogenfungus strains is not widely well investıgated. The current study was therefore initiated to test the pathogenicity of some entomopathogenic fungus isolates against Tutaabsoluta. The experiment was conducted at Bursa Uludag University, Agiculutre faculty, horticulture department glasshouse in 2020/2021. Tutabasoluta adult were collected, and masslarvae were reared in a growth chamber. Then, ten third instar larvae were inoculated with four entomopathogen fungi isolates (Beuaveriabassania Ak-10, Beuaveriabassania Ak-14, Metarhziumanisoplai Ak-11, and Metarhziumanisoplai Ak-12) with different inoculum suspension (0, 1x10⁶, 1x10⁷,,4 × 10⁸, 4× 10⁹ and 1×10¹⁰ conidia /ml) in a factorial experiment arranged in randomized complete block design with three replication. Mortality data assessment was done on the 3rd, 5thand 7th days after treatment and analyzed. The analysis of variance for mortality rate revealed significant variations (p<0.05) among entomoptahogen fungi isolates and conidia concentrations. The results revealed thatMetarhziumanisoplai Ak-12was found to show the lowest mortality percentage80.77%, highest LC50 2.3x108, and the longest incubation period, LT50, 4.9 and LT90, 9.9daysand considered to be less pathogenic fungi. On the other hand, Beuaveriabassania Ak-10 isolate showed the highest mortality percentage, 91%, and the lowest LT50, 4, and LT90, 7.6 values at 1×10¹⁰ conidia /ml, followed by Beuaveriabassania Ak-14 and being considered as the most aggressive bio-agent. Metarhziumanisoplai Ak-11 was determined as moderately virulent, having a mortality rate 27-81%. Results also revealed that among conidia concentrations, 1x10⁹ and 1x10¹⁰ suspensions is the most effective, while 1x10⁶ conidia/ml concentration is the least effective. Hence, results indicated that EPF tested were effective against T. absoluta larvae. As the current work revealed the potential variation among entomopathogen fungi isolates and concentration against third instar larvae.

Keywords: tuta absoluta, tomato, metarhizium anisopliae, beauveria bassiana, biological control

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Authors: H. Scholz, P. Kuehne, G. Heckenberger

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Dietary Anion-Cation Balance (DCAB) in grazing systems under German conditions has a tendency to decrease from May until September and often are measured DCAB lower than 100 meq per kg dry matter. Lower DCAB in grass feeding system can change the metabolic status of suckler cows and often are results in acidotic metabolism. Measurement of acid-base excretion in dairy cows has been proved to a method to evaluate the acid-base status. The hypothesis was that metabolic imbalances could be identified by urine measurement in suckler cows. The farm study was conducted during the grazing seasons 2017 and 2018 and involved 7 suckler cow farms in Germany. Suckler cows were grazing during the whole time of the investigation and had no access to other feeding components. Cows had free access to water and salt block and free access to minerals (loose). The dry matter of the grass was determined at 60 °C and were then analysed for energy and nutrient content and for the Dietary Cation-Anion Balance (DCAB). Urine was collected in 50 ml-glasses and analysed for net acid-base excretion (NSBA) and the concentration of creatinine and urea in the laboratory. Statistical analysis took place with ANOVA with fixed effects of farms (1-7), month (May until September), and number of lactations (1, 2, and ≥ 3 lactations) using SPSS Version 25.0 for windows. An alpha of 0.05 was used for all statistical tests. During the grazing periods of years 2017 and 2018, an average DCAB was observed in the grass of 167 meq per kg DM. A very high mean variation could be determined from -42 meq/kg to +439 meq/kg. Reference values in relation to DCAB were described between 150 meq and 400 meq per kg DM. It was found the high chlorine content with reduced potassium level led to this reduction in DCAB at the end of the grazing period. Between the DCAB of the grass and the NSBA in urine of suckler cows was a correlation according to PEARSON of r = 0.478 (p ≤ 0.001) or after SPEARMAN of r = 0.601 (p ≤ 0.001) observed. For the control of urine values of grazing suckler cows, the wide spread of the values poses a challenge of the interpretation, especially since the DCAB is unknown. The influence of several feeding components such as chlorine, sulfur, potassium, and sodium (ions for the DCAB) and dry matter feed intake during the grazing period of suckler cows should be taken into account in further research. The results obtained show that up a decrease in the DCAB is related to a decrease in NSBA in urine of suckler cows. Monitoring of metabolic disturbances should include analysis of urine, blood, milk, and ruminal fluid.

Keywords: dietary anion-cation balance, DCAB, net acid-base excretion, NSBA, suckler cow, grazing period

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Authors: Marcelo Lopes Dias Kolling, Felipe Ferreira Laranjeira, Guilherme Augusto Hettwer, Pedro Salomão Piccinini, Marwan Masri, Carlos Oscar Uebel

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Introduction: Injectable fillers are among the most common nonsurgical cosmetic procedures, with significant growth yearly. Knowledge of rheological and mechanical characteristics of fillers, facial anatomy, and injection technique is essential for safety. Concepts such as the use of cannula versus needle, aspiration before injection, and facial danger zones have been well discussed. In case of an accidental intravascular puncture, the pressure inside the vessel may not be sufficient to push blood into the syringe due to the characteristics of the filler product; this is especially true for calcium hydroxyapatite (CaHA) or hyaluronic acid (HA) fillers with high G’. Since viscoelastic properties of normal saline are much lower than those of fillers, aspiration with saline prior to filler injection may decrease the risk of a false negative aspiration and subsequent catastrophic effects. We discuss a technique to add an additional safety step to the procedure with saline aspiration prior to injection, a ‘’reverse Seldinger’’ technique for intravascular access, which we term SANIT: Saline Aspiration Negative Intravascular Test. Objectives: To demonstrate the author’s (PSP) technique which adds an additional safety step to the process of filler injection, with both CaHA and HA, in order to decrease the risk of intravascular injection. Materials and Methods: Normal skin cleansing and topical anesthesia with prilocaine/lidocaine cream are performed; the facial subunits to be treated are marked. A 3mL Luer lock syringe is filled with 2mL of 0.9% normal saline and a 27G needle, which is turned one half rotation. When a cannula is to be used, the Luer lock syringe is attached to a 27G 4cm single hole disposable cannula. After skin puncture, the 3mL syringe is advanced with the plunger pulled back (negative pressure). Progress is made to the desired depth, all the while aspirating. Once the desired location of filler injection is reached, the syringe is exchanged for the syringe containing a filler, securely grabbing the hub of the needle and taking care to not dislodge the needle tip. Prior to this, we remove 0.1mL of filler to allow for space inside the syringe for aspiration. We again aspirate and inject retrograde. SANIT is especially useful for CaHA, since the G’ is much higher than HA, and thus reflux of blood into the syringe is less likely to occur. Results: The technique has been used safely for the past two years with no adverse events; the increase in cost is negligible (only the cost of 2mL of normal saline). Over 100 patients (over 300 syringes) have been treated with this technique. The risk of accidental intravascular puncture has been calculated to be between 1:6410 to 1:40882 syringes among expert injectors; however, the consequences of intravascular injection can be catastrophic even with board-certified physicians. Conclusions: While the risk of intravascular filler injection is low, the consequences can be disastrous. We believe that adding the SANIT technique can help further mitigate risk with no significant untoward effects and could be considered by all performing injectable fillers. Further follow-up is ongoing.

