Search results for: relative biological effectiveness

Authors: Simona Dostalova, Pavel Kopel, Marketa Vaculovicova, Vojtech Adam, Rene Kizek

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Protein apoferritin seems to be a very promising structure for use as a nanocarrier. It is prepared from intracellular ferritin protein naturally found in most organisms. The role of ferritin proteins is to store and transport ferrous ions. Apoferritin is a hollow protein cage without ferrous ions that can be prepared from ferritin by reduction with thioglycolic acid or dithionite. The structure of apoferritin is composed of 24 protein subunits, creating a sphere with 12 nm in diameter. The inner cavity has a diameter of 8 nm. The drug encapsulation process is based on the response of apoferritin structure to the pH changes of surrounding solution. In low pH, apoferritin is disassembled into individual subunits and its structure is “opened”. It can then be mixed with any desired cytotoxic drug and after adjustment of pH back to neutral the subunits are reconnected again and the drug is encapsulated within the apoferritin particles. Excess drug molecules can be removed by dialysis. The receptors for apoferritin, SCARA5 and TfR1 can be found in the membrane of both healthy and cancer cells. To enhance the specific targeting of apoferritin nanocarrier, it is possible to modify its surface with targeting moieties, such as antibodies. To ensure sterically correct complex, we used a a peptide linker based on a protein G with N-terminus affinity towards Fc region of antibodies. To connect the peptide to the surface of apoferritin, the C-terminus of peptide was made of cysteine with affinity to gold. The surface of apoferritin with encapsulated doxorubicin (ApoDox) was coated either with gold nanoparticles (ApoDox-Nano) or gold (III) chloride hydrate reduced with sodium borohydride (ApoDox-HAu). The applied amount of gold in form of gold (III) chloride hydrate was 10 times higher than in the case of gold nanoparticles. However, after removal of the excess unbound ions by electrophoretic separation, the concentration of gold on the surface of apoferritin was only 6 times higher for ApoDox-HAu in comparison with ApoDox-Nano. Moreover, the reduction with sodium borohydride caused a loss of doxorubicin fluorescent properties (excitation maximum at 480 nm with emission maximum at 600 nm) and thus its biological activity. Fluorescent properties of ApoDox-Nano were similar to the unmodified ApoDox, therefore it was more suited for the intended use. To evaluate the specificity of apoferritin modified with antibodies, we used ELISA-like method with the surface of microtitration plate wells coated by the antigen (goat anti-human IgG antibodies). To these wells, we applied ApoDox without targeting antibodies and ApoDox-Nano modified with targeting antibodies (human IgG antibodies). The amount of unmodified ApoDox on antigen after incubation and subsequent rinsing with water was 5 times lower than in the case of ApoDox-Nano modified with targeting antibodies. The modification of non-gold ApoDox with antibodies caused no change in its targeting properties. It can therefore be concluded that the demonstrated procedure allows us to create nanocarrier with enhanced targeting properties, suitable for nanomedicine.

Keywords: apoferritin, doxorubicin, nanocarrier, targeting antibodies

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Authors: Ren-Jy Ben, Jo-Chi Jao, Chiu-Ya Liao, Ya-Ru Tsai, Lain-Chyr Hwang, Po-Chou Chen

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Cancer is still one of the serious diseases threatening the lives of human beings. How to have an early diagnosis and effective treatment for tumors is a very important issue. The animal carcinoma model can provide a simulation tool for the study of pathogenesis, biological characteristics and therapeutic effects. Recently, drug delivery systems have been rapidly developed to effectively improve the therapeutic effects. Liposome plays an increasingly important role in clinical diagnosis and therapy for delivering a pharmaceutic or contrast agent to the targeted sites. Liposome can be absorbed and excreted by the human body, and is well known that no harm to the human body. This study aimed to compare the therapeutic effects between encapsulated (doxorubicin liposomal, LipoDox) and un-encapsulated (doxorubicin, Dox) anti-tumor drugs using Magnetic Resonance Imaging (MRI). Twenty-four New Zealand rabbits implanted with VX2 carcinoma at left thigh were classified into three groups: control group (untreated), Dox-treated group and LipoDox-treated group, 8 rabbits for each group. MRI scans were performed three days after tumor implantation. A 1.5T GE Signa HDxt whole body MRI scanner with a high resolution knee coil was used in this study. After a 3-plane localizer scan was performed, Three-Dimensional (3D) Fast Spin Echo (FSE) T2-Weighted Images (T2WI) was used for tumor volumetric quantification. And Two-Dimensional (2D) spoiled gradient recalled echo (SPGR) dynamic Contrast-enhanced (DCE) MRI was used for tumor perfusion evaluation. DCE-MRI was designed to acquire four baseline images, followed by contrast agent Gd-DOTA injection through the ear vein of rabbits. Afterwards, a series of 32 images were acquired to observe the signals change over time in the tumor and muscle. The MRI scanning was scheduled on a weekly basis for a period of four weeks to observe the tumor progression longitudinally. The Dox and LipoDox treatments were prescribed 3 times in the first week immediately after VX2 tumor implantation. ImageJ was used to quantitate tumor volume and time course signal enhancement on DCE images. The changes of tumor size showed that the growth of VX2 tumors was effectively inhibited for both LipoDox-treated and Dox-treated groups. Furthermore, the tumor volume of LipoDox-treated group was significantly lower than that of Dox-treated group, which implies that LipoDox has better therapeutic effect than Dox. The signal intensity of LipoDox-treated group is significantly lower than that of the other two groups, which implies that targeted therapeutic drug remained in the tumor tissue. This study provides a radiation-free and non-invasive MRI method for therapeutic monitoring of targeted liposome on an animal tumor model.

Keywords: doxorubicin, dynamic contrast-enhanced MRI, lipodox, magnetic resonance imaging, VX2 tumor model

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Authors: Sara R. Knigge, Sugat R. Tuladhar, Alexander Becker, Tobias Schilling, Birgit Glasmacher

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Commercially available mechanical or biological heart valve prostheses both tend to fail long-term due to thrombosis, calcific degeneration, infection, or immunogenic rejection. Moreover, these prostheses are non-viable and do not grow with the patients, which is a problem for young patients. As a result, patients often need to undergo redo-operations. Tissue-engineered (TE) heart valves based on degradable electrospun fiber scaffolds represent a promising approach to overcome these limitations. Such scaffolds need sufficient mechanical properties to withstand the hydrodynamic stress of intracardiac hemodynamics. Additionally, the scaffolds should be colonized by autologous or homologous cells to facilitate the in vivo remodeling of the scaffolds to a viable structure. This study investigates how process parameters of electrospinning and degradation affect the mechanical properties of electrospun scaffolds made of FDA-approved, biodegradable polymer polycaprolactone (PCL). Fiber mats were produced from a PCL/tetrafluoroethylene solution by electrospinning. The e-spinning process was varied in terms of scaffold thickness, fiber diameter, fiber orientation, and fiber interconnectivity. The morphology of the fiber mats was characterized with a scanning electron microscope (SEM). The mats were degraded in different solutions (cell culture media, SBF, PBS and 10 M NaOH-Solution). At different time points of degradation (2, 4 and 6 weeks), tensile and cyclic loading tests were performed. Fresh porcine pericardium and heart valves served as a control for the mechanical assessment. The progression of polymer degradation was quantified by SEM and differential scanning calorimetry (DSC). Primary Human aortic endothelial cells (HAECs) and Human induced pluripotent stem cell-derived endothelial cells (iPSC-ECs) were seeded on the fiber mats to investigate the cell colonization potential. The results showed that both the electrospinning parameters and the degradation significantly influenced the mechanical properties. Especially the fiber orientation has a considerable impact and leads to a pronounced anisotropic behavior of the scaffold. Preliminary results showed that the polymer became strongly more brittle over time. However, the embrittlement can initially only be detected in the mechanical test. In the SEM and DSC investigations, neither morphological nor thermodynamic changes are significantly detectable. Live/Dead staining and SEM imaging of the cell-seeded scaffolds showed that HAECs and iPSC-ECs were able to grow on the surface of the polymer. In summary, this study's results indicate a promising approach to the development of a TE aortic heart valve based on an electrospun scaffold.

Keywords: electrospun scaffolds, long-term polymer degradation, mechanical behavior of electrospun PCL, tissue engineered aortic heart valve

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Authors: Hakimullah Hakim, Chiharu Toyofuku, Mari Ota, Mayuko Suzuki, Miyuki Komura, Masashi Yamada, Md. Shahin Alam, Natthanan Sangsriratanakul, Dany Shoham, Kazuaki Takehara

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Disinfectants and their application are essential part of infection control strategies and enhancement of biosecurity at farms, worldwide. Alkaline agents are well known for their strong and long term antimicrobial capacities and most frequently are applied at farms for control and prevention of biological hazards. However, inadequate information regarding such materials’ capacities to inactivate pathogens and their improper applications fail farmers to achieve the mentioned goal. Thus, this requires attention to further evaluate their efficacies, under different conditions and in different ways. Here in this study we evaluated bactericidal efficacies of food additive grade of calcium hydroxide (FdCa(OH)2) powder derived from natural calcium carbonates obtained from limestone (Fine Co., Ltd., Tokyo, Japan), and bioceramic powder (BCX) derived from chicken feces at pH 13 (NMG environmental development Co., Ltd., Tokyo, Japan), for their efficacies to inactivate bacteria in feces. [Materials & Methods] Chicken feces were inoculated by 100 µl Escherichia coli and Salmonella Infantis in falcon tubes, individually, then FdCa(OH)2 or BCX powders were individually added to make final concentration of 0, 5, 10, 20 and 30% (w/w) in total weight of 0.5g, followed by properly mixing and incubating at room temperature for certain periods of time, in a dark place. Afterwards, 10 ml 1M Tris-HCl (pH 7.2) was added onto them to reduce their pH, in order to stop powders’ activities and to harvest the remained viable bacteria, whereas using normal medium or dW2 to recover bacteria increases the mixture pH, and as a result bacteria would be inactivated soon; therefore, the latter practice brings about incorrect and misleading results. Samples were then inoculated on DHL agar plates in order to calculate colony forming units (CFU)/ml of viable bacteria. [Results and Discussion] FdCa(OH)2 powder at 10% and 5% required 3 hr and 6 hr exposure times, respectively, while BCX powder at 20% concentrations required 6 hr exposure time to kill the mentioned bacteria in feces down to lower than detectable level (≤ 3.6 log10 CFU/ml). This study confirmed capacities of FdCa(OH)2 and BCX powders to inactivate bacteria in feces, and both materials are environment friendly materials, with no risk to human or animal’s health. This finding helps farmers to properly apply alkaline agents in appropriate concentrations and exposure times in their farms, in order to prevent and control infectious diseases outbreaks and to enhance biosecurity. Finally, this finding may help farmers to implement better strategies for infections control in their livestock farms.

