Search results for: Izabela Zielinska

Authors: Joanna E. Owczarek, Izabela Zielinska

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The incidence of temporomandibular disorders is 50% up to 80%. Kinesio taping (KT) is treatment method for musculoskeletal disorders. The aim of our study was to assess the impact of KT on masseter muscles’ tone evaluated by electromyography. 30 adults (aged 22±2.1) were examined. The tone of masseters before and after 4 days KT application on sternocleidomastoideus muscle was measured during resting mandibular position and clenching. Noraxon DTS device was used. Masseter muscles’ tone during clenching after KT application was relevently lower in comparison to its tone before the KT.

Keywords: electromyography, kinesio taping, masseter muscle, TMD

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Authors: Izabela Zielinska

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Kinesio Taping is classified as physiotherapy method supporting rehabilitation and modulating some physiological processes. It is commonly used in sports medicine and orthopedics. This sensory method has influence on muscle function, pain sensation, intensifies lymphatic system as well as improves microcirculation. The aim of this study was to assess the effect of Kinesio Taping in patients with ongoing treatment of peripheral artery disease (PAD). The study group comprised 60 patients (stadium II B at Fontain's scale). All patients were divided into two groups (30 person/each), where 12 weeks long treadmill training was administrated. In the second group, the Kinesio Taping was applied to support the function of the gastrocnemius muscle. The measurements of distance and time until claudication pain, blood flow of arteries in lower limbs and ankle brachial index were taken under evaluation. Examination performed after Kinesio Taping therapy showed statistically significant increase in gait parameters and muscle strength in patients with intermittent claudication. The Kinesio Taping method has clinically significant effects on enhancement of pain-free distance and time until claudication pain in patients with peripheral artery disease. Kinesio Taping application can be used to support non-invasive treatment in patients with intermittent claudication. Kinesio Taping can be employed as an alternative way of therapy for patients with orthopedic or cardiac contraindications to be treated with treadmill training.

Keywords: intermittent claudication, kinesiotaping, peripheral artery disease, treadmill training

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Authors: Krzysztof Plesiewicz, Izabela Plesiewicz, Krzysztof Chiżyński, Marzenna Zielińska

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Background: Obesity is an independent risk factor for cardiovascular diseases. There are several anthropometric parameters proposed to estimate the level of obesity, but until now there is no agreement which one is the best predictor of cardiometabolic risk. Scientists defined metabolically obese normal weight, who suffer from metabolic abnormalities, the same as obese individuals, and defined this syndrome as normal weight obesity (NWO). Aim of the study: The aim of our study was to determine the occurrence of overweight and obesity in a cohort of young, adult women, using standard and complementary methods of obesity assessment and to indicate those, who are at risk of obesity. The second aim of our study was to test additional methods of obesity assessment and proof that body mass index using alone is not sufficient parameter of obesity assessment. Materials and methods: 384 young women, aged 18-32, were enrolled into the study. Standard anthropometric parameters (waist to hips ratio (WTH), waist to height ratio (WTHR)) and two other methods of body fat percentage measurement (BFPM) were used in the study: electrical bioimpendance analysis (BIA) and skinfold measurement test by digital fat body mass clipper (SFM). Results: In the study group 5% and 7% of participants had waist to hips ratio and accordingly waist to height ratio values connected with visceral obesity. According to BMI 14% participants were overweight and obese. Using additional methods of body fat assessment, there were 54% and 43% of obese for BIA and SMF method. In the group of participants with normal BMI and underweight (not overweight, n =340) there were individuals with the level of BFPM above the upper limit, for the BIA 49% (n =164) and for the SFM 36 % (n=125). Statistical analysis revealed strong correlation between BIA and SFM methods. Conclusion: BMI using alone is not a sufficient parameter of obesity assessment. High percentage of young women with normal BMI values seem to be normal weight obese.

Keywords: electrical bioimpedance, normal weight obesity, skin-fold measurement test, women

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Authors: Izabela L. Ciesielska-Wrόbel

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This paper presents finite element models of the fingertip skin which have been created to simulate the contact of textile objects with the skin to gain a better understanding of the perception of textiles through the skin, so-called Hand of Textiles (HoT). Many objective and subjective techniques have been developed to analyze HoT, however none of them provide exact overall information concerning the sensation of textiles through the skin. As the human skin is a complex heterogeneous hyperelastic body composed of many particles, some simplifications had to be made at the stage of building the models. The same concerns models of woven structures, however their utilitarian value was maintained. The models reflect only friction between skin and woven textiles, deformation of the skin and fabrics when “touching” textiles and heat transfer from the surface of the skin into direction of textiles.

Keywords: fingertip skin models, finite element models, modelling of textiles, sensation of textiles through the skin

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Authors: Izabela Nowicka, Jacek Dworzecki

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The paper will be presented to selected legal and organizational solutions for the prevention and combating of terrorism by the police in Poland. Development will include information on the organization and functioning of the police anti-terrorist sub-units, whose officers are on the front line of the fight against terrorism. They will be presented to the conditions and cases of use of firearms by police officers in the course of special operations aimed against organizations and terrorist groups, and the perpetrators of criminal acts of terrorism as well as the legal foundation for the Polish police to take immediate counterterrorism operations. Article will be prepared in the context of an international research project entitled. Understand the Dimensions of Organised Crime and Terrorist Networks for Developing Effective and Efficient Security Solutions for First-line-practitioners and Professionals [Project: H2020-FCT-2015, No: 700688].

Keywords: the fight against terrorism, police, Poland, takedown

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Authors: Iman Taraghi, Sandra Paszkiewicz, Daria Pawlikowska, Anna Szymczyk, Izabela Irska, Rafal Stanik, Amelia Linares, Tiberio A. Ezquerra, Elżbieta Piesowicz

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Currently, the massive use of thermoplastic materials in industrial applications causes huge amounts of polymer waste. The poly (ethylene glycol-co-1,4-cyclohexanedimethanol terephthalate) (PET-G) has been widely used in food packaging and polymer foils. In this research, the PET-G foils have been recycled and reused as a matrix to combine with different types of nanofillers such as carbon nanotubes, graphene nanoplatelets, and nanosized carbon black. The mechanical and electrical properties, as well as thermal stability and thermal conductivity of the PET-G, improved along with the addition of the aforementioned nanofillers and hybrid system of them.

Keywords: polymer hybrid nanocomposites, carbon nanofillers, recycling, physical performance

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Authors: Maciej Major, Izabela Major

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The aim of this paper is a numerical analysis of three-layered block design to walls construction subjected to the dynamic load. The block consists of the layers: concrete with rubber pads in shape of crosses, space filled with air and concrete with I-shape rubber pads. The main purpose of rubber inserts embedded during the production process is additional protection against the transversal dynamic load. For the analysis, as rubber, the Zahorski hyperelastic incompressible material model was assumed. A concentrated force as dynamic load applied to the external block surface was investigated. The results for the considered block observed as the stress distribution plot were compared to the results obtained for the solid concrete block. In order to estimate the percentage damping of proposed composite, rubber-concrete block in relation to the solid block the numerical analysis with the use of finite element method based on ADINA software was performed.

