Search results for: mitochondrial membrane

Authors: Majid Ali, Xinfang Jin, Victor Eniola, Henning Hoene

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The performance of proton exchange membrane (PEM) fuel cells is sensitive to several factors, including power fluctuations, hydrogen utilization, and the quality orientation of the fuel cell stack. In this study, we investigate the impact of these factors on the performance of a PEM fuel cell. We start by analyzing the power fluctuations that are typical in renewable energy systems and their effects on the 50 Watt fuel cell's performance. Next, we examine the hydrogen utilization rate (0-1000 mL/min) and its impact on the cell's efficiency and durability. Finally, we investigate the quality orientation (three different positions) of the fuel cell stack, which can significantly affect the cell's lifetime and overall performance. The basis of our analysis is the utilization of experimental results, which have been further validated by comparing them with simulations and manufacturer results. Our results indicate that power fluctuations can cause significant variations in the fuel cell's voltage and current, leading to a reduction in its performance. Moreover, we show that increasing the hydrogen utilization rate beyond a certain threshold can lead to a decrease in the fuel cell's efficiency. Finally, our analysis demonstrates that the orientation of the fuel cell stack can affect its performance and lifetime due to non-uniform distribution of reactants and products. In summary, our study highlights the importance of considering power fluctuations, hydrogen utilization, and quality orientation in designing and optimizing PEM fuel cell systems. The findings of this study can be useful for researchers and engineers working on the development of fuel cell systems for various applications, including transportation, stationary power generation, and portable devices.

Keywords: fuel cell, proton exchange membrane, renewable energy, power fluctuation, experimental

Procedia PDF Downloads 135

Authors: Leila Karshenas, Hamid Reza Khodaei, Behnaz Mahdavi

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We know that nitric oxide (NO) plays an important role in all sexual activities of animals. It is made in body from NO synthase enzyme and L-arginin molecule. NO can bound with sulfur-iron complexes and because production of steroid sexual hormones is related to enzymes which have this complex, NO can change the activity of these enzymes. NO affects many cells including endothelial cells of veins, macrophages and mast cells. These cells are found in testis leydig cells and therefore are important source of NO in testis tissue. Minimizing damages to sperm at the time of sperm freezing and thawing is really important. The goal of this study was to determine the function of NO before freezing and its effects on quality and viability of sperms after thawing and incubation. 4 Holstein bulls were selected from the age of 4, and artificial insemination was done for 3 weeks (2 times a week). Treatments were 0, 10, 50 and 100 nm of sodium nitroprusside (SNP). Data analysis was performed by SAS98 program. Also, mean comparison was done using Duncan's multiple ranges test (P<0.05). Concentrations used was found to increase motility and viability of spermatozoa at 1, 2 and 3 hours after thawing significantly (P<0.05), but there was no significant difference at zero time. SNP levels reduced the amount of lipid peroxidation in sperm membrane, increased acrosome health and improved sample membranes especially in 50 and 100 nm treatments. According to results, adding SNP to semen diluents increases motility and viability of spermatozoa. Also, it reduces lipid peroxidation in sperm membrane and improves sperm function.

Keywords: sperm motility, nitric oxide, lipid peroxidation, spermatozoa

Procedia PDF Downloads 359

Authors: Azza A. Ali, Doaa M. Abd El-Latif, Amany M. Gad, Yasser M. A. Elnahas, Karema Abu-Elfotuh

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Background: Exposure to Aluminium (Al) has been increased recently. It is found in food products, food additives, drinking water, cosmetics and medicines. Chronic consumption of Al causes oxidative stress and has been implicated in several chronic disorders. Liver is considered as the major site for detoxification while kidney is involved in the elimination of toxic substances and is a target organ of metal toxicity. Social isolation (SI) or protein malnutrition (PM) also causes oxidative stress and has negative impact on Al-induced nephrotoxicity as well as hepatotoxicity. Coenzyme Q10 (CoQ10) is a powerful intracellular antioxidant with mitochondrial membrane stabilizing ability while wheat grass is a natural product with antioxidant, anti-inflammatory and different protective activities, cocoa is also potent antioxidants and can protect against many diseases. They provide different degrees of protection from the impact of oxidative stress. Objective: To study the impact of social isolation together with Protein malnutrition on nephro- and hepato-toxicity induced by chronic Al exposure in rats as well as to investigate the postulated protection using a combination of Co Q10, wheat grass and cocoa. Methods: Eight groups of rats were used; four served as protected groups and four as un-protected. Each of them received daily for five weeks AlCl3 (70 mg/kg, IP) for Al-toxicity model groups except one group served as control. Al-toxicity model groups were divided to Al-toxicity alone, SI- associated PM (10% casein diet) and Al- associated SI&PM groups. Protection was induced by oral co-administration of CoQ10 (200mg/kg), wheat grass (100mg/kg) and cocoa powder (24mg/kg) combination together with Al. Biochemical changes in total bilirubin, lipids, cholesterol, triglycerides, glucose, proteins, creatinine and urea as well as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate deshydrogenase (LDH) were measured in serum of all groups. Specimens of kidney and liver were used for assessment of oxidative parameters (MDA, SOD, TAC, NO), inflammatory mediators (TNF-α, IL-6β, nuclear factor kappa B (NF-κB), Caspase-3) and DNA fragmentation in addition to evaluation of histopathological changes. Results: SI together with PM severely enhanced nephro- and hepato-toxicity induced by chronic Al exposure. Co Q10, wheat grass and cocoa combination showed clear protection against hazards of Al exposure either alone or when associated with SI&PM. Their protection were indicated by the significant decrease in Al-induced elevations in total bilirubin, lipids, cholesterol, triglycerides, glucose, creatinine and urea levels as well as ALT, AST, ALP, LDH. Liver and kidney of the treated groups also showed significant decrease in MDA, NO, TNF-α, IL-6β, NF-κB, caspase-3 and DNA fragmentation, together with significant increase in total proteins, SOD and TAC. Biochemical results were confirmed by the histopathological examinations. Conclusion: SI together with PM represents a risk factor in enhancing nephro- and hepato-toxicity induced by Al in rats. CoQ10, wheat grass and cocoa combination provide clear protection against nephro- and hepatotoxicity as well as the consequent degenerations induced by chronic Al-exposure even when associated with the risk of SI together with PM.

Keywords: aluminum, nephrotoxicity, hepatotoxicity, isolation and protein malnutrition, coenzyme Q10, wheatgrass, cocoa, nutrients combinations

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Authors: Ayesha Imtiaz, Darlina Md. Naim

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Family Nemipteridae is among the most abundantly distributed family in Malaysian fish markets due to its high contribution to landing sites of Malaysia. Using an advanced molecular approach that used two mitochondrial (Cytochrome oxidase c I and Cytochrome oxidase b) and one nuclear gene (Recombination activating gene, RAGI) to expose cryptic diversity and phylogenetic relationships among commercially important species of family Nemipteridae. Our research covered all genera (including 31 species out total 45 species) of family Nemipteridae, distributed in Malaysia. We also found certain type of geographical barriers in the South China sea that reduces dispersal and stops a few species to intermix. Northside of the South China Sea (near Vietnam) does not allow genetic diversity to mix with the Southern side of the South China sea (Sarawak) and reduces dispersal. Straits of Malacca reduce the intermixing genetic diversity of South China Sea and the Indian Ocean.

