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Authors: Mirosław Krzemieniewski, Marcin Zieliński, Marcin Dębowski

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Electrokinetic disintegration is one of the high-voltage electric methods. The design of systems is exceptionally simple. Biomass flows through a system of pipes with alongside mounted electrodes that generate an electric field. Discharges in the electric field deform cell walls and lead to their successive perforation, thereby making their contents easily available to bacteria. The spark-over occurs between electrode surface and pipe jacket which is the second pole and closes the circuit. The value of voltage ranges from 10 to 100kV. Electrodes are supplied by normal “power grid” monophase electric current (230V, 50Hz). Next, the electric current changes into direct current of 24V in modules serving for particular electrodes, and this current directly feeds the electrodes. The installation is completely safe because the value of generated current does not exceed 250mA and because conductors are grounded. Therefore, there is no risk of electric shock posed to the personnel, even in the case of failure or incorrect connection. Low values of the electric current mean small energy consumption by the electrode which is extremely low – only 35W per electrode – compared to other methods of disintegration. Pipes with electrodes with diameter of DN150 are made of acid-proof steel and connected from both sides with 90º elbows ended with flanges. The available S and U types of pipes enable very convenient fitting with system construction in the existing installations and rooms or facilitate space management in new applications. The system of pipes for electrokinetic disintegration may be installed horizontally, vertically, askew, on special stands or also directly on the wall of a room. The number of pipes and electrodes is determined by operating conditions as well as the quantity of substrate, type of biomass, content of dry matter, method of disintegration (single or circulatory), mounting site etc. The most effective method involves pre-treatment of substrate that may be pumped through the disintegration system on the way to the fermentation tank or recirculated in a buffered intermediate tank (substrate mixing tank). Biomass structure destruction in the process of electrokinetic disintegration causes shortening of substrate retention time in the tank and acceleration of biogas production. A significant intensification of the fermentation process was observed in the systems operating in the technical scale, with the greatest increase in biogas production reaching 18%. The secondary, but highly significant for the energetic balance, effect is a tangible decrease of energy input by agitators in tanks. It is due to reduced viscosity of the biomass after disintegration, and may result in energy savings reaching even 20-30% of the earlier noted consumption. Other observed phenomena include reduction in the layer of surface scum, reduced sewage capability for foaming and successive decrease in the quantity of bottom sludge banks. Considering the above, the system for electrokinetic disintegration seems a very interesting and valuable solutions meeting the offer of specialist equipment for the processing of plant biomass, including Virginia fanpetals, before the process of methane fermentation.

Keywords: electrokinetic disintegration, biomass, biogas production, fermentation, Virginia fanpetals

Procedia PDF Downloads 343

Authors: Linara Salikhova, Elmira Nizamova, Aleksandra Katasonova, Gleb Vitkov, Olga Sarapulova.

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While most European cities conduct research on heat-related risks, there is a research gap in the Caucasus region, particularly in Yerevan, Armenia. This study aims to test the method of establishing a correlation between urban heat islands (UHI) and urban greenery distribution for prioritization of heat-vulnerable areas for revegetation. Armenia has failed to consider measures to mitigate UHI in urban development strategies despite a 2.1°C increase in average annual temperature over the past 32 years. However, planting vegetation in the city is commonly used to deal with air pollution and can be effective in reducing UHI if it prioritizes heat-vulnerable areas. The research focuses on establishing such priorities while considering the distribution of urban greenery across the city. The lack of spatially explicit air temperature data necessitated the use of satellite images to achieve the following objectives: (1) identification of land surface temperatures (LST) and quantification of temperature variations across districts; (2) classification of massifs of land surface types using normalized difference vegetation index (NDVI); (3) correlation of land surface classes with LST. Examination of the heat-vulnerable city areas (in this study, the proportion of individuals aged 75 years and above) is based on demographic data (Census 2011). Based on satellite images (Sentinel-2) captured on June 5, 2021, NDVI calculations were conducted. The massifs of the land surface were divided into five surface classes. Due to capacity limitations, the average LST for each district was identified using one satellite image from Landsat-8 on August 15, 2021. In this research, local relief is not considered, as the study mainly focuses on the interconnection between temperatures and green massifs. The average temperature in the city is 3.8°C higher than in the surrounding non-urban areas. The temperature excess ranges from a low in Norq Marash to a high in Nubarashen. Norq Marash and Avan have the highest tree and grass coverage proportions, with 56.2% and 54.5%, respectively. In other districts, the balance of wastelands and buildings is three times higher than the grass and trees, ranging from 49.8% in Quanaqer-Zeytun to 76.6% in Nubarashen. Studies have shown that decreased tree and grass coverage within a district correlates with a higher temperature increase. The temperature excess is highest in Erebuni, Ajapnyak, and Nubarashen districts. These districts have less than 25% of their area covered with grass and trees. On the other hand, Avan and Norq Marash districts have a lower temperature difference, as more than 50% of their areas are covered with trees and grass. According to the findings, a significant proportion of the elderly population (35%) aged 75 years and above reside in the Erebuni, Ajapnyak, and Shengavit neighborhoods, which are more susceptible to heat stress with an LST higher than in other city districts. The findings suggest that the method of comparing the distribution of green massifs and LST can contribute to the prioritization of heat-vulnerable city areas for revegetation. The method can become a rationale for the formation of an urban greening program.

Keywords: heat-vulnerability, land surface temperature, urban greenery, urban heat island, vegetation

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Authors: Prabhjot Kaur, B.K. Kang, Balwinder Singh

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The tobacco caterpillar, Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae) is an agricultural important pest species. S. litura has a wide host range of approximately recorded 150 plant species worldwide. In Punjab, this pest attains sporadic status primarily on cauliflower, Brassica oleracea (L.). This pest destroys vegetable crop and particularly prefers the cruciferae family. However, it is also observed feeding on other crops such as arbi, Colocasia esculenta (L.), mung bean, Vigna radiata (L.), sunflower, Helianthus annuus (L.), cotton, Gossypium hirsutum (L.), castor, Ricinus communis (L.), etc. Larvae of this pest completely devour the leaves of infested plant resulting in huge crop losses which ranges from 50 to 70 per cent. Indiscriminate and continuous use of insecticides has contributed in development of insecticide resistance in insects and caused the environmental degradation as well. Moreover, a base line data regarding the toxicity of the newer insecticides would help in understanding the level of resistance developed in this pest and any possible cross-resistance there in, which could be assessed in advance. Therefore, present studies on development of resistance in S. litura against four new chemistry insecticides (emamectin benzoate, chlorantraniliprole, indoxacarb and spinosad) were carried out in the Toxicology laboratory, Department of Entomology, Punjab Agricultural University, Ludhiana, Punjab, India during the year 2011-12. Various stages of S. litura (eggs, larvae) were collected from four different locations (Malerkotla, Hoshiarpur, Amritsar and Samrala) of Punjab. Resistance is developed in third instars of lepidopterous pests. Therefore, larval bioassays were conducted to estimate the response of field populations of thirty third-instar larvae of S. litura under laboratory conditions at 25±2°C and 65±5 per cent relative humidity. Leaf dip bioassay technique with diluted insecticide formulations recommended by Insecticide Resistance Action Committee (IRAC) was performed in the laboratory with seven to ten treatments depending on the insecticide class, respectively. LC50 values were estimated by probit analysis after correction to record control mortality data which was used to calculate the resistance ratios (RR). The LC50 values worked out for emamectin benzoate, chlorantraniliprole, indoxacarb, spinosad are 0.081, 0.088, 0.380, 4.00 parts per million (ppm) against pest populations collected from Malerkotla; 0.051, 0.060, 0.250, 3.00 (ppm) of Amritsar; 0.002, 0.001, 0.0076, 0.10 ppm for Samrala and 0.000014, 0.00001, 0.00056, 0.003 ppm against pest population of Hoshiarpur, respectively. The LC50 values for populations collected from these four locations were in the order Malerkotla>Amritsar>Samrala>Hoshiarpur for the insecticides (emamectin benzoate, chlorantraniliprole, indoxacarb and spinosad) tested. Based on LC50 values obtained, emamectin benzoate (0.000014 ppm) was found to be the most toxic among all the tested populations, followed by chlorantraniliprole (0.00001 ppm), indoxacarb (0.00056 ppm) and spinosad (0.003 ppm), respectively. The pairwise correlation coefficients of LC50 values indicated that there was lack of cross resistance for emamectin benzoate, chlorantraniliprole, spinosad, indoxacarb in populations of S. litura from Punjab. These insecticides may prove to be promising substitutes for the effective control of insecticide resistant populations of S. litura in Punjab state, India.