Keywords: injectable fillers, safety, saline aspiration, injectable filler complications, hyaluronic acid, calcium hydroxyapatite

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Authors: Lise P. Labéjof, Ivna Mororó, Raquel G. Bastos, Maria Isabel G. Severo, Arno H. de Oliveira

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This work presents an alternative mode of intestine mucosa renewal that may allow to better understand the total loss of villi after irradiation. It was tested a morphological method of 3d reconstruction using micrographs of serial sections of rat duodenum. We used hundreds of sections of each specimen of duodenum placed on glass slides and examined under a light microscope. Those containing the detachment, approximately a dozen, were chosen for observation under a transmission electron microscope (TEM). Each of these sections was glued on a block of epon resin and recut into a hundred of 60 nm-thick sections. Ribbons of these ultrathin sections were distributed on a series of copper grids in the same order of appearance than during the process of microstomia. They were then stained by solutions of uranyl and lead salts and observed under a TEM. The sections were pictured and the electron micrographs showing signs of cells detachment were transferred into two softwares, ImageJ to align the cellular structures and Reconstruct to realize the 3d reconstruction. It has been detected epithelial cells that exhibited all signs of programmed cell death and localized at the villus-crypt junction. Their nucleus was irregular in shape with a condensed chromatin in clumps. Their cytoplasm was darker than that of neighboring cells, containing many swollen mitochondria. In some places of the sections, we could see intercellular spaces enlarged by the presence of shrunk cells which displayed a plasma membrane with an irregular shape in thermowell as if the cell interdigitations would distant from each other. The three-dimensional reconstruction of the crypts has allowed observe gradual loss of intercellular contacts of crypt cells in the longitudinal plan of the duodenal mucosa. In the transverse direction, there was a gradual increase of the intercellular space as if these cells moved away from one another. This observation allows assume that the gradual remoteness of the cells at the villus-crypt junction is the beginning of the mucosa detachment. Thus, the shrinking of cells due to apoptosis is the way that they detach from the mucosa and progressively the villi also. These results are in agreement with our initial hypothesis and thus have demonstrated that the villi become detached from the mucosa at the villus-crypt junction by the programmed cell death process. This type of loss of entire villus helps explain the rapid denudation of the intestinal mucosa in case of irradiation.

Keywords: 3dr, transmission electron microscopy, ionizing radiations, rat small intestine, apoptosis

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Authors: A. G. Gashua, T. T. Bello, I. Alhassan, K. K. Gwiokura

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Weeds are known to interfere seriously with crop growth, thereby affecting the productivity and quality of crops. Crops are also known to compete for natural growth resources if they are not adequately spaced, also affecting the performance of the growing crop. Farmers grow cowpea in mixtures with cereals and this is known to affect its yield. For this reason, a field experiment was conducted at Yobe State College of Agriculture Gujba, Damaturu station in the 2014 and 2015 rainy seasons to determine the appropriate intra row spacing and weeding regime for optimum growth and yield of cowpea (Vigna unguiculata L.) in pure stand in Sudan Savanna ecology. The treatments consist of three levels of spacing within rows (20 cm, 30 cm and 40 cm) and four weeding regimes (none, once at 3 weeks after sowing (WAS), twice at 3 and 6WAS, thrice at 3WAS, 6WAS and 9WAS); arranged in a Randomized Complete Block Design (RCBD) and replicated three times. The variety used was the local cowpea variety (white, early and spreading) commonly grown by farmers. The growth and yield data were collected and subjected to analysis of variance using SAS software, and the significant means were ranked by Students Newman Keul’s test (SNK). The findings of this study revealed better crop performance in 2015 than in 2014 despite poor soil condition. Intra row spacing significantly influenced vegetative growth especially the number of main branches, leaves and canopy spread at 6WAS and 9WAS with the highest values obtained at wider spacing (40 cm). The values obtained in 2015 doubled those obtained in 2014 in most cases. Spacing also significantly affected the number of pods in 2015, seed weight in both years and grain yield in 2014 with the highest values obtained when the crop was spaced at 30-40 cm. Similarly, weeding regime significantly influenced almost all the growth attributes of cowpea with higher values obtained from where cowpea was weeded three times at 3-week intervals, though statistically similar results were obtained even from where cowpea was weeded twice. Weeding also affected the entire yield and yield components in 2015 with the highest values obtained with increase weeding. Based on these findings, it is recommended that spreading cowpea varieties should be grown at 40 cm (or wider spacing) within rows and be weeded twice at three-week intervals for better crop performance in related ecologies.

Keywords: intra-row spacing, local cowpea, Nigeria, weeding

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Authors: Michio Kurosu

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Alterations in glycosylation not only directly impact cell growth and survival but also facilitate tumor-induced immunomodulation and eventual metastasis. Identification of cell type-specific glycoconjugates (tumor markers) has led to the discovery of new assay systems for certain cancers via immunodetection reagents. N- and O-linked glycans are the most abundant forms of glycoproteins. Recent studies of cancer immunotherapy are based on the immunogenicity of truncated O-glycan chains (e.g., Tn, sTn, T, and sLea/x). The prevalence of N-linked glycan changes in the development of tumor cells is known; however, therapeutic antibodies against N-glycans have not yet been developed. This is due to the lack of specificity of N-linked glycans between normal/healthy and cancer cells. Abnormal branching of N-linked glycans has been observed, particularly in solid cancer cells. While the discovery of drug-like glycosyltransferase inhibitors that block the biosynthesis of specific branching has a very low likelihood of success, altered glycosylation levels can be exploited by suppressing N-glycan biosynthesis through the inhibition of dolichyl-phosphate N-acetylglucosaminephosphotransferase1 (DPAGT1) activity. Inhibition of DPAGT1 function leads to changes of O-glycosylation on proteins associated with mitochondria and zinc finger binding proteins (indirect effects). On the basis of dynamic crosstalk between DPAGT1 and Snail/Slung/ZEB1 (a family of transcription factors that promote the repression of the adhesion molecules), we have developed pharmacologically acceptable selective DPAGT1 inhibitors. Tunicamycin kills a wide range of cancer and healthy cells in a non-selective manner. In sharp contrast, our DPAGT1 inhibitors display strong cytostatic effects against 16 solid cancers, which require the overexpression of DPAGT1 in their progression but do not affect the cell viability of healthy cells. The identified DPAGT1 inhibitors possess impressive anti-metastatic ability in various solid cancer cell lines and induce their mitochondrial structural changes, resulting in apoptosis. A prototype DPAGT1 inhibitor, APPB has already been proven to shrink solid tumors (e.g., pancreatic cancers, triple-negative breast cancers) in vivo while suppressing metastases and has strong synergistic effects when combined with current cytotoxic drugs (e.g., paclitaxel). At this conference, our discovery of selective DPAGT1 inhibitors with drug-like properties and proof-of-pharmaceutical concept studies of a novel DPAGT1 inhibitor are presented.

Keywords: DPAGT1 inhibitors, anti-metastatic drugs, natural product based drug designs, cytostatic effects

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Authors: Luz Stella Cadavid-Rodriguez, Viviana E. Castro-Lopez

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Fish waste (FW) has a high content of potentially biodegradable components, so it is amenable to be digested anaerobically. In this line, anaerobic digestion (AD) of FW has been studied for biogas production. Nevertheless, intermediate products such as volatile fatty acids (VFA), generated during the acidogenic stage, have been scarce investigated, even though they have a high potential as a renewable source of carbon. In the literature, there are few studies about the Inoculum-Substrate (I/S) ratio on acidogenesis. On the other hand, it is well known that pH is a critical factor in the production of VFA. The optimum pH for the production of VFA seems to change depending on the substrate and can vary in a range between 5.25 and 11. Nonetheless, the literature about VFA production from protein-rich waste, such as FW, is scarce. In this context, it is necessary to deepen on the determination of the optimal operating conditions of acidogenic fermentation for VFA production from protein-rich waste. Therefore, the aim of this research was to optimize the volatile fatty acid production from artisanal fishing waste, studying the effect of pH and the I/S ratio on the acidogenic process. For this research, the inoculum used was a methanogenic sludge (MS) obtained from a UASB reactor treating wastewater of a slaughterhouse plant, and the FW was collected in the port of Tumaco (Colombia) from the local artisanal fishers. The acidogenic fermentation experiments were conducted in batch mode, in 500 mL glass bottles as anaerobic reactors, equipped with rubber stoppers provided with a valve to release biogas. The effective volume used was 300 mL. The experiments were carried out for 15 days at a mesophilic temperature of 37± 2 °C and constant agitation of 200 rpm. The effect of 3 pH levels: 5, 7, 9, coupled with five I/S ratios, corresponding to 0.20, 0.15, 0.10, 0.05, 0.00 was evaluated taking as a response variable the production of VFA. A complete randomized block design was selected for the experiments in a 5x3 factorial arrangement, with two repetitions per treatment. At the beginning and during the process, pH in the experimental reactors was adjusted to the corresponding values of 5, 7, and 9 using 1M NaOH or 1M H2SO4, as was appropriated. In addition, once the optimum I/S ratio was determined, the process was evaluated at this condition without pH control. The results indicated that pH is the main factor in the production of VFA, obtaining the highest concentration with neutral pH. By reducing the I/S ratio, as low as 0.05, it was possible to maximize VFA production. Thus, the optimum conditions found were natural pH (6.6-7.7) and I/S ratio of 0.05, with which it was possible to reach a maximum total VFA concentration of 70.3 g Ac/L, whose major components were acetic acid (35%) and butyric acid (32%). The findings showed that the acidogenic fermentation of FW is an efficient way of producing VFA and that the operating conditions can be simple and economical.