Keywords: bacterial inactivation, bioceramic, biosecurity at livestock farms, chicken feces

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Authors: Anna Drabczyk, Sonia Kudlacik-Kramarczyk, Dagmara Malina, Bozena Tyliszczak, Agnieszka Sobczak-Kupiec

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Nowadays, nano-size materials are very popular group of materials among scientists. What is more, these materials find an application in a wide range of various areas. Therefore constantly increasing demand for nanomaterials including metallic nanoparticles such as silver of gold ones is observed. Therefore, new routes of their preparation are sought. Considering potential application of nanoparticles, it is important to select an adequate methodology of their preparation because it determines their size and shape. Among the most commonly applied methods of preparation of nanoparticles chemical and electrochemical techniques are leading. However, currently growing attention is directed into the biological or biochemical aspects of syntheses of metallic nanoparticles. This is associated with a trend of developing of new routes of preparation of given compounds according to the principles of green chemistry. These principles involve e.g. the reduction of the use of toxic compounds in the synthesis as well as the reduction of the energy demand or minimization of the generated waste. As a result, a growing popularity of the use of such components as natural plant extracts, infusions or essential oils is observed. Such natural substances may be used both as a reducing agent of metal ions and as a stabilizing agent of formed nanoparticles therefore they can replace synthetic compounds previously used for the reduction of metal ions or for the stabilization of obtained nanoparticles suspension. Methods that proceed in the presence of previously mentioned natural compounds are environmentally friendly and proceed without the application of any toxic reagents. Methodology: Presented research involves preparation of silver nanoparticles using selected plant extracts, e.g. artichoke extract. Extracts of natural origin were used as reducing and stabilizing agents at the same time. Furthermore, syntheses were carried out in the presence of additional polymeric stabilizing agent. Next, such features of obtained suspensions of nanoparticles as total antioxidant activity as well as content of phenolic compounds have been characterized. First of the mentioned studies involved the reaction with DPPH (2,2-Diphenyl-1-picrylhydrazyl) radical. The content of phenolic compounds was determined using Folin-Ciocalteu technique. Furthermore, an essential issue was also the determining of the stability of formed suspensions of nanoparticles. Conclusions: In the research it was demonstrated that metallic nanoparticles may be obtained using plant extracts or infusions as stabilizing or reducing agent. The methodology applied, i.e. a type of plant extract used during the synthesis, had an impact on the content of phenolic compounds as well as on the size and polydispersity of obtained nanoparticles. What is more, it is possible to prepare nano-size particles that will be characterized by properties desirable from the viewpoint of their potential application and such an effect may be achieved with the use of non-toxic reagents of natural origin. Furthermore, proposed methodology stays in line with the principles of green chemistry.

Keywords: green chemistry principles, metallic nanoparticles, plant extracts, stabilization of nanoparticles

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Authors: Natalia V. Beloglazova, Pieterjan Lenain, Martin Hedstrom, Dietmar Knopp, Sarah De Saeger

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In recent years polycyclic aromatic hydrocarbons (PAHs), which represent a hazard to humans and entire ecosystem, have been receiving an increased interest due to their mutagenic, carcinogenic and endocrine disrupting properties. They are formed in all incomplete combustion processes of organic matter and, as a consequence, ubiquitous in the environment. Benzo(a)pyrene (BaP) is on the priority list published by the Environmental Agency (US EPA) as the first PAH to be identified as a carcinogen and has often been used as a marker for PAHs contamination in general. It can be found in different types of water samples, therefore, the European Commission set up a limit value of 10 ng L–1 (10 ppt) for BAP in water intended for human consumption. Generally, different chromatographic techniques are used for PAHs determination, but these assays require pre-concentration of analyte, create large amounts of solvent waste, and are relatively time consuming and difficult to perform on-site. An alternative robust, stand-alone, and preferably cheap solution is needed. For example, a sensing unit which can be submerged in a river to monitor and continuously sample BaP. An affinity sensor based on capacitive transduction was developed. Natural antibodies or their synthetic analogues can be used as ligands. Ideally the sensor should operate independently over a longer period of time, e.g. several weeks or months, therefore the use of molecularly imprinted polymers (MIPs) was discussed. MIPs are synthetic antibodies which are selective for a chosen target molecule. Their robustness allows application in environments for which biological recognition elements are unsuitable or denature. They can be reused multiple times, which is essential to meet the stand-alone requirement. BaP is a highly lipophilic compound and does not contain any functional groups in its structure, thus excluding non-covalent imprinting methods based on ionic interactions. Instead, the MIPs syntheses were based on non-covalent hydrophobic and π-π interactions. Different polymerization strategies were compared and the best results were demonstrated by the MIPs produced using electropolymerization. 4-vinylpyridin (VP) and divinylbenzene (DVB) were used as monomer and cross-linker in the polymerization reaction. The selectivity and recovery of the MIP were compared to a non-imprinted polymer (NIP). Electrodes were functionalized with natural receptor (monoclonal anti-BaP antibody) and with MIPs selective towards BaP. Different sets of electrodes were evaluated and their properties such as sensitivity, selectivity and linear range were determined and compared. It was found that both receptor can reach the cut-off level comparable to the established ML, and despite the fact that the antibody showed the better cross-reactivity and affinity, MIPs were more convenient receptor due to their ability to regenerate and stability in river till 7 days.

Keywords: antibody, benzo(a)pyrene, capacitive sensor, MIPs, river water

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Authors: Nasreen Abdulmannan

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Fetal abnormalities are categorized as a structural abnormality, non-structural abnormality, or a combination of both. Fetal structural abnormalities (FSA) include, but are not limited, to Down syndrome, congenital diaphragmatic hernia, and cleft lip and palate. These abnormalities can be detected in the first weeks of pregnancy, which is almost around 9 - 20 weeks gestational. Etiological factors for FSA are unknown; however, transmitted genetic risk can be one of these factors. Consanguineous marriage often referred to as inbreeding, represents a significant risk factor for FSA due to the increased likelihood of deleterious genetic traits shared by both biological parents. In a country such as the Kingdom of Saudi Arabia (KSA), consanguineous marriage is high, which creates a significant risk of children being born with congenital abnormalities. Historically, the practice of consanguinity occurred commonly among European royalty. For example, Great Britain’s Queen Victoria married her German first cousin, Prince Albert of Coburg. Although a distant blood relationship, the United Kingdom’s Queen Elizabeth II married her cousin, Prince Philip of Greece and Denmark—both of them direct descendants of Queen Victoria. In Middle Eastern countries, a high incidence of consanguineous unions still exists, including in the KSA. Previous studies indicated that a significant gap exists in understanding the lived experiences of Saudi women dealing with an FSA-complicated pregnancy. Eleven participants were interviewed using a semi-structured interview format for this qualitative phenomenological study investigating the lived experiences of pregnant Saudi women carrying a child with FSA. This study explored the gaps in current literature regarding the lived experiences of pregnant Saudi women whose pregnancies were complicated by FSA. In addition, the researcher acquired knowledge about the available support and resources as well as the Saudi cultural perspective on FSA. This research explored the lived experiences of pregnant Saudi women utilizing Giorgi’s (2009) approach to data collection and data management. Findings for this study cover five major themes: (1) initial maternal reaction to the FSA diagnosis per ultrasound screening; (2) strengthening of the maternal relationship with God; (3) maternal concern for their child’s future; (4) feeling supported by their loved ones; and (5) lack of healthcare provider support and guidance. Future research in the KSA is needed to explore the network support for these mothers. This study recommended further clinical nursing research, nursing education, clinical practice, and healthcare policy/procedures to provide opportunities for improvement in nursing care and increase awareness in KSA society.

Keywords: fetal structural abnormalities, psychological distress, health provider, health care

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Authors: Alexandria Carey, Angela Starkweather, Ann Horgas, Hwayoung Cho, Jason Beneciuk

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Background/Significance: Over 3.4 million acute axial low back pain (aLBP) cases are treated annually in the United States (US) emergency departments (ED). ED patients with aLBP receive varying verbal and written discharge routine care (RC), leading to ineffective patient self-management. Ineffective self-management increase chronic low back pain (cLPB) transition risks, a chief cause of worldwide disability, with associated costs >$60 million annually. This research addresses this significant problem by evaluating an ED digital pain self-management intervention (EDPSI) focused on improving self-management through improved knowledge retainment, skills, and self-efficacy (confidence) (KSC) thus reducing aLBP to cLBP transition in ED patients discharged with aLBP. The research has significant potential to increase self-efficacy, one of the most potent mechanisms of behavior change and improve health outcomes. Focusing on accessibility and usability, the intervention may reduce discharge disparities in aLBP self-management, especially with low health literacy. Study Questions: This research will answer the following questions: 1) Will an EDPSI focused on improving KSC progress patient self-management behaviors and health status?; 2) Is the EDPSI sustainable to improve pain severity, interference, and pain recurrence?; 3) Will an EDPSI reduce aLBP to cLBP transition in patients discharged with aLBP? Aims: The pilot randomized-controlled trial (RCT) study’s objectives assess the effects of a 12-week digital self-management discharge tool in patients with aLBP. We aim to 1) Primarily assess the feasibility [recruitment, enrollment, and retention], and [intervention] acceptability, and sustainability of EDPSI on participant’s pain self-management; 2) Determine the effectiveness and sustainability of EDPSI on pain severity/interference among participants. 3) Explore patient preferences, health literacy, and changes among participants experiencing the transition to cLBP. We anticipate that EDPSI intervention will increase likelihood of achieving self-management milestones and significantly improve pain-related symptoms in aLBP. Methods: The study uses a two-group pilot RCT to enroll 30 individuals who have been seen in the ED with aLBP. Participants are randomized into RC (n=15) or RC + EDPSI (n=15) and receive follow-up surveys for 12-weeks post-intervention. EDPSI innovative content focuses on 1) highlighting discharge education; 2) provides self-management treatment options; 3) actor demonstration of ergonomics, range of motion movements, safety, and sleep; 4) complementary alternative medicine (CAM) options including acupuncture, yoga, and Pilates; 5) combination therapies including thermal application, spinal manipulation, and PT treatments. The intervention group receives Booster sessions via Zoom to assess and reinforce their knowledge retention of techniques and provide return demonstration reinforcing ergonomics, in weeks two and eight. Outcome Measures: All participants are followed for 12-weeks, assessing pain severity/ interference using the Brief Pain Inventory short-form (BPI-sf) survey, self-management (measuring KSC) using the short 13-item Patient Activation Measure (PAM), and self-efficacy using the Pain Self-Efficacy Questionnaire (PSEQ) weeks 1, 6, and 12. Feasibility is measured by recruitment, enrollment, and retention percentages. Acceptability and education satisfaction are measured using the Education-Preference and Satisfaction Questionnaire (EPSQ) post-intervention. Self-management sustainment is measured including PSEQ, PAM, and patient satisfaction and healthcare utilization (PSHU) requesting patient overall satisfaction, additional healthcare utilization, and pain management related to continued back pain or complications post-injury.