Keywords: Spodoptera litura, insecticides, toxicity, resistance

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Authors: Biao Hu, Mark E. J. Cutler, Alexandra C. Morel

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Global climate change introduces both opportunities and challenges for crop productivity, with differences in temperature stress across latitudes and crop types, one of the most important meteorological factors impacting crop productivity. The development and productivity of crops are particularly impacted when temperatures occur outwith their preferred ranges, which has implications for global agri-food sector. This study investigated the spatiotemporal dynamics of heat stress and cold stress on UK arable lands for rapeseed cropping between 1961 and 2020, using a 1 km spatial resolution temperature dataset. Stress indices, including heat stress index (fHS) defined as the ratio of “Tmax - Tcrit_h” to “Tlimit_h - Tcrit_h” where Tmax, Tcrit_h and Tlimit_h represent the daily maximum temperature (°C), critical high temperature threshold (°C) and limiting high temperature threshold (°C) of rapeseed crop respectively; cold degree days (CDD) as the difference between daily Tmin (minimum temperature) and Tcrit_l (critical low temperature threshold); and a normalized rapeseed production loss index (fRPL) as the product of fHS and attainable rapeseed yield in the same land pixel were established. The values of fHS and CDD, percentages of days experiencing each stress and fRPL were investigated. Results found increasing fHS and the areas impacted by heat stress during flowering (from April to May) and reproductive (from April to July) stages over time, with the mean fHS being negatively correlated with latitude. This pattern of increased heat stress agrees with previous research on rapeseed cropping, which have been noted at global scale in response to changes in climate. The decreasing number of CDD and frequency of cold stress suggest cold stress decreased during flowering, vegetative (from September to March next year) and reproductive stages, and the magnitude of cold stress in the south of the UK was smaller to that compared to northern regions over the studied periods. The decreasing CDD matches observed declining cold stress of global rapeseed and of other crops such as rice in the northern hemisphere. Notably, compared with previous studies which mainly tracked the trends of heat stress and cold stress individually, this study conducted a comparative analysis of the rate of their changes and found heat stress of rapeseed crops in the UK was increasing at a faster rate than cold stress, which was seen to decrease during flowering. The increasing values of fRPL, with statistically significant differences (p < 0.05) between regions of the UK, suggested an increasing loss in rapeseed due to heat stress in the studied period. The largest increasing trend in heat stress was observed in South-eastern England, where a decreasing cold stress was taking place. While the present study observed a relatively slowly increasing heat stress, there is a worrying trend of increasing heat stress for rapeseed cropping into the future, as the cases of other main rapeseed cropping systems in the northern hemisphere including China, European counties, the US, and Canada. This study demonstrates the negative impact of global warming on rapeseed cropping, highlighting the adaptation and mitigations strategies for sustainable rapeseed cultivation across the globe.

Keywords: rapeseed, UK, heat stress, cold stress, global climate change, spatiotemporal analysis, production loss index

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Authors: Silvia D. Vaca, Benjamin J. Kuo, Joao Ricardo N. Vissoci, Catherine A. Staton, Linda W. Xu, Michael Muhumuza, Hussein Ssenyonjo, John Mukasa, Joel Kiryabwire, Henry E. Rice, Gerald A. Grant, Michael M. Haglund

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Background: While delays to care exist in resource rich settings, greater delays are seen along the care continuum in low- and middle-income countries (LMICs) largely due to limited healthcare capacity to address the disproportional rates of traumatic brain injury (TBI) in Sub Saharan Africa (SSA). While many LMICs have government subsidized systems to offset surgical costs, the burden of securing funds by the patients for medications, supplies, and CT diagnostics poses a significant challenge to timely surgical interventions. In Kampala Uganda, the challenge of obtaining timely CT scans is twofold. First, due to a lack of a functional CT scanner at the tertiary hospital, patients need to arrange their own transportation to the nearby private facility for CT scans. Second, self-financing for the private CT scans ranges from $80 - $130, which is near the average monthly income in Kampala. These bottlenecks contribute significantly to the care continuum delays and are associated with poor TBI outcomes. Objective: The objectives of this study are to 1) describe the temporal delays through a modified three delays model that fits the context of neurosurgical interventions for TBI patients in Kampala and 2) investigate the association between delays and mortality. Methods: Prospective data were collected for 563 TBI patients presenting to a tertiary hospital in Kampala from 1 June – 30 November 2016. Four time intervals were constructed along five time points: injury, hospital arrival, neurosurgical evaluation, CT results, and definitive surgery. Time interval differences among mild, moderate and severe TBI and their association with mortality were analyzed. Results: The mortality rate of all TBI patients presenting to MNRH was 9.6%, which ranged from 4.7% for mild and moderate TBI patients receiving surgery to 81.8% for severe TBI patients who failed to receive surgery. The duration from injury to surgery varied considerably across TBI severity with the largest gap seen between mild TBI (174 hours) and severe TBI (69 hours) patients. Further analysis revealed care continuum differences for interval 3 (neurosurgical evaluation to CT result) and 4 (CT result to surgery) between severe TBI patients (7 hours for interval 3 and 24 hours for interval 4) and mild TBI patients (19 hours for interval 3, and 96 hours for interval 4). These post-arrival delays were associated with mortality for mild (p=0.05) and moderate TBI (p=0.03) patients. Conclusions: To our knowledge, this is the first analysis using a modified ‘three delays’ framework to analyze the care continuum of TBI patients in Uganda from injury to surgery. We found significant associations between delays and mortality for mild and moderate TBI patients. As it currently stands, poorer outcomes were observed for these mild and moderate TBI patients who were managed non-operatively or failed to receive surgery while surgical services were shunted to more severely ill patients. While well intentioned, high mortality rates were still observed for the severe TBI patients managed surgically. These results suggest the need for future research to optimize triage practices, understand delay contributors, and improve pre-hospital logistical referral systems.