Keywords: dynamics, composite, rubber, Zahorski

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Authors: Aneta Nitsch-Osuch, Damian Okrucinski, Magdalena Dawgialło, Izabela Gołębiak, Ernest Kuchar

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The Hospital Antibiotic Policy (HAP) should be implemented to rationalize the antibiotic use and to decrease the risk of spreading of spreading of resistant bacteria. The aim of our study was to describe the antibiotic consumption patterns at the single oncosurgery ward before and after implementation of the HAP. We conducted a retrospective analysis of the antibiotic use at the Oncosurgery Ward in Warsaw (Poland) in years 2011-2016. Calculations were based on daily defined doses (DDDs), DDDs/100 hospitalizations and DDDs/100 person-days, drug utilization rates (DU 90% and DU 100%) were also analysed. After implementation of the HAP, the total antibiotic consumption increased (365.35 DDD in 2011 vs. 1359,22 DDD in 2016). The significant change was observed in antibiotic consumption patterns: the use of amoxicillin clavulanate and carbapenems or glycopeptides decreased significantly (p < 0,05), while the use of ciprofloxacin and aminoglycosides increased (p < 0,05). The DU100% rate varied from 6 in 2011 to 12 in 2016; while DU 90% rate varied from 2 in 2011 to 3-5 in 2013-2016. Although the implementation of the HAP did not result in the decreased total antibiotic consumption, it provided favorable changes in the antibiotic consumption patterns.

Keywords: antibiotics, hospital, policy, stewardship

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Authors: M. Zielinska, B. Daniels, J. Gabel, A. Paletko

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A common nickel based superalloy Inconel625 was brazed with Ni-base braze filler material (AMS4777) containing melting-point-depressants such as B and Si. Different braze gaps, brazing times and forms of braze filler material were tested. It was determined that the melting point depressants B and Si tend to form hard and brittle phases in the joint during the braze cycle. Brittle phases significantly reduce mechanical properties (e. g. tensile strength) of the joint. Therefore, it is important to define optimal process parameters to achieve high strength joints, free of brittle phases. High ultimate tensile strength (UTS) values can be obtained if the joint area is free of brittle phases, which is equivalent to a complete isothermal solidification of the joint. Isothermal solidification takes place only if the concentration of the melting point depressant in the braze filler material of the joint is continuously reduced by diffusion into the base material. For a given brazing temperature, long brazing times and small braze filler material volumes (small braze gaps) are beneficial for isothermal solidification. On the base of the obtained results it can be stated that the form of the braze filler material has an additional influence on the joint quality. Better properties can be achieved by the use of braze-filler-material in form of foil instead of braze-filler-material in form of paste due to a reduced amount of voids and a more homogeneous braze-filler-material-composition in the braze-gap by using foil.

Keywords: diffusion brazing, microstructure, superalloy, tensile strength

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Authors: Dawid Stawski, Silviya Halacheva, Dorota Zielińska

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The surface properties of many materials can be readily and predictably modified by the controlled deposition of thin layers containing appropriate functional groups and this research area is now a subject of widespread interest. The layer-by-layer (lbl) method involves depositing oppositely charged layers of polyelectrolytes onto the substrate material which are stabilized due to strong electrostatic forces between adjacent layers. This type of modification affords products that combine the properties of the original material with the superficial parameters of the new external layers. Through an appropriate selection of the deposited layers, the surface properties can be precisely controlled and readily adjusted in order to meet the requirements of the intended application. In the presented paper a variety of anionic (poly(acrylic acid)) and cationic (linear poly(ethylene imine), polymers were successfully deposited onto the polypropylene nonwoven using the lbl technique. The chemical structure of the surface before and after modification was confirmed by reflectance FTIR spectroscopy, volumetric analysis and selective dyeing tests. As a direct result of this work, new materials with greatly improved properties have been produced. For example, following a modification process significant changes in the electrostatic activity of a range of novel nanocomposite materials were observed. The deposition of polyelectrolyte nanolayers was found to strongly accelerate the loss of electrostatically generated charges and to increase considerably the thermal resistance properties of the modified fabric (the difference in T50% is over 20°C). From our results, a clear relationship between the type of polyelectrolyte layer deposited onto the flat fabric surface and the properties of the modified fabric was identified.

Keywords: layer-by-layer technique, polypropylene nonwoven, surface modification, surface properties

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Authors: Dorota Zielińska, Monika Trząskowska, Anna Łepecka, Katarzyna Neffe-Skocińska, Beata Bilska, Marzena Tomaszewska, Danuta Kołożyn-Krajewska

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Minimum durability date (MDD) labeled food is known to have a long shelf life. A properly stored or transported food retains its physical, chemical, microbiological, and sensory properties up to MDD. The aim of the study was to assess the sensory quality and microbiological safety of selected thermally processed products,i.e., mayonnaise, jam, and canned tuna within and after MDD. The scope of the study was to determine the markers of microbiological quality, i.e., the total viable count (TVC), the Enterobacteriaceae count and the total yeast and mold (TYMC) count on the last day of MDD and after 1 and 3 months of storage, after the MDD expired. In addition, the presence of Salmonella and Listeria monocytogenes was examined on the last day of MDD. The sensory quality of products was assessed by quantitative descriptive analysis (QDA), the intensity of differentiators (quality features), and overall quality were defined and determined. It was found that during three months storage of tested food products, after the MDD expired, the microbiological quality slightly decreased, however, regardless of the tested sample, TVC was at the level of <3 log cfu/g, similarly, the Enterobacretiaceae, what indicates the good microbiological quality of the tested foods. The TYMC increased during storage but did not exceed 2 logs cfu/g of product. Salmonella and Listeria monocytogenes were not found in any of the tested food samples. The sensory quality of mayonnaise negatively changed during storage. After three months from the expiry of MDD, a decrease in the "fat" and "egg" taste and aroma intensity, as well as the "density" were found. The "sour" taste intensity of blueberry jam after three months of storage was slightly higher, compared to the jam tested on the last day of MDD, without affecting the overall quality. In the case of tuna samples, an increase in the "fishy" taste and aroma intensity was observed during storage, and the overall quality did not change. Tested thermally processed products (mayonnaise, jam, and canned tuna) were characterized by good microbiological and sensory quality on the last day of MDD, as well as after three months of storage under conditions recommended by the producer. These findings indicate the possibility of reducing food losses by extending or completely abolishing the MDD of selected thermal processed food products.