Keywords: acidogenesis, artisanal fishing waste, inoculum to substrate ratio, volatile fatty acids

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Authors: Masood Otarod, Ronald M. Supkowski

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This abstract discusses a method that reduces the general dispersion model in cylindrical coordinates to a second order linear ordinary differential equation with constant coefficients so that it can be utilized to conduct kinetic studies in packed bed tubular catalytic reactors at a broad range of Reynolds numbers. The model was tested by 13CO isotope transient tracing of the CO adsorption of Boudouard reaction in a differential reactor at an average Reynolds number of 0.2 over Pd-Al2O3 catalyst. Detailed experimental results have provided evidence for the validity of the theoretical framing of the model and the estimated parameters are consistent with the literature. The solution of the general dispersion model requires the knowledge of the radial distribution of axial velocity. This is not always known. Hence, up until now, the implementation of the dispersion model has been largely restricted to the plug-flow regime. But, ideal plug-flow is impossible to achieve and flow regimes approximating plug-flow leave much room for debate as to the validity of the results. The reduction of the general dispersion model transpires as a result of the application of a factorization theorem. Factorization theorem is derived from the observation that a cross section of a catalytic bed consists of a solid phase across which the reaction takes place and a void or porous phase across which no significant measure of reaction occurs. The disparity in flow and the heterogeneity of the catalytic bed cause the concentration of reacting compounds to fluctuate radially. These variabilities signify the existence of radial positions at which the radial gradient of concentration is zero. Succinctly, factorization theorem states that a concentration function of axial and radial coordinates in a catalytic bed is factorable as the product of the mean radial cup-mixing function and a contingent dimensionless function. The concentration of adsorbed compounds are also factorable since they are piecewise continuous functions and suffer the same variability but in the reverse order of the concentration of mobile phase compounds. Factorability is a property of packed beds which transforms the general dispersion model to an equation in terms of the measurable mean radial cup-mixing concentration of the mobile phase compounds and mean cross-sectional concentration of adsorbed species. The reduced model does not require the knowledge of the radial distribution of the axial velocity. Instead, it is characterized by new transport parameters so denoted by Ωc, Ωa, Ωc, and which are respectively denominated convection coefficient cofactor, axial dispersion coefficient cofactor, and radial dispersion coefficient cofactor. These cofactors adjust the dispersion equation as compensation for the unavailability of the radial distribution of the axial velocity. Together with the rest of the kinetic parameters they can be determined from experimental data via an optimization procedure. Our data showed that the estimated parameters Ωc, Ωa Ωr, are monotonically correlated with the Reynolds number. This is expected to be the case based on the theoretical construct of the model. Computer generated simulations of methanation reaction on nickel provide additional support for the utility of the newly conceptualized dispersion model.

Keywords: factorization, general dispersion model, isotope transient kinetic, partial differential equations

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Authors: Hamza Benzerrouk, Alexander Nebylov

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In classical GNSS/INS integration designs, the loosely coupled approach uses the GNSS derived position and the velocity as the measurements vector. This design is suboptimal from the standpoint of preventing GNSSoutliers/outages. The tightly coupled GPS/INS navigation filter mixes the GNSS pseudo range and inertial measurements and obtains the vehicle navigation state as the final navigation solution. The ultra‐tightly coupled GNSS/INS design combines the I (inphase) and Q(quadrature) accumulator outputs in the GNSS receiver signal tracking loops and the INS navigation filter function intoa single Kalman filter variant (EKF, UKF, SPKF, CKF and HCKF). As mentioned, EKF and UKF are the most used nonlinear filters in the literature and are well adapted to inertial navigation state estimation when integrated with GNSS signal outputs. In this paper, it is proposed to move a step forward with more accurate filters and modern approaches called Cubature and High Degree cubature Kalman Filtering methods, on the basis of previous results solving the state estimation based on INS/GNSS integration, Cubature Kalman Filter (CKF) and High Degree Cubature Kalman Filter with (HCKF) are the references for the recent developed generalized Cubature rule based Kalman Filter (GCKF). High degree cubature rules are the kernel of the new solution for more accurate estimation with less computational complexity compared with the Gauss-Hermite Quadrature (GHQKF). Gauss-Hermite Kalman Filter GHKF which is not selected in this work because of its limited real-time implementation in high-dimensional state-spaces. In ultra tightly or a deeply coupled GNSS/INS system is dynamics EKF is used with transition matrix factorization together with GNSS block processing which is well described in the paper and assumes available the intermediary frequency IF by using a correlator samples with a rate of 500 Hz in the presented approach. GNSS (GPS+GLONASS) measurements are assumed available and modern SPKF with Cubature Kalman Filter (CKF) are compared with new versions of CKF called high order CKF based on Spherical-radial cubature rules developed at the fifth order in this work. Estimation accuracy of the high degree CKF is supposed to be comparative to GHKF, results of state estimation are then observed and discussed for different initialization parameters. Results show more accurate navigation state estimation and more robust GNSS receiver when Ultra Tightly Coupled approach applied based on High Degree Cubature Kalman Filter.

Keywords: GNSS, INS, Kalman filtering, ultra tight integration

Procedia PDF Downloads 279

Authors: Kusmartono, Siti Chuzaemi, Hartutik dan Mashudi

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Two studies were conducted involving an in vitro (Expt 1) and in vivo (Expt 2) measurements. Expt 1. aimed to evaluate effects of adding CT extracts on gas production and efficiency of microbial protein synthesis (EMPS), Expt 2 aimed to evaluate effects of supplementing shrub/tree leaves as CT source on feed consumption, digestibility, N retention, body weight gain and dressing percentage of growing sheep fed on elephant grass (EG) as a basal diet.Ten shrub and tree leaves used as CT sources were wild sunflower (Tithonia diversifolia), mulberry (Morus macroura), cassava (Manihot utilissima), avicienna (Avicennia marina), calliandra (Calliandra calothyrsus), sesbania (Sesbania grandiflora), acacia (acacia vilosa), glyricidia (Glyricidia sepium), jackfruit (Artocarpus heterophyllus), moringa (Moringa oleifera). The treatments applied in Expt 1 were: T1=Elephant grass (60%)+concentrate (40%); T2 = T1 + CT (3% DM); T3= T2 + PEG; T4 = T1 + CT (3.5% DM); T5 = T4 + PEG; T6 = T1 + CT (4% DM) and T7 = T6 + PEG. Data obtained were analysed using Randomized Block Design. Statistical analyses showed that treatments significanty affected (P<0.05) total gas production and EMPS. The lowest values of total gas production (45.9 ml/500 mg DM) and highest value of EMPS (64.6 g/kg BOTR) were observed in the treatment T4 (3.5% CT from cassava leave extract). Based on this result it was concluded that this treatment was the best and was chosen for further investigation using in vivo method. The treatmets applied for in vivo trial were: T1 = EG (60%) + concentrate (40%); T2 = T1 + dried cassava leave (equivalent to 3.5% CT); T3 = T2 + PEG. 18 growing sheep aging of 8-9 months and weighing of 23.67kg ± 1.23 were used in Expt 2. Results of in vivo study showed that treatments significanty affected (P<0.05) nutrients intake and digestibility (DM, OM and CP). N retention for sheep receiving treatment T2 were significantly higher (P<0.05; 15.6 g/d) than T1 (9.1 g/d) and T3 (8.53 g/d). Similar results were obtained for daily weight gain where T2 were the highest (62.79 g/d), followed by T1 (51.9 g/d) and T3 (52.85 g/d). Dressing percentage of T2 was the highest (51.54%) followed by T1 (49.61%) and T3 (49.32%). It can be concluded that adding adding dried cassava leaves did not reduce palatability due to CT, but rather increased OM digestibility and hence feed consumption was improved. N retention was increased due to the action of CT in the cassava leaves and this may have explained a higher input of N into duodenum which was further led to higer daily weight gain and dressing percentage.