Keywords: care continuum, global neurosurgery, Kampala Uganda, LMIC, Mulago, prospective registry, traumatic brain injury

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Authors: Philipp Wenger, Michael Beltle, Stefan Tenbohlen, Uwe Riechert

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The integration of renewable energy introduces new challenges to the transmission grid, as the power generation is located far from load centers. The associated necessary long-range power transmission increases the demand for high voltage direct current (HVDC) transmission lines and DC distribution grids. HVDC gas-insulated switchgears (GIS) are considered being a key technology, due to the combination of the DC technology and the long operation experiences of AC-GIS. To ensure long-term reliability of such systems, insulation defects must be detected in an early stage. Operational experience with AC systems has proven evidence, that most failures, which can be attributed to breakdowns of the insulation system, can be detected and identified via partial discharge (PD) measurements beforehand. In AC systems the identification of defects relies on the phase resolved partial discharge pattern (PRPD). Since there is no phase information within DC systems this method cannot be transferred to DC PD diagnostic. Furthermore, the behaviour of e.g. free-moving particles differs significantly at DC: Under the influence of a constant direct electric field, charge carriers can accumulate on particles’ surfaces. As a result, a particle can lift-off, oscillate between the inner conductor and the enclosure or rapidly bounces at just one electrode, which is known as firefly motion. Depending on the motion and the relative position of the particle to the electrodes, broadband electromagnetic PD pulses are emitted, which can be recorded by ultra-high frequency (UHF) measuring methods. PDs are often accompanied by light emissions at the particle’s tip which enables optical detection. This contribution investigates PD characteristics of free moving metallic particles in a commercially available 300 kV SF6-insulated HVDC-GIS. The influences of various defect parameters on the particle motion and the PD characteristic are evaluated experimentally. Several particle geometries, such as cylinder, lamella, spiral and sphere with different length, diameter and weight are determined. The applied DC voltage is increased stepwise from inception voltage up to UDC = ± 400 kV. Different physical detection methods are used simultaneously in a time-synchronized setup. Firstly, the electromagnetic waves emitted by the particle are recorded by an UHF measuring system. Secondly, a photomultiplier tube (PMT) detects light emission with a wavelength in the range of λ = 185…870 nm. Thirdly, a high-speed camera (HSC) tracks the particle’s motion trajectory with high accuracy. Furthermore, an electrically insulated electrode is attached to the grounded enclosure and connected to a current shunt in order to detect low frequency ion currents: The shunt measuring system’s sensitivity is in the range of 10 nA at a measuring bandwidth of bw = DC…1 MHz. Currents of charge carriers, which are generated at the particle’s tip migrate through the gas gap to the electrode and can be recorded by the current shunt. All recorded PD signals are analyzed in order to identify characteristic properties of different particles. This includes e.g. repetition rates and amplitudes of successive pulses, characteristic frequency ranges and detected signal energy of single PD pulses. Concluding, an advanced understanding of underlying physical phenomena particle motion in direct electric field can be derived.

Keywords: current shunt, free moving particles, high-speed imaging, HVDC-GIS, UHF

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Authors: Christelle Rabbat, Carole Vouebou, Sary Awad, Alan Jean-Marie

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Hydrogen has been proven to be a viable alternative to standard fuels as it is easy to produce and only generates water vapour and zero carbon emissions. However, despite the hydrogen benefits, the widespread adoption of hydrogen fuel cell vehicles and internal combustion engine vehicles is impeded by several challenges. The lack of refueling infrastructures remains one of the main hindering factors due to the high costs associated with their design, construction, and operation. Besides, the lack of hydrogen vehicles on the road diminishes the economic viability of investing in refueling infrastructure. Simultaneously, the absence of accessible refueling stations discourages consumers from adopting hydrogen vehicles, perpetuating a cycle of limited market uptake. To address these challenges, the implementation of adequate policies incentivizing the use of hydrogen vehicles and the reduction of the investment and operation costs of hydrogen refueling stations (HRS) are essential to put both investors and customers at ease. Even though the transition to hydrogen cars has been rather slow, public transportation companies have shown a keen interest in this highly promising fuel. Besides, their hydrogen demand is easier to predict and regulate than personal vehicles. Due to the reduced complexity of designing a suitable hydrogen supply chain for public vehicles, this sub-sector could be a great starting point to facilitate the adoption of hydrogen vehicles. Consequently, this study will focus on designing a chain of on-site green HRS for the public transportation network in Nantes Metropole leveraging the latest relevant technological advances aiming to reduce the costs while ensuring reliability, safety, and ease of access. To reduce the cost of HRS and encourage their widespread adoption, a network of 7 H35-T40 HRS has been designed, replacing the conventional J-T valves with turbo-expanders. Each station in the network has a daily capacity of 1,920 kg. Thus, the HRS network can produce up to 12.5 tH2 per day. The detailed cost analysis has revealed a CAPEX per station of 16.6 M euros leading to a network CAPEX of 116.2 M euros. The proposed station siting prioritized Nantes metropole’s 5 bus depots and included 2 city-centre locations. Thanks to the turbo-expander technology, the cooling capacity of the proposed HRS is 19% lower than that of a conventional station equipped with J-T valves, resulting in significant CAPEX savings estimated at 708,560 € per station, thus nearly 5 million euros for the whole HRS network. Besides, the turbo-expander power generation ranges from 7.7 to 112 kW. Thus, the power produced can be used within the station or sold as electricity to the main grid, which would, in turn, maximize the station’s profit. Despite the substantial initial investment required, the environmental benefits, cost savings, and energy efficiencies realized through the transition to hydrogen fuel cell buses and the deployment of HRS equipped with turbo-expanders offer considerable advantages for both TAN and Nantes Metropole. These initiatives underscore their enduring commitment to fostering green mobility and combatting climate change in the long term.

Keywords: green hydrogen, refueling stations, turbo-expander, heavy-duty vehicles

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Authors: Milos Travar, Jovana Nikolov, Andrej Vranicar, Natasa Todorovic

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Over the years, High Purity Germanium (HPGe) detectors proved to be an excellent practical tool and, as such, have established their today's wide use in low background γ-spectrometry. One of the advantages of gamma-ray spectrometry is its easy sample preparation as chemical processing and separation of the studied subject are not required. Thus, with a single measurement, one can simultaneously perform both qualitative and quantitative analysis. One of the most prominent features of HPGe detectors, besides their excellent efficiency, is their superior resolution. This feature virtually allows a researcher to perform a thorough analysis by discriminating photons of similar energies in the studied spectra where otherwise they would superimpose within a single-energy peak and, as such, could potentially scathe analysis and produce wrongly assessed results. Naturally, this feature is of great importance when the identification of radionuclides, as well as their activity concentrations, is being practiced where high precision comes as a necessity. In measurements of this nature, in order to be able to reproduce good and trustworthy results, one has to have initially performed an adequate full-energy peak (FEP) efficiency calibration of the used equipment. However, experimental determination of the response, i.e., efficiency curves for a given detector-sample configuration and its geometry, is not always easy and requires a certain set of reference calibration sources in order to account for and cover broader energy ranges of interest. With the goal of overcoming these difficulties, a lot of researches turned towards the application of different software toolkits that implement the Monte Carlo method (e.g., MCNP, FLUKA, PENELOPE, Geant4, etc.), as it has proven time and time again to be a very powerful tool. In the process of creating a reliable model, one has to have well-established and described specifications of the detector. Unfortunately, the documentation that manufacturers provide alongside the equipment is rarely sufficient enough for this purpose. Furthermore, certain parameters tend to evolve and change over time, especially with older equipment. Deterioration of these parameters consequently decreases the active volume of the crystal and can thus affect the efficiencies by a large margin if they are not properly taken into account. In this study, the optimisation method of two HPGe detectors through the implementation of the Geant4 toolkit developed by CERN is described, with the goal of further improving simulation accuracy in calculations of FEP efficiencies by investigating the influence of certain detector variables (e.g., crystal-to-window distance, dead layer thicknesses, inner crystal’s void dimensions, etc.). Detectors on which the optimisation procedures were carried out were a standard traditional co-axial extended range detector (XtRa HPGe, CANBERRA) and a broad energy range planar detector (BEGe, CANBERRA). Optimised models were verified through comparison with experimentally obtained data from measurements of a set of point-like radioactive sources. Acquired results of both detectors displayed good agreement with experimental data that falls under an average statistical uncertainty of ∼ 4.6% for XtRa and ∼ 1.8% for BEGe detector within the energy range of 59.4−1836.1 [keV] and 59.4−1212.9 [keV], respectively.