Keywords: food wastes, food quality and safety, mayonnaise, jam, tuna

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Authors: André C. Silva, Izabela L. A. Moraes, Elenice M. S. Silva, Carlos M. Silva Filho

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In Brazil, most soils are acidic and low in essential nutrients required for the growth and development of plants, making fertilizers essential for agriculture. As the biggest producer of soy in the world and a major producer of coffee, sugar cane and citrus fruits, Brazil is a large consumer of phosphate. Brazilian’s phosphate ores are predominantly from igneous rocks showing a complex mineralogy, associated with carbonites and oxides, typically iron, silicon and barium. The adopted industrial concentration circuit for this type of ore is a mix between magnetic separation (both low and high field) to remove the magnetic fraction and a froth flotation circuit composed by a reverse flotation of apatite (barite’s flotation) followed by direct flotation circuit (rougher, cleaner and scavenger circuit). Since the 70’s fatty acids obtained from vegetable oils are widely used as lower-cost collectors in apatite froth flotation. This is a very effective approach to the apatite family of minerals, being that this type of collector is both selective and efficient (high recovery). This paper presents Jatropha curcas L. oil (JCO) as a renewable and sustainable source of fatty acids with high selectivity in froth flotation of apatite. JCO is considerably rich in fatty acids such as linoleic, oleic and palmitic acid. The experimental campaign involved 216 tests using a modified Hallimond tube and two different minerals (apatite and quartz). In order to be used as a collector, the oil was saponified. The results found were compared with the synthetic collector, Fotigam 5806 produced by Clariant, which is composed mainly by soy oil. JCO showed the highest selectivity for apatite flotation with cold saponification at pH 8 and concentration of 2.5 mg/L. In this case, the mineral recovery was around 95%.

Keywords: froth flotation, jatropha curcas, microflotation, selectivity

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Authors: Aneta Nitsch-Osuch, Sylwia Dyk, Izabela Gołebiak

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Background. Since 2014 the pertussis vaccine is recommended to Polish health care workers who have close contacts with infants. Although this recommendation is implemented into the National Immunization Programme, its realization has remained unknown. The Purpose: The aim of the study, conducted at the department of Social Medicine and Public Health (Medical University of Warsaw, Poland), was to describe a perception, knowledge and coverage rates regarding pertussis vaccination among nursing staff. According to the authors' knowledge, it was the first study related to this topic in our country. Material and Methods: A total number of 543 nurses who work at pediatric or neonatal wards was included into the study (501 women and 42 men), average age was 47 years. All nurses were asked to fulfill the anonymous survey, previously validated. Results: 1. Coverage rates: The analysis of results revealed that only 4% of responders reported they were vaccinated with Tdpa within past 10 years, while 8% declared they would plan the vaccine in the future. 35% of responders would consider the Tdpa vaccine whether there is some kind of the reimbursement. 2. Perception and knowledge of the disease and vaccination: The majority (82%) of nurses did not recognize pertussis as a re-emerging infectious disease. 54% of them believed that obligatory vaccinations in the childhood protect against the disease and the protection is a life-long one. Only 15% of nurses considered pertussis as a possible nosocomial infection. The current epidemiology of the disease was known to 6% of responders, while 24% of them were familiar with pertussis vaccination schedules for infants, children and adolescents, but only 9% of responders knew that adults older than 19 years are recommended to be vaccinated with Tdpa every 10 years. Many nurses (82%) would expect more educational activities related to pertussis and methods of its prophylaxis. Conclusions: The pertussis vaccine coverage rate among Polish nurses is extremely low. This is a result of not enough knowledge about the disease and its prevention. Educational activities addressed to health care workers and reimbursement of the pertussis vaccine are required to improve awareness and increase of vaccine coverage rates in the future.

Keywords: coverage, nurse, pertussis, vaccine

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Authors: Katarzyna Neffe-Skocinska, Marzena Tomaszewska, Beata Bilska, Dorota Zielinska, Monika Trzaskowska, Anna Lepecka, Danuta Kolozyn-Krajewska

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The aim of the study was to assess Polish consumer behavior towards food waste, including knowledge of information on food labels. The survey was carried out using the CAPI (computer assisted personal interview) method, which involves interviewing the respondent using mobile devices. The research group was a representative sample for Poland due to demographic variables: gender, age, place of residence. A total of 1.115 respondents participated in the study (51.1% were women and 48.9% were men). The questionnaire included questions on five thematic aspects: 1. General knowledge and sources of information on the phenomenon of food waste; 2. Consumption of food after the date of minimum durability; 3. The meanings of the phrase 'best before ...'; 4. Indication of the difference between the meaning of the words 'best before ...' and 'use by'; 5. Indications products marked with the phrase 'best before ...'. It was found that every second surveyed Pole met with the topic of food waste (54.8%). Among the respondents, the most popular source of information related to the research topic was television (89.4%), radio (26%) and the Internet (24%). Over a third of respondents declared that they consume food after the date of minimum durability. Only every tenth (9.8%) respondent does not pay attention to the expiry date and type of consumed products (durable and perishable products). Correctly 39.8% of respondents answered the question: How do you understand the phrase 'best before ...'? In the opinion of 42.8% of respondents, the statements 'best before ...' and 'use by' mean the same thing, while 36% of them think differently. In addition, more than one-fifth of respondents could not respond to the questions. In the case of products of the indication information 'best before ...', more than 40% of the respondents chosen perishable products, e.g., yoghurts and durable, e.g., groats. A slightly lower percentage of indications was recorded for flour (35.1%), sausage (32.8%), canned corn (31.8%), and eggs (25.0%). Based on the assessment of the behavior of Polish consumers towards the phenomenon of food waste, it can be concluded that respondents have elementary knowledge of the study subject. Noteworthy is the good conduct of most respondents in terms of compliance with shelf life and dates of minimum durability of food products. The publication was financed on the basis of an agreement with the National Center for Research and Development No. Gospostrateg 1/385753/1/NCBR/2018 for the implementation and financing of the project under the strategic research and development program social and economic development of Poland in the conditions of globalizing markets – GOSPOSTRATEG - acronym PROM.