Keywords: Nemipteridae, RAG I, south east Asia, Malaysia

Procedia PDF Downloads 143

Authors: Laura Canagueral-Pellice, Antonio Munar-Frau, Adaia Valls-Ontanon, Joao Carames, Federico Hernandez-Alfaro, Jordi Caballe-Serrano

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Objective: Guided bone regeneration (GBR) aims to replace the missing bone with a new structure to achieve long-term stability of rehabilitations. The aim of the present systematic review and meta-analysis is to determine the effect of barrier membranes on histological outcomes after GBR procedures. Moreover, the effect of the grafting material and tissue gain were analyzed. Materials & methods: Two independent reviewers performed an electronic search in Pubmed and Scopus, identifying all eligible publications up to March 2020. Only randomized controlled trials (RCTs) assessing a histological analysis of augmented areas were included. Results: A total of 6 publications were included for the present systematic review. A total of 110 biopsied sites were analysed; 10 corresponded to vertical bone augmentation procedures, whereas 100 analysed horizontal regeneration procedures. A mean tissue gain of 3 ± 1.48mm was obtained for horizontal defects. Histological assessment of new bone formation, residual particle and sub-epithelial connective tissue (SCT) was reported. The four main barrier membranes used were natural collagen membranes, e-PTFE, polylactic resorbable membranes and acellular dermal matrix membranes (AMDG). The analysis demonstrated that resorbable membranes result in higher values of new bone formation and lower values of residual particles and SCT. Xenograft resulted in lower new bone formation compared to allograft; however, no statistically significant differences were observed regarding residual particle and SCT. Overall, regeneration procedures adding autogenous bone, plasma derivate or growth factors achieved in general greater new bone formation and tissue gain. Conclusions: There is limited evidence favoring the effect of a certain type of barrier membrane in GBR. Data needs to be evaluated carefully; however, resorbable membranes are correlated with greater new bone formation values, especially when combined with allograft materials and/or the addition of autogenous bone, platelet reach plasma (PRP) or growth factors in the regeneration area. More studies assessing the histological outcomes of different GBR protocols and procedures testing different biomaterials are needed to maximize the clinical and histological outcomes in bone regeneration science.

Keywords: barrier membrane, graft material, guided bone regeneration, implant surgery, histology

Procedia PDF Downloads 152

Authors: Thiago E. Goto, Carla C. Lopes, Helena B. Nader, Anielle C. A. Silva, Noelio O. Dantas, José R. Siqueira Jr., Luciano Caseli

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Quantum dots (QD) are semiconductor nanocrystals that can be employed in biological research as a tool for fluorescence imagings, having the potential to expand in vivo and in vitro analysis as cancerous cell biomarkers. Particularly, cadmium selenide (CdSe) magic-sized quantum dots (MSQDs) exhibit stable luminescence that is feasible for biological applications, especially for imaging of tumor cells. For these facts, it is interesting to know the mechanisms of action of how such QDs mark biological cells. For that, simplified models are a suitable strategy. Among these models, Langmuir films of lipids formed at the air-water interface seem to be adequate since they can mimic half a membrane. They are monomolecular films formed at liquid-gas interfaces that can spontaneously form when organic solutions of amphiphilic compounds are spread on the liquid-gas interface. After solvent evaporation, the monomolecular film is formed, and a variety of techniques, including tensiometric, spectroscopic and optic can be applied. When the monolayer is formed by membrane lipids at the air-water interface, a model for half a membrane can be inferred where the aqueous subphase serve as a model for external or internal compartment of the cell. These films can be transferred to solid supports forming the so-called Langmuir-Blodgett (LB) films, and an ampler variety of techniques can be additionally used to characterize the film, allowing for the formation of devices and sensors. With these ideas in mind, the objective of this work was to investigate the specific interactions of CdSe MSQDs with tumorigenic and non-tumorigenic cells using Langmuir monolayers and LB films of lipids and specific cell extracts as membrane models for diagnosis of cancerous cells. Surface pressure-area isotherms and polarization modulation reflection-absorption spectroscopy (PM-IRRAS) showed an intrinsic interaction between the quantum dots, inserted in the aqueous subphase, and Langmuir monolayers, constructed either of selected lipids or of non-tumorigenic and tumorigenic cells extracts. The quantum dots expanded the monolayers and changed the PM-IRRAS spectra for the lipid monolayers. The mixed films were then compressed to high surface pressures and transferred from the floating monolayer to solid supports by using the LB technique. Images of the films were then obtained with atomic force microscopy (AFM) and confocal microscopy, which provided information about the morphology of the films. Similarities and differences between films with different composition representing cell membranes, with or without CdSe MSQDs, was analyzed. The results indicated that the interaction of quantum dots with the bioinspired films is modulated by the lipid composition. The properties of the normal cell monolayer were not significantly altered, whereas for the tumorigenic cell monolayer models, the films presented significant alteration. The images therefore exhibited a stronger effect of CdSe MSQDs on the models representing cancerous cells. As important implication of these findings, one may envisage for new bioinspired surfaces based on molecular recognition for biomedical applications.

Keywords: biomembrane, langmuir monolayers, quantum dots, surfaces

Procedia PDF Downloads 196

Authors: Ayodeji Sowale, Athanasios Kolios, Beatriz Fidalgo, Tosin Somorin, Aikaterini Anastasopoulou, Alison Parker, Leon Williams, Ewan McAdam, Sean Tyrrel

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Recently, energy recovery systems are thriving and raising attention in the power generation sector, due to the request for cleaner forms of energy that are friendly and safe for the environment. This has created an avenue for cogeneration, where Combined Heat and Power (CHP) technologies have been recognised for their feasibility, and use in homes and small-scale businesses. The efficiency of combustors and the advantages of the free piston Stirling engines over other conventional engines in terms of output power and efficiency, have been observed and considered. This study presents the numerical analysis of a micro-combustor with a free piston Stirling engine in an integrated model of a Nano Membrane Toilet (NMT) unit. The NMT unit will use the micro-combustor to produce waste heat of high energy content from the combustion of human waste and the heat generated will power the free piston Stirling engine which will be connected to a linear alternator for electricity production. The thermodynamic influence of the combustor on the free piston Stirling engine was observed, based on the heat transfer from the flue gas to working gas of the free piston Stirling engine. The results showed that with an input of 25 MJ/kg of faecal matter, and flue gas temperature of 773 K from the micro-combustor, the free piston Stirling engine generates a daily output power of 428 W, at thermal efficiency of 10.7% with engine speed of 1800 rpm. An experimental investigation into the integration of the micro-combustor and free piston Stirling engine with the NMT unit is currently underway.