Keywords: HPGe detector, γ spectrometry, efficiency, Geant4 simulation, Monte Carlo method

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Authors: Saeed Abbas, Rahilla Tabassum, Haider Abbas, Babar Khan, Shahid Hussain, Muhammad Zafar Khan, Fazal Karim, Yawar Abbas, Rizwan Karim

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Human–carnivore conflict is considered to be a major conservation and rural livelihood concern because many carnivore species have been heavily victimized due to elevated conflict levels with communities. Like other snow leopard range countries, this situation prevails in Pakistan, where WWF is currently working under Asia High Mountain Project (AHMP) in Gilgit-Baltistan of Pakistan. To mitigate such conflicts requires a firm understanding of grazing and predation pattern including human-carnivore interaction. For this purpose we conducted a survey in Hoper valley (one of the AHMP project sites in Pakistan), during August, 2013 through a questionnaire based survey and unstructured interviews covering 647 households, permanently residing in the project area out of the total 900 households. The valley, spread over 409 km2 between 36°7'46" N and 74°49'2"E, at 2900m asl in Karakoram ranges is considered to be one of an important habitat of snow leopard and associated prey species such as Himalayan ibex. The valley is home of 8100 Brusho people (ancient tribe of Northern Pakistan) dependent on agro-pastoral livelihoods including farming and livestock rearing. The total number of livestock reported were (N=15,481) out of which 8346 (53.91%) were sheep, 3546 (22.91%) goats, 2193 (14.16%) cows, 903 (5.83%) yaks, 508 (3.28%) bulls, 28 (0.18%) donkeys, 27 (0.17%) zo/zomo (cross breed of yak and cow), and 4 (0.03%) horses. 83 percent respondent (n=542 households) confirmed loss of their livestock during the last one year July, 2012 to June, 2013 which account for 2246 (14.51%) animals. The major reason of livestock loss include predation by large carnivores such as snow leopards and wolf (1710, 76.14%) followed by diseases (536, 23.86%). Of the total predation cases snow leopard is suspected to kill 1478 animals (86.43%). Among livestock sheep were found to be the major prey of snow leopard (810, 55%) followed by goats (484, 32.7%) cows (151, 10.21%), yaks (15, 1.015%), zo/zomo (7, 0.5%) and donkey (1, 0.07%). The reason for the mass depredation of sheep and goats is that they tend to browse on twigs of bushes and graze on soft grass near cliffs. They are also considered to be very active as compared to other species in moving quickly and covering more grazing area. This makes them more vulnerable to snow leopard attack. The majority (1283, 75%) of livestock killed by predators occurred during the warm season (May-September) in alpine and sub-alpine pastures and remaining (427, 25%) occurred in the winter season near settlements in valley. It was evident from the recent study that Snow leopard kills outside the pen were (1351, 79.76%) as compared to inside pen (359, 20.24%). Assessing the economic loss of livestock predation we found that the total loss of livestock predation in the study area is equal to PKR 11,230,000 (USD 105,797), which is about PRK 17, 357 (USD 163.51) per household per year. Economic loss incurred by the locals due to predation is quite significant where the average cash income per household per year is PKR 85,000 (USD 800.75).

Keywords: carnivores, conflict, predation, livelihood, conservation, rural, snow leopard, livestock

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Authors: Lionel Trovalet, Lisa Liu, Dimitri Bigot, Nadia Hammami, Jean-Pierre Habas, Bruno Malet-Damour

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The MCP-iBAT1 project is carried out to study the behavior of Phase Change Materials (PCM) integrated in building envelopes in a tropical environment. Through the phase transitions (melting and freezing) of the material, thermal energy can be absorbed or released. This process enables the regulation of indoor temperatures and the improvement of thermal comfort for the occupants. Most of the commercially available PCMs are more suitable to temperate climates than to tropical climates. The case of Reunion Island is noteworthy as there are multiple micro-climates. This leads to our key question: developing one or multiple bio-based PCMs that cover the thermal needs of the different locations of the island. The present paper focuses on the numerical approach to select the PCM properties relevant to tropical areas. Numerical simulations have been carried out with two softwares: EnergyPlusTM and Isolab. The latter has been developed in the laboratory, with the implicit Finite Difference Method, in order to evaluate different physical models. Both are Thermal Dynamic Simulation (TDS) softwares that predict the building’s thermal behavior with one-dimensional heat transfers. The parameters used in this study are the construction’s characteristics (dimensions and materials) and the environment’s description (meteorological data and building surroundings). The building is modeled in accordance with the experimental setup. It is divided into two rooms, cells A and B, with same dimensions. Cell A is the reference, while in cell B, a layer of commercial PCM (Thermo Confort of MCI Technologies) has been applied to the inner surface of the North wall. Sensors are installed in each room to retrieve temperatures, heat flows, and humidity rates. The collected data are used for the comparison with the numerical results. Our strategy is to implement two similar buildings at different altitudes (Saint-Pierre: 70m and Le Tampon: 520m) to measure different temperature ranges. Therefore, we are able to collect data for various seasons during a condensed time period. The following methodology is used to validate the numerical models: calibration of the thermal and PCM models in EnergyPlusTM and Isolab based on experimental measures, then numerical testing with a sensitivity analysis of the parameters to reach the targeted indoor temperatures. The calibration relies on the past ten months’ measures (from September 2020 to June 2021), with a focus on one-week study on November (beginning of summer) when the effect of PCM on inner surface temperatures is more visible. A first simulation with the PCM model of EnergyPlus gave results approaching the measurements with a mean error of 5%. The studied property in this paper is the melting temperature of the PCM. By determining the representative temperature of winter, summer and inter-seasons with past annual’s weather data, it is possible to build a numerical model of multi-layered PCM. Hence, the combined properties of the materials will provide an optimal scenario for the application on PCM in tropical areas. Future works will focus on the development of bio-based PCMs with the selected properties followed by experimental and numerical validation of the materials. 1Materiaux ´ a Changement de Phase, une innovation pour le B ` ati Tropical

Keywords: energyplus, multi-layer of PCM, phase changing materials, tropical area

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Authors: Eleni Akritopoulou, Katerina Topouzoglou