Keywords: food waste, shelf life, dates of durability, consumer knowledge and behavior

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Authors: Katarzyna Kwiatkowska, Izabela Walasik, Katarzyna Kosińska-Kaczyńska, Olga Płaza, Kinga Żebrowska

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Introduction Adequate physical activity of a pregnant woman has been proven to decrease the risk of pregnancy complications. The knowledge of women regarding physical exercise in pregnancy is a part of conscious motherhood, while a lack of it may lead to not taking up any form of physical activity during pregnancy. Aim: The aim of the study was to assess the knowledge and experience of women regarding physical activity during their latest pregnancy. Material and methodology: An anonymous questionnaire, consisting of 57 questions, was completed electronically in 2018 by women who gave birth at least once. The respondents were qualified as 'physically active during pregnancy' if they performed physical exercises such as regular walks, marching, jogging, working out at a gym, swimming, yoga, pilates, fitness, exercise-ball workouts or home gymnastics. Results: The study group consisted of 9345 women. 52% of them performed exercises during pregnancy. The main reasons for the lack of physical activity were: lack of interest in physical activity (45%), lack of energy (40%), lack of knowledge regarding proper exercise during pregnancy (34%), lack of time (27%) and medical contraindications (25%). Non-active respondents suffered from gestational hypertension (6,7% vs 9,2%; p<00,1) and gave birth prematurely (11% vs 15%; p < 001) to newborns with a lower birth weight significantly more often ( < 2500g vs > 2500g; p < 0,001). Physically active women reported suffering from pregnancy-related ailments such as fatigue, back pain or constipation significantly less often. 22% of all respondents were unable to identify reliable sources of information regarding exercise during pregnancy. A majority of the exercising women used the Internet to obtain gain information on physical activity during pregnancy (69,1%). 4% of women thought that exercising during pregnancy is forbidden, while 20% thought it is not allowed in the 3rd trimester. Physically active women had vaginal delivery more often (61% vs 55%; p < 0,05). Episiotomy was performed most often on non-active primiparous respondents (77,5% vs 71% active primiparous, p < 0,001). 13% of women felt discriminated due to their physical activity during pregnancy. 22% of respondents’ physical activity was not accepted by their environment. 39,1% of the women were told by others to stop physical exercise because it was bad for the baby’s health. Conclusion: The knowledge of Polish women regarding proper physical activity during pregnancy is insufficient, which may influence a lack of will to initiate such activity among pregnant women. Physical activity of a pregnant woman may have an impact on the course of pregnancy and birth.

Keywords: childbirth, discrimination, physical activity, pregnancy

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Authors: Marcin Debowski, Marcin Zielinski, Magdalena Zielinska, Paulina Rusanowska

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The problem with digestate management is one of the most important factors influencing on the development and operation of biogas plant. Turbidity and bacterial contamination negatively affect the growth of algae, which can limit the use of the effluent in the production of algae biomass on a large scale. These problems can be overcome by cultivating of algae species resistant to environmental factors, such as Chlorella sp., Scenedesmus sp., or reducing load of organic compounds to prevent bacterial contamination. The effluent requires dilution and/or purification. One of the methods of effluent treatment is the use of a membrane technology such as microfiltration (MF), ultrafiltration (UF), nanofiltration (NF) and reverse osmosis (RO), depending on the membrane pore size and the cut off point. Membranes are a physical barrier to solids and particles larger than the size of the pores. MF membranes have the largest pores and are used to remove turbidity, suspensions, bacteria and some viruses. UF membranes remove also color, odor and organic compounds with high molecular weight. In treatment of wastewater or other waste streams, MF and UF can provide a sufficient degree of purification. NF membranes are used to remove natural organic matter from waters, water disinfection products and sulfates. RO membranes are applied to remove monovalent ions such as Na⁺ or K⁺. The effluent was used in UF for medium to cultivation of two microalgae: Chlorella sp. and Phaeodactylum tricornutum. Growth rates of Chlorella sp. and P. tricornutum were similar: 0.216 d⁻¹ and 0.200 d⁻¹ (Chlorella sp.); 0.128 d⁻¹ and 0.126 d⁻¹ (P. tricornutum), on synthetic medium and permeate from UF, respectively. The final biomass composition was also similar, regardless of the medium. Removal of nitrogen was 92% and 71% by Chlorella sp. and P. tricornutum, respectively. The fermentation effluents after UF and dilution were also used for cultivation of algae Scenedesmus sp. that is resistant to environmental conditions. The authors recommended the development of biorafinery based on the production of algae for the biogas production. There are examples of using a multi-stage membrane system to purify the liquid fraction from digestate. After the initial UF, RO is used to remove ammonium nitrogen and COD. To obtain a permeate with a concentration of ammonium nitrogen allowing to discharge it into the environment, it was necessary to apply three-stage RO. The composition of the permeate after two-stage RO was: COD 50–60 mg/dm³, dry solids 0 mg/dm³, ammonium nitrogen 300–320 mg/dm³, total nitrogen 320–340 mg/dm³, total phosphorus 53 mg/dm³. However compostion of permeate after three-stage RO was: COD < 5 mg/dm³, dry solids 0 mg/dm³, ammonium nitrogen 0 mg/dm³, total nitrogen 3.5 mg/dm³, total phosphorus < 0,05 mg/dm³. Last stage of RO might be replaced by ion exchange process. The negative aspect of membrane filtration systems is the fact that the permeate is about 50% of the introduced volume, the remainder is the retentate. The management of a retentate might involve recirculation to a biogas plant.

Keywords: digestate, membrane filtration, microalgae cultivation, Chlorella sp.

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Authors: M. Zielinski, M. Debowski, P. Rusanowska, A. Glowacka-Gil, M. Zielinska, A. Cydzik-Kwiatkowska, J. Kazimierowicz

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Technologies for the micro-biogas plant with heating and mixing systems are presented as a part of the Research Coordination for a Low-Cost Biomethane Production at Small and Medium Scale Applications (Record Biomap). The main objective of the Record Biomap project is to build a network of operators and scientific institutions interested in cooperation and the development of promising technologies in the sector of small and medium-sized biogas plants. The activities carried out in the project will bridge the gap between research and market and reduce the time of implementation of new, efficient technological and technical solutions. Reactor with simultaneously mixing and heating system is a concrete tank with a rectangular cross-section. In the reactor, heating is integrated with the mixing of substrate and anaerobic sludge. This reactor is solution dedicated for substrates with high solids content, which cannot be introduced to the reactor with pumps, even with positive displacement pumps. Substrates are poured to the reactor and then with a screw pump, they are mixed with anaerobic sludge. The pumped sludge, flowing through the screw pump, is simultaneously heated by a heat exchanger. The level of the fermentation sludge inside the reactor chamber is above the bottom edge of the cover. Cover of the reactor is equipped with the screw pump driver. Inside the reactor, an electric motor is installed that is driving a screw pump. The heated sludge circulates in the digester. The post-fermented sludge is collected using a drain well. The inlet to the drain well is below the level of the sludge in the digester. The biogas is discharged from the reactor by the biogas intake valve located on the cover. The technology is very useful for fermentation of lignocellulosic biomass and substrates with high content of dry mass (organic wastes). The other technology is a reactor for micro-biogas plant with a pressure mixing system. The reactor has a form of plastic or concrete tank with a circular cross-section. The effective mixing of sludge is ensured by profiled at 90° bottom of the tank. Substrates for fermentation are supplied by an inlet well. The inlet well is equipped with a cover that eliminates odour release. The introduction of a new portion of substrates is preceded by pumping of digestate to the disposal well. Optionally, digestate can gravitationally flow to digestate storage tank. The obtained biogas is discharged into the separator. The valve supplies biogas to the blower. The blower presses the biogas from the fermentation chamber in such a way as to facilitate the introduction of a new portion of substrates. Biogas is discharged from the reactor by valve that enables biogas removal but prevents suction from outside the reactor.