Keywords: in vitro gas production, sheep, shrub and tree leaves, condensed tannin

Procedia PDF Downloads 264

Authors: Timothy O. Adekunle, Sigridur Bjarnadottir

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Several studies have investigated thermal behavior of buildings with limited studies focusing on high-rise buildings. Of the limited investigations that have considered thermal performance of high-rise buildings, only a few studies have considered thermal behavior of high-rise structural sustainable buildings. As a result, this study investigates the thermal behavior of a high-rise structural timber building. The study aims to understand the thermal environment of a high-rise structural timber block of apartments located in East London, UK by comparing the indoor environmental conditions at different floors (ground and upper floors) of the building. The environmental variables (temperature and relative humidity) were measured at 15-minute intervals for a few weeks in the summer of 2012 to generate data that was considered for calibration and validation of the simulated results. The study employed mainly dynamic thermal building simulation using DesignBuilder by EnergyPlus and supplemented with environmental monitoring as major techniques for data collection and analysis. The weather file (Test Reference Years- TRYs) for the 2000s from the weather generator carried out by the Prometheus Group was considered for the simulation since the study focuses on investigating thermal behavior of high-rise structural timber buildings in the summertime and not in extreme summertime. In this study, the simulated results (May-September of the 2000s) will be the focus of discussion, but the results will be briefly compared with the environmental monitoring results. The simulated results followed a similar trend with the findings obtained from the short period of the environmental monitoring at the building. The results revealed lower temperatures are often predicted (at least 1.1°C lower) at the ground floor than the predicted temperatures at the upper floors. The simulated results also showed that higher temperatures are predicted in spaces at southeast facing (at least 0.5°C higher) than spaces in other orientations across the floors considered. There is, however, a noticeable difference between the thermal environment of spaces when the results obtained from the environmental monitoring are compared with the simulated results. The field survey revealed higher temperatures were recorded in the living areas (at least 1.0°C higher) while higher temperatures are predicted in bedrooms (at least 0.9°C) than living areas for the simulation. In addition, the simulated results showed spaces on lower floors of high-rise structural timber buildings are predicted to provide more comfortable thermal environment than spaces on upper floors in summer, but this may not be the same in wintertime due to high upward movement of hot air to spaces on upper floors.

Keywords: building simulation, high-rise, structural timber buildings, sustainable, temperatures, thermal behavior

Procedia PDF Downloads 175

Authors: Susan Mwai, Mary Muchane, Peter Wachira, Sheila Okoth, Muchai Muchane, Halima Saado

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Tropical forests harbor a wide range of biodiversity and rich macro-fungi diversity compared to the temperate regions in the World. However, biodiversity is facing the threat of extinction following the rate of forest loss taking place before proper study and documentation of macrofungi is achieved. The present study was undertaken to determine the effect of converting indigenous habitat to plantation forest on macrofungi diversity. To achieve the objective of this study, an inventory focusing on macro-fungi diversity was conducted within Kereita block in Kikuyu Escarpment forest which is on the southern side of Aberdare mountain range. The macrofungi diversity was conducted in the indigenous forest and in more than 15 year old Patula plantation forest , during the wet (long rain season, December 2014) and dry (Short rain season, May, 2015). In each forest type, 15 permanent (20m x 20m) sampling plots distributed across three (3) forest blocks were used. Both field and laboratory methods involved recording abundance of fruiting bodies, taxonomic identity of species and analysis of diversity indices and measures in terms of species richness, density and diversity. R statistical program was used to analyze for species diversity and Canoco 4.5 software for species composition. A total number of 76 genera in 28 families and 224 species were encountered in both forest types. The most represented taxa belonged to the Agaricaceae (16%), Polyporaceae (12%), Marasmiaceae, Mycenaceae (7%) families respectively. Most of the recorded macro-fungi were saprophytic, mostly colonizing the litter 38% and wood 34% based substrates, which was followed by soil organic dwelling species (17%). Ecto-mycorrhiza fungi (5%) and parasitic fungi (2%) were the least encountered. The data established that indigenous forests (native ecosystems) hosts a wide range of macrofungi assemblage in terms of density (2.6 individual fruit bodies / m2), species richness (8.3 species / plot) and species diversity (1.49/ plot level) compared to the plantation forest. The Conversion of native forest to plantation forest also interfered with species composition though did not alter species diversity. Seasonality was also shown to significantly affect the diversity of macro-fungi and 61% of the total species being present during the wet season. Based on the present findings, forested ecosystems in Kenya hold diverse macro-fungi community which warrants conservation measures.

Keywords: diversity, Indigenous forest, macro-fungi, plantation forest, season

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Authors: Seyed Ali Mohammad, Modarres Sanavy, Hamed Keshavarz, Ali Mokhtassi-Bidgoli

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One of the biggest challenges for the current and future generations is to produce sufficient food for the world population with the existing limited available water resources. Peppermint is a specialty crop used for food and medicinal purposes. Its main component is menthol. It is used predominantly for oral hygiene, pharmaceuticals, and foods. Although drought stress is considered as a negative factor in agriculture, being responsible for severe yield losses; medicinal plants grown under semi-arid conditions usually produce higher concentrations of active substances than same species grown under moderate climates. Nitrogen (N) fertilizer management is central to the profitability and sustainability of forage crop production. Sub-optimal N supply will result in poor yields, and excess N application can lead to nitrate leaching and environmental pollution. In order to determine the response of peppermint to drought stress and different fertilizer treatments, a field experiment with peppermint was conducted in a sandy loam soil at a site of the Tarbiat Modares University, Agriculture Faculty, Tehran, Iran. The experiment used a complete randomized block design, with six rates of fertilizer strategies (F1: control, F2: Urea, F3: 75% urea + 25% vermicompost, F4: 50% urea + 50% vermicompost, F5: 25% urea + 75% vermicompost and F6: vermicompost) and three irrigation regime (S1: 45%, S2: 60% and S3: 75% FC) with three replication. The traits such as nitrogen, chlorophyll, carotenoids, anthocyanin, flavonoid and fresh biomass were studied. The results showed that the treatments had a significant effect on the studied traits as drought stress reduced photosynthetic pigment concentration. Also, drought stress reduced fresh yield of peppermint. Non stress condition had the greater amount of chlorophyll and fresh yield more than other irrigation treatments. The highest concentration of chlorophyll and the fresh biomass was obtained in F2 fertilizing treatments. Sever water stress (S1) produced decreased photosynthetic pigment content fresh yield of peppermint. Supply of N could improve photosynthetic capacity by enhancing photosynthetic pigment content. Perhaps application of vermicompost significantly improved the organic carbon, available N, P and K content in soil over urea fertilization alone. To get sustainable production of peppermint, application of vermicompost along with N through synthetic fertilizer is recommended for light textured sandy loam soils.

Keywords: fresh yield, peppermint, synthetic nitrogen, vermicompost, water stress