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The last 20 years marine debris has been identified as one of the main marine pollution sources caused by anthropogenic activities. Plastics has reached the farthest marine areas of the planet affecting all marine trophic levels including the, recently discovered, amphipoda Eurythenes plasticus inhabiting Mariana Trench to large cetaceans, marine reptiles and sea birds causing immunodeficiency disorders, deteriorating health and death overtime. For the time period 2016-20, in the framework of the national initiative ‘Keep Aegean Blue”, All for Blue team has been collecting marine debris (coastline and underwater) following a modified in situ MEDSEALITTER monitoring protocol from eight Greek islands. After collection, marine debris was weighted, sorted and categorised according to material; plastic (PL), glass (G), metal (M), wood (W), rubber (R), cloth (CL), paper (P), mixed (MX). The goal of the project included the documentation of marine debris sources, human trends, waste management and public marine environmental awareness. Waste management was focused on plastics recycling and utilisation into daily useful products. This research is focused on the island of Mykonos due to its continuous touristic activity and lack of scientific information. In overall, a field work area of 1.832.856 m2 was cleaned up yielding 5092 kg of marine debris. The preliminary results indicated PL as main source of marine debris (62,8%) followed by M (15,5%), GL (13,2%) and MX (2,8%). Main items found were fishing tools (lines, nets), disposable cutlery, cups and straws, cigarette butts, flip flops and other items like plastic boat compartments. In collaboration with a local company for plastic management and the Circular Economy and Eco Innovation Institute (Sweden), all plastic debris was recycled. Granulation process was applied transforming plastic into building materials used for refugees’ houses, litter bins bought by municipalities and schools and, other items like shower components. In terms of volunteering and attendance in public awareness seminars, there was a raise of interest by 63% from different age ranges and professions. Regardless, the research being fairly new for Mykonos island and logistics issues potentially affected systemic sampling, it appeared that plastic debris is the main littering source attributed, possibly to the intense touristic activity of the island all year around. However, marine environmental awareness activities were pointed out to be an effective tool in forming public perception against marine debris and, alter the daily habits of local society. Since the beginning of this project, three new local environmental teams were formed against marine pollution supported by the local authorities and stakeholders. The continuous need and request for the production of items made by recycled marine debris appeared to be beneficial socio-economically to the local community and actions are taken to expand the project nationally. Finally, as an ongoing project and whilst, new scientific information is collected, further funding and research is needed.

Keywords: Greece, marine debris, marine environmental awareness, Mykonos island, plastics debris, plastic granulation, recycled plastic, tourism, waste management

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Authors: Layla Haidar, Shari Esquenazi-Karonika

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Introduction: Mental health includes one’s emotional, psychological, and interpersonal well-being; it ranges from “good” to “poor” on a continuum. At the individual-level, it affects how a person thinks, feels, and acts. Moreover, it determines how they cope with stress, relate to others, and interface with their surroundings. Research has yielded that mental health is directly related with short- and long-term physical health (including chronic disease), health risk behaviors, education-level, employment, and social relationships. As is the case with physical conditions like diabetes, heart disease, and cancer, mitigating the behavioral and genetic risks of debilitating mental health conditions like anxiety and depression can nurture a healthier quality of mental health throughout one’s life. In order to maximize the benefits of prevention, it is important to identify modifiable risks and develop protective habits earlier in life. Methods: The Adolescent Behaviors and Experiences Survey (ABES) dataset was used for this study. The ABES survey was administered to high school students (9th-12th grade) during January 2021- June 2021 by the Centers for Disease Control and Prevention (CDC). The data was analyzed to identify any associations between feelings of sadness, hopelessness, or increased suicidality among high school students with relation to their participation on one or more sports teams and their average daily consumed screen time. Data was analyzed using descriptive and multivariable analytic techniques. A multinomial logistic regression of each variable was conducted to examine if there was an association, while controlling for grade-level, sex, and race. Results: The findings from this study are insightful for administrators and policymakers who wish to address mounting concerns related to student mental health. The study revealed that compared to a student who participated on zero sports teams, students who participated in 1 or more sports teams showed a significantly increased risk of depression (p<0.05). Conversely, the rate of depression in students was significantly less in those who consumed 5 or more hours of screen time per day, compared to those who consumed less than 1 hour per day of screen time (p<0.05). Conclusion: These findings are informative and highlight the importance of understanding the nuances of student participation on sports teams (e.g., physical exertion, social dynamics of team, and the level of competitiveness within the sport). Likewise, the context of an individual’s screen time (e.g., social media, engaging in team-based video games, or watching television) can inform parental or school-based policies about screen time activity. Although physical activity has been proven to be important for emotional and physical well-being of youth, playing on multiple teams could have negative consequences on the emotional state of high school students potentially due to fatigue, overtraining, and injuries. Existing literature has highlighted the negative effects of screen time; however, further research needs to consider the type of screen-based consumption to better understand its effects on mental health.

Keywords: behavioral science, mental health, adolescents, prevention

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Authors: K. Ghosh, S. N. Tripathi, Manish Joshi, Y. S. Mayya, Arshad Khan, B. K. Sapra

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We have developed a CFD coupled aerosol microphysics model in the context of aerosol generation from a glowing wire. The governing equations can be solved implicitly for mass, momentum, energy transfer along with aerosol dynamics. The computationally efficient framework can simulate temporal behavior of total number concentration and number size distribution. This formulation uniquely couples standard K-Epsilon scheme with boundary layer model with detailed aerosol dynamics through residence time. This model uses measured temperatures (wire surface and axial/radial surroundings) and wire compositional data apart from other usual inputs for simulations. The model predictions show that bulk fluid motion and local heat distribution can significantly affect the aerosol behavior when the buoyancy effect in momentum transfer is considered. Buoyancy generated turbulence was found to be affecting parameters related to aerosol dynamics and transport as well. The model was validated by comparing simulated predictions with results obtained from six controlled experiments performed with a laboratory-made hot wire nanoparticle generator. Condensation particle counter (CPC) and scanning mobility particle sizer (SMPS) were used for measurement of total number concentration and number size distribution at the outlet of reactor cell during these experiments. Our model-predicted results were found to be in reasonable agreement with observed values. The developed model is fast (fully implicit) and numerically stable. It can be used specifically for applications in the context of the behavior of aerosol particles generated from glowing wire technique and in general for other similar large scale domains. Incorporation of CFD in aerosol microphysics framework provides a realistic platform to study natural convection driven systems/ applications. Aerosol dynamics sub-modules (nucleation, coagulation, wall deposition) have been coupled with Navier Stokes equations modified to include buoyancy coupled K-Epsilon turbulence model. Coupled flow-aerosol dynamics equation was solved numerically and in the implicit scheme. Wire composition and temperature (wire surface and cell domain) were obtained/measured, to be used as input for the model simulations. Model simulations showed a significant effect of fluid properties on the dynamics of aerosol particles. The role of buoyancy was highlighted by observation and interpretation of nucleation zones in the planes above the wire axis. The model was validated against measured temporal evolution, total number concentration and size distribution at the outlet of hot wire generator cell. Experimentally averaged and simulated total number concentrations were found to match closely, barring values at initial times. Steady-state number size distribution matched very well for sub 10 nm particle diameters while reasonable differences were noticed for higher size ranges. Although tuned specifically for the present context (i.e., aerosol generation from hotwire generator), the model can also be used for diverse applications, e.g., emission of particles from hot zones (chimneys, exhaust), fires and atmospheric cloud dynamics.