Keywords: free piston stirling engine, micro-combustor, nano membrane toilet, thermodynamics

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Authors: Suhair Saleh, Ahmad Aljaberi

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Cationic lipid-mediated delivery of nucleic acids represents an exciting approach for developing therapeutically realistic gene medicines. Elucidation of the molecular and formulation requirements for efficient lipofection is a prerequisite to enhance the biological activity of such delivery systems. To this end, the in vitro lipofection activity of the ionizable asymmetric 1,2-dialkoylamidopropane-based derivatives bearing single primary amine group as the cationic head group was evaluated. The electrostatic interactions of these cationic lipids with plasmid DNA in physiologically relevant medium were investigated by means of gel electrophoresis retardation and Eth-Br quenching assays. The effect of the presence of the helper lipid on these interactions was evaluated. The physicochemical properties of these lipids in terms of bilayer fluidity and extent of ionization were investigated using fluorescence anisotropy and surface potential techniques, respectively. The results showed that only the active lipid, 1,2lmp[5], existed in a liquid crystalline state at physiological temperature. Moreover, the extent of ionization of this lipid in assemblies was significantly higher that it's saturated analogues. Inclusion of the helper lipid DOPE improved the encapsulation and association between 1,2lmp[5] and plasmid DNA, which was reflected by the significant boost of lipofection activity of the 1,2lmp[5]/DOPE formulation as compared to the lipid alone. In conclusion, membrane fluidity and sufficient protonation of ionizable cationic lipid are required for efficient association and encapsulation of plasmid DNA and promoting improved in vitro lipofection activity.

Keywords: cationic lipids, gene delivery, lipofection, membrane fluidity, helper lipids, surface potential

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Authors: Evgen Prokhorov, Gabriel Luna-Barcenas

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The polysaccharide chitosan (CS) is an attractive biopolymer for the stabilization of several nanoparticles in acidic aqueous media. This is due in part to the presence of abundant primary NH2 and OH groups which may lead to steric or chemical stabilization. Applications of most CS nanocomposites are based upon the interaction of high surface area nanoparticles (NPs) with different substance. Therefore, agglomeration of NPs leads to decreasing effective surface area such that it may decrease the efficiency of nanocomposites. The aim of this work is to measure nanocomposite’s electrical conductivity phenomena that will allow one to formulate optimal concentrations of conductivity NPs in CS-based nanocomposites. Additionally, by comparing the efficiency of such nanocomposites, one can guide applications in the biomedical (antibacterial properties and tissue regeneration) and sensor fields (detection of copper and nitrate ions in aqueous solutions). It was shown that the best antibacterial (CS-AgNPs, CS-AgNPs-carbon nanotubes) and would healing properties (CS-AuNPs) are observed in nanocomposites with concentrations of NPs near the percolation threshold. In this regard, the best detection limit in potentiometric and impedimetric sensors for detection of copper ions (using CS-AuNPs membrane) and nitrate ions (using CS-clay membrane) in aqueous solutions have been observed for membranes with concentrations of NPs near percolation threshold. It is well known that at the percolation concentration of NPs an abrupt increasing of conductivity is observed due to the presence of physical contacts between NPs; above this concentration, agglomeration of NPs takes place such that a decrease in the effective surface and performance of nanocomposite appear. The obtained relationship between electrical percolation threshold and performance of polymer nanocomposites with conductivity NPs is important for the design and optimization of polymer-based nanocomposites for different applications.

Keywords: chitosan, conductivity nanoparticles, percolation threshold, polymer nanocomposites

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Authors: Ibrahim Rassoul, E-K. Si Ahmed

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The growth, deformation, and detachment of fluid droplets adherent to solid substrates is a problem of fundamental interest with numerous practical applications. Specific interest in this proposal is the problem of a droplet on a fibrous, hydrophobic substrate subjected to body or external forces (gravity, convection). The past decade has seen tremendous advances in proton exchange membrane fuel cell (PEMFC) technology. However, there remain many challenges to bring commercially viable stationary PEMFC products to the market. PEMFCs are increasingly emerging as a viable alternative clean power source for automobile and stationary applications. Before PEMFCs can be employed to power automobiles and homes, several key technical challenges must be properly addressed. One technical challenge is elucidating the mechanisms underlying water transport in and removal from PEMFCs. On the one hand, sufficient water is needed in the polymer electrolyte membrane or PEM to maintain sufficiently high proton conductivity. On the other hand, too much liquid water present in the cathode can cause 'flooding' (that is, pore space is filled with excessive liquid water) and hinder the transport of the oxygen reactant from the gas flow channel (GFC) to the three-phase reaction sites. The aim of this work is to investigate the stability of a liquid water droplet emerging form a GDL pore, to gain fundamental insight into the instability process leading to detachment. The approach will combine analytical and numerical modeling with experimental visualization and measurements.

Keywords: polymer electrolyte fuel cell, water droplet, gas diffusion layer, contact angle, surface tension

Procedia PDF Downloads 251

Authors: Hafiza Mamoona Khalid, Afshan Mujahid, Asif Ali, Asim Laeeq Khan, Mahmood Saleem, Rafael M. Santos

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The ever-escalating CO₂ concentration in the atmosphere calls for accelerated development and deployment of carbon capture processes to reduce emissions. Mixed matrix membranes (MMMs), which are fabricated by incorporating the beneficial properties of highly selective inorganic fillers into a polymer matrix, have exhibited significant progress and the ability to enhance the performance of a membrane for gas separation. In this research, an amine-based ionic liquid (IL) [APTMS][AC] was prepared, which has greater CO₂ affinity and greater solubility due to its amine moiety. The metal–organic framework (MOF) UiO-66 with a multidimensional crystalline structure was used as a filler due to its appropriate porosity and tunable properties, and it was functionalized with NH₂. MOFs were further modified with an IL to prepare UiO-66@IL and UiO-66-NH₂@IL, and MMMs incorporating each MOF were fabricated with the polymer Pebax-1657. All the prepared membranes and MOFs were characterized to predict their separation efficiency. Several characterization techniques, namely, FTIR spectroscopy, XRD, and SEM, were used to successfully synthesize UiO-66@IL and UiO-66-NH₂@IL composites and confirmed proper dispersion and excellent polymer‒ filler compatibility at filler loadings ranging from 0 to 30 wt.%. The separation performances were investigated, and the results showed that the incorporation of RTIL with the highly crystalline structure and large surface area of UiO-66 enhanced the separation efficiency of the membrane. The permeability of CO₂ for all fabricated membranes continuously increased with increasing filler concentration, wherein the permeability was comparatively high for the UiO-66-NH₂ MMMs. The CO₂/CH₄ selectivity improved by 35%, 54%, and 60%, respectively, for UiO-66@IL, UiO-66-NH₂, and UiO-66-NH₂@IL MMMs compared to simple UiO-66 for CO₂/CH₄ and by 28%, 36%, and 63%, respectively, for CO₂/N₂, with an increase in filler loading in the MMMs.

Keywords: gas separation, mixed matrix membranes, CO₂ sequestration, climate change, global warming

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Authors: Ching-Yi Huang, Pi-Fen Cheng, Hui-Zhu Chen, Shi Ting Huang, Heng-Hua Wang

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This is a case of oral cancer after extensive resection and modified right lateral neck lymph node dissection followed by reconstruction with a skin flap. The nursing period lasted From September 25 to October 3, 2017, through observation, interview, physical assessment, and medical record review, the author identified the following nursing problems: acute pain, impaired oral mucous membrane, and body image change. During the nursing period, the author provided individual and overall nursing care and established mutual trust through the use of empathy. Author listened and eased the patient's physical indisposition, such as wound pain, we use medications and acupuncture massage to relieve pain. However, for oral mucosa change caused by surgery, provide continuous and complete oral care and oral exercise training to improve oral mucosal healing and restore swallowing function. In the body-image changes, guided him to express his feeling after the body-image change, and enhanced support and from the family, and encouraged him to attend head and neck cancer survivor alliance which allowed the patient to accept the altered body image and reaffirm self-worth. Hopefully, through sharing this nursing experience will help to the nursing care quality of nursing care for oral cancer patients after extensive resection and modified right lateral neck lymph node dissection followed by reconstruction with a skin flap.