Keywords: digital, pain self-management, education, tool

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Authors: M. I. Tyletc, O. I. Podgornya, T. G. Shaposhnikova, S. V. Shabelnikov, A. G. Mittenberg, M. A. Daugavet

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Ascidiacea is the most numerous class of the Tunicata subtype. These chordates' distinctive feature of the anatomical structure is a tunic consisting of cellulose fibrils, protein molecules, and single cells. The mechanisms of the tunic formation are not known in detail; tunic formation could be used as the model system for studying the interaction of cells with the extracellular matrix. Our model species is the ascidian Styela rustica, which is prevalent in benthic communities of the White Sea. As previously shown, the tunic formation involves morula blood cells, which contain the major 48 kDa protein p48. P48 participation in the tunic formation was proved using antibodies against the protein. The nature of the protein and its function remains unknown. The current research aims to determine the amino acid sequence of p48, as well as to clarify its role in the tunic formation. The peptides that make up the p48 amino acid sequence were determined by mass spectrometry. A search for peptides in protein sequence databases identified sequences homologous to p48 in Styela clava, Styela plicata, and Styela canopus. Based on sequence alignment, their level of similarity was determined as 81-87%. The correspondent sequence of ascidian Styela canopus was used for further analysis. The Styela rustica p48 sequence begins with a signal peptide, which could indicate that the protein is secretory. This is consistent with experimentally obtained data: the contents of morula cells secreted in the tunic matrix. The isoelectric point of p48 is 9.77, which is consistent with the experimental results of acid electrophoresis of morula cell proteins. However, the molecular weight of the amino acid sequence of ascidian Styela canopus is 103 kDa, so p48 of Styela rustica is a shorter homolog. The search for conservative functional domains revealed the presence of two Ca-binding EGF-like domains, thrombospondin (TSP1) and tyrosinase domains. The p48 peptides determined by mass spectrometry fall into the region of the sequence corresponding to the last two domains and have amino acid substitutions as compared to Styela canopus homolog. The tyrosinase domain (pfam00264) is known to be part of the phenoloxidase enzyme, which participates in melanization processes and the immune response. The thrombospondin domain (smart00209) interacts with a wide range of proteins, and is involved in several biological processes, including coagulation, cell adhesion, modulation of intercellular and cell-matrix interactions, angiogenesis, wound healing and tissue remodeling. It can be assumed that the tyrosinase domain in p48 plays the role of the phenoloxidase enzyme, and TSP1 provides a link between the extracellular matrix and cell surface receptors, and may also be responsible for the repair of the tunic. The results obtained are consistent with experimental data on p48. The domain organization of protein suggests that p48 is an enzyme involved in the tunic tunning and is an important regulator of the organization of the extracellular matrix.

Keywords: ascidian, p48, thrombospondin, tyrosinase, tunic, tunning

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Authors: Simran Baweja, Bhavika Kalal, Surajit Maity

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Photoinduced tautomerization reactions have been the centre of attention among scientific community over past several decades because of their significance in various biological systems. 7-azaindole (7AI) is considered as a model system for DNA base pairing and to understand the role of such tautomerization reactions in mutations. To the best of our knowledge, extensive studies have been carried on 7-azaindole and its solvent clusters exhibiting proton/ hydrogen transfer in both solution as well as gas phase. Derivatives of above molecule, like 2,7- and 2,6-diazaindoles are proposed to have even better photophysical properties due to the presence of -aza group on the 2nd position. However, there are a few studies in the solution phase which suggest the relevance of these molecules, but there are no experimental studies reported in the gas phase yet. In our current investigation, we present the first gas phase spectroscopic data of 2,7-diazaindole (2,7-DAI) and its solvent cluster (2,7-DAI-H2O). In this, we have employed state-of-the-art laser spectroscopic methods such as fluorescence excitation (LIF), dispersed fluorescence (DF), resonant two-photon ionization time of flight mass spectrometry (2C-R2PI), photoionization efficiency spectroscopy (PIE), IR-UV double resonance spectroscopy i.e. fluorescence-dip infrared spectroscopy (FDIR) and resonant ion-dip infrared spectroscopy (IDIR) to understand the electronic structure of the molecule. The origin band corresponding to S1 ← S0 transition of the bare 2,7-DAI is found to be positioned at 33910 cm-1 whereas the origin band corresponding to S1 ← S0 transition of the 2,7-DAI-H2O is positioned at 33074 cm-1. The red shifted transition in case of solvent cluster suggests the enhanced feasibility of excited state hydrogen/ proton transfer. The ionization potential for the 2,7-DAI molecule is found to be 8.92 eV, which is significantly higher that the previously reported 7AI (8.11 eV) molecule, making it a comparatively complex molecule to study. The ionization potential is reduced by 0.14 eV in case of 2,7-DAI-H2O (8.78 eV) cluster compared to that of 2,7-DAI. Moreover, on comparison with the available literature values of 7AI, we found the origin band of 2,7-DAI and 2,7-DAI-H2O to be red shifted by -729 and -280 cm-1 respectively. The ground and excited state N-H stretching frequencies of the 27DAI molecule were determined using fluorescence-dip infrared spectra (FDIR) and resonant ion dip infrared spectroscopy (IDIR), obtained at 3523 and 3467 cm-1, respectively. The lower value of vNH in the electronic excited state of 27DAI implies the higher acidity of the group compared to the ground state. Moreover, we have done extensive computational analysis, which suggests that the energy barrier in excited state reduces significantly as we increase the number of catalytic solvent molecules (S= H2O, NH3) as well as the polarity of solvent molecules. We found that the ammonia molecule is a better candidate for hydrogen transfer compared to water because of its higher gas-phase basicity. Further studies are underway to understand the excited state dynamics and photochemistry of such N-rich chromophores.

Keywords: photoinduced tautomerization reactions, gas phse spectroscopy, ), IR-UV double resonance spectroscopy, resonant two-photon ionization time of flight mass spectrometry (2C-R2PI)

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Authors: Yakup Ulusu, Faruk Ozel, Numan Eczacioglu, Abdurrahman Ozen, Sabriye Acikgoz

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GFP discovered in the mid-1970s, has been used as a marker after replicated genetic study by scientists. In biotechnology, cell, molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms, it has been used to make biosensors. Many animals have been created that express GFP as an evidence that a gene can be expressed throughout a given organism. Proteins labeled with GFP identified locations are determined. And so, cell connections can be monitored, gene expression can be reported, protein-protein interactions can be observed and signals that create events can be detected. Additionally, monitoring GFP is noninvasive; it can be detected by under UV-light because of simply generating fluorescence. Moreover, GFP is a relatively small and inert molecule, that does not seem to treat any biological processes of interest. The synthesis of GFP has some steps like, to construct the plasmid system, transformation in E. coli, production and purification of protein. GFP carrying plasmid vector pBAD–GFPuv was digested using two different restriction endonuclease enzymes (NheI and Eco RI) and DNA fragment of GFP was gel purified before cloning. The GFP-encoding DNA fragment was ligated into pET28a plasmid using NheI and Eco RI restriction sites. The final plasmid was named pETGFP and DNA sequencing of this plasmid indicated that the hexa histidine-tagged GFP was correctly inserted. Histidine-tagged GFP was expressed in an Escherichia coli BL21 DE3 (pLysE) strain. The strain was transformed with pETGFP plasmid and grown on LuiraBertoni (LB) plates with kanamycin and chloramphenicol selection. E. coli cells were grown up to an optical density (OD 600) of 0.8 and induced by the addition of a final concentration of 1mM isopropyl-thiogalactopyranoside (IPTG) and then grown for additional 4 h. The amino-terminal hexa-histidine-tag facilitated purification of the GFP by using a His Bind affinity chromatography resin (Novagen). Purity of GFP protein was analyzed by a 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of protein was determined by UV absorption at 280 nm (Varian Cary 50 Scan UV/VIS spectrophotometer). Synthesis of GFP-Polymer composite nanofibers was produced by using GFP solution (10mg/mL) and polymer precursor Polyvinylpyrrolidone, (PVP, Mw=1300000) as starting materials and template, respectively. For the fabrication of nanofibers with the different fiber diameter; a sol–gel solution comprising of 0.40, 0.60 and 0.80 g PVP (depending upon the desired fiber diameter) and 100 mg GFP in 10 mL water: ethanol (3:2) mixtures were prepared and then the solution was covered on collecting plate via electro spinning at 10 kV with a feed-rate of 0.25 mL h-1 using Spellman electro spinning system. Results show that GFP-based nano-fiber can be used plenty of biomedical applications such as bio-imaging, bio-mechanic, bio-material and tissue engineering.