Keywords: nanoparticles, k-epsilon model, buoyancy, CFD, hot wire generator, aerosol dynamics

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Authors: Mohammad R. Aghakashkooli, Milutin G. Jovanovic

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The brushless doubly-fed reluctance generator (BDFRG) is an emerging, medium-speed alternative to a conventional wound rotor slip-ring doubly-fed induction generator (DFIG) in wind energy conversion systems (WECS). It can provide competitive overall performance and similar low failure rates of a typically 30% rated back-to-back power electronics converter in 2:1 speed ranges but with the following important reliability and cost advantages over DFIG: the maintenance-free operation afforded by its brushless structure, 50% synchronous speed with the same number of rotor poles (allowing the use of a more compact, and more efficient two-stage gearbox instead of a vulnerable three-stage one), and superior grid integration properties including simpler protection for the low voltage ride through compliance of the fractional converter due to the comparatively higher leakage inductances and lower fault currents. Vector controlled pulse-width-modulated converters generally feature a much lower total harmonic distortion relative to hysteresis counterparts with variable switching rates and as such have been a predominant choice for BDFRG (and DFIG) wind turbines. Eliminating a shaft position sensor, which is often required for control implementation in this case, would be desirable to address the associated reliability issues. This fact has largely motivated the recent growing research of sensorless methods and developments of various rotor position and/or speed estimation techniques for this purpose. The main limitation of all the observer-based control approaches for grid-connected wind power applications of the BDFRG reported in the open literature is the requirement for pre-commissioning procedures and prior knowledge of the machine inductances, which are usually difficult to accurately identify by off-line testing. A model reference adaptive system (MRAS) based sensor-less vector control scheme to be presented will overcome this shortcoming. The true machine parameter independence of the proposed field-oriented algorithm, offering robust, inherently decoupled real and reactive power control of the grid-connected winding, is achieved by on-line estimation of the inductance ratio, the underlying rotor angular velocity and position MRAS observer being reliant upon. Such an observer configuration will be more practical to implement and clearly preferable to the existing machine parameter dependent solutions, and especially bearing in mind that with very little modifications it can be adapted for commercial DFIGs with immediately obvious further industrial benefits and prospects of this work. The excellent encoder-less controller performance with maximum power point tracking in the base speed region will be demonstrated by realistic simulation studies using large-scale BDFRG design data and verified by experimental results on a small laboratory prototype of the WECS emulation facility.

Keywords: brushless doubly fed reluctance generator, model reference adaptive system, sensorless vector control, wind energy conversion

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Authors: Mohd Zaimmudin Mohd Zain, Patsy Perry, Lee Quinn

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This study explores how fashion consumers are influenced by fashion bloggers towards pre-purchase decision for online fashion products in a non-Western context. Malaysians rank among the world’s most avid online shoppers, with apparel the third most popular purchase category. However, extant research on fashion blogging focuses on the developed Western market context. Numerous international fashion retailers have entered the Malaysian market from luxury to fast fashion segments of the market; however Malaysian fashion consumers must balance religious and social norms for modesty with their dress style and adoption of fashion trends. Consumers increasingly mix and match Islamic and Western elements of dress to create new styles enabling them to follow Western fashion trends whilst paying respect to social and religious norms. Social media have revolutionised the way that consumers can search for and find information about fashion products. For online fashion brands with no physical presence, social media provide a means of discovery for consumers. By allowing the creation and exchange of user-generated content (UGC) online, they provide a public forum that gives individual consumers their own voices, as well as access to product information that facilitates their purchase decisions. Social media empower consumers and brands have important roles in facilitating conversations among consumers and themselves, to help consumers connect with them and one another. Fashion blogs have become an important fashion information sources. By sharing their personal style and inspiring their followers with what they wear on popular social media platforms such as Instagram, fashion bloggers have become fashion opinion leaders. By creating UGC to spread useful information to their followers, they influence the pre-purchase decision. Hence, successful Western fashion bloggers such as Chiara Ferragni may earn millions of US dollars every year, and some have created their own fashion ranges and beauty products, become judges in fashion reality shows, won awards, and collaborated with high street and luxury brands. As fashion blogging has become more established worldwide, increasing numbers of fashion bloggers have emerged from non-Western backgrounds to promote Islamic fashion styles, such as Hassanah El-Yacoubi and Dian Pelangi. This study adopts a qualitative approach using netnographic content analysis of consumer comments on two famous Malaysian fashion bloggers’ Instagram accounts during January-March 2016 and qualitative interviews with 16 Malaysian Generation Y fashion consumers during September-October 2016. Netnography adapts ethnographic techniques to the study of online communities or computer-mediated communications. Template analysis of the data involved coding comments according to the theoretical framework, which was developed from the literature review. Initial data analysis shows the strong influence of Malaysian fashion bloggers on their followers in terms of lifestyle and morals as well as fashion style. Followers were guided towards the mix and match trend of dress with Western and Islamic elements, for example, showing how vivid colours or accessories could be worked into an outfit whilst still respecting social and religious norms. The blogger’s Instagram account is a form of online community where followers can communicate and gain guidance and support from other followers, as well as from the blogger.

Keywords: fashion bloggers, Malaysia, qualitative, social media

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Authors: Dilek Ozturk, Narin Ozturk, Zeliha Pala Kara, Engin Kaptan, Serap Sancar Bas, Nurten Ozsoy, Alper Okyar

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Most physiological processes oscillate in a rhythmic manner in mammals including metabolism and energy homeostasis, locomotor activity, hormone secretion, immune and endocrine system functions. Endocrine body rhythms are tightly regulated by the circadian timing system. The hypothalamic-pituitary-thyroid (HPT) axis is under circadian control at multiple levels from hypothalamus to thyroid gland. Since circadian timing system controls a variety of biological functions in mammals, circadian rhythms of biological functions may modify the drug tolerability/toxicity depending on the dosing time. Selective mTOR (mammalian target of rapamycin) inhibitor everolimus is an immunosuppressant and anticancer agent that is active against many cancers. It was also found to be active in medullary thyroid cancer. The aim of this study was to investigate the dosing time-dependent toxicity of everolimus on the thyroid gland and hormones in mice. Healthy C57BL/6J mice were synchronized with 12h:12h Light-Dark cycle (LD12:12, with Zeitgeber Time 0 – ZT0 – corresponding to Light onset). Everolimus was administered to male (5 mg/kg/day) and female mice (15 mg/kg/day) orally at ZT1-rest period- and ZT13-activity period- for 4 weeks; body weight loss, clinical signs and possible changes in serum thyroid hormone levels (TSH and free T4) were examined. Histological alterations in the thyroid gland were evaluated according to the following criteria: follicular size, colloid density and viscidity, height of the follicular epithelium and the presence of necrotic cells. The statistical significance between differences was analyzed with ANOVA. Study findings included everolimus-related diarrhea, decreased activity, decreased body weight gains, alterations in serum TSH levels, and histopathological changes in thyroid gland. Decreases in mean body weight gains were more evident in mice treated at ZT1 as compared to ZT13 (p < 0.001, for both sexes). Control tissue sections of thyroid glands exhibited well-organized histoarchitecture when compared to everolimus-treated groups. Everolimus caused histopathological alterations in thyroid glands in male (5 mg/kg, slightly) and female mice (15 mg/kg; p < 0.01 for both ZT as compared to their controls) irrespective of dosing-time. TSH levels were slightly decreased upon everolimus treatment at ZT13 in both males and females. Conversely, increases in TSH levels were observed when everolimus treated at ZT1 in both males (5 mg/kg; p < 0.05) and females (15 mg/kg; slightly). No statistically significant alterations in serum free T4 levels were observed. TSH and free T4 is clinically important thyroid hormones since a number of disease states have been linked to alterations in these hormones. Serum free T4 levels within the normal ranges in the presence of abnormal serum TSH levels in everolimus treated mice may suggest subclinical thyroid disease which may have repercussions on the cardiovascular system, as well as on other organs and systems. Our study has revealed the histological damage on thyroid gland induced by subacute everolimus administration, this effect was irrespective of dosing time. However, based on the body weight changes and clinical signs upon everolimus treatment, tolerability for the drug was best following dosing at ZT13 in both male and females. Yet, effects of everolimus on thyroid functions may deserve further studies regarding their clinical importance and chronotoxicity.