Keywords: biogas, digestion, heating system, mixing system

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Authors: Carlos Alberto Rivera-corredor, Angie Dayana Vargas-Ceballos, Edison Gilpavas, Izabela Dobrosz-Gómez, Miguel Ángel Gómez-García

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Hexavalent chromium, Cr (VI) is present in the effluents from different industries such as electroplating, mining, leather tanning, etc. This compound is of great academic and industrial concern because of its toxic and carcinogenic behavior. Its dumping to both environmental and public health for animals and humans causes serious problems in water sources. The amount of Cr (VI) in industrial wastewaters ranges from 0.5 to 270,000 mgL-1. According to the Colombian standard for water quality (NTC-813-2010), the maximum allowed concentration for the Cr (VI) in drinking water is 0.05 mg L-1. To comply with this limit, it is essential that industries treat their effluent to reduce the Cr (VI) to acceptable levels. Numerous methods have been reported for the treatment removing metal ions from aqueous solutions such as: reduction, ion exchange, electrodialysis, etc. Adsorption has become a promising method for the purification of metal ions in water, since its application corresponds with an economic and efficient technology. The absorbent selection and the kinetic and thermodynamic study of the adsorption conditions are key to the development of a suitable adsorption technology. The Ce0.25Zr0.75O2.nH2O presents higher adsorption capacity between a series of hydrated mixed oxides Ce1-xZrxO2 (x = 0, 0.25, 0.5, 0.75, 1). This work presents the kinetic and thermodynamic study of Cr (VI) adsorption on Ce0.25Zr0.75O2.nH2O. Experiments were performed under the following experimental conditions: initial Cr (VI) concentration = 25, 50 and 100 mgL-1, pH = 2, adsorbent charge = 4 gL-1, stirring time = 60 min, temperature=20, 28 and 40 °C. The Cr (VI) concentration was spectrophotometrically estimated by the method of difenilcarbazide with monitoring the absorbance at 540 nm. The Cr (VI) adsorption over hydrated Ce0.25Zr0.75O2.nH2O models was analyzed using pseudo-first and pseudo-second order kinetics. The Langmuir and Freundlich models were used to model the experimental data. The convergence between the experimental values and those predicted by the model, is expressed as a linear regression correlation coefficient (R2) and was employed as the model selection criterion. The adsorption process followed the pseudo-second order kinetic model and obeyed the Langmuir isotherm model. The thermodynamic parameters were calculated as: ΔH°=9.04 kJmol-1,ΔS°=0.03 kJmol-1 K-1, ΔG°=-0.35 kJmol-1 and indicated the endothermic and spontaneous nature of the adsorption process, governed by physisorption interactions.

Keywords: adsorption, hexavalent chromium, kinetics, thermodynamics

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Authors: Monika Trzaskowska, Dorota Zielinska, Anna Lepecka, Katarzyna Neffe-Skocinska, Beata Bilska, Marzena Tomaszewska, Danuta Kolozyn-Krajewska

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Microbiological quality and food safety are important food characteristics. Regulation (EU) No 1169/2011 of the European Parliament and of the Council on the provision of food information to consumers introduces the obligation to provide information on the 'use-by' date or the date of minimum durability (DMD). The second term is the date until which the properly stored or transported foodstuff retains its physical, chemical, microbiological and organoleptic properties. The date should be preceded by 'best before'. It is used for durable products, e.g., pasta. In relation to reducing food losses, the question may be asked whether products with the date of minimum durability currently declared retain quality and safety beyond this. The aim of the study was to assess the sensory quality and microbiological safety of selected cereal products, i.e., pasta and millet after DMD. The scope of the study was to determine the markers of microbiological quality, i.e., the total viable count (TVC), the number of bacteria from the Enterobacteriaceae family and the number of yeast and mold (TYMC) on the last day of DMD and after 1 and 3 months of storage. In addition, the presence of Salmonella and Listeria monocytogenes was examined on the last day of DMD. The sensory quality of products was assessed by quantitative descriptive analysis (QDA), the intensity of 14 differentiators and overall quality were defined and determined. In the tested samples of millet and pasta, no pathogenic bacteria Salmonella and Listeria monocytogenes were found. The value of the distinguishing features of selected quality and microbiological safety indicators on the last DMD day was in the range of about 3-1 log cfu/g. This demonstrates the good microbiological quality of the tested food. Comparing the products, a higher number of microorganisms was found in the samples of millet. After 3 months of storage, TVC decreased in millet, while in pasta, it was found to increase in value. In both products, the number of bacteria from the Enterobacretiaceae family decreased. In contrast, the number of TYMCs increased in samples of millet, and in pasta decreased. The intensity of sensory characteristic in the studied period varied. It remained at a similar level or increased. Millet was found to increase the intensity and flavor of 'cooked porridge' 3 months after DMD. Similarly, in the pasta, the smell and taste of 'cooked pasta' was more intense. To sum up, the researched products on the last day of the minimum durability date were characterized by very good microbiological and sensory quality, which was maintained for 3 months after this date. Based on these results, the date of minimum durability of tested products could be extended. The publication was financed on the basis of an agreement with the National Center for Research and Development No. Gospostrateg 1/385753/1/NCBR/2018 for the implementation and financing of the project under the strategic research and development program 'social and economic development of Poland in the conditions of globalizing markets – GOSPOSTRATEG - acronym PROM'.