Keywords: biomaterial, GFP, nano-fibers, protein expression

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Authors: Deinzel R. Uezono, Vivien Fe F. Fadrilan-Camacho, Bianca Margarita L. Medina, Antonio Domingo R. Reario, Trisha M. Salcedo, Luke Wesley P. Borromeo

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The environment of health care workers is considered one of the most hazardous settings due to the nature of their work. In developing countries especially, the Philippines, this continues to be overlooked in terms of programs and services on occupational health and safety (OHS). One possible reason for this is the existing information gap on OHS which limits data comparability and impairs effective monitoring and assessment of interventions. To address this gap, there is a need to determine the current conditions of Filipino health care workers in their workplace. This descriptive cross-sectional study assessed the occupational health and safety conditions of health care workers in barangay health centers in a selected city in Metro Manila, Philippines by: (1) determining the hazards present in the workplace; (2) determining the most common self-reported medical problems; and (3) describing the elements of an OHS system based on the six building blocks of health system. Assessment was done through walkthrough survey, self-administered questionnaire, and key informant interview. Data analysis was done using Epi Info 7 and NVivo 11. Results revealed different health hazards present in the workplace particularly biological hazards (exposure to sick patients and infectious specimens), physical hazards (inadequate space and/or lighting), chemical hazards (toxic reagents and flammable chemicals), and ergonomic hazards (activities requiring repetitive motion and awkward posture). Additionally, safety hazards (improper capping of syringe and lack of fire safety provisions) were also observed. Meanwhile, the most commonly self-reported chronic diseases among health care workers (N=336) were hypertension (20.24%, n=68) and diabetes (12.50%, n=42). Top commonly self-reported symptoms were colds (66.07%, n=222), coughs (63.10%, n=212), headache (55.65%, n=187), and muscle pain (50.60%, n=170) while other diseases were influenza (16.96%, n=57) and UTI (15.48%, n=52). In terms of the elements of the OHS system, a general policy on occupational health and safety was found to be lacking and in effect, an absence of health and safety committee overseeing the implementing and monitoring of the policy. No separate budget specific for OHS programs and services was also found to be a limitation. As a result, no OHS personnel and trainings/seminar were identified. No established information system for OHS was in place. In conclusion, health and safety hazards were observed to be present across the barangay health centers visited in a selected city in Metro Manila. Medical conditions identified as most commonly self-reported were hypertension and diabetes for chronic diseases; colds, coughs, headache, and muscle pain for medical symptoms; and influenza and UTI for other diseases. As for the elements of the occupational health and safety system, there was a lack in the general components of the six building blocks of the health system.

Keywords: health hazards, occupational health and safety, occupational health and safety system, safety hazards

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Authors: Mitali Saha, Soma Das

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The fabrication of nanoscale materials for use in chemical sensing, biosensing and biological analyses has proven a promising avenue in the last few years. Cholesterol has aroused considerable interest in recent years on account of its being an important parameter in clinical diagnosis. There is a strong positive correlation between high serum cholesterol level and arteriosclerosis, hypertension, and myocardial infarction. Enzyme-based electrochemical biosensors have shown high selectivity and excellent sensitivity, but the enzyme is easily denatured during its immobilization procedure and its activity is also affected by temperature, pH, and toxic chemicals. Besides, the reproducibility of enzyme-based sensors is not very good which further restrict the application of cholesterol biosensor. It has been demonstrated that carbon nanotubes could promote electron transfer with various redox active proteins, ranging from cytochrome c to glucose oxidase with a deeply embedded redox center. In continuation of our earlier work on the synthesis and applications of carbon and metal based nanoparticles, we have reported here the synthesis of carbon nanotubes (CCNT) by burning coconut oil under insufficient flow of air using an oil lamp. The soot was collected from the top portion of the flame, where the temperature was around 6500C which was purified, functionalized and then characterized by SEM, p-XRD and Raman spectroscopy. The SEM micrographs showed the formation of tubular structure of CCNT having diameter below 100 nm. The XRD pattern indicated the presence of two predominant peaks at 25.20 and 43.80, which corresponded to (002) and (100) planes of CCNT respectively. The Raman spectrum (514 nm excitation) showed the presence of 1600 cm-1 (G-band) related to the vibration of sp2-bonded carbon and at 1350 cm-1 (D-band) responsible for the vibrations of sp3-bonded carbon. A nonenzymatic cholesterol biosensor was then fabricated on an insulating Teflon material containing three silver wires at the surface, covered by CCNT, obtained from coconut oil. Here, CCNTs worked as working as well as counter electrodes whereas reference electrode and electric contacts were made of silver. The dimensions of the electrode was 3.5 cm×1.0 cm×0.5 cm (length× width × height) and it is ideal for working with 50 µL volume like the standard screen printed electrodes. The voltammetric behavior of cholesterol at CCNT electrode was investigated by cyclic voltammeter and differential pulse voltammeter using 0.001 M H2SO4 as electrolyte. The influence of the experimental parameters on the peak currents of cholesterol like pH, accumulation time, and scan rates were optimized. Under optimum conditions, the peak current was found to be linear in the cholesterol concentration range from 1 µM to 50 µM with a sensitivity of ~15.31 μAμM−1cm−2 with lower detection limit of 0.017 µM and response time of about 6s. The long-term storage stability of the sensor was tested for 30 days and the current response was found to be ~85% of its initial response after 30 days.

Keywords: coconut oil, CCNT, cholesterol, biosensor

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Authors: Juliana Chiarini Balbino Fernandes

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This article aims to investigate the Spanish didactic book, entitled ‘El Calculo y La Medida en El Primer Grado de La Escuela Decroly’, written by Dr. O. Decroly and A. Hamaide, published in Madrid, in the year 1934. In addition to analyzing how mathematical knowledge is present in the proposed Centers of Interest. The textbooks, in addition to pedagogical tools, reflect a certain moment in society and allow the analysis of the theoretical-methodological proposal that can be implemented by the teacher. The study proposed here will be carried out by the lens of Cultural History, supported by Roger Chartier (1991) and by the concepts on textbooks, based on Alain Choppin (2004). The textbook selected for this study exposes a program of ideas associated with the method of Centers of Interest and arithmetic is linked to these interests. In the first courses (six to eight years), most centers can be considered to correspond to occasional calls, as they take advantage of events that arise spontaneously to work with observation, measurement, association and expression exercises. The program of ideas associated with Centers of Interest addresses the biological and social aspects of children, as long as they can express their needs for activities and games, satisfying the natural curiosity. Still, the program of associated ideas offers occasions for problems whose data are taken in observation exercises and concrete expressions (manuals, drawings). In the method applied at the school of L'Ermitage, school created by Decroly in Belgium in 1907, observation, is the basis of each center of interest. It offers the chance to compare and measure. To observe is more than to perceive; it is also to establish relations between the graded aspects of the same object, to seek relations between different intensities; is to verify successions, special and temporary relationships; is to make comparisons, to notice differences and similarities in block or datable (analysis), is to establish a bridge between the world and the thought. To make the observation more precise, it is important to compare, measure, and resort to considered objects as natural units of measure. Measurement and calculation are, therefore, quite naturally subject to observation. Thus, it is possible to make the child enter into the interest in the calculation, linking it to the observation. It was observed that the Centers of Interest, according to Decroly, should respond to the concerns and attend to the motivations of the students and the teaching of arithmetical must obey a logical seriation, considering the interest and the experience of the children. The teaching of arithmetical should not be limited to the schedule, it should cover every quantitative aspect that arises in the other disciplines. The feeling of unity is established in observation, association and expression, which coordinate a whole program of cultural activities, concentrating it around a central idea.

Keywords: didactic book, centers of interest, mathematical knowledge, primary education

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Authors: Saloni Khandelwal

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The Aravalli hills are one of the oldest and most distinctive mountain chains of peninsular India spanning in around 692 Km. More than 60% of it falls in the state of Rajasthan and influences ecological equilibrium in about 30% of the state. Because of natural and human-induced activities, physical gaps in the Aravallis are increasing, new gaps are coming up, and its physical structure is changing. There are no strict regulations to protect and monitor the Aravallis and no comprehensive research and study has been done for the enhancement of ecosystem functions of these ranges. Through this study, various factors leading to Aravalli’s degradation are identified and its impacts on selected areas are analyzed. A literature study is done to identify factors responsible for the degradation. To understand the severity of the problem at the lowest level, two tehsils from different districts in Rajasthan, which are the most affected due to illegal mining and increasing physical gaps are selected for the study. Case-1 of three-gram panchayats in Kishangarh Tehsil of Ajmer district focuses on the expanding physical gaps in the Aravalli range, and case-2 of three-gram panchayats in Tijara Tehsil of Alwar district focuses on increasing illegal mining in the Aravalli range. For measuring the degradation, physical, biological and social indicators are identified through literature review and for both the cases analysis is done on the basis of these indicators. Primary survey and focus group discussions are done with villagers, mining owners, illegal miners, and various government officials to understand dependency of people on the Aravalli and its importance to them along with the impact of degradation on their livelihood and environment. From the analysis, it has been found that green cover is continuously decreasing in both cases, dense forest areas do not exist now, the groundwater table is depleting at a very fast rate, soil is losing its moisture resulting in low yield and shift in agriculture. Wild animals which were easily seen earlier are now extinct. Cattles of villagers are dependent on the forest area in the Aravalli range for food, but with a decrease in fodder, their cattle numbers are decreasing. There is a decrease in agricultural land and an increase in scrub and salt-affected land. Analysis of various national and state programmes, acts which were passed to conserve biodiversity has been done showing that none of them is helping much to protect the Aravalli. For conserving the Aravalli and its forest areas, regional level and local level initiatives are required and are proposed in this study. This study is an attempt to formulate conservation and management strategies for the Aravalli range. These strategies will help in improving biodiversity which can lead to the revival of its ecosystem functions. It will also help in curbing the pollution at the regional and local level. All this will lead to the sustainable development of the region.