Keywords: circadian rhythm, chronotoxicity, everolimus, thyroid gland, thyroid hormones

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Authors: F. Madani, M. Doudi, L. Rahimzadeh Torabi

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The irregular consumption of current antibiotics contributes to an escalation in antibiotic resistance among urinary pathogens on a global scale. The objective of this research was to investigate the process of biologically synthesized silver nanoparticles through the utilization of Zataria multiflora extract. Additionally, the study aimed to evaluate the efficacy of these synthesized nanoparticles in inhibiting the growth of multi-drug resistant negative bacillus bacteria, which commonly contribute to urinary tract infections. The botanical specimen utilized in the current research investigation was Z. multiflora, and its extract was produced employing the Soxhlet extraction technique. The study examined the green synthesis conditions of silver nanoparticles by considering three key parameters: the quantity of extract used, the concentration of silver nitrate salt, and the temperature. The particle dimensions were ascertained using the Zetasizer technique. In order to identify synthesized Silver nanoparticles TEM, XRD, and FTIR methods were used. For evaluating the antibacterial effects of nanoparticles synthesized through a biological method, different concentrations of silver nanoparticles were studied on 140 cases of Multiple drug resistance (MDR) bacteria strains Escherichia coli, Klebsiella pneumoniae, Enterobacter aerogenes, Proteus vulgaris,Citrobacter freundii, Acinetobacter bumanii and Pseudomonas aeruginosa, (each genus of bacteria, 20 samples), which all were MDR and cause urinary tract infections, for identification of bacteria were used of PCR test and laboratory methods (Agar well diffusion and Microdilution methods) to assess their sensitivity to Nanoparticles. The data were subjected to analysis using the statistical software SPSS, specifically employing nonparametric Kruskal-Wallis and Mann-Whitney tests. This study yielded noteworthy findings regarding the impacts of varying concentrations of silver nitrate, different quantities of Z. multiflora extract, and levels of temperature on nanoparticles. Specifically, it was observed that an increase in the concentration of silver nitrate, extract amount, and temperature resulted in a reduction in the size of the nanoparticles synthesized. However, the impact of the aforementioned factors on the index of particle diffusion was found to be statistically non-significant. According to the transmission electron microscopy (TEM) findings, the particles exhibited predominantly spherical morphology, with a diameter spanning from 25 to 50 nanometers. Nanoparticles in the examined sample. Nanocrystals of silver. FTIR method illustrated that the spectrums of Z. multiflora and synthesized nanoparticles had clear peaks in the ranges of 1500-2000, and 3500 - 4000. The obtained results of antibacterial effects of different concentrations of silver nanoparticles on according to agar well diffusion and microdilution method, biologically synthesized nanoparticles showed 1000 mg /ml highest and lowest mean inhibition zone diameter in E. coli, A. bumanii 23 and 15mm, respectively. MIC was observed for all of bacteria 125 mg/ml and for A. bumanii 250 mg/ml. Comparing the growth inhibitory effect of chemically synthesized the results obtained from the experiment indicated that both nanoparticles and biologically synthesized nanoparticles exhibit a notable growth inhibition effect. Specifically, the chemical method of synthesizing nanoparticles demonstrated the highest level of growth inhibition at a concentration of 62.5 mg/mL The present study demonstrated an inhibitory effect on bacterial growth, facilitating the causative factors of urine infection and multidrug resistance (MDR).

Keywords: multiple drug resistance, negative bacillus bacteria, urine infection, Zataria multiflora

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Authors: Nkechi V. Enwuru, Bullum Nkeki, Elizabeth A. Adekoya, Olumide A. Adebesin, Rebecca F. Peters, Victoria A. Aikhomu, Mendie E. U.

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Background: Probiotics are live microbial feed supplement beneficial for host. Probiotics and their postbiotic products have been used to prevent or treat various health conditions. However, the products cell viability is often low due to harsh conditions subjected during processing, handling, storage, and gastrointestinal transit. These strongly influence probiotics’ benefits; thus, viability is essential for probiotics to produce health benefits for the host. Microencapsulation is a promising technique with considerable effects on probiotic survival. The study is aimed to formulate a microencapsulated probiotic and evaluate its viability, antimicrobial efficacy, and cytotoxic activity of its postbiotic on the MCF-7 breast cancer cell line. Method: Human and animal raw milk were sampled for lactic acid bacteria. The isolated bacteria were identified using conventional and VITEK 2 systems. The identified lactic acid bacterium was encapsulated using spray-dried and extrusion methods. The free, encapsulated, and chitosan-coated encapsulated probiotics were tested for viability in simulated-gastric intestinal (SGI) fluid and different storage conditions at refrigerated (4oC) and room (25oC) temperatures. The disintegration time and weight uniformity of the spray-dried hard gelatin capsules were tested. The antimicrobial property of free and encapsulated probiotics was tested against enteric pathogenic isolates from antiretroviral therapy (ART) treated HIV-positive patients. The postbiotic of the free cells was extracted, and its cytotoxic effect on the MCF-7 breast cancer cell line was tested through an MTT assay. Result: The Lactobacillus plantarum was isolated from animal raw milk. Zero-size hard gelatin L. plantarum capsules with granules within a size range of 0.71–1.00 mm diameter was formulated. The disintegration time ranges from 2.14±0.045 to 2.91±0.293 minutes, while the average weight is 502.1mg. Simulated gastric solution significantly affected viability of both free and microcapsules. However, the encapsulated cells were more protected and viable due to impermeability in the microcapsules. Furthermore, the viability of free cells stored at 4oC and 25oC were less than 4 log CFU/g and 6 log CFU/g respectively after 12 weeks. However, the microcapsules stored at 4oC achieved the highest viability among the free and microcapsules stored at 25oC and the free cells stored at 4oC. Encapsulated cells were released in the simulated gastric fluid, viable and effective against the enteric pathogens tested. However, chitosan-coated calcium alginate encapsulated probiotics significantly inhibited Shigella flexneri, Candida albicans, and Escherichia coli. The Postbiotic Metabolites (PM) of L. plantarum produced a cytotoxic effect on the MCF-7 breast cancer cell line. The postbiotic showed significant cytotoxic activity similar to 5FU, a standard antineoplastic agent. The inhibition concentration of 50% growth (IC50) of postbiotic metabolite K3 is low and consistent with the IC50 of the positive control (Cisplatin). Conclusions: Lactobacillus plantarum postbiotic exhibited a cytotoxic effect on the MCF-7 breast cancer cell line and could be used as combined adjuvant therapy in breast cancer management. The microencapsulation technique protects the probiotics, improving their viability and delivery to the gastrointestinal tract. Chitosan enhances antibacterial efficacy; thus, chitosan-coated microencapsulated L. plantarum probiotics could be more effective and used as a combined therapy in HIV management of opportunistic enteric infection.