Procedia PDF Downloads 215

Authors: Utpal Das, Sona Das

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InAs/GaSb type-II superlattice (T2SL) Mid-wave infrared (MWIR) focal plane arrays (FPAs) have recently seen rapid development. However, in small pixel size large format FPAs, the occurrence of high mesa sidewall surface leakage current is a major constraint necessitating proper surface passivation. A simple pixel isolation technique in InAs/GaSb T2SL detector arrays without the conventional mesa etching has been proposed to isolate the pixels by forming a more resistive higher band gap material from the SL, in the inter-pixel region. Here, a single step femtosecond (fs) laser anneal of the T2SL structure of the inter-pixel T2SL regions, have been used to increase the band gap between the pixels by QW-intermixing and hence increase isolation between the pixels. The p-i-n photodiode structure used here consists of a 506nm, (10 monolayer {ML}) InAs:Si (1x10¹⁸cm⁻³)/(10ML) GaSb SL as the bottom n-contact layer grown on an n-type GaSb substrate. The undoped absorber layer consists of 1.3µm, (10ML)InAs/(10ML)GaSb SL. The top p-contact layer is a 63nm, (10ML)InAs:Be(1x10¹⁸cm⁻³)/(10ML)GaSb T2SL. In order to improve the carrier transport, a 126nm of graded doped (10ML)InAs/(10ML)GaSb SL layer was added between the absorber and each contact layers. A 775nm 150fs-laser at a fluence of ~6mJ/cm² is used to expose the array where the pixel regions are masked by a Ti(200nm)-Au(300nm) cap. Here, in the inter-pixel regions, the p+ layer have been reactive ion etched (RIE) using CH₄+H₂ chemistry and removed before fs-laser exposure. The fs-laser anneal isolation improvement in 200-400μm pixels due to spatially selective quantum well intermixing for a blue shift of ~70meV in the inter-pixel regions is confirmed by FTIR measurements. Dark currents are measured between two adjacent pixels with the Ti(200nm)-Au(300nm) caps used as contacts. The T2SL quality in the active photodiode regions masked by the Ti-Au cap is hardly affected and retains the original quality of the detector. Although, fs-laser anneal of p+ only etched p-i-n T2SL diodes show a reduction in the reverse dark current, no significant improvement in the full RIE-etched mesa structures is noticeable. Hence for a 128x128 array fabrication of 8μm square pixels and 10µm pitch, SU8 polymer isolation after RIE pixel delineation has been used. X-n+ row contacts and Y-p+ column contacts have been used to measure the optical response of the individual pixels. The photo-response of these 8μm and other 200μm pixels under a 2ns optical pulse excitation from an Optical-Parametric-Oscillator (OPO), shows a peak responsivity of ~0.03A/W and 0.2mA/W, respectively, at λ~3.7μm. Temporal response of this detector array is seen to have a fast response ~10ns followed typical slow decay with ringing, attributed to impedance mismatch of the connecting co-axial cables. In conclusion, response times of a few ns have been measured in 8µm pixels of a 128x128 array. Although fs-laser anneal has been found to be useful in increasing the inter-pixel isolation in InAs/GaSb T2SL arrays by QW inter-mixing, it has not been found to be suitable for passivation of full RIE etched mesa structures with vertical walls on InAs/GaSb T2SL.

Keywords: band-gap blue-shift, fs-laser-anneal, InAs/GaSb T2SL, Inter-pixel isolation, ns-Response, photodiode array

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Authors: Julien G. Chenet, Mario A. Hernandez, U. Leonardo Rodriguez

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The tropical areas are places where there is scarcity of access to potable water and where renewable energies need further development. They also display high undernourishment levels, even though they are one of the resources-richest areas in the world. In these areas, it is common to count on great extension of soils, high solar radiation and raw water from rain, groundwater, surface water or even saltwater. Even though resources are available, access to them is limited, and the low-density habitat makes central solutions expensive and investments not worthy. In response to this lack of investment, rural inhabitants use fossil fuels and timber as an energy source and import agrochemical for soils fertilization, which increase GHG emissions. The OASIS project brings an answer to this situation. It supplies renewable energy, potable water and organic food. The first step is the determination of the needs of the communities in terms of energy, water quantity and quality, food requirements and soil characteristics. Second step is the determination of the available resources, such as solar energy, raw water and organic residues on site. The pilot OASIS project is located in the Vichada department, Colombia, and ensures the sustainable use of natural resources to meet the community needs. The department has roughly 70% of indigenous people. They live in a very scattered landscape, with no access to clean water and energy. They use polluted surface water for direct consumption and diesel for energy purposes. OASIS pilot will ensure basic needs for a 400-students education center. In this case, OASIS will provide 20 kW of solar energy potential and 40 liters per student per day. Water will be treated form groundwater, with two qualities. A conventional one with chlorine, and as the indigenous people are not used to chlorine for direct consumption, second train is with reverse osmosis to bring conservable safe water without taste. OASIS offers a solution to supply basic needs, shifting from fossil fuels, timber, to a no-GHG-emission solution. This solution is part of the mitigation strategy against Climate Change for the communities in low-density areas of the tropics. OASIS is a learning center to teach how to convert natural resources into utilizable ones. It is also a meeting point for the community with high pedagogic impact that promotes the efficient and sustainable use of resources. OASIS system is adaptable to any tropical area and competes technically and economically with any conventional solution, that needs transport of energy, treated water and food. It is a fully automatic, replicable and sustainable solution to sort out the issue of access to basic needs in rural areas. OASIS is also a solution to undernourishment, ensuring a responsible use of resources, to prevent long-term pollution of soils and groundwater. It promotes the closure of the nutrient cycle, and the optimal use of the land whilst ensuring food security in depressed low-density regions of the tropics. OASIS is under optimization to Vichada conditions, and will be available to any other tropical area in the following months.

Keywords: climate change adaptation and mitigation, rural development, sustainable access to clean and renewable resources, social inclusion

Procedia PDF Downloads 248

Authors: Mala Tankam Carine, Kekeunou Sévilor, Nukenine Elias

Abstract:

Storage and preservation of agricultural products remain the only conditions ensuring the almost permanent availability of foodstuffs. However, infestations due to insects and microorganisms often occur. Callosobruchus maculatus is a pest that causes a lot of damage to cowpea stocks in the tropics. Several methods are adopted to limit their damage, but the use of synthetic chemical insecticides is the most widespread. Biopesticides in sustainable agriculture respond to several environmental, economic and social concerns while offering innovative opportunities that are ecologically and economically viable for producers, workers, consumers and ecosystems. Our main objective is to evaluate the insecticidal efficacy of binary combinations of Fossilshield with root-bark powder of Securidaca longepedunculata against Callosobruchus maculatus in stored cowpea Vigna unguiculata. Laboratory bioassays were conducted in stored grains to evaluate the toxicity of root-bark powder of Securidaca longepedunculata alone or combined with diatomaceous earth Fossil-Shield ® against C. maculatus. Twenty-hour-old adults of C. maculatus were exposed to 50g of cowpea seeds treated with four doses (10, 20, 30, and 40g/kg) of root-bark powder of S. longepedunculata, on the one hand, and (0.5, 1, 1.5, and 2 g/kg) on DE and binary combinations on the other hand. 0g/kg corresponded to untreated control. Adult mortality was recorded up to 7 days (d) post-treatment, whereas the number of F1 progeny was assessed after 30 d. Weight loss and germinative ability were conducted after 120 d. All treatments were arranged according to a completely randomized block with four replicates. The combined mixture of S. longepedunculata and DE controlled the beetle faster compared to the root-bark powder of S. longepedunculata alone. According to the Co-toxicity coefficient, additive effect of binary combinations was recorded at 3-day post-exposure time with the mixture 25% FossilShield + 75% S. longepedunculata. A synergistic action was observed after 3-d post-exposure at mixture 50% FossilShield + 50% S. longepedunculata and at 1-d and 3-d post-exposure periods at mixture 75% FossilShield + 25% S. longepedunculata. The mixture 25% FossilShield + 75% S. longepedunculata induced a decreased progeny of 6 times fewer individuals for 4.5 times less weight loss and 2, 9 times more sprouted grains than with root-bark powder of S. longepedunculata. The combination of FossilShield + S. longepedunculata was more potent than root-bark powder of S. longepedunculata alone, although the root-bark powder of S. longepedunculata caused significant reduction of F1 adults compared to the control. Combined action of botanical insecticides with FossilShield as a grain protectant in an integrated pest management approach is discussed.

Keywords: diatomaceous earth, cowpea, callosobruchus maculatus, securidaca longepedunculata, combined action, co-toxicity coefficient

Procedia PDF Downloads 69

Authors: Bao-Fang Wen, Meng-Na Dai, Gao-Pei Zhu, Chen-Xi Zhang, Jing Sun, Xun-Bao Yin, Yu-Han Zhao, Hong-Wei Sun, Wei-Fen Zhang

Abstract:

A drug-loaded nanoparticles containing berberine hydrochloride (BH/FA-CTS-NPs) was prepared. The physicochemical characterizations of BH/FA-CTS-NPs and the inhibitory effect on the HeLa cells were investigated. Folic acid-conjugated chitosan (FA-CTS) was prepared by amino reaction of folic acid active ester and chitosan molecules; BH/FA-CTS-NPs were prepared using ionic cross-linking technique with BH as a model drug. The morphology and particle size were determined by Transmission Electron Microscope (TEM). The average diameters and polydispersity index (PDI) were evaluated by Dynamic Light Scattering (DLS). The interaction between various components and the nanocomplex were characterized by Fourier Transform Infrared Spectroscopy (FT-IR). The entrapment efficiency (EE), drug-loading (DL) and in vitro release were studied by UV spectrophotometer. The effect of cell anti-migratory and anti-invasive actions of BH/FA-CTS-NPs were investigated using MTT assays, wound healing assays, Annexin-V-FITC single staining assays, and flow cytometry, respectively. HeLa nude mice subcutaneously transplanted tumor model was established and treated with different drugs to observe the effect of BH/FA-CTS-NPs in vivo on HeLa bearing tumor. The BH/FA-CTS-NPs prepared in this experiment have a regular shape, uniform particle size, and no aggregation phenomenon. The results of DLS showed that mean particle size, PDI and Zeta potential of BH/FA-CTS NPs were (249.2 ± 3.6) nm, 0.129 ± 0.09, 33.6 ± 2.09, respectively, and the average diameter and PDI were stable in 90 days. The results of FT-IR demonstrated that the characteristic peaks of FA-CTS and BH/FA-CTS-NPs confirmed that FA-CTS cross-linked successfully and BH was encapsulated in NPs. The EE and DL amount were (79.3 ± 3.12) % and (7.24 ± 1.41) %, respectively. The results of in vitro release study indicated that the cumulative release of BH/FA-CTS NPs was (89.48±2.81) % in phosphate-buffered saline (PBS, pH 7.4) within 48h; these results by MTT assays and wund healing assays indicated that BH/FA-CTS NPs not only inhibited the proliferation of HeLa cells in a concentration and time-dependent manner but can induce apoptosis as well. The subcutaneous xenograft tumor formation rate of human cervical cancer cell line HeLa in nude mice was 98% after inoculation for 2 weeks. Compared with BH group and BH/CTS-NPs group, the xenograft tumor growth of BH/FA-CTS-NPs group was obviously slower; the result indicated that BH/FA-CTS-NPs could significantly inhibit the growth of HeLa xenograft tumor. BH/FA-CTS NPs with the sustained release effect could be prepared successfully by the ionic crosslinking method. Considering these properties, block proliferation and impairing the migration of the HeLa cell line, BH/FA-CTS NPs could be an important compound for consideration in the treatment of cervical cancer.

Keywords: folic-acid, chitosan, berberine hydrochloride, nanoparticles, cervical cancer

Procedia PDF Downloads 121

Authors: Ayoti Banerjee, Meenakshi Banerjee

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The study of metamorphic reaction textures is important in contributing to our understanding of the evolution of metamorphic terranes. Where preserved, they provide information on changes in the P-T conditions during the metamorphic history of the rock, and thus allow us to speculate on the P-T-t evolution of the terrane. Mafic dykes have attracted the attention of petrologists because they act as window to mantle. This rock represents a mafic dyke of doleritic composition. It is fine to medium grained in which clinopyroxene are enclosed by the lath shaped plagioclase grains to form spectacular ophitic texture. At places, sub ophitic texture was also observed. Grains of pyroxene and plagioclase show very less deformation typically plagioclase showing deformed lamella along with plagioclase-clinopyroxene-phyric granoblastic fabric within a groundmass of feldspar microphenocrysts and Fe–Ti oxides. Both normal and reverse zoning were noted in the plagioclase laths. The clinopyroxene grains contain exsolved phases such as orthopyroxene, plagioclase, magnetite, ilmenite along the cleavage traces and the orthopyroxene lamella form granules in the periphery of the clinopyroxene grains. Garnet corona also develops preferentially around plagioclase at the contact of clinopyroxene, ilmenite or magnetite. Tiny quartz and K-fs grains showed symplectic intergrowth with garnet at a few places. The product quartz formed along with garnet rims the coronal garnet and the reacting clinopyroxene. Thin amphibole corona formed along the periphery of deformed plagioclase and clinopyroxene occur as patches over the magmatic minerals. The amphibole coronas cannot be assigned to a late magmatic stage and are interpreted as reactive being restricted to the contact between clinopyroxene and plagioclase, thus postdating the crystallization of both. The amphibole and garnet do not share grain boundary in the entire rock and is thus pointing towards simultaneous crystallization. Olivine is absent. Spectacular myrmekitic growth of orthoclase and quartz rimming the plagioclase is consistent with the potash metasomatic effects that is also found in other rocks of this region. These textural features are consistent with a phase of fluid induced metamorphism (retrogression). But the appearance of coronal garnet and amphibole exclusive of each other reflects the participation if Ti as the prime reason. Presence of Ti as a reactant phase is a must for amphibole forming reactions whereas it is not so in case of garnet forming reactions although the reactants are the same plagioclase and clinopyroxene in both cases. These findings are well validated by petrographical and textural analysis. In order to obtain balanced chemical reactions that explain formation of amphibole and garnet in the mafic dyke rocks a matrix operation technique called Singular Value Decomposition (SVD) was adopted utilizing the measured chemical compositions of the minerals. The computer program C-Space was used for this purpose and the required compositional matrix. Data fed to C-Space was after doing cation-calculation of the oxide percentages obtained from EPMA analysis. The Garnet-Clinopyroxene geothermometer yielded a temperature of 650 degrees Celsius. The Garnet-Clinopyroxene-Plagioclase geobarometer and Al-in amphibole yielded roughly 7.5 kbar pressure.

Keywords: corona, dolerite, geothermometer, metasomatism, metamorphic reaction texture, retrogression

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Authors: Avyarthana Dey, Naren P. Rao, Arpitha Jacob, Chaitra V. Hiremath, Shivarama Varambally, Ganesan Venkatasubramanian, Rose Dawn Bharath, Bangalore N. Gangadhar

Abstract:

Objective: Impaired facial emotion recognition is widely reported in schizophrenia. Neuropeptide oxytocin is known to modulate brain regions involved in facial emotion recognition, namely amygdala, in healthy volunteers. However, its effect on facial emotion recognition deficits seen in schizophrenia is not well explored. In this study, we examined the effect of intranasal OXT on processing facial emotions and its neural correlates in patients with schizophrenia. Method: 12 male patients (age= 31.08±7.61 years, education= 14.50±2.20 years) participated in this single-blind, counterbalanced functional magnetic resonance imaging (fMRI) study. All participants underwent three fMRI scans; one at baseline, one each after single dose 24IU intranasal OXT and intranasal placebo. The order of administration of OXT and placebo were counterbalanced and subject was blind to the drug administered. Participants performed a facial emotion recognition task presented in a block design with six alternating blocks of faces and shapes. The faces depicted happy, angry or fearful emotions. The images were preprocessed and analyzed using SPM 12. First level contrasts comparing recognition of emotions and shapes were modelled at individual subject level. A group level analysis was performed using the contrasts generated at the first level to compare the effects of intranasal OXT and placebo. The results were thresholded at uncorrected p < 0.001 with a cluster size of 6 voxels. Neuropeptide oxytocin is known to modulate brain regions involved in facial emotion recognition, namely amygdala, in healthy volunteers. Results: Compared to placebo, intranasal OXT attenuated activity in inferior temporal, fusiform and parahippocampal gyri (BA 20), premotor cortex (BA 6), middle frontal gyrus (BA 10) and anterior cingulate gyrus (BA 24) and enhanced activity in the middle occipital gyrus (BA 18), inferior occipital gyrus (BA 19), and superior temporal gyrus (BA 22). There were no significant differences between the conditions on the accuracy scores of emotion recognition between baseline (77.3±18.38), oxytocin (82.63 ± 10.92) or Placebo (76.62 ± 22.67). Conclusion: Our results provide further evidence to the modulatory effect of oxytocin in patients with schizophrenia. Single dose oxytocin resulted in significant changes in activity of brain regions involved in emotion processing. Future studies need to examine the effectiveness of long-term treatment with OXT for emotion recognition deficits in patients with schizophrenia.