Keywords: Aravalli, ecosystem, LULC, Rajasthan

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Authors: Jeet Chakraborty, Sanjay Dutta

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Antibiotic resistance by bacteria has proved to be a severe threat to mankind in recent times, and this fortifies an urgency to design and develop potent antibacterial small molecules/compounds with nonconventional mechanisms than the conventional ones. DNA carries the genetic signature of any organism, and bacteria maintain their genomic DNA inside the cell in a well-regulated compact form with the help of various nucleoid associated proteins like HU, HNS, etc. These proteins control various fundamental processes like gene expression, replication, etc., inside the cell. Alteration of the native DNA structure of bacteria can lead to severe consequences in cellular processes inside the bacterial cell that ultimately result in the death of the organism. The change in the global DNA structure by small molecules initiates a plethora of cellular responses that have not been very well investigated. Echinomycin and Triostin-A are biologically active Quinoxaline small molecules that typically consist of a quinoxaline chromophore attached with an octadepsipeptide ring. They bind to double-stranded DNA in a sequence-specific way and have high activity against a wide variety of bacteria, mainly against Gram-positive ones. To date, few synthetic quinoxaline scaffolds were synthesized, displaying antibacterial potential against a broad scale of pathogenic bacteria. QNOs (Quinoxaline N-oxides) are known to target DNA and instigate reactive oxygen species (ROS) production in bacteria, thereby exhibiting antibacterial properties. The divergent role of Quinoxaline small molecules in medicinal research qualifies them for the evaluation of their antimicrobial properties as a potential candidate. The previous study from our lab has given new insights on a 6-nitroquinoxaline derivative 1d as an intercalator of DNA, which induces conformational changes in DNA upon binding.7 The binding event observed was dependent on the presence of a crucial benzyl substituent on the quinoxaline moiety. This was associated with a large induced CD (ICD) appearing in a sigmoidal pattern upon the interaction of 1d with dsDNA. The induction of DNA superstructures by 1d at high Drug:DNA ratios was observed that ultimately led to DNA condensation. Eviction of invitro-assembled nucleosome upon treatment with a high dose of 1d was also observed. In this work, monoquinoxaline derivatives of 1d were synthesized by various modifications of the 1d scaffold. The set of synthesized 6-nitroquinoxaline derivatives along with 1d were all subjected to antibacterial evaluation across five different bacteria species. Among the compound set, 3a displayed potent antibacterial activity against Staphylococcus aureus bacteria. 3a was further subjected to various biophysical studies to check whether the DNA structural alteration potential was still intact. The biological response of S. aureus cells upon treatment with 3a was studied using various cell biology processes, which led to the conclusion that 3d can initiate DNA damage in the S. aureus cells. Finally, the potential of 3a in disrupting preformed S.aureus and S.epidermidis biofilms was also studied.

Keywords: DNA structural change, antibacterial, intercalator, DNA superstructures, biofilms

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Authors: Lina Paola Orozco Marin, Yuliet Montoya Osorio, John Bustamante Osorno

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Ischemic events can culminate in acute myocardial infarction by irreversible cardiac lesions that cannot be restored due to the limited regenerative capacity of the heart. Cell therapy seeks to replace these injured or necrotic cells by transplanting healthy and functional cells. The therapeutic alternatives proposed by tissue engineering and cardiovascular regenerative medicine are the use of biomaterials to mimic the native extracellular medium, which is full of proteins, proteoglycans, and glycoproteins. The selected biomaterials must provide structural support to the encapsulated cells to avoid their migration and death in the host tissue. In this context, the present research work focused on developing a natural thermosensitive hydrogel, its physical and chemical characterization, and the determination of its biocompatibility in vitro. The hydrogel was developed by mixing hydrolyzed bovine and porcine collagen at 2% w/v, chitosan at 2.5% w/v, and beta-glycerolphosphate at 8.5% w/w and 10.5% w/w in magnetic stirring at 4°C. Once obtained, the thermosensitivity and gelation time were determined, incubating the samples at 37°C and evaluating them through the inverted tube method. The morphological characterization of the hydrogels was carried out through scanning electron microscopy. Chemical characterization was carried out employing infrared spectroscopy. The biocompatibility was determined using the MTT cytotoxicity test according to the ISO 10993-5 standard for the hydrogel’s precursors using the fetal human ventricular cardiomyocytes cell line RL-14. The RL-14 cells were also seeded on the top of the hydrogels, and the supernatants were subculture at different periods to their observation under a bright field microscope. Four types of thermosensitive hydrogels were obtained, which differ in their composition and concentration, called A1 (chitosan/bovine collagen/beta-glycerolphosphate 8.5%w/w), A2 (chitosan/porcine collagen/beta-glycerolphosphate 8.5%), B1 (chitosan/bovine collagen/beta-glycerolphosphate 10.5%) and B2 (chitosan/porcine collagen/beta-glycerolphosphate 10.5%). A1 and A2 had a gelation time of 40 minutes, and B1 and B2 had a gelation time of 30 minutes at 37°C. Electron micrographs revealed a three-dimensional internal structure with interconnected pores for the four types of hydrogels. This facilitates the exchange of nutrients, oxygen, and the exit of metabolites, allowing to preserve a microenvironment suitable for cell proliferation. In the infrared spectra, it was possible to observe the interaction that occurs between the amides of polymeric compounds with the phosphate groups of beta-glycerolphosphate. Finally, the biocompatibility tests indicated that cells in contact with the hydrogel or with each of its precursors are not affected in their proliferation capacity for a period of 16 days. These results show the potential of the hydrogel to increase the cell survival rate in the cardiac cell therapies under investigation. Moreover, the results lay the foundations for its characterization and biological evaluation in both in vitro and in vivo models.

Keywords: cardiac cell therapy, cardiac ischemia, natural polymers, thermosensitive hydrogel

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Authors: Lolwa Alansari, Hanen Mrabet, Kholoud Khaled, Abdelhamid Azhaghdani, Sufia Athar, Aska Kaima, Zaineb Mhamdia, Zubaria Altaf, Almunzer Zakaria, Tamara Alshadafat

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Introduction: The Obstetrics and Gynecology Emergency Department at Alwakra Hospital has faced significant challenges, which have been further worsened by the impact of the COVID-19 pandemic. These challenges involve issues such as overcrowding, extended wait times, and a notable surge in demand for emergency care services. Moreover, prolonged waiting times have emerged as a primary factor contributing to situations where patients leave without receiving attention, known as left without being seen (LWBS), and unexpectedly abscond. Addressing the issue of insufficient patient mobility in the obstetrics and gynecology emergency department has brought about substantial improvements in patient care, healthcare administration, and overall departmental efficiency. These changes have not only alleviated overcrowding but have also elevated the quality of emergency care, resulting in higher patient satisfaction, better outcomes, and operational rewards. Methodology: The COVID-19 pandemic has served as a catalyst for substantial transformations in the obstetrics and gynecology emergency, aligning seamlessly with the strategic direction of Hamad Medical Corporation (HMC). The fundamental aim of this initiative is to revolutionize the operational efficiency of the OB-GYN ED. To accomplish this mission, a range of transformations has been initiated, focusing on essential areas such as digitizing systems, optimizing resource allocation, enhancing budget efficiency, and reducing overall costs. The project utilized the Plan-Do-Study-Act (PDSA) model, involving a diverse team collecting baseline data and introducing throughput improvements. Post-implementation data and feedback were analysed, leading to the integration of effective interventions into standard procedures. These interventions included optimized space utilization, real-time communication, bedside registration, technology integration, pre-triage screening, enhanced communication and patient education, consultant presence, and a culture of continuous improvement. These strategies significantly reduced waiting times, enhancing both patient care and operational efficiency. Results: Results demonstrated a substantial reduction in overall average waiting time, dropping from 35 to approximately 14 minutes by August 2023. The wait times for priority 1 cases have been reduced from 22 to 0 minutes, and for priority 2 cases, the wait times have been reduced from 32 to approximately 13.6 minutes. The proportion of patients spending less than 8 hours in the OB ED observation beds rose from 74% in January 2022 to over 98% in 2023. Notably, there was a remarkable decrease in LWBS and absconded patient rates from 2020 to 2023. Conclusion: The project initiated a profound change in the department's operational environment. Efficiency became deeply embedded in the unit's culture, promoting teamwork among staff that went beyond the project's original focus and had a positive influence on operations in other departments. This effectiveness not only made processes more efficient but also resulted in significant cost reductions for the hospital. These cost savings were achieved by reducing wait times, which in turn led to fewer prolonged patient stays and reduced the need for additional treatments. These continuous improvement initiatives have now become an integral part of the Obstetrics and Gynecology Division's standard operating procedures, ensuring that the positive changes brought about by the project persist and evolve over time.

Keywords: overcrowding, waiting time, person centered care, quality initiatives

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Authors: Karim Hrratha, Khaled Essalahb, Christophe Morellc, Henry Chermettec, Salima Boughdiria

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Among the carbon-carbon bond formation types, allylation of active methylene compounds with cyclic Baylis-Hillman (BH) alcohols is a reliable and widely used method. This reaction is a very attractive tool in organic synthesis of biological and biodiesel compounds. Thus, in view of an insistent and peremptory request for an efficient and straightly method for synthesizing the desired product, a thorough analysis of various aspects of the reaction processes is an important task. The product afforded by the reaction of active methylene with BH alcohols depends largely on the experimental conditions, notably on the catalyst properties. All experiments reported that catalysis is needed for this reaction type because of the poor ability of alcohol hydroxyl group to be as a suitable leaving group. Within the catalysts, several transition- metal based have been used such as palladium in the presence of acid or base and have been considered as reliable methods. Furthemore, acid catalysts such as BF3.OEt2, BiX3 (X= Cl, Br, I, (OTf)3), InCl3, Yb(OTf)3, FeCl3, p-TsOH and H-montmorillonite have been employed to activate the C-C bond formation through the alkylation of active methylene compounds. Interestingly a report of a smoothly process for the ability of 4-imethyaminopyridine(DMAP) to catalyze the allylation reaction of active methylene compounds with cyclic Baylis-Hillman (BH) alcohol appeared recently. However, the reaction mechanism remains ambiguous, since the C- allylation process leads to an unexpected product (noted P1), corresponding to a direct allylation instead of conjugate allylation, which involves the most electrophilic center according to the electron withdrawing group CO effect. The main objective of the present theoretical study is to better understand the role of the DMAP catalytic activity as well as the process leading to the end- product (P1) for the catalytic reaction of a cyclic BH alcohol with active methylene compounds. For that purpose, we have carried out computations of a set of active methylene compounds varying by R1 and R2 toward the same alcohol, and we have attempted to rationalize the mechanisms thanks to the acid–base approach, and conceptual DFT tools such as chemical potential, hardness, Fukui functions, electrophilicity index and dual descriptor, as these approaches have shown a good prediction of reactions products.The present work is then organized as follows: In a first part some computational details will be given, introducing the reactivity indexes used in the present work, then Section 3 is dedicated to the discussion of the prediction of the selectivity and regioselectivity. The paper ends with some concluding remarks. In this work, we have shown, through DFT method at the B3LYP/6-311++G(d,p) level of theory that: The allylation of active methylene compounds with cyclic BH alcohol is governed by orbital control character. Hence the end- product denoted P1 is generated by direct allylation.