Keywords: probiotics, encapsulation, gastrointestinal conditions, antimicrobial effect, postbiotic, cytotoxicity effect

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Authors: Denis V. Axenov-Gribanov, Irina V. Voytsekhovskaya, Evgenii S. Protasov, Maxim A. Timofeyev

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Isolated ecosystems existing under specific environmental conditions have been shown to be promising sources of new strains of actinobacteria. The taiga forest of Baikal Siberia has not been well studied, and its actinobacterial population remains uncharacterized. The proximity between the huge water mass of Lake Baikal and high mountain ranges influences the structure and diversity of the plant world in Siberia. Here, we report the isolation of eighteen actinobacterial strains from male cones of Pinus sylvestris trees growing on the shore of the ancient Lake Baikal in Siberia. The actinobacterial strains were isolated on solid nutrient MS media and Czapek agar supplemented with cycloheximide and phosphomycin. Identification of actinobacteria was carried out by 16S rRNA gene sequencing and further analysis of the evolutionary history. Four different liquid and solid media (NL19, DNPM, SG and ISP) were tested for metabolite production. The metabolite extracts produced by the isolated strains were tested for antibacterial and antifungal activities. Also, antiradical activity of crude extracts was carried out. Strain Streptomyces sp. IB 2014 I 74-3 that active against Gram-negative bacteria was selected for dereplication analysis with using the high-yield liquid chromatography with mass-spectrometry. Mass detection was performed in both positive and negative modes, with the detection range set to 160–2500 m/z. Data were collected and analyzed using Bruker Compass Data Analysis software, version 4.1. Dereplication was performed using the Dictionary of Natural Products (DNP) database version 6.1 with the following search parameters: accurate molecular mass, absorption spectra and source of compound isolation. Thus, in addition to more common representative strains of Streptomyces, several species belonging to the genera Rhodococcus, Amycolatopsis, and Micromonospora were isolated. Several of the selected strains were deposited in the Russian Collection of Agricultural Microorganisms (RCAM), St. Petersburg, Russia. All isolated strains exhibited antibacterial and antifungal activities. We identified several strains that inhibited the growth of the pathogen Candida albicans but did not hinder the growth of Saccharomyces cerevisiae. Several isolates were active against Gram-positive and Gram-negative bacteria. Moreover, extracts of several strains demonstrated high antioxidant activity. The high proportion of biologically active strains producing antibacterial and specific antifungal compounds may reflect their role in protecting pollen against phytopathogens. Dereplication of the secondary metabolites of the strain Streptomyces sp. IB 2014 I 74-3 was resulted in the fact that a total of 59 major compounds were detected in the culture liquid extract of strain cultivated in ISP medium. Eight compounds were preliminarily identified based on characteristics described in the Dictionary of Natural Products database, using the search parameters Streptomyces sp. IB 2014 I 74-3 was found to produce saframycin A, Y3 and S; 2-amino-3-oxo-3H-phenoxazine-1,8-dicarboxylic acid; galtamycinone; platencin A4-13R and A4-4S; ganefromycin d1; the antibiotic SS 8201B; and streptothricin D, 40-decarbamoyl, 60-carbamoyl. Moreover, forty-nine of the 59 compounds detected in the extract examined in the present study did not result in any positive hits when searching within the DNP database and could not be identified based on available mass-spec data. Thus, these compounds might represent new findings.

Keywords: actinobacteria, Baikal Lake, biodiversity, male cones, Pinus sylvestris

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Authors: Rowan Moore, Kai Scheffler, Eugen Damoc, Jennifer Sutton, Aaron Bailey, Stephane Houel, Simon Cubbon, Jonathan Josephs

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Purpose: MS analysis of monoclonal antibodies (mAbs) at the protein and peptide levels is critical during development and production of biopharmaceuticals. The compositions of current generation therapeutic proteins are often complex due to various modifications which may affect efficacy. Intact proteins analyzed by MS are detected in higher charge states that also provide more complexity in mass spectra. Protein analysis in native or native-like conditions with zero or minimal organic solvent and neutral or weakly acidic pH decreases charge state value resulting in mAb detection at higher m/z ranges with more spatial resolution. Methods: Three commercially available mAbs were used for all experiments. Intact proteins were desalted online using size exclusion chromatography (SEC) or reversed phase chromatography coupled on-line with a mass spectrometer. For streamlined use of the LC- MS platform we used a single SEC column and alternately selected specific mobile phases to perform separations in either denaturing or native-like conditions: buffer A (20 % ACN, 0.1 % FA) with Buffer B (100 mM ammonium acetate). For peptide analysis mAbs were proteolytically digested with and without prior reduction and alkylation. The mass spectrometer used for all experiments was a commercially available Thermo Scientific™ hybrid Quadrupole-Orbitrap™ mass spectrometer, equipped with the new BioPharma option which includes a new High Mass Range (HMR) mode that allows for improved high mass transmission and mass detection up to 8000 m/z. Results: We have analyzed the profiles of three mAbs under reducing and native conditions by direct infusion with offline desalting and with on-line desalting via size exclusion and reversed phase type columns. The presence of high salt under denaturing conditions was found to influence the observed charge state envelope and impact mass accuracy after spectral deconvolution. The significantly lower charge states observed under native conditions improves the spatial resolution of protein signals and has significant benefits for the analysis of antibody mixtures, e.g. lysine variants, degradants or sequence variants. This type of analysis requires the detection of masses beyond the standard mass range ranging up to 6000 m/z requiring the extended capabilities available in the new HMR mode. We have compared each antibody sample that was analyzed individually with mixtures in various relative concentrations. For this type of analysis, we observed that apparent native structures persist and ESI is benefited by the addition of low amounts of acetonitrile and formic acid in combination with the ammonium acetate-buffered mobile phase. For analyses on the peptide level we analyzed reduced/alkylated, and non-reduced proteolytic digests of the individual antibodies separated via reversed phase chromatography aiming to retrieve as much information as possible regarding sequence coverage, disulfide bridges, post-translational modifications such as various glycans, sequence variants, and their relative quantification. All data acquired were submitted to a single software package for analysis aiming to obtain a complete picture of the molecules analyzed. Here we demonstrate the capabilities of the mass spectrometer to fully characterize homogeneous and heterogeneous therapeutic proteins on one single platform. Conclusion: Full characterization of heterogeneous intact protein mixtures by improved mass separation on a quadrupole-Orbitrap™ mass spectrometer with extended capabilities has been demonstrated.

Keywords: disulfide bond analysis, intact analysis, native analysis, mass spectrometry, monoclonal antibodies, peptide mapping, post-translational modifications, sequence variants, size exclusion chromatography, therapeutic protein analysis, UHPLC

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