Keywords: date of minimum durability, food losses, food quality and safety, millet, pasta

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Authors: Agnieszka Feliczak-Guzik, Paulina Szczyglewska, Izabela Nowak

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A catalyst-assisted oxidation reaction is one of the key reactions exploited by various industries. Their conductivity yields essential compounds and intermediates, such as alcohols, epoxides, aldehydes, ketones, and organic acids. Researchers are devoting more and more attention to developing active and selective materials that find application in many catalytic reactions, such as cyclohexene epoxidation. This reaction yields 1,2-epoxycyclohexane and 1,2-diols as the main products. These compounds are widely used as intermediates in the perfume industry and synthesizing drugs and lubricants. Hence, our research aimed to use hierarchical zeolites modified with transition metal ions, e.g., Nb, V, and Ta, in the epoxidation reaction of cyclohexene using microwaveheating. Hierarchical zeolites are materials with secondary porosity, mainly in the mesoporous range, compared to microporous zeolites. In the course of the research, materials based on two commercial zeolites, with Faujasite (FAU) and Zeolite Socony Mobil-5 (ZSM-5) structures, were synthesized and characterized by various techniques, such as X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM), and low-temperature nitrogen adsorption/desorption isotherms. The materials obtained were then used in a cyclohexene epoxidation reaction, which was carried out as follows: catalyst (0.02 g), cyclohexene (0.1 cm3), acetonitrile (5 cm3) and dihydrogen peroxide (0.085 cm3) were placed in a suitable glass reaction vessel with a magnetic stirrer inside in a microwave reactor. Reactions were carried out at 45° C for 6 h (samples were taken every 1 h). The reaction mixtures were filtered to separate the liquid products from the solid catalyst and then transferred to 1.5 cm3 vials for chromatographic analysis. The test techniques confirmed the acquisition of additional secondary porosity while preserving the structure of the commercial zeolite (XRD and low-temperature nitrogen adsorption/desorption isotherms). The results of the activity of the hierarchical catalyst modified with niobium in the cyclohexene epoxidation reaction indicate that the conversion of cyclohexene, after 6 h of running the process, is about 70%. As the main product of the reaction, 2-cyclohexanediol was obtained (selectivity > 80%). In addition to the mentioned product, adipic acid, cyclohexanol, cyclohex-2-en-1-one, and 1,2-epoxycyclohexane were also obtained. Furthermore, in a blank test, no cyclohexene conversion was obtained after 6 h of reaction. Acknowledgments The work was carried out within the project “Advanced biocomposites for tomorrow’s economy BIOG-NET,” funded by the Foundation for Polish Science from the European Regional Development Fund (POIR.04.04.00-00-1792/18-00.

Keywords: epoxidation, oxidation reactions, hierarchical zeolites, synthesis

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Authors: Marcin Debowski, Marcin Zielinski, Miroslaw Krzemieniewski, Agata Glowacka-Gil, Paulina Rusanowska, Magdalena Zielinska, Agnieszka Cydzik-Kwiatkowska

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Technologies of biogas upgrading are now perceived as competitive solution combustion and production of electricity and heat. Biomethane production will ensure broader application as energy carrier than biogas. Biomethane can be used as fuel in internal combustion engines or introduced into the natural gas transmission network. Therefore, there is a need to search for innovative, economically and technically justified methods for biogas enrichment. The aim of this paper is to present a technology solution for biogas upgrading with immobilized algae biomass. Reactor for biogas upgrading with immobilized algae biomass can be used for removing CO₂ from the biogas, flue gases and the waste gases especially coming from different industry sectors, e.g. from the food industry from yeast production process, biogas production systems, liquid and gaseous fuels combustion systems, hydrocarbon processing technology. The basis for the technological assumptions of presented technology were laboratory works and analyses that tested technological variants of biogas upgrading. The enrichment of biogas with a methane content of 90-97% pointed to technological assumptions for installation on a technical scale. Reactor for biogas upgrading with algae biomass is characterized by a significantly lower cubature in relation to the currently used solutions which use CO₂ removal processes. The invention, by its structure, assumes achieving a very high concentration of biomass of algae through its immobilization in capsules. This eliminates the phenomenon of lowering the pH value, i.e. acidification of the environment in which algae grow, resulting from the introduction of waste gases at a high CO₂ concentration. The system for introducing light into algae capsules is characterized by a higher degree of its use, due to lower losses resulting from the phenomenon of absorption of light energy by water. The light from the light source is continuously supplied to the formed biomass of algae or cyanobacteria in capsules by the light tubes. The light source may be sunlight or a light generator of a different wavelength of light from 300 nm to 800 nm. A portion of gas containing CO₂, accumulated in the tank and conveyed by the pump is periodically introduced into the housing of the photobioreactor tank. When conveying the gas that contains CO₂, it penetrates the algal biomass in capsules through the outer envelope, displacing, from the algal biomass, gaseous metabolic products which are discharged by the outlet duct for gases. It contributes to eliminating the negative impact of this factor on CO₂ binding processes. As a result of the cyclic dosing of gases containing carbon dioxide, gaseous metabolic products of algae are displaced and removed outside the technological system. Technology for biogas upgrading with immobilized algae biomass is suitable for the small biogas plant. The advantages of this technology are high efficiency as well as useful algae biomass which can be used mainly as animal feed, fertilizers and in the power industry. The construction of the device allows effective removal of carbon dioxide from gases at a high CO₂ concentration.

Keywords: biogas, carbon dioxide, immobilised biomass, microalgae, upgrading

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Authors: Ewelina Musielak, Agnieszka Feliczak-Guzik, Izabela Nowak

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Based on the latest data, it is expected that the substances of therapeutic interest used will be as natural as possible. Therefore, active substances with the highest possible efficacy and low toxicity are sought. Among natural substances with therapeutic effects, those of plant origin stand out. Curcumin isolated from the Curcuma longa plant has proven to be particularly important from a medical point of view. Due to its ability to regulate many important transcription factors, cytokines, and protein kinases, curcumin has found use as an anti-inflammatory, antioxidant, antiproliferative, antiangiogenic, and anticancer agent. The unfavorable properties of curcumin, such as low solubility, poor bioavailability, and rapid degradation under neutral or alkaline pH conditions, limit its clinical application. These problems can be solved by combining curcumin with suitable carriers such as hierarchical zeolites. This is a new class of materials that exhibit several advantages. Hierarchical zeolites used as drug carriers enable delayed release of the active ingredient and promote drug transport to the desired tissues and organs. In addition, hierarchical zeolites play an important role in regulating micronutrient levels in the body and have been used successfully in cancer diagnosis and therapy. To apply curcumin to hierarchical zeolites synthesized from commercial FAU zeolite, solutions containing curcumin, carrier and acetone were prepared. The prepared mixtures were then stirred on a magnetic stirrer for 24 h at room temperature. The curcumin-filled hierarchical zeolites were drained into a glass funnel, where they were washed three times with acetone and distilled water, after which the obtained material was air-dried until completely dry. In addition, the effect of piperine addition to zeolite carrier containing a sufficient amount of curcumin was studied. The resulting products were weighed and the percentage of pure curcumin in the hierarchical zeolite was calculated. All the synthesized materials were characterized by several techniques: elemental analysis, transmission electron microscopy (TEM), Fourier transform infrared spectroscopy, Fourier transform infrared (FT-IR), N2 adsorption, and X-ray diffraction (XRD) and thermogravimetric analysis (TGA). The aim of the presented study was to improve the biological activity of curcumin by applying it to hierarchical zeolites based on FAU zeolite. The results showed that the loading efficiency of curcumin into hierarchical zeolites based on commercial FAU-type zeolite is enhanced by modifying the zeolite carrier itself. The hierarchical zeolites proved to be very good and efficient carriers of plant-derived active ingredients such as curcumin.