Keywords: recognition, functional connectivity, oxytocin, schizophrenia, social cognition

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Authors: Zhe Li, Tao Ju, Liguo Zhang, Zehong Zhang, Baoshun Zhang

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In-grow stacking-faults (IGSFs) in 4H-SiC epilayers can cause increased leakage current and reduce the blocking voltage of 4H-SiC power devices. Double Shockley stacking fault (2SSF) is a common type of IGSF with double slips on the basal planes. In this study, a 2SSF in the 4H-SiC epilayer grown by chemical vaper deposition (CVD) is characterized. The nucleation site of the 2SSF is discussed, and a model for the 2SSF nucleation is proposed. Homo-epitaxial 4H-SiC is grown on a commercial 4 degrees off-cut substrate by a home-built hot-wall CVD. Defect-selected-etching (DSE) is conducted with melted KOH at 500 degrees Celsius for 1-2 min. Room temperature cathodoluminescence (CL) is conducted at a 20 kV acceleration voltage. Low-temperature photoluminescence (LTPL) is conducted at 3.6 K with the 325 nm He-Cd laser line. In the CL image, a triangular area with bright contrast is observed. Two partial dislocations (PDs) with a 20-degree angle in between show linear dark contrast on the edges of the IGSF. CL and LTPL spectrums are conducted to verify the IGSF’s type. The CL spectrum shows the maximum photoemission at 2.431 eV and negligible bandgap emission. In the LTPL spectrum, four phonon replicas are found at 2.468 eV, 2.438 eV, 2.420 eV and 2.410 eV, respectively. The Egx is estimated to be 2.512 eV. A shoulder with a red-shift to the main peak in CL, and a slight protrude at the same wavelength in LTPL are verified as the so called Egx- lines. Based on the CL and LTPL results, the IGSF is identified as a 2SSF. Back etching by neutral loop discharge and DSE are conducted to track the origin of the 2SSF, and the nucleation site is found to be a threading screw dislocation (TSD) in this sample. A nucleation mechanism model is proposed for the formation of the 2SSF. Steps introduced by the off-cut and the TSD on the surface are both suggested to be two C-Si bilayers height. The intersections of such two types of steps are along [11-20] direction from the TSD, while a four-bilayer step at each intersection. The nucleation of the 2SSF in the growth is proposed as follows. Firstly, the upper two bilayers of the four-bilayer step grow down and block the lower two at one intersection, and an IGSF is generated. Secondly, the step-flow grows over the IGSF successively, and forms an AC/ABCABC/BA/BC stacking sequence. Then a 2SSF is formed and extends by the step-flow growth. In conclusion, a triangular IGSF is characterized by CL approach. Base on the CL and LTPL spectrums, the estimated Egx is 2.512 eV and the IGSF is identified to be a 2SSF. By back etching, the 2SSF nucleation site is found to be a TSD. A model for the 2SSF nucleation from an intersection of off-cut- and TSD- introduced steps is proposed.

Keywords: cathodoluminescence, defect-selected-etching, double Shockley stacking fault, low-temperature photoluminescence, nucleation model, silicon carbide

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Authors: Elia Favarelli, Enrico Testi, Andrea Giorgetti

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Autonomous structural health monitoring (SHM) of many structures and bridges became a topic of paramount importance for maintenance purposes and safety reasons. This paper proposes a set of machine learning (ML) tools to perform automatic feature selection and detection of anomalies in a bridge from vibrational data and compare different feature extraction schemes to increase the accuracy and reduce the amount of data collected. As a case study, the Z-24 bridge is considered because of the extensive database of accelerometric data in both standard and damaged conditions. The proposed framework starts from the first four fundamental frequencies extracted through operational modal analysis (OMA) and clustering, followed by density-based time-domain filtering (tracking). The fundamental frequencies extracted are then fed to a dimensionality reduction block implemented through two different approaches: feature selection (intelligent multiplexer) that tries to estimate the most reliable frequencies based on the evaluation of some statistical features (i.e., mean value, variance, kurtosis), and feature extraction (auto-associative neural network (ANN)) that combine the fundamental frequencies to extract new damage sensitive features in a low dimensional feature space. Finally, one class classifier (OCC) algorithms perform anomaly detection, trained with standard condition points, and tested with normal and anomaly ones. In particular, a new anomaly detector strategy is proposed, namely one class classifier neural network two (OCCNN2), which exploit the classification capability of standard classifiers in an anomaly detection problem, finding the standard class (the boundary of the features space in normal operating conditions) through a two-step approach: coarse and fine boundary estimation. The coarse estimation uses classics OCC techniques, while the fine estimation is performed through a feedforward neural network (NN) trained that exploits the boundaries estimated in the coarse step. The detection algorithms vare then compared with known methods based on principal component analysis (PCA), kernel principal component analysis (KPCA), and auto-associative neural network (ANN). In many cases, the proposed solution increases the performance with respect to the standard OCC algorithms in terms of F1 score and accuracy. In particular, by evaluating the correct features, the anomaly can be detected with accuracy and an F1 score greater than 96% with the proposed method.

Keywords: anomaly detection, frequencies selection, modal analysis, neural network, sensor network, structural health monitoring, vibration measurement

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Authors: Alexander David Natanson

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This paper focuses on the effect of Sanctuary and Immigration and Customs Enforcement (ICE) policies on local economies. "Sanctuary cities" refers to municipal jurisdictions that limit their cooperation with the federal government's efforts to enforce immigration. Using county-level data from the American Community Survey and ICE data on economic indicators from 2006 to 2018, this study isolates the effects of local immigration policies on U.S. counties. The investigation is accomplished by simultaneously studying the policies' effects in counties where immigrants' families are persecuted via collaboration with Immigration and Customs Enforcement (ICE), in contrast to counties that provide protections. The analysis includes a difference-in-difference & two-way fixed effect model. Results are robust to nearest-neighbor matching, after the random assignment of treatment, after running estimations using different cutoffs for immigration policies, and with a regression discontinuity model comparing bordering counties with opposite policies. Results are also robust after restricting the data to a single-year policy adoption, using the Sun and Abraham estimator, and with event-study estimation to deal with the staggered treatment issue. In addition, the study reverses the estimation to understand what drives the decision to choose policies to detect the presence of reverse causality biases in the estimated policy impact on economic factors. The evidence demonstrates that providing protections to undocumented immigrants increases economic activity. The estimates show gains in per capita income ranging from 3.1 to 7.2, median wages between 1.7 to 2.6, and GDP between 2.4 to 4.1 percent. Regarding labor, sanctuary counties saw increases in total employment between 2.3 to 4 percent, and the unemployment rate declined from 12 to 17 percent. The data further shows that ICE policies have no statistically significant effects on income, median wages, or GDP but adverse effects on total employment, with declines from 1 to 2 percent, mostly in rural counties, and an increase in unemployment of around 7 percent in urban counties. In addition, results show a decline in the foreign-born population in ICE counties but no changes in sanctuary counties. The study also finds similar results for sanctuary counties when separating the data between urban, rural, educational attainment, gender, ethnic groups, economic quintiles, and the number of business establishments. The takeaway from this study is that institutional inclusion creates the dynamic nature of an economy, as inclusion allows for economic expansion due to the extension of fundamental freedoms to newcomers. Inclusive policies show positive effects on economic outcomes with no evident increase in population. To make sense of these results, the hypothesis and theoretical model propose that inclusive immigration policies play an essential role in conditioning the effect of immigration by decreasing uncertainties and constraints for immigrants' interaction in their communities, decreasing the cost from fear of deportation or the constant fear of criminalization and optimize their human capital.

Keywords: inclusive and exclusive institutions, post matching, fixed effect, time trend, regression discontinuity, difference-in-difference, randomization inference and sun, Abraham estimator

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Authors: A. Newman-Tancredi, R. Depoortère, P. Gruca, E. Litwa, M. Lason, M. Papp