Keywords: DFT calculation, gas phase pKa, theoretical mechanism, orbital control, charge control, Fukui function, transition state

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Authors: Mikko Salminen, Niklas Ravaja

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Emotional intelligence (EI) consists of skills for monitoring own emotions and emotions of others, skills for discriminating different emotions, and skills for using this information in thinking and actions. EI enhances, for example, work outcomes and organizational climate. We suggest that the role and manifestations of EI should also be studied in real leadership situations, especially during the emotional, social interaction. Leadership is essentially a process to influence others for reaching a certain goal. This influencing happens by managerial processes and computer-mediated communication (e.g. e-mail) but also by face-to-face, where facial expressions have a significant role in conveying emotional information. Persons with high EI are typically perceived more positively, and they have better social skills. We hypothesize, that during social interaction high EI enhances the ability to detect other’s emotional state and controlling own emotional expressions. We suggest, that emotionally intelligent leader’s experience less stress during social leadership situations, since they have better skills in dealing with the related emotional work. Thus the high-EI leaders would be more able to enjoy these situations, but also be more efficient in choosing appropriate expressions for building constructive dialogue. We suggest, that emotionally intelligent leaders show more positive emotional expressions than low-EI leaders. To study these hypotheses we observed performance review discussions of 40 leaders (24 female) with 78 (45 female) of their followers. Each leader held a discussion with two followers. Psychophysiological methods were chosen because they provide objective and continuous data from the whole duration of the discussions. We recorded sweating of the hands (electrodermal activation) by electrodes placed to the fingers of the non-dominant hand to assess the stress-related physiological arousal of the leaders. In addition, facial electromyography was recorded from cheek (zygomaticus major, activated during e.g. smiling) and periocular (orbicularis oculi, activated during smiling) muscles using electrode pairs placed on the left side of the face. Leader’s trait EI was measured with a 360 questionnaire, filled by each leader’s followers, peers, managers and by themselves. High-EI leaders had less sweating of the hands (p = .007) than the low-EI leaders. It is thus suggested that the high-EI leaders experienced less physiological stress during the discussions. Also, high scores in the factor “Using of emotions” were related to more facial muscle activation indicating positive emotional expressions (cheek muscle: p = .048; periocular muscle: p = .076, almost statistically significant). The results imply that emotionally intelligent managers are positively relaxed during s social leadership situations such as a performance review discussion. The current study also highlights the importance of EI in face-to-face social interaction, given the central role facial expressions have in interaction situations. The study also offers new insight to the biological basis of trait EI. It is suggested that the identification, forming, and intelligently using of facial expressions are skills that could be trained during leadership development courses.

Keywords: emotional intelligence, leadership, performance review discussion, psychophysiology, social interaction

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Authors: Claudiu Mazilu, Ion Robu, Radu Deju

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At the end of 2011 worldwide there were 124 power reactors shut down, from which: 16 fully decommissioned, 50 power reactors in a decommissioning process, 49 reactors in “safe enclosure mode”, 3 reactors “entombed”, for other 6 reactors it was not yet have specified the decommissioning strategy. The concrete radioactive waste that will be generated from dismantled structures of VVR-S nuclear research reactor from Magurele (e.g.: biological shield of the reactor core and hot cells) represents an estimated amount of about 70 tons. Until now the solid low activity radioactive waste (LLW) was pre-placed in containers and cementation with mortar made from cement and natural fine aggregates, providing a fill ratio of the container of approximately 50 vol. % for concrete. In this paper is presented an innovative technology in which radioactive concrete is crushed and the mortar made from recycled radioactive sand, cement, water and superplasticizer agent is poured in container with radioactive rubble (that is pre-placed in container) for cimentation. Is achieved a radioactive waste package in which the degree of filling of radioactive waste increases substantially. The tests were carried out on non-radioactive material because the radioactive concrete was not available in a good time. Waste concrete with maximum size of 350 mm were crushed in the first stage with a Liebhher type jaw crusher, adjusted to nominal size of 50 mm. Crushed concrete less than 50 mm was sieved in order to obtain useful sort for preplacement, 10 to 50 mm. The rest of the screening > 50 mm obtained from primary crushing of concrete was crushed in the second stage, with different working principles crushers at size < 2.5 mm, in order to produce recycled fine aggregate (sand) for the filler mortar and which fulfills the technical specifications proposed: –jaw crusher, Retsch type, model BB 100; –hammer crusher, Buffalo Shuttle model WA-12-H; presented a series of characteristics of recycled concrete aggregates by predefined class (the granulosity, the granule shape, the absorption of water, behavior to the Los Angeles test, the content of attached mortar etc.), most in comparison with characteristics of natural aggregates. Various mortar recipes were used in order to identify those that meet the proposed specification (flow-rate: 16-50s, no bleeding, min. 30N/mm2 compressive strength of the mortar after 28 days, the proportion of recycled sand used in mortar: min. 900kg/m3) and allow obtaining of the highest fill ratio for mortar. In order to optimize the mortars following compositional factors were varied: aggregate nature, water/cement (W/C) ratio, sand/cement (S/C) ratio, nature and proportion of additive. To confirm the results obtained on a small scale, it made an attempt to fill the mortar in a container that simulates the final storage drums. Was measured the mortar fill ratio (98.9%) compared with the results of laboratory tests and targets set out in the proposed specification. Although fill ratio obtained on the mock-up is lower by 0.8 vol. % compared to that obtained in the laboratory tests (99.7%), the result meets the specification criteria.

Keywords: characteristics, radioactive recycled concrete aggregate, mortars, fill ratio

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Authors: Danute Karcauskiene, Dalia Ambrazaitiene, Regina Skuodiene, Monika Vilkiene, Regina Repsiene, Ieva Jokubauskaite

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The main ambition for nowadays agriculture is to get the economically effective yield and to secure the soil ecological sustainability. According to the effect on the main soil quality indexes, tillage systems may be separated into two types, conventional and conservation tillage. The goal of this study was to determine the impact of conservation and conventional primary soil tillage methods and soil fertility improvement measures on soil properties and crop productivity. Methods: The soil of the experimental site is Dystric Glossic Retisol (WRB 2014) with texture of sandy loam. The trial was established in 2003 in the experimental field of crop rotation of Vėžaičiai Branch of Lithuanian Research Centre for Agriculture and Forestry. Trial factors and treatments: factor A- primary soil tillage in (autumn): deep ploughing (20-25cm), shallow ploughing (10-12cm), shallow ploughless tillage (8-10cm); factor B – soil fertility improvement measures: plant residues, plant residues + straw, green manure 1st cut + straw, farmyard manure 40tha-1 + straw. The four - course crop rotation consisted of red clover, winter wheat, spring rape and spring barley with undersown. Results: The tillage had no statistically significant effect on topsoil (0-10 cm) pHKCl level, it was 5.5 - 5.7. During all experiment period, the highest soil pHKCl level (5.65) was in the shallow ploughless tillage. The organic fertilizers particularly the biomass of grass and farmyard manure had tendency to increase the soil pHKCl. The content of plant - available phosphorus and potassium significantly increase in the shallow ploughing compared with others tillage systems. The farmyard manure increases those elements in whole arable layer. The dissolved organic carbon concentration was significantly higher in the 0 - 10 cm soil layer in the shallow ploughless tillage compared with deep ploughing. After the incorporation of clover biomass and farmyard manure the concentration of dissolved organic carbon increased in the top soil layer. During all experiment period the largest amount of water stable aggregates was determined in the soil where the shallow ploughless tillage was applied. It was by 12% higher compared with deep ploughing. During all experiment time, the soil moisture was higher in the shallow ploughing and shallow ploughless tillage (9-27%) compared to deep ploughing. The lowest emission of CO2 was determined in the deep ploughing soil. The highest rate of CO2 emission was in shallow ploughless tillage. The addition of organic fertilisers had a tendency to increase the CO2 emission, but there was no statistically significant effect between the different types of organic fertilisers. The crop yield was larger in the deep ploughing soil compared to the shallow and shallow ploughless tillage.

Keywords: reduced tillage, soil structure, soil pH, biological activity, crop productivity

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Authors: Varsha Puri, Sharon Hudson, Ian Kim

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Background: Incest-related childhood sexual abuse (IRCSA) is challenging to study due to the shame and secrecy experienced by its survivors. Ramifications of IRCSA worsen when it is unidentified, and interventions are not made. IRCSA perspectives are essential for future prevention and intervention strategies. However, there is limited understanding of this population’s experiences, perspectives, and long-term struggles. To date, research for IRCSA has utilized data from treatment programs and qualitative research with cohorts of 10-20 people, much of the data is from 10-40 years prior. Methods. In June 2018, an anonymous online survey was posted to multiple social media sites (e.g., Facebook IRCSA groups) and sexual abuse resource sites. Survey responses were collected for a year. The survey collected non-identifying demographics, IRCSA experiences, and outcomes data. Results: We obtained 1310 completed surveys. Demographics of all ages, racial backgrounds, financial backgrounds, and genders were obtained; the majority identified as white (81%) and female (76%). Childhood sexual abuse (CSA) started before the age of 6 in 49% and was endured for more than one year in 84% of respondents, and 39% reported ten or more years of abuse. CSA by multiple perpetrators occurred in 58%, while 8% had ten or more perpetrators. CSA by perpetrators under 21 years old was reported by 46%. Female perpetrators were reported by 28% of respondents. Fathers were the highest reported sexual abusers at 60%, and mothers were reported at 17%. Only 16% reported that at least one of their perpetrators was prosecuted for sexual abuse of a minor. Respondents confirmed that 54% of the time, they informed an adult of the abuse; only 2% agreed that “an intervention was made by the family that protected me.” A majority reported that IRCSA has negatively impacted their intimate/sexual relationships (96%) and mental health (96%). A majority reported negative impacts on biological family relationships (88%), physical health (73%), finances (59%), educational achievement (57%), and employment (56%). When asked about suffering from addiction, 85% of respondents answered yes. Prevention strategies selected most by respondents include early school education around CSA prevention (67%), removing the statute of limitations for reporting CSA (69%), and improved laws protecting IRCSA survivors (63%). Conclusion: The data document that IRCSA can be pervasive, and the dearth of intervention and support for survivors have major lasting impacts. Survivors have a unique and valuable perspective on what interventions are needed to prevent IRCSA and support survivors; their voice has long been unheard in crafting prevention and intervention policies and services. These results thus provide an important call to action from these critical stakeholders. Pediatricians should recognize that perpetrators can be pediatric patients, women, and parents. Pediatricians can advocate for more early CSA prevention education and policy changes that remove the statute of limitations for reporting CSA.