Keywords: carriers of active substances, curcumin, hierarchical zeolites, incorporation

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Authors: Izabela Kapsa

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Civic participation is an important aspect of democracy. The contemporary model of democracy is based on citizens' participation in political decision-making (deliberative democracy, participatory democracy). This participation takes many forms of activities like display of slogans and symbols, voting, social consultations, political demonstrations, membership in political parties or organizing civil disobedience. The countries of Central and Eastern Europe after 1989 are characterized by great social, economic and political diversity. Civil society is also part of the process of democratization. Civil society, funded by the rule of law, civil rights, such as freedom of speech and association and private ownership, was to play a central role in the development of liberal democracy. Among the many interpretations of concepts, defining the concept of contemporary democracy, one can assume that the terms civil society and democracy, although different in meaning, nowadays overlap. In the post-communist countries, the process of shaping and maturing societies took place in the context of a struggle with a state governed by undemocratic power. State fraud or repudiation of the institution is a representative state, which in the past was the only way to manifest and defend its identity, but after the breakthrough became one of the main obstacles to the development of civil society. In Central and Eastern Europe, there are many obstacles to the development of civil society, for example, the elimination of economic poverty, the implementation of educational campaigns, consciousness-related obstacles, the formation of social capital and the deficit of social activity. Obviously, civil society does not only entail an electoral turnout but a broader participation in the decision-making process, which is impossible without direct and participative democratic institutions. This article considers such broad forms of civic participation and their characteristics in Central and Eastern Europe. The paper is attempts to analyze the functioning of electronic forms of civic participation in Central and Eastern European states. This is not accompanied by a referendum or a referendum initiative, and other forms of political participation, such as public consultations, participative budgets, or e-Government. However, this paper will broadly present electronic administration tools, the application of which results from both legal regulations and increasingly common practice in state and city management. In the comparative analysis, the experiences of post-communist bloc countries will be summed up to indicate the challenges and possible goals for further development of this form of citizen participation in the political process. The author argues that for to function efficiently and effectively, states need to involve their citizens in the political decision-making process, especially with the use of electronic tools.

Keywords: Central and Eastern Europe, e-participation, e-government, post-communism

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Authors: Izabela Poprawa, Alina Chachulska-Zymelka, Lukasz Chajec, Grazyna Wilczek, Piotr Wilczek, Sebastian Student, Magdalena Rost-Roszkowska

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Heavy metals polluting the environment, especially soil, have a harmful effect on organisms, because they can damage the organ structure, disturb their function and cause developmental disorders. They can affect not only the somatic tissues but also the germinal tissues. In the natural environment, plants and animals are exposed to short- and long-term exposure to these stressors, which have a major influence on the functioning of these organisms. Numerous animals have been treated as the bioindicators of the environment. Therefore, studies on any alterations caused by, e.g., heavy metals are in the center of interests of not only environmental but also medical and biological science. Myriapods are invertebrates which are bioindicators of the environment. One of the species which lives in the upper layers of soil, particularly under stones and rocks is Lithobius forficatus (Chilopoda), commonly known as the brown centipede or stone centipede. It is a European species of the family Lithobiidae. This centipede living in the soil is exposed to, e.g., heavy metals such as cadmium, lead, arsenic. The main goal of our project was to analyze the impact of long-term exposure to cadmium on the structure of ovary with the emphasis on the course of oogenesis. As the material for analysis of cadmium exposure to ovaries, we chose the centipede species, L. forficatus. Animals were divided into two experimental groups: C – the control group, the animals cultured in laboratory conditions in a horticultural soil; Cd2 – the animals cultured in a horticultural soil supplemented with 80 mg/kg (dry weight) of CdCl2 for 45 days – long-term exposure. Animals were fed with Acheta and Chironomus larvae maintained in tap water. The analyzes were carried out using transmission electron microscopy (TEM), flow cytometry and laser scanning (confocal) microscopy. Here we present the results of long-term exposure to cadmium concentration in soil on the organ responsible for female germ cell formation. Analysis with the use of the transmission electron microscope showed changes in the ultrastructure of both somatic and germ cells in the ovary. Moreover, quantitative analysis revealed the decrease in the percentage of cells viability, the increase in the percentage of cells with depolarized mitochondria and increasing the number of early apoptotic cells. All these changes were statistically significant compared to the control. Additionally, an increase in the ADP/ATP index was recorded. However, changes were not statistically significant to the control. Acknowledgment: The study has been financed by the National Science Centre, Poland, grant no 2017/25/B/NZ4/00420.

Keywords: cadmium, centipede, ovary, ultrastructure

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Authors: Izabela P. Vatanabe, Rafaela Peron, Patricia Manzine, Marcia R. Cominetti

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Introduction: Considering the increase in life expectancy of world population, there is an emerging concern in health services to allocate better care and care to elderly, through promotion, prevention and treatment of health. It has been observed that frailty syndrome is prevalent in elderly people worldwide and this complex and heterogeneous clinical syndrome consist of the presence of physical frailty associated with cognitive dysfunction, though in absence of dementia. This can be characterized by exhaustion, unintentional weight loss, decreased walking speed, weakness and low level of physical activity, in addition, each of these symptoms may be a predictor of adverse outcomes such as hospitalization, falls, functional decline, institutionalization, and death. Cognitive frailty is a recent concept in literature, which is defined as the presence of physical frailty associated with mild cognitive impairment (MCI) however in absence of dementia. This new concept has been considered as a subtype of frailty, which along with aging process and its interaction with physical frailty, accelerates functional declines and can result in poor quality of life of the elderly. MCI represents a risk factor for Alzheimer's disease (AD) in view of high conversion rate for this disease. Comorbidities and physical frailty are frequently found in AD patients and are closely related to heterogeneity and clinical manifestations of the disease. The decreased platelets ADAM10 levels in AD patients, compared to cognitively healthy subjects, matched by sex, age and education. Objective: Based on these previous results, this study aims to evaluate whether ADAM10 platelet levels of could act as a biomarker of cognitive frailty. Methods: The study was approved by Ethics Committee of Federal University of São Carlos (UFSCar) and conducted in the municipality of São Carlos, headquarters of Federal University of São Carlos (UFSCar). Biological samples of subjects were collected, analyzed and then stored in a biorepository. ADAM10 platelet levels were analyzed by western blotting technique in subjects with MCI and compared to subjects without cognitive impairment, both with and without presence of frailty. Statistical tests of association, regression and diagnostic accuracy were performed. Results: The results have shown that ADAM10/β-actin ratio is decreased in elderly individuals with cognitive frailty compared to non-frail and cognitively healthy controls. Previous studies performed by this research group, already mentioned above, demonstrated that this reduction is still higher in AD patients. Therefore, the ADAM10/β-actin ratio appears to be a potential biomarker for cognitive frailty. The results bring important contributions to an accurate diagnosis of cognitive frailty from the perspective of ADAM10 as a biomarker for this condition, however, more experiments are being conducted, using a high number of subjects, and will help to understand the role of ADAM10 as biomarker of cognitive frailty and contribute to the implementation of tools that work in the diagnosis of cognitive frailty. Such tools can be used in public policies for the diagnosis of cognitive frailty in the elderly, resulting in a more adequate planning for health teams and better quality of life for the elderly.