Keywords: oral cancer, acute pain, impaired oral mucous membrane, body image change

Procedia PDF Downloads 187

Authors: M. Maier, I. Dedigama, J. Majasan, Y. Wu, Q. Meyer, L. Castanheira, G. Hinds, P. R. Shearing, D. J. L. Brett

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Increasing the efficiency of electrolyser technology is commonly seen as one of the main challenges on the way to the Hydrogen Economy. There is a significant lack of understanding of the different states of operation of polymer electrolyte membrane water electrolysers (PEMWE) and how these influence the overall efficiency. This in particular means the two-phase flow through the membrane, gas diffusion layers (GDL) and flow channels. In order to increase the efficiency of PEMWE and facilitate their spread as commercial hydrogen production technology, new analytic approaches have to be found. Acoustic emission (AE) offers the possibility to analyse the processes within a PEMWE in a non-destructive, fast and cheap in-situ way. This work describes the generation and analysis of AE data coming from a PEM water electrolyser, for, to the best of our knowledge, the first time in literature. Different experiments are carried out. Each experiment is designed so that only specific physical processes occur and AE solely related to one process can be measured. Therefore, a range of experimental conditions is used to induce different flow regimes within flow channels and GDL. The resulting AE data is first separated into different events, which are defined by exceeding the noise threshold. Each acoustic event consists of a number of consequent peaks and ends when the wave diminishes under the noise threshold. For all these acoustic events the following key attributes are extracted: maximum peak amplitude, duration, number of peaks, peaks before the maximum, average intensity of a peak and time till the maximum is reached. Each event is then expressed as a vector containing the normalized values for all criteria. Principal Component Analysis is performed on the resulting data, which orders the criteria by the eigenvalues of their covariance matrix. This can be used as an easy way of determining which criteria convey the most information on the acoustic data. In the following, the data is ordered in the two- or three-dimensional space formed by the most relevant criteria axes. By finding spaces in the two- or three-dimensional space only occupied by acoustic events originating from one of the three experiments it is possible to relate physical processes to certain acoustic patterns. Due to the complex nature of the AE data modern machine learning techniques are needed to recognize these patterns in-situ. Using the AE data produced before allows to train a self-learning algorithm and develop an analytical tool to diagnose different operational states in a PEMWE. Combining this technique with the measurement of polarization curves and electrochemical impedance spectroscopy allows for in-situ optimization and recognition of suboptimal states of operation.

Keywords: acoustic emission, gas diffusion layers, in-situ diagnosis, PEM water electrolyser

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Authors: Welder A. Baldassini, Jon J. Ramsey, Marcos R. Chiaratti, Amália S. Chaves, Renata H. Branco, Sarah F. M. Bonilha, Dante P. D. Lanna

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With increased production costs there is a need for animals that are more efficient in terms of meat production. In this context, the role of mitochondrial DNA (mtDNA) on physiological processes in liver, muscle and adipose tissues may account for inter-animal variation in energy expenditures and heat production. The purpose this study was to investigate if the amounts of mtDNA in liver, muscle and adipose tissue (subcutaneous and visceral depots) of Nellore bulls are associated with residual feed intake (RFI) and estimated heat production (EHP). Eighteen animals were individually fed in a feedlot for 90 days. RFI values were obtained by regression of dry matter intake (DMI) in relation to average daily gain (ADG) and mid-test metabolic body weight (BW). The animals were classified into low (more efficient) and high (less efficient) RFI groups. The bulls were then randomly distributed in individual pens where they were given excess feed twice daily to result in 5 to 10% orts for 90 d with diet containing 15% crude protein and 2.7 Mcal ME/kg DM. The heart rate (HR) of bulls was monitored for 4 consecutive days and used for calculation of EHP. Electrodes were fitted to bulls with stretch belts (POLAR RS400; Kempele, Finland). To calculate oxygen pulse (O2P), oxygen consumption was obtained using a facemask connected to the gas analyzer (EXHALYZER, ECOMedics, Zurich, Switzerland) and HR were simultaneously measured for 15 minutes period. Daily oxygen (O2) consumption was calculated by multiplying the volume of O2 per beat by total daily beats. EHP was calculated multiplying O2P by the average HR obtained during the 4 days, assuming 4.89 kcal/L of O2 to measure daily EHP that was expressed in kilocalories/day/kilogram metabolic BW (kcal/day/kg BW0.75). Blood samples were collected between days 45 and 90th after the beginning of the trial period in order to measure the concentration of hemoglobin and hematocrit. The bulls were slaughtered in an experimental slaughter house in accordance with current guidelines. Immediately after slaughter, a section of liver, a portion of longissimus thoracis (LT) muscle, plus a portion of subcutaneous fat (surrounding LT muscle) and portions of visceral fat (kidney, pelvis and inguinal fat) were collected. Samples of liver, muscle and adipose tissues were used to quantify mtDNA copy number per cell. The number of mtDNA copies was determined by normalization of mtDNA amount against a single copy nuclear gene (B2M). Mean of EHP, hemoglobin and hematocrit of high and low RFI bulls were compared using two-sample t-tests. Additionally, the one-way ANOVA was used to compare mtDNA quantification considering the mains effects of RFI groups. We found lower EHP (83.047 vs. 97.590 kcal/day/kgBW0.75; P < 0.10), hemoglobin concentration (13.533 vs. 15.108 g/dL; P < 0.10) and hematocrit percentage (39.3 vs. 43.6 %; P < 0.05) in low compared to high RFI bulls, respectively, which may be useful traits to identify efficient animals. However, no differences were observed between the mtDNA content in liver, muscle and adipose tissue of Nellore bulls with high and low RFI.

Keywords: bioenergetics, Bos indicus, feed efficiency, mitochondria

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Authors: Sihyun Chae, Ryoto Arai, Waldo Concepcion, Paula Popescu

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The kidneys perform an important role in the human hormones that regulate the blood pressure, produce an active form of vitamin D and control the production of red blood cells. Kidney disease can cause health problems, such as heart disease. Also, increase the chance of having a stroke or heart attack. There are mainly to types of treatments for kidney disease, dialysis, and kidney transplant. For a better quality of life, the kidney transplant is desirable. However, kidney transplant can cause antibody reaction and patients’ body would be attacked by immune system of their own. For solving that issue, patients with transplanted kidney always take immunosuppressive drugs which can hurt kidney as side effects. Patients willing to do a kidney transplant have a waiting time of 3.6 years in average searching to find an appropriate kidney, considering there are almost 96,380 patients waiting for kidney transplant. There is a promising method to solve these issues: bioartificial kidney. Our membrane is specially designed with unique perforations capable to filter the blood cells separating the white blood cells from red blood cells. White blood cells will not pass through the encapsulated kidney preventing the immune system to attack the new organ and eliminating the need of a matching donor. It is possible to construct life-time long encapsulation without needing pumps or a power supply on the cell’s separation method preventing futures surgeries due the Cross-Channel Flow inside the device. This technology allows the possibility to use an animal kidney, prevent cancer cells to spread through the body, arm and leg transplants in the future. This project aims to improve the quality of life of patients with kidney disease.