Keywords: incest, childhood sexual abuse, incest-related childhood sexual abuse, incest survivor

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Authors: Dimitra C. Banti, Gesthimani Liona, Petros Samaras, Manasis Mitrakas

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Municipal and industrial wastewaters are often treated biologically, by the activated sludge process (ASP). The ASP not only requires large aeration and sedimentation tanks, but also generates large quantities of excess sludge. An alternative technology is the membrane bioreactor (MBR), which replaces two stages of the conventional ASP—clarification and settlement—with a single, integrated biotreatment and clarification step. The advantages offered by the MBR over conventional treatment include reduced footprint and sludge production through maintaining a high biomass concentration in the bioreactor. Notwithstanding these advantages, the widespread application of the MBR process is constrained by membrane fouling. Fouling leads to permeate flux decline, making more frequent membrane cleaning and replacement necessary and resulting to increased operating costs. In general, membrane fouling results from the interaction between the membrane material and the components in the activated sludge liquor. The latter includes substrate components, cells, cell debris and microbial metabolites, such as Extracellular Polymeric Substances (EPS) and Sludge Microbial Products (SMPs). The challenge for effective MBR operation is to minimize the rate of Transmembrane Pressure (TMP) increase. This can be achieved by several ways, one of which is the addition of specific additives, that enhance the coagulation and flocculation of compounds, which are responsible for fouling, hence reducing biofilm formation on the membrane surface and limiting the fouling rate. In this project the effectiveness of a non-commercial composite coagulant was studied as an agent for fouling control in a lab scale MBR system consisting in two aerated tanks. A flat sheet membrane module with 0.40 um pore size was submerged into the second tank. The system was fed by50 L/d of municipal wastewater collected from the effluent of the primary sedimentation basin. The TMP increase rate, which is directly related to fouling growth, was monitored by a PLC system. EPS, MLSS and MLVSS measurements were performed in samples of mixed liquor; in addition, influent and effluent samples were collected for the determination of physicochemical characteristics (COD, BOD5, NO3-N, NH4-N, Total N and PO4-P). The coagulant was added in concentrations 2, 5 and 10mg/L during a period of 2 weeks and the results were compared with the control system (without coagulant addition). EPS fractions were extracted by a three stages physical-thermal treatment allowing the identification of Soluble EPS (SEPS) or SMP, Loosely Bound EPS (LBEPS) and Tightly Bound EPS (TBEPS). Proteins and carbohydrates concentrations were measured in EPS fractions by the modified Lowry method and Dubois method, respectively. Addition of 2 mg/L coagulant concentration did not affect SEPS proteins in comparison with control process and their values varied between 32 to 38mg/g VSS. However a coagulant dosage of 5mg/L resulted in a slight increase of SEPS proteins at 35-40 mg/g VSS while 10mg/L coagulant further increased SEPS to 44-48mg/g VSS. Similar results were obtained for SEPS carbohydrates. Carbohydrates values without coagulant addition were similar to the corresponding values measured for 2mg/L coagulant; the addition of mg/L coagulant resulted to a slight increase of carbohydrates SEPS to 6-7mg/g VSS while a dose of 10 mg/L further increased carbohydrates content to 9-10mg/g VSS. Total LBEPS and TBEPS, consisted of proteins and carbohydrates of LBEPS and TBEPS respectively, presented similar variations by the addition of the coagulant. Total LBEPS at 2mg/L dose were almost equal to 17mg/g VSS, and their values increased to 22 and 29 mg/g VSS during the addition of 5 mg/L and 10 mg/L of coagulant respectively. Total TBEPS were almost 37 mg/g VSS at a coagulant dose of 2 mg/L and increased to 42 and 51 mg/g VSS at 5 mg/L and 10 mg/L doses, respectively. Therefore, it can be concluded that coagulant addition could potentially affect microorganisms activities, excreting EPS in greater amounts. Nevertheless, EPS increase, mainly SEPS increase, resulted to a higher membrane fouling rate, as justified by the corresponding TMP increase rate. However, the addition of the coagulant, although affected the EPS content in the reactor mixed liquor, did not change the filtration process: an effluent of high quality was produced, with COD values as low as 20-30 mg/L.

Keywords: extracellular polymeric substances, MBR, membrane fouling, EPS

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Authors: K. K. Balavenkatraman, B. Bertschi, K. Bigot, A. Grevot, A. Doelemeyer, S. D. Chibout, A. Wolf, F. Pognan, N. Manevski, O. Kretz, P. Swart, K. Litherland, J. Ashton-Chess, B. Ling, R. Wettstein, D. J. Schaefer

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Skin toxicity is poorly detected during preclinical studies, and drug-induced side effects in humans such as rashes, hyperplasia or more serious events like bullous pemphigus or toxic epidermal necrolysis represent an important hurdle for clinical development. In vitro keratinocyte-based epidermal skin models are suitable for the detection of chemical-induced irritancy, but do not recapitulate the biological complexity of full skin and fail to detect potential serious side-effects. Normal healthy skin explants may represent a valuable complementary tool, having the advantage of retaining the full skin architecture and the resident immune cell diversity. This study investigated several conditions for the maintenance of good morphological structure after several days of culture and the retention of phase II metabolism for 24 hours in skin explants in vitro. Human skin samples were collected with informed consent from patients undergoing plastic surgery and immediately transferred and processed in our laboratory by removing the underlying dermal fat. Punch biopsies of 4 mm diameter were cultured in an air-liquid interface using transwell filters. Different cultural conditions such as the effect of calcium, temperature and cultivation media were tested for a period of 14 days and explants were histologically examined after Hematoxylin and Eosin staining. Our results demonstrated that the use of Williams E Medium at 32°C maintained the physiological integrity of the skin for approximately one week. Upon prolonged incubation, the upper layers of the epidermis become thickened and some dead cells are present. Interestingly, these effects were prevented by addition of EGFR inhibitors such as Afatinib or Erlotinib. Phase II metabolism of the skin such as glucuronidation (4-methyl umbeliferone), sulfation (minoxidil), N-acetyltransferase (p-toluidene), catechol methylation (2,3-dehydroxy naphthalene), and glutathione conjugation (chlorodinitro benzene) were analyzed by using LCMS. Our results demonstrated that the human skin explants possess metabolic activity for a period of at least 24 hours for all the substrates tested. A time course for glucuronidation with 4-methyl umbeliferone was performed and a linear correlation was obtained over a period of 24 hours. Longer-term culture studies will indicate the possible evolution of such metabolic activities. In summary, these results demonstrate that human skin explants maintain a normal structure for several days in vitro and are metabolically active for at least the first 24 hours. Hence, with further characterisation, this model may be suitable for the study of drug-induced toxicity.

Keywords: human skin explant, phase II metabolism, epidermal growth factor receptor, toxicity

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Authors: L. P. Gomes, G. F. Araújo, Y. M. L. Cordeiro, C. T. Andrade, E. M. Del Aguila, V. M. F. Paschoalin

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The application of nanoscale materials and nanostructures is an emerging area, these since materials may provide solutions to technological and environmental challenges in order to preserve the environment and natural resources. To reach this goal, the increasing demand must be accompanied by 'green' synthesis methods. Chitosan is a natural, nontoxic, biopolymer derived by the deacetylation of chitin and has great potential for a wide range of applications in the biological and biomedical areas, due to its biodegradability, biocompatibility, non-toxicity and versatile chemical and physical properties. Chitosan also presents high antimicrobial activities against a wide variety of pathogenic and spoilage microorganisms. Ultrasonication is a common tool for the preparation and processing of polymer nanoparticles. It is particularly effective in breaking up aggregates and in reducing the size and polydispersity of nanoparticles. High-intensity ultrasonication has the potential to modify chitosan molecular weight and, thus, alter or improve chitosan functional properties. The aim of this study was to evaluate the influence of sonication intensity and time on the changes of commercial chitosan characteristics, such as molecular weight and its potential antibacterial activity against Gram-negative bacteria. The nanoparticles (NPs) were produced from two commercial chitosans, of medium molecular weight (CS-MMW) and low molecular weight (CS-LMW) from Sigma-Aldrich®. These samples (2%) were solubilized in 100 mM sodium acetate pH 4.0, placed on ice and irradiated with an ultrasound SONIC ultrasonic probe (model 750 W), equipped with a 1/2" microtip during 30 min at 4°C. It was used on constant duty cycle and 40% amplitude with 1/1s intervals. The ultrasonic degradation of CS-MMW and CS-LMW were followed up by means of ζ-potential (Brookhaven Instruments, model 90Plus) and dynamic light scattering (DLS) measurements. After sonication, the concentrated samples were diluted 100 times and placed in fluorescence quartz cuvettes (Hellma 111-QS, 10 mm light path). The distributions of the colloidal particles were calculated from the DLS and ζ-potential are measurements taken for the CS-MMW and CS-LMW solutions before and after (CS-MMW30 and CS-LMW30) sonication for 30 min. Regarding the results for the chitosan sample, the major bands can be distinguished centered at Radius hydrodynamic (Rh), showed different distributions for CS-MMW (Rh=690.0 nm, ζ=26.52±2.4), CS-LMW (Rh=607.4 and 2805.4 nm, ζ=24.51±1.29), CS-MMW30 (Rh=201.5 and 1064.1 nm, ζ=24.78±2.4) and CS-LMW30 (Rh=492.5, ζ=26.12±0.85). The minimal inhibitory concentration (MIC) was determined using different chitosan samples concentrations. MIC values were determined against to E. coli (106 cells) harvested from an LB medium (Luria-Bertani BD™) after 18h growth at 37 ºC. Subsequently, the cell suspension was serially diluted in saline solution (0.8% NaCl) and plated on solid LB at 37°C for 18 h. Colony-forming units were counted. The samples showed different MICs against E. coli for CS-LMW (1.5mg), CS-MMW30 (1.5 mg/mL) and CS-LMW30 (1.0 mg/mL). The results demonstrate that the production of nanoparticles by modification of their molecular weight by ultrasonication is simple to be performed and dispense acid solvent addition. Molecular weight modifications are enough to provoke changes in the antimicrobial potential of the nanoparticles produced in this way.