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The N-methyl-D-aspartic acid (NMDA) receptor antagonist, ketamine, can elicit rapid-acting antidepressant (RAAD) effects in treatment-resistant patients, but it requires parenteral co-administration with a classical antidepressant under medical supervision. In addition, ketamine can also produce serious side effects that limit its long-term use, and there is much interest in identifying RAADs based on ketamine’s mechanism of action but with safer profiles. Ketamine elicits GABAergic interneuron inhibition, glutamatergic neuron stimulation, and, notably, activation of serotonin 5-HT1A receptors in the prefrontal cortex (PFC). Direct activation of the latter receptor subpopulation with selective ‘biased agonists’ may therefore be a promising strategy to identify novel RAADs and, consistent with this hypothesis, the prototypical cortical biased agonist, NLX-101, exhibited robust RAAD-like activity in the chronic mild stress model of depression (CMS). The present study compared the effects of a novel, selective 5-HT1A receptor-biased agonist, NLX-204, with those of ketamine and NLX-101. Materials and methods: CMS procedure was conducted on Wistar rats; drugs were administered either intraperitoneally (i.p.) or by bilateral intracortical microinjection. Ketamine: 10 mg/kg i.p. or 10 µg/side in PFC; NLX-204 and NLX-101: 0.08 and 0.16 mg/kg i.p. or 16 µg/side in PFC. In addition, interaction studies were carried out with systemic NLX-204 or NLX-101 (each at 0.16 mg/kg i.p.) in combination with intracortical WAY-100635 (selective 5-HT1A receptor antagonist; 2 µg/side) or muscimol (GABA-A receptor agonist, 12.5 ng/side). Anhedonia was assessed by CMS-induced decrease in sucrose solution consumption; anxiety-like behavior was assessed using the Elevated Plus Maze (EPM), and cognitive impairment was assessed by the Novel Object Recognition (NOR) test. Results: A single administration of NLX-204 was sufficient to reverse the CMS-induced deficit in sucrose consumption, similarly to ketamine and NLX-101. NLX-204 also reduced CMS-induced anxiety in the EPM and abolished CMS-induced NOR deficits. These effects were maintained (EPM and NOR) or enhanced (sucrose consumption) over a subsequent 2-week period of treatment. The anti-anhedonic response of the drugs was also maintained for several weeks Following treatment discontinuation, suggesting that they had sustained effects on neuronal networks. A single PFC administration of NLX-204 reversed deficient sucrose consumption, similarly to ketamine and NLX-101. Moreover, the anti-anhedonic activities of systemic NLX-204 and NLX 101 were abolished by coadministration with intracortical WAY-100635 or muscimol. Conclusions: (i) The antidepressant-like activity of NLX-204 in the rat CMS model was as rapid as that of ketamine or NLX-101, supporting targeting cortical 5-HT1A receptors with selective, biased agonists to achieve RAAD effects. (ii)The anti-anhedonic activity of systemic NLX-204 was mimicked by local administration of the compound in the PFC, confirming the involvement of cortical circuits in its RAAD-like effects. (iii) Notably, the effects of systemic NLX-204 and NLX-101 were abolished by PFC administration of muscimol, indicating that they act by (indirectly) eliciting a reduction in cortical GABAergic neurotransmission. This is consistent with ketamine’s mechanism of action and suggests that there are converging NMDA and 5-HT1A receptor signaling cascades in PFC underlying the RAAD-like activities of ketamine and NLX-204. Acknowledgements: The study was financially supported by NCN grant no. 2019/35/B/NZ7/00787.

Keywords: depression, ketamine, serotonin, 5-HT1A receptor, chronic mild stress

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Authors: S. M. Ashikur Elahee, Md. Zahidur Rahman, Md. Shofiqur Rahman

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This study evaluated the impact of different water interventions under WASH program on access of household's to safe drinking water. Following survey method, the study was carried out in two Upazila of South-west coastal region of Bangladesh namely Koyra from Khulna and Shymnagar from Satkhira district. Being an explanatory study, a total of 200 household's selected applying random sampling technique were interviewed using a structured interview schedule. The predicted probability suggests that around 62 percent household's are out of year-round access to safe drinking water whereby, only 25 percent household's have access at SPHERE standard (913 Liters/per person/per year). Besides, majority (78 percent) of the household's have not accessed at both indicators simultaneously. The distance from household residence to the water source varies from 0 to 25 kilometer with an average distance of 2.03 kilometers. The study also reveals that the increase in monthly income around BDT 1,000 leads to additional 11 liters (coefficient 0.01 at p < 0.1) consumption of safe drinking water for a person/year. As expected, lining up time has significant negative relationship with dependent variables i.e., for higher lining up time, the probability of getting access for both SPHERE standard and year round access variables becomes lower. According to ordinary least square (OLS) regression results, water consumption decreases at 93 liters for per person/year of a household if one member is added to that household. Regarding water consumption intensity, ordered logistic regression (OLR) model shows that one-minute increase of lining up time for water collection tends to reduce water consumption intensity. On the other hand, as per OLS regression results, for one-minute increase of lining up time, the water consumption decreases by around 8 liters. Considering access to Deep Tube Well (DTW) as a reference dummy, in OLR, the household under Pond Sand Filter (PSF), Shallow Tube Well (STW), Reverse Osmosis (RO) and Rainwater Harvester System (RWHS) are respectively 37 percent, 29 percent, 61 percent and 27 percent less likely to ensure year round access of water consumption. In line of health impact, different type of water born diseases like diarrhea, cholera, and typhoid are common among the coastal community caused by microbial impurities i.e., Bacteria, Protozoa. High turbidity and TDS in pond water caused by reduction of water depth, presence of suspended particle and inorganic salt stimulate the growth of bacteria, protozoa, and algae causes affecting health hazard. Meanwhile, excessive growth of Algae in pond water caused by excessive nitrate in drinking water adversely effects on child health. In lieu of ensuring access at SPHERE standard, we need to increase the number of water interventions at reasonable distance, preferably a half kilometer away from the dwelling place, ensuring community peoples involved with its installation process where collectively owned water intervention is found more effective than privately owned. In addition, a demand-responsive approach to supply of piped water should be adopted to allow consumer demand to guide investment in domestic water supply in future.

Keywords: access, impact, safe drinking water, Sphere standard, water interventions

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Authors: Ricardo Alberto Chiong Zevallos, Eduardo Moraes Rego Reis

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Pancreatic adenocarcinoma (PDAC) patients have a less than 10% 5-year survival rate. PDAC has no defined diagnostic and prognostic biomarkers. Gemcitabine is the first-line drug in PDAC and several other cancers. Long non-coding RNAs (lncRNAs) contribute to the tumorigenesis and are potential biomarkers for PDAC. Although lncRNAs aren’t translated into proteins, they have important functions. LncRNAs can decoy or recruit proteins from the epigenetic machinery, act as microRNA sponges, participate in protein translocation through different cellular compartments, and even promote chemoresistance. The chromatin remodeling enzyme EZH2 is a histone methyltransferase that catalyzes the methylation of histone 3 at lysine 27, silencing local expression. EZH2 is ambivalent, it can also activate gene expression independently of its histone methyltransferase activity. EZH2 is overexpressed in several cancers and interacts with lncRNAs, being recruited to a specific locus. EZH2 can be recruited to activate an oncogene or silence a tumor suppressor. The lncRNAs misregulation in cancer can result in the differential recruitment of EZH2 and in a distinct epigenetic landscape, promoting chemoresistance. The relevance of the EZH2-lncRNAs interaction to chemoresistant PDAC was assessed by Real Time quantitative PCR (RT-qPCR) and RNA Immunoprecipitation (RIP) experiments with naïve and gemcitabine-resistant PDAC cells. The expression of several lncRNAs and EZH2 gene targets was evaluated contrasting naïve and resistant cells. Selection of candidate genes was made by bioinformatic analysis and literature curation. Indeed, the resistant cell line showed higher expression of chemoresistant-associated lncRNAs and protein coding genes. RIP detected lncRNAs interacting with EZH2 with varying intensity levels in the cell lines. During RIP, the nuclear fraction of the cells was incubated with an antibody for EZH2 and with magnetic beads. The RNA precipitated with the beads-antibody-EZH2 complex was isolated and reverse transcribed. The presence of candidate lncRNAs was detected by RT-qPCR, and the enrichment was calculated relative to INPUT (total lysate control sample collected before RIP). The enrichment levels varied across the several lncRNAs and cell lines. The EZH2-lncRNA interaction might be responsible for the regulation of chemoresistance-associated genes in multiple cancers. The relevance of the lncRNA-EZH2 interaction to PDAC was assessed by siRNA knockdown of a lncRNA, followed by the analysis of the EZH2 target expression by RT-qPCR. The chromatin immunoprecipitation (ChIP) of EZH2 and H3K27me3 followed by RT-qPCR with primers for EZH2 targets also assess the specificity of the EZH2 recruitment by the lncRNA. This is the first report of the interaction of EZH2 and lncRNAs HOTTIP and PVT1 in chemoresistant PDAC. HOTTIP and PVT1 were described as promoting chemoresistance in several cancers, but the role of EZH2 is not clarified. For the first time, the lncRNA LINC01133 was detected in a chemoresistant cancer. The interaction of EZH2 with LINC02577, LINC00920, LINC00941, and LINC01559 have never been reported in any context. The novel lncRNAs-EZH2 interactions regulate chemoresistant-associated genes in PDAC and might be relevant to other cancers. Therapies targeting EZH2 alone weren’t successful, and a combinatorial approach also targeting the lncRNAs interacting with it might be key to overcome chemoresistance in several cancers.

Keywords: epigenetics, chemoresistance, long non-coding RNAs, pancreatic cancer, histone modification

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