Keywords: kidney encapsulation, immunosuppression filter, leukocyte filter, leukocyte

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Authors: Benedetta Isella, Aleksander Drinic, Alissa Heim, Phillip Czichowski, Lisa Lauts, Hans Leemhuis

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Introduction: Membranes for guided bone regeneration are essential to perform a barrier function between the soft and the regenerating bone tissue. Bioabsorbable membranes are desirable in this field as they do not require a secondary surgery for removal, decreasing patient surgical risk. Collagen was the first bioabsorbable alternative introduced on the market, but its degradation time may be too fast to guarantee bone regeneration, and optimisation is needed. Silk fibroin, being biocompatible, slowly bioabsorbable, and processable into different scaffold types, could be a promising alternative. Objectives: The objective is to compare the general performance of a silk fibroin membrane for guided bone regeneration to current collagen alternatives developing suitable standardized tests for the mechanical and morphological characterization. Methods: Silk fibroin and collagen-based membranes were compared from the morphological and chemical perspective, with techniques such as SEM imaging and from the mechanical point of view with techniques such as tensile and suture retention strength (SRS) tests. Results: Silk fibroin revealed a high degree of reproducibility in surface density. The SRS of silk fibroin (0.76 ± 0.04 N), although lower than collagen, was still comparable to native tissues such as the internal mammary artery (0.56 N), and the same can be extended to general mechanical behaviour in tensile tests. The SRS could be increased by an increase in thickness. Conclusion: Silk fibroin is a promising material in the field of guided bone regeneration, covering the interesting position of not being considered a product containing cells or tissues of animal origin from the regulatory perspective and having longer degradation times with respect to collagen.

Keywords: guided bone regeneration, mechanical characterization, membrane, silk fibroin

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Authors: Muhammad Solikhudin Nafi, Wahyu Afif Mufida, Mita Erna Wati, Fitri Setyani Rokim, M. Al-Rizqi Dharma Fauzi

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High dose activity without any pre-exercise will impact Delayed Onset Muscle Soreness (DOMS). DOMS known as delayed pain post-exercise and induce skeletal injury which will decrease athletes’ performances. From now on, post-exercise muscle damage can be detected by measuring skeletal troponin I (sTnI) concentration in serum using ELISA but this method needs more time and cost. To prevent decreased athletes performances, screening need to be done rapidly. We want to introduce our new prototype to detect DOMS acutely. Rapid detection tests are based on immunological reaction between skeletal troponin I antibodies and sTnI in human serum or whole blood. Chemical methods that are used in the manufacture of diagnostic test is lateral flow immunoassay. The material used is rat monoclonal antibody sTnI, colloidal gold, anti-mouse IgG, nitrocellulose membrane, conjugate pad, sample pad, wick and backing card. The procedure are made conjugate (colloidal gold and mAb sTnI) and insert into the conjugate pad, gives spray sTnI mAb and anti-mouse IgG into nitrocellulose membrane, and assemble RDT. RDT had been evaluated by measuring the sensitivity of positive human serum (n = 30) and negative human serum (n = 30). Overall sensitivity value was 93% and specificity value was 90%. ADCOR as the first rapid detection test qualitatively showed antigen-antibody reaction and showed good overall performances for screening of muscle damage. Furthermore, these finding still need more improvements to get best results.

Keywords: DOMS, sTnI, rapid detection test, ELISA

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Authors: Jale Çatak, Bayram Yılmaz, Mustafa Ozilgen

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The second law muscle work efficiency is obtained by multiplying the metabolic and mechanical work efficiencies. Thermodynamic analyses are carried out with 19 sets of arms and legs exercise data which were obtained from the healthy young people. These data are used to simulate the changes occurring during aging. The muscle work efficiency decreases with aging as a result of the reduction of the metabolic energy generation in the mitochondria. The reduction of the mitochondrial energy efficiency makes it difficult to carry out the maintenance of the muscle tissue, which in turn causes a decline of the muscle work efficiency. When the muscle attempts to produce more work, entropy generation and exergy destruction increase. Increasing exergy destruction may be regarded as the result of the deterioration of the muscles. When the exergetic efficiency is 0.42, exergy destruction becomes 1.49 folds of the work performance. This proportionality becomes 2.50 and 5.21 folds when the exergetic efficiency decreases to 0.30 and 0.17 respectively.

Keywords: aging mitochondria, entropy generation, exergy destruction, muscle work performance, second law efficiency

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Authors: Ibrahim Rassoul, A. Serir, E-K. Si Ahmed, J. Legrand

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The acronym PEM refers to Proton Exchange Membrane or alternatively Polymer Electrolyte Membrane. Due to its high efficiency, low operating temperature (30–80 °C), and rapid evolution over the past decade, PEMFCs are increasingly emerging as a viable alternative clean power source for automobile and stationary applications. Before PEMFCs can be employed to power automobiles and homes, several key technical challenges must be properly addressed. One technical challenge is elucidating the mechanisms underlying water transport in and removal from PEMFCs. On one hand, sufficient water is needed in the polymer electrolyte membrane or PEM to maintain sufficiently high proton conductivity. On the other hand, too much liquid water present in the cathode can cause “flooding” (that is, pore space is filled with excessive liquid water) and hinder the transport of the oxygen reactant from the gas flow channel (GFC) to the three-phase reaction sites. The experimental transparent fuel cell used in this work was designed to represent actual full scale of fuel cell geometry. According to the operating conditions, a number of flow regimes may appear in the microchannel: droplet flow, blockage water liquid bridge /plug (concave and convex forms), slug/plug flow and film flow. Some of flow patterns are new, while others have been already observed in PEMFC microchannels. An algorithm in MATLAB was developed to automatically determine the flow structure (e.g. slug, droplet, plug, and film) of detected liquid water in the test microchannels and yield information pertaining to the distribution of water among the different flow structures. A video processing algorithm was developed to automatically detect dynamic and static liquid water present in the gas channels and generate relevant quantitative information. The potential benefit of this software allows the user to obtain a more precise and systematic way to obtain measurements from images of small objects. The void fractions are also determined based on images analysis. The aim of this work is to provide a comprehensive characterization of two-phase flow in an operating fuel cell which can be used towards the optimization of water management and informs design guidelines for gas delivery microchannels for fuel cells and its essential in the design and control of diverse applications. The approach will combine numerical modeling with experimental visualization and measurements.