Keywords: antimicrobial agent, chitosan, green production, nanoparticles

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Authors: Moustafa Osman Mohammed

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This paper introduces topological order in descried social systems starting with the original concept of autopoiesis by biologists and scientists, including the modification of general systems based on socialized medicine. Topological order is important in describing the physical systems for exploiting optical systems and improving photonic devices. The stats of topological order have some interesting properties of topological degeneracy and fractional statistics that reveal the entanglement origin of topological order, etc. Topological ideas in photonics form exciting developments in solid-state materials, that being; insulating in the bulk, conducting electricity on their surface without dissipation or back-scattering, even in the presence of large impurities. A specific type of autopoiesis system is interrelated to the main categories amongst existing groups of the ecological phenomena interaction social and medical sciences. The hypothesis, nevertheless, has a nonlinear interaction with its natural environment 'interactional cycle' for exchange photon energy with molecules without changes in topology. The engineering topology of a biosensor is based on the excitation boundary of surface electromagnetic waves in photonic band gap multilayer films. The device operation is similar to surface Plasmonic biosensors in which a photonic band gap film replaces metal film as the medium when surface electromagnetic waves are excited. The use of photonic band gap film offers sharper surface wave resonance leading to the potential of greatly enhanced sensitivity. So, the properties of the photonic band gap material are engineered to operate a sensor at any wavelength and conduct a surface wave resonance that ranges up to 470 nm. The wavelength is not generally accessible with surface Plasmon sensing. Lastly, the photonic band gap films have robust mechanical functions that offer new substrates for surface chemistry to understand the molecular design structure and create sensing chips surface with different concentrations of DNA sequences in the solution to observe and track the surface mode resonance under the influences of processes that take place in the spectroscopic environment. These processes led to the development of several advanced analytical technologies: which are; automated, real-time, reliable, reproducible, and cost-effective. This results in faster and more accurate monitoring and detection of biomolecules on refractive index sensing, antibody-antigen reactions with a DNA or protein binding. Ultimately, the controversial aspect of molecular frictional properties is adjusted to each other in order to form unique spatial structure and dynamics of biological molecules for providing the environment mutual contribution in investigation of changes due to the pathogenic archival architecture of cell clusters.

Keywords: autopoiesis, photonics systems, quantum topology, molecular structure, biosensing

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Authors: Jie Lin, Yiwen Pan, Li Zhang, Zhangyong Xia

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Atherosclerosis is a chronic inflammatory disease characterized by the accumulation of lipids, immune cells, and extracellular matrix in the arterial walls. This pathological process can lead to the formation of plaques that can obstruct blood flow and trigger various cardiovascular diseases such as heart attack and stroke. The underlying molecular mechanisms still remain unclear, although many studies revealed the dysfunction of endothelial cells, recruitment and activation of monocytes and macrophages, and the production of pro-inflammatory cytokines and chemokines in atherosclerosis. This study aimed to identify hub genes involved in the progression of atherosclerosis and to analyze their biological function in silico, thereby enhancing our understanding of the disease’s molecular mechanisms. Through the analysis of microarray data, we examined the gene expression in media and neo-intima from plaques, as well as distant macroscopically intact tissue, across a cohort of 32 hypertensive patients. Initially, 112 differentially expressed genes (DEGs) were identified. Subsequent immune infiltration analysis indicated a predominant presence of 27 immune cell types in the atherosclerosis group, particularly noting an increase in monocytes and macrophages. In the Weighted gene co-expression network analysis (WGCNA), 10 modules with a minimum of 30 genes were defined as key modules, with blue, dark, Oliver green and sky-blue modules being the most significant. These modules corresponded respectively to monocyte, activated B cell, and activated CD4 T cell gene patterns, revealing a strong morphological-genetic correlation. From these three gene patterns (modules morphology), a total of 2509 key genes (Gene Significance >0.2, module membership>0.8) were extracted. Six hub genes (CD36, DPP4, HMOX1, PLA2G7, PLN2, and ACADL) were then identified by intersecting 2509 key genes, 102 DEGs with lipid-related genes from the Genecard database. The bio-functional analysis of six hub genes was estimated by a robust classifier with an area under the curve (AUC) of 0.873 in the ROC plot, indicating excellent efficacy in differentiating between the disease and control group. Moreover, PCA visualization demonstrated clear separation between the groups based on these six hub genes, suggesting their potential utility as classification features in predictive models. Protein-protein interaction (PPI) analysis highlighted DPP4 as the most interconnected gene. Within the constructed key gene-drug network, 462 drugs were predicted, with ursodeoxycholic acid (UDCA) being identified as a potential therapeutic agent for modulating DPP4 expression. In summary, our study identified critical hub genes implicated in the progression of atherosclerosis through comprehensive bioinformatic analyses. These findings not only advance our understanding of the disease but also pave the way for applying similar analytical frameworks and predictive models to other diseases, thereby broadening the potential for clinical applications and therapeutic discoveries.

Keywords: atherosclerosis, hub genes, drug prediction, bioinformatics

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Authors: Rawan Ashraf, Ahmed E. Gomaa, Gehan Safwat, Ayman Diab

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Background: Three-dimensional (3D) bio-printing has become a versatile and powerful method for generating a variety of biological constructs, including bone or extracellular matrix scaffolds endo- or epithelial, muscle tissue, as well as organoids. Aim of the study: Fabricate a low cost DIY 3D bio-printer to produce 3D bio-printed products such as anti-microbial packaging or multi-organs on chips. We demonstrate the alignment between two types of 3D printer technology (3D Bio-printer and DLP) on Multi-organ-on-a-chip (multi-OoC) devices fabrication. Methods: First, Design and Fabrication of the Syringe Unit for Modification of an Off-the-Shelf 3D Printer, then Preparation of Hydrogel based on natural polymers Sodium Alginate and Gelatin, followed by acquisition of the cell suspension, then modeling the desired 3D structure. Preparation for 3D printing, then Cell-free and cell-laden hydrogels went through the printing process at room temperature under sterile conditions and finally post printing curing process and studying the printed structure regards physical and chemical characteristics. The hard scaffold of the Organ on chip devices was designed and fabricated using the DLP-3D printer, following similar approaches as the Microfluidics system fabrication. Results: The fabricated Bio-Ink was based onHydrogel polymer mix of sodium alginate and gelatin 15% to 0.5%, respectively. Later the 3D printing process was conducted using a higher percentage of alginate-based hydrogels because of it viscosity and the controllable crosslinking, unlike the thermal crosslinking of Gelatin. The hydrogels were colored to simulate the representation of two types of cells. The adaption of the hard scaffold, whether for the Microfluidics system or the hard-tissues, has been acquired by the DLP 3D printers with fabricated natural bioactive essential oils that contain antimicrobial activity, followed by printing in Situ three complex layers of soft-hydrogel as a cell-free Bio-Ink to simulate the real-life tissue engineering process. The final product was a proof of concept for a rapid 3D cell culturing approaches that uses an engineered hard scaffold along with soft-tissues, thus, several applications were offered as products of the current prototype, including the Organ-On-Chip as a successful integration between DLP and 3D bioprinter. Conclusion: Multiple designs for the organ-on-a-chip (multi-OoC) devices have been acquired in our study with main focus on the low cost fabrication of such technology and the potential to revolutionize human health research and development. We describe circumstances in which multi-organ models are useful after briefly examining the requirement for full multi-organ models with a systemic component. Following that, we took a look at the current multi-OoC platforms, such as integrated body-on-a-chip devices and modular techniques that use linked organ-specific modules.

Keywords: 3d bio-printer, hydrogel, multi-organ on chip, bio-inks

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Authors: Rikke Lybæk, Tyge Kjær

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The Baltic Sea is suffering from nitrogen and phosphorus pollution, which causes eutrophication of the maritime environment and hence threatens the biodiversity of the Baltic Sea area. The intensified quantity of nutrients in the water has created challenges with the growth of seaweed being discarded on beaches around the sea. The cast seaweed has led to odor problems hampering the use of beach areas around the Bay of Køge in Denmark. This is the case in, e.g., Solrød Municipality, where recreational activities have been disrupted when cast seaweed pile up on the beach. Initiatives have, however, been introduced within the municipality to remove the cast seaweed from the beach and utilize it for renewable energy production at the nearby Solrød Biogas Plant, thus being co-digested with animal manure for power and heat production. This paper investigates which type of technology application’s have been applied in the effort to optimize the collection of cast seaweed, and will further reveal, how the seaweed has been pre-treated at the biogas plant to be utilized for energy production the most efficient, hereunder the challenges connected with the content of sand. Heavy metal contents in the seaweed and how it is managed will also be addressed, which is vital as the digestate is utilized as soil fertilizer on nearby farms. Finally, the paper will outline the energy production scheme connected to the use of seaweed as feedstock for biogas production, as well as the amount of nitrogen-rich fertilizer produced. The theoretical approach adopted in the paper relies on the thinking of Circular Bio-Economy, where biological materials are cascaded and re-circulated etc., to increase and extend their value and usability. The data for this research is collected as part of the EU Interreg project “Cluster On Anaerobic digestion, environmental Services, and nuTrients removAL” (COASTAL Biogas), 2014-2020. Data gathering consists of, e.g., interviews with relevant stakeholders connected to seaweed collection and operation of the biogas plant in Solrød Municipality. It further entails studies of progress and evaluation reports from the municipality, analysis of seaweed digestion results from scholars connected to the research, as well as studies of scientific literature to supplement the above. Besides this, observations and photo documentation have been applied in the field. This paper concludes, among others, that the seaweed harvester technology currently adopted is functional in the maritime environment close to the beachfront but inadequate in collecting seaweed directly on the beach. New technology hence needs to be developed to increase the efficiency of seaweed collection. It is further concluded that the amount of sand transported to Solrød Biogas Plant with the seaweed continues to pose challenges. The seaweed is pre-treated for sand in a receiving tank with a strong stirrer, washing off the sand, which ends at the bottom of the tank where collected. The seaweed is then chopped by a macerator and mixed with the other feedstock. The wear down of the receiving tank stirrer and the chopper are, however, significant, and new methods should be adopted.

Keywords: biogas, circular bio-economy, Denmark, maritime technology, cast seaweed, solrød municipality

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