Keywords: ADAM10, biomarkers, cognitive frailty, elderly

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Authors: Magdalena Rost-Roszkowska, Izabela Poprawa, Alina Chachulska-Zymelka, Lukasz Chajec, Grazyna Wilczek, Piotr Wilczek, Malgorzata Lesniewska

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Lithobius forficatus, commonly known as the brown centipede, is a widespread European species, which lives in the upper layers of soil, under stones, litter, rocks, and leaves. As the soil organism, it is exposed to numerous stressors such as xenobiotics, including heavy metals, temperature, starvation, pathogens, etc. Heavy metals are treated as the environmental pollutants of the soil because of their toxic effects on plants, animals and human being. One of the heavy metals which is xenobiotic and can be taken up by plants or animals from the soil is cadmium. The digestive system of centipedes is composed of three distinct regions: fore-, mid- and hindgut. The salivary glands of centipedes are the organs which belong to the anterior region of the digestive system and take part in the synthesis, accumulation, and secretion of many substances. The middle region having contact with the food masses is treated as one of the barriers which protect the organism against any stressors which originate from the external environment, e.g., toxic metals. As the material for our studies, we chose two organs of the digestive system in brown centipede, the organs which take part in homeostasis maintenance: the salivary glands and the midgut. The main purpose of the project was to investigate the relationship between the percentage of depolarized mitochondria, mitophagy and ATP level in cells of mentioned above organs. The animals were divided into experimental groups: K – the control group, the animals cultured in a laboratory conditions in a horticultural soil and fed with Acheta domesticus larvae; Cd1 – the animals cultured in a horticultural soil supplemented with 80 mg/kg (dry weight) of CdCl2, fed with A. domesticus larvae maintained in tap water, 12 days – short-term exposure; Cd2 – the animals cultured in a horticultural soil supplemented with 80 mg/kg (dry weight) of CdCl2, fed with A. domesticus larvae maintained in tap water, 45 days – long-term exposure. The studies were conducted using transmission electron microscopy (TEM), flow cytometry and confocal microscopy. Quantitative analysis revealed that regardless of the organ, a progressive increase in the percentage of cells with depolarized mitochondria was registered, but only in the salivary glands. These were statistically significant changes from the control. In both organs, there were no differences in the level of the analyzed parameter depending on the duration of exposure of individuals to cadmium. Changes in the ultrastructure of mitochondria have been observed. With the extension of the body's exposure time to metal, an increase in the ADP/ATP index was recorded. However, changes statistically significant to the control were demonstrated in the intestine and salivary glands. The size of this intestinal index and salivary glands in the Cd2 group was about thirty and twenty times higher, respectively than in control. Acknowledgment: The study has been financed by the National Science Centre, Poland, grant no 2017/25/B/NZ4/00420.

Keywords: cadmium, digestive system, ultrastructure, centipede

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Authors: Michal Kielbik, Izabela Szulc-Kielbik, Anna Brzostek, Jaroslaw Dziadek, Magdalena Klink

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The goal of many research groups in the world is to find new components that are important for survival of mycobacteria in the host cells. Mycobacterium tuberculosis (Mtb) possesses a number of enzymes degrading cholesterol that are considered to be an important factor for its survival and persistence in host macrophages. One of them - cholesterol oxidase (ChoD), although not being essential for cholesterol degradation, is discussed as a virulence compound, however its involvement in macrophages’ response to Mtb is still not sufficiently determined. The recognition of tubercle bacilli antigens by pathogen recognition receptors is crucial for the initiation of the host innate immune response. An important receptor that has been implicated in the recognition and/or uptake of Mtb is Toll-like receptor type 2 (TLR2). Engagement of TLR2 results in the activation and phosphorylation of intracellular signaling proteins including IRAK-1 and -4, TRAF-6, which in turn leads to the activation of target kinases and transcription factors responsible for bactericidal and pro-inflammatory response of macrophages. The aim of these studies was a detailed clarification of the role of Mtb cholesterol oxidase as a virulence factor affecting the TLR2 signaling pathway in human macrophages. As human macrophages the THP-1 differentiated cells were applied. The virulent wild-type Mtb strain (H37Rv), its mutant lacking a functional copy of gene encoding cholesterol oxidase (∆choD), as well as complimented strain (∆choD–choD) were used. We tested the impact of Mtb strains on the expression of TLR2-depended signaling proteins (mRNA level, cytosolic level and phosphorylation status). The cytokine and bactericidal response of THP-1 derived macrophages infected with Mtb strains in relation to TLR2 signaling pathway dependence was also determined. We found that during the 24-hours of infection process the wild-type and complemented Mtb significantly reduced the cytosolic level and phosphorylation status of IRAK-4 and TRAF-6 proteins in macrophages, that was not observed in the case of ΔchoD mutant. Decreasement of TLR2-dependent signaling proteins, induced by wild-type Mtb, was not dependent on the activity of proteasome. Blocking of TLR2 expression, before infection, effectively prevented the induced by wild-type strain reduction of cytosolic level and phosphorylation of IRAK-4. None of the strains affected the surface expression of TLR2. The mRNA level of IRAK-4 and TRAF-6 genes were significantly increased in macrophages 24 hours post-infection with either of tested strains. However, the impact of wild-type Mtb strain on both examined genes was significantly stronger than its ΔchoD mutant. We also found that wild-type strain stimulated macrophages to release high amount of immunosuppressive IL-10, accompanied by low amount of pro-inflammatory IL-8 and bactericidal nitric oxide in comparison to mutant lacking cholesterol oxidase. The influence of wild-type Mtb on this type of macrophages' response strongly dependent on fully active IRAK-1 and IRAK-4 signaling proteins. In conclusion, Mtb using cholesterol oxidase causes the over-activation of TLR2 signaling proteins leading to the reduction of their cytosolic level and activity resulting in the modulation of macrophages response to allow its intracellular survival. Supported by grant: 2014/15/B/NZ6/01565, National Science Center, Poland

Keywords: Mycobacterium tuberculosis, cholesterol oxidase, macrophages, TLR2-dependent signaling pathway

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