Keywords: polymer electrolyte fuel cell, air-water two phase flow, gas diffusion layer, microchannels, advancing contact angle, receding contact angle, void fraction, surface tension, image processing

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Authors: Imdad Ullah Khan, Yongseok Yoon, Kyeung Uk Choi, Kwang Rae Jo, Namyul Kim, Eunbee Lee, Wan Hee Kim, Oh-Kyeong Kweon

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The primary spinal cord injury (SCI) causes mechanical damage to the neurons and blood vessels. It leads to secondary SCI, which activates multiple pathological pathways, which expand neuronal damage at the injury site. It is characterized by vascular disruption, ischemia, excitotoxicity, oxidation, inflammation, and apoptotic cell death. It causes nerve demyelination and disruption of axons, which perpetuate a loss of impulse conduction through the injured spinal cord. It also leads to the production of myelin inhibitory molecules, which with a concomitant formation of an astroglial scar, impede axonal regeneration. The pivotal role regarding the neuronal necrosis is played by oxidation and inflammation. During an early stage of spinal cord injury, there occurs an abundant expression of reactive oxygen species (ROS) due to defective mitochondrial metabolism and abundant migration of phagocytes (macrophages, neutrophils). ROS cause lipid peroxidation of the cell membrane, and cell death. Abundant migration of neutrophils, macrophages, and lymphocytes collectively produce pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-1beta (IL-1β), matrix metalloproteinase, superoxide dismutase, and myeloperoxidases which synergize neuronal apoptosis. Therefore, it is crucial to control inflammation and oxidation injury to minimize the nerve cell death during secondary spinal cord injury. Therefore, in response to oxidation and inflammation, heme oxygenase-1 (HO-1) is induced by the resident cells to ameliorate the milieu. In the meanwhile, neurotrophic factors are induced to promote neuroregeneration. However, it seems that anti-stress enzyme (HO-1) and neurotrophic factor (BDNF) do not significantly combat the pathological events during secondary spinal cord injury. Therefore, optimum healing can be induced if anti-inflammatory and neurotrophic factors are administered in a higher amount through an exogenous source. During the first experiment, the inflammation and neuroregeneration were selectively targeted. HO-1 expressing MSCs (HO-1 MSCs) and BDNF expressing MSCs (BDNF MSC) were co-transplanted in one group (combination group) of dogs with subacute spinal cord injury to selectively control the expression of inflammatory cytokines by HO-1 and induce neuroregeneration by BDNF. We compared the combination group with the HO-1 MSCs group, BDNF MSCs group, and GFP MSCs group. We found that the combination group showed significant improvement in functional recovery. It showed increased expression of neural markers and growth-associated proteins (GAP-43) than in other groups, which depicts enhanced neuroregeneration/neural sparing due to reduced expression of pro-inflammatory cytokines such as TNF-alpha, IL-6 and COX-2; and increased expression of anti-inflammatory markers such as IL-10 and HO-1. Histopathological study revealed reduced intra-parenchymal fibrosis in the injured spinal cord segment in the combination group than in other groups. Thus it was concluded that selectively targeting the inflammation and neuronal growth with the combined use of HO-1 MSCs and BDNF MSCs more favorably promote healing of the SCI. HO-1 MSCs play a role in controlling the inflammation, which favors the BDNF induced neuroregeneration at the injured spinal cord segment of dogs.

Keywords: HO-1 MSCs, BDNF MSCs, neuroregeneration, inflammation, anti-inflammation, spinal cord injury, dogs

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Authors: Kien Xuan Ngo

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Cassiicolin (Cas), a toxin produced by Corynespora cassiicola, is responsible for corynespora leaf fall (CLF) disease in rubber trees. Currently, the molecular mechanism of the cytotoxicity of Cas isoforms (i.e., Cas1, Cas2) on rubber leaves and its host selectivity have not been fully elucidated. This study analyzed the binding of Cas1 and Cas2 to membranes consisting of different plant lipids and their membrane-disruption activities. Using high-speed atomic force microscopy and confocal microscopy, this study reveals that the binding and disruption activities of Cas1 and Cas2 on lipid membranes are strongly dependent on the specific plant lipids. The negative phospholipids, glycerolipids, and sterols are more susceptible to membrane damage caused by Cas1 and Cas2 than neutral phospholipids and betaine lipids. In summary, This study unveils that (i) Cas1 and Cas2 directly damage and cause necrosis in the leaves of specific rubber clones; (ii) Cas1 and Cas2 can form biofilm-like structures on specific lipid membranes (negative phospholipids, glycerolipids, and sterols). The biofilm-like formation of Cas toxin plays an important role in selective disruption on lipid membranes; (iii) Vulnerability of the specific cytoplasmic membranes to the selective Cas toxin is the most remarkable feature of cytotoxicity of Cas toxin on plant cells. Finally, researcher’s exploration is crucial to understand the basic molecular mechanism underlying the host-selective toxic interaction of Cas toxin with cytoplasmic membranes in plant cells.

Keywords: cassiicolin, corynespora leaf fall disease, high-speed AFM, giant liposome vesicles

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Authors: Zeyu Zhou, Ziyu Zhao, Xiaochen Yang, Ling AI, Heng Zhai, Stuart Holmes

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As a promising sustainable alternative to traditional fossil fuels, fuel cell technology is highly favoured due to its enhanced working efficiency and reduced emissions. In the context of high-temperature fuel cells (operating above 100 °C), the most commonly used proton exchange membrane (PEM) is the Polybenzimidazole (PBI) doped phosphoric acid (PA) membrane. Grafting is a promising strategy to advance PA-doped PBI PEM technology. The existing grafting modification on PBI PEMs mainly focuses on grafting phosphate-containing or alkaline groups onto the PBI molecular chains. However, quaternary ammonium-based grafting approaches face a common challenge. To initiate the N-alkylation reaction, deacidifying agents such as NaH, NaOH, KOH, K2CO3, etc., can lead to ionic crosslinking between the quaternary ammonium group and PBI. Polyvinylpyrrolidone (PVP) is another widely used polymer, the N-heterocycle groups within PVP endow it with a significant ability to absorb PA. Recently, PVP has attracted substantial attention in the field of fuel cells due to its reduced environmental impact and impressive fuel cell performance. However, due to the the poor compatibility of PVP in PBI, few research apply PVP in PA-doped PBI PEMs. This work introduces an innovative strategy to graft PVP onto PBI to form a network-like polymer. Due to the absence of quaternary ammonium groups, PVP does not pose issues related to crosslinking with PBI. Moreover, the nitrogen-containing functional groups on PVP provide PBI with a robust phosphoric acid retention ability. The nuclear magnetic resonance (NMR) hydrogen spectrum analysis results indicate the successful completion of the grafting reaction where N-alkylation reactions happen on both sides of the grafting agent 1,4-bis(chloromethyl)benzene. On one side, the reaction takes place with the hydrogen atoms on the imidazole groups of PBI, while on the other side, it reacts with the terminal amino group of PVP. The XPS results provide additional evidence from the perspective of the element. On synthesized PBI-g-PVP surfaces, there is an absence of chlorine (chlorine in grafting agent 1,4-bis(chloromethyl)benzene is substituted) element but a presence of sulfur element (sulfur element in terminal amino PVP appears in PBI), which demonstrates the occurrence of the grafting reaction and PVP is successfully grafted onto PBI. Prepare these modified membranes into MEA. It was found that during the fuel cell operation, all the grafted membranes showed substantial improvement in maximum current density and peak power density compared to unmodified one. For PBI-g-PVP 30, with a grafting degree of 22.4%, the peak power density reaches 1312 mW cm⁻², marking a 59.6% enhancement compared to the pristine PBI membrane. The improvement is caused by the improved PA binding ability of the membrane after grafting. The AST test result shows that the grafting membranes have better long-term durability and performance than unmodified membranes attributed to the presence of added PA binding sites, which can effectively prevent the PA leaching caused by proton migration. In conclusion, the test results indicate that grafting PVP onto PBI is a promising strategy which can effectively improve the fuel cell performance.

Keywords: fuel cell, grafting modification, PA doping ability, PVP

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Authors: Lubena Achmad, Herman Kristanto, Julian Dewantiningrum

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Background: The Bishop score is a standard method used to predict the success of induction. This examination tends to be subjective with high inter and intraobserver variability, so it was presumed to have a low predictive value in terms of the outcome of labor induction. Cervical length measurement using transvaginal ultrasound is considered to be more objective to assess the cervical length. Meanwhile, this examination is not a complicated procedure and less invasive than vaginal touché. Objective: To compare transvaginal ultrasound and Bishop score in predicting successful induction. Methods: This study was a prospective cohort study. One hundred and twenty women with singleton pregnancies undergoing induction of labor at 37 – 42 weeks and met inclusion and exclusion criteria were enrolled in this study. Cervical assessment by both transvaginal ultrasound and Bishop score were conducted prior induction. The success of labor induction was defined as an ability to achieve active phase ≤ 12 hours after induction. To figure out the best cut-off point of cervical length and Bishop score, receiver operating characteristic (ROC) curves were plotted. Logistic regression analysis was used to determine which factors best-predicted induction success. Results: This study showed significant differences in terms of age, premature rupture of the membrane, the Bishop score, cervical length and funneling as significant predictors of successful induction. Using ROC curves found that the best cut-off point for prediction of successful induction was 25.45 mm for cervical length and 3 for Bishop score. Logistic regression was performed and showed only premature rupture of membranes and cervical length ≤ 25.45 that significantly predicted the success of labor induction. By excluding premature rupture of the membrane as the indication of induction, cervical length less than 25.3 mm was a better predictor of successful induction. Conclusion: Compared to Bishop score, cervical length using transvaginal ultrasound was a better predictor of successful induction.

Keywords: Bishop Score, cervical length, induction, successful induction, transvaginal sonography

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Authors: Boris Melnikov, Ye Zhang, Dmitrii Chaikovskii

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We continue to consider one of the cybernetic methods in computational biology related to the study of DNA chains. Namely, we are considering the problem of reconstructing the not fully filled distance matrix of DNA chains. When applied in a programming context, it is revealed that with a modern computer of average capabilities, creating even a small-sized distance matrix for mitochondrial DNA sequences is quite time-consuming with standard algorithms. As the size of the matrix grows larger, the computational effort required increases significantly, potentially spanning several weeks to months of non-stop computer processing. Hence, calculating the distance matrix on conventional computers is hardly feasible, and supercomputers are usually not available. Therefore, we started publishing our variants of the algorithms for calculating the distance between two DNA chains; then, we published algorithms for restoring partially filled matrices, i.e., the inverse problem of matrix processing. In this paper, we propose an algorithm for restoring the distance matrix for DNA chains, and the primary focus is on enhancing the algorithms that shape the greedy function within the branches and boundaries method framework.

Keywords: DNA chains, distance matrix, optimization problem, restoring algorithm, greedy algorithm, heuristics

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Authors: Ziyue Zeng

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Bacteriophages are viruses which infect bacteria specifically. Over the past decades, phage λ has been extensively studied, especially its interaction with the Escherichia coli LamB (EcLamB) protein receptor. Nonetheless, despite the enormous numbers and near-ubiquity of environmental phages, aside from phage λ, there is a paucity of information on other phages which target EcLamB as a receptor. In this study, to answer the question of whether there are other EcLamB-targeting phages in the natural environment, a simple and convenient method was developed and used for isolating environmental phages which target a particular surface structure of a particular bacterium; in this case, the EcLamB outer membrane protein. From the enrichments with the engineered bacterial hosts, a collection of EcLamB-targeting phages (ΦZZ phages) were easily isolated. Intriguingly, unlike phage λ, an obligate EcLamB-dependent phage in the Siphoviridae family, the newly isolated ΦZZ phages alternatively recognised EcLamB or E. coli OmpC (EcOmpC) as a receptor when infecting E. coli. Furthermore, ΦZZ phages were suggested to represent new species in the Tequatrovirus genus in the Myoviridae family, based on phage morphology and genomic sequences. Most phages are thought to have a narrow host range due to their exquisite specificity in receptor recognition. With the ability to optionally recognise two receptors, ΦZZ phages were considered relatively promiscuous. Via the heterologous expression of EcLamB on the bacterial cell surface, the host range of ΦZZ phages was further extended to three different enterobacterial genera. Besides, an interesting selection of evolved phage mutants with a broader host range was isolated, and the key mutations involved in their evolution to adapt to new hosts were investigated by genomic analysis. Finally, and importantly, two ΦZZ phages were found to be putative generalised transducers, which could be exploited as tools for DNA manipulations.

Keywords: environmental microbiology, phage, microbe-host interactions, microbial ecology

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Authors: Suman Chaudhary, Rupinder K. Kanwar, Jagat R. Kanwar

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Azadirachta indica, also known as Neem belonging to the mahogany family is actively gaining interest in the era of modern day medicine due to its extensive applications in homeopathic medicine such as Ayurveda and Unani. More than 140 phytochemicals have been extracted from neem leaves, seed, bark and flowers for agro-medicinal applications. Among the various components, neem leaf protein (NLP) is currently the most investigated active ingredient, due to its immunomodulatory activities against tumor growth. However, these therapeutic ingredients of neem are susceptible to degradation and cannot withstand the drastic pH changes under physiological environment, and therefore, there is an urgent need of an alternative strategy such as a nano-delivery system to exploit its medicinal benefits. This study hypothesizes that guar gum (GG) derived biodegradable nano-carrier based encapsulation of NLP will improve its stability, specificity and sensitivity, thus facilitating targeted anti-cancer therapeutics. GG is a galactomannan derived from the endosperm of the guar beans seeds. Synthesis of guar nanocapsules (NCs) was performed using nanoprecipitation technique where the GG was encapsulated with NLP. Preliminary experiments conducted to characterize the NCs confirmed spherical morphology with a narrow size distribution of 30-40 nm. Differential scanning colorimetric analysis (DSC) validated the stability of these NCs even at a temperature range of 50-60°C which was well within the physiological and storage conditions. Thermogravimetric (TGA) analysis indicated high decomposition temperature of these NCs ranging upto 350°C. Additionally, Fourier Transform Infrared spectroscopy (FTIR) and the SDS-PAGE data acquired confirmed the successful encapsulation of NLP in the NCs. The anti-cancerous therapeutic property of this NC was tested on colon cancer cells (caco-2) as they are one of the most prevalent form of cancer. These NCs (both NLP loaded and void) were also tested on human intestinal epithelial cells (FHs 74) cells to evaluate their effect on normal cells. Cytotoxicity evaluation of the NCs in the cell lines confirmed that the IC50 for NLP in FHs 74 cells was ~2 fold higher than in caco-2 cells, indicating that this nanoformulation system possessed biocompatible anti-cancerous properties Immunoconfocal microscopy analysis confirmed the time dependent internalization of the NCs within 6h. Recent findings performed using Annexin V and PI staining indicated a significant increase (p ≤ 0.001) in the early and late apoptotic cell population when treated with the NCs signifying the role of NLP in inducing apoptosis in caco-2 cells. This was further validated using Western blot, Polymerase chain reaction (PCR) and Fluorescence activated cell sorter (FACS) aided protein expressional analysis which presented a downregulation of survivin, an anti-apoptotic cell marker and upregulation of Bax/Bcl-2 ratio (pro-apoptotic indicator). Further, both the NLP NC and unencapsulated NLP treatment destabilized the mitochondrial membrane potential subsequently facilitating the release of the pro-apoptotic caspase cascade initiator, cytochrome-c. Future studies will be focused towards granting specificity to these NCs towards cancer cells, along with a comprehensive analysis of the anti-cancer potential of this naturally occurring compound in different cancer and in vivo animal models, will validate the clinical application of this unprecedented protein therapeutic.

Keywords: anti-tumor, guar gum, nanocapsules, neem leaf protein

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Authors: C. I. Boshoff, S. Steenkamp-Jonker

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Cystic echinococcosis (CE), caused by Echinococcus granulosus is an important parasitic infection in livestock worldwide, with severe zoonotic potential. It is important to understand the variability of Echinococcus granulosus, as genotype variations may influence lifecycle patterns, development rate, and transmission. Cystic Echinococcus samples were collected from domestic animals in Eastern Cape Province, South Africa. A molecular study was performed on 14 hydatid cysts obtained from caprine, ovine and bovine livers in order to determine the Echinococcus granulosus strain present in these hosts. The sequencing of the mitochondrial cytochrome C oxidase subunit I (coxI) gene of the hydatid cysts produced sequences of 400 bp for each sample analysed. These sequences were aligned with those present in GenBank and a phylogenetic tree was constructed. Based on coxI genotype the isolates could be grouped into E. granulosus sensu stricto. The findings of the study represent a pilot molecular study on Echinococcus from domestic animals undertaken in South Africa.

Keywords: Echinococcus granulosus, genotypes, livestock, South Africa

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

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

Keywords: coumarin, MTT, apoptosis, cytotoxicity

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Authors: Qing Zhang, Janak L. Pathak, Macro N. Helder, Richard T. Jaspers, Yin Xiao

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Introduction: Nanomaterial-based bone regeneration is greatly influenced by the immune microenvironment. Tissue-engineered nanomaterials mediate the inflammatory response of macrophages to regulate bone regeneration. Silica nanoparticles have been widely used in tissue engineering-related preclinical studies. However, the effect of topological features on the surface of silica nanoparticles on the immune response of macrophages remains unknown. Purposes: The aims of this research are to compare the influences of normal and pollen-like silica nano-surface topography on macrophage immune responses and to obtain insight into their potential regulatory mechanisms. Method: Macrophages (RAW 264.7 cells) were exposed to mesoporous silica nanoparticles with normal morphology (MSNs) and pollen-like morphology (PMSNs). RNA-seq, RT-qPCR, and LSCM were used to assess the changes in expression levels of immune response-related genes and proteins. SEM and TEM were executed to evaluate the contact and adherence of silica nanoparticles by macrophages. For the assessment of the immunomodulation-mediated osteogenic potential, BMSCs were cultured with conditioned medium (CM) from LPS pre-stimulated macrophage cultures treated with MSNs or PMSNs. Osteoimmunomodulatory potential of MSNs and PMSNs in vivo was tested in a mouse cranial bone osteolysis model. Results: The results of the RNA-seq, RT-qPCR, and LSCM assays showed that PMSNs inhibited the expression of pro-inflammatory genes and proteins in macrophages. SEM images showed distinct macrophage membrane surface binding patterns of MSNs and PMSNs. MSNs were more evenly dispersed across the macrophage cell membrane, while PMSNs were aggregated. PMSNs-induced macrophage anti-inflammatory response was associated with upregulation of the cell surface receptor CD28 and inhibition of ERK phosphorylation. TEM images showed that both MSNs and PMSNs could be phagocytosed by macrophages, and inhibiting nanoparticle phagocytosis did not affect the expression of anti-inflammatory genes and proteins. Moreover, PMSNs-induced conditioned medium from macrophages enhanced BMP-2 expression and osteogenic differentiation mBMSCs. Similarly, PMSNs prevented LPS-induced bone resorption via downregulation of inflammatory reaction. Conclusions: PMSNs can promote bone regeneration by modulating osteoimmunological processes through surface topography. The study offers insights into how surface physical contact cues can modulate the regulation of osteoimmunology and provides a basis for the application of nanoparticles with pollen-like morphology to affect immunomodulation in bone tissue engineering and regeneration.

Keywords: physical contact, osteoimmunology, macrophages, silica nanoparticles, surface morphology, membrane receptor, osteogenesis, inflammation

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Authors: Hsiao-Chien Tsai, Yu-Pin Chen, Ruei-Ming Chen

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Introduction: Osteoarthritis (OA) is characterized by joint destruction and causes chronic disability. One of the prominent symptoms is pain. Alleviating the pain is necessary and urgent for the therapy of OA patients. However, currently, understanding the mechanisms that drive OA-induced pain remains challenging, which hampers the optimistic management of pain in OA patients. Semaphorin 4D (Sema4D) participates in axon guidance pathway and bone remodeling, thus, may play a role in the regulation of pain in OA. In this study, we have established a rat model of OA to find out the mechanisms of OA-induced pain and to deliberate the roles of Sema4D. Methods: Behavioral changes and the pro-inflammatory cytokines (IL-1β, TNF-α, and IL-17) associated with pain were measured during the development of OA. Sema4D expression in cartilage and synovial membrane at 1, 4, and 12 weeks after inducing OA was analyzed. To assess if Sema4D is related to the neurogenesis in OA as an axon repellant, we analyzed the expression of PGP9.5 as well. Results: Synovitis and cartilage degradation were evident histologically during the development of OA. Mechanical hyperalgesia was most severe at week 1, then persisted thereafter. It was associated with stress coping strategies. Similar to the pain behavioral results, levels of IL-1β and TNF-α in synovial lavage fluid were significantly elevated in the OA group at weeks 1 and 4, respectively. Sema4D expression in cartilage and the synovial membrane was also enhanced in the OA group and was correlated with pain and pro-inflammatory cytokines. The marker of neurogenesis, PGP9.5, was also enhanced during the development of OA. Discussion: OA induced mechanical hyperalgesia, which might be through upregulating IL-1β/TNF-α-mediated Sema4D expressions. If anti-Sema4D treatment could reduce OA-induced mechanical hyperalgesia and prevent the subsequent progression of OA needs to be further investigated. Significance: OA can induce mechanical hyperalgesia through upregulation of IL-1β/TNF-α-mediated Sema4D and PGP9.5 expressions. And the upregulation of Sema4D may indicate the severity or active status of OA and OA-induced pain.

Keywords: traumatic osteoarthritis, mechanical hyperalgesia, Sema4D, inflammatory cytokines

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