Search results for: membrane protein

Authors: Anamarel Medina-Hernandez, Teresa Ponce-Noyola, Ileana Vera-Reyes, Ana C. Ramos-Valdivia

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Somatic embryos reproduce original seed characteristics and could be implemented in biotechnological studies. Jatropha curcas L. is an important plant for biodiesel production, but also is used in traditional medicine. Seeds from J. curcas are toxic because contain diterpenoids called phorbol esters, but in Mexico exist a non-toxic variety. Therefore, somatic embryos suspension cultures from non-toxic J. curcas variety were induced. In order to investigate the characteristics of somatic embryos, a differential proteomic analysis was made between pre-globular and globular stages by 2-D gel electrophoresis. 108 spots were differentially expressed (p<0.02), and 20 spots from globular somatic embryos were sequenced by MALDI-TOF-TOF mass spectrometry. A comparative analysis of terpenoids production between the two stages was made by RP-18 TLC plates. The sequenced proteins were related to energy production (68%), protein destination and storage (9%), secondary metabolism (9%), signal transduction (5%), cell structure (5%) and aminoacid metabolism (4%). Regarding terpenoid production, in pre-globular and globular somatic embryos were identified sterols and triterpenes of pharmacological interest (alpha-amyrin and betulinic acid) but also it was found compounds that were unique to each stage. The results of this work are the basis to characterize at different levels the J. curcas somatic embryos so that this system can be used efficiently in biotechnological processes.

Keywords: Jatropha curcas, proteomics, somatic embryo, terpenoids

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Authors: Harish Rajak, Swati Singh

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A series of novel pyrimidine based semicarbazone were designed and synthesized on the basis of semicarbazone based pharmacophoric model to satisfy the structural prerequisite crucial for antiepileptic activity. The semicarbazones based pharmacophoric model consists of following four essential binding sites: (i) An aryl hydrophobic binding site with halo substituent; (ii) A hydrogen bonding domain; (iii) An electron donor group and (iv) Another hydrophobic-hydrophilic site controlling the pharmacokinetic features of the anticonvulsant. The aryl semicarbazones has been recognized as a structurally novel class of compounds with remarkable anticonvulsant activity. In the present study, all the test semicarbazones were subjected to molecular docking using Glide v5.8. Some of the compounds were found to interact with ARG192, GLU270 and THR353 residues of 1OHV protein, present in GABA-AT receptor. The chemical structures of the synthesized molecules were characterized by elemental and spectral (IR, 1H NMR, 13C NMR and MS) analysis. The anticonvulsant activities of the compounds were investigated using maximal electroshock seizure (MES) and subcutaneous pentylenetrtrazole (scPTZ) models. The neurotoxicity was evaluated in mice by the rotorod test. The attempts were also made to establish structure-activity relationships among synthesized compounds. The results of the present study confirmed that the pharmacophore model with four binding sites is essential for antiepileptic activity.

Keywords: pyrimidine, semicarbazones, anticonvulsant activity, neurotoxicity

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Authors: Ntokozo Dambuza, Maryna Van De Venter, Trevor Koekemoer

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Oxidative stress contributes to the pathogenesis of many diseases and consequently antioxidant therapy has attracted much attention as a potential therapeutic strategy. Regardless of the quantities ingested, antioxidants need to reach the diseased tissues at concentrations sufficient to combat oxidative stress. Bioavailability is thus a defining criterion for the therapeutic efficacy of antioxidants. In addition, therapeutic antioxidants must possess biologically relevant characteristics which can target the specific molecular mechanisms responsible for disease related oxidative stress. While many chemical antioxidant assays are available to quantify antioxidant capacity, they relate poorly to the biological environment and provide no information as to the bioavailability. The present comparative study thus aims to characterise green and fermented rooibos extracts, well recognized for their exceptional antioxidant capacity, in terms of antioxidant bioavailability and efficacy in a disease relevant cellular setting. Chinese green tea antioxidant activity was also evaluated. Chemical antioxidant assays (FRAP, DPPH and ORAC) confirmed the potent antioxidant capacity of both green and fermented rooibos, with green rooibos possessing antioxidant activity superior to that of fermented rooibos and Chinese green tea. Bioavailability was assessed using the PAMPA assay and the results indicate that green and fermented rooibos have a permeation coefficient of 5.7 x 10-6 and 6.9 x 10-6 cm/s, respectively. Chinese green tea permeability coefficient was 8.5 x 10-6 cm/s. These values were comparable to those of rifampicin, which is known to have a high permeability across intestinal epithelium with a permeability coefficient of 5 x 10 -6 cm/s. To assess the antioxidant efficacy in a cellular context, U937 and red blood cells were pre-treated with rooibos and Chinese green tea extracts in the presence of a dye DCFH-DA and then exposed to oxidative stress. Green rooibos exhibited highest activity with an IC50 value of 29 μg/ml and 70 μg/ml, when U937 and red blood cells were exposed oxidative stress, respectively. Fermented rooibos and Chinese green tea had IC50 values of 61 μg/ml and 57 μg/ml for U937, respectively, and 221 μg/ml and 405 μg/ml for red blood cells, respectively. These results indicate that fermented and green rooibos extracts were able to permeate the U937 cells and red blood cell membrane and inhibited oxidation of DCFH-DA to a fluorescent DCF within the cells.

Keywords: rooibos, antioxidants, permeability, bioavailability

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Authors: Belen Carrillo, Mauricio Mosquera

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Fatty acids are bioactive compounds widely used as nutritional supplements in the food and pharmaceutical industry. Bluefish such as sardines have a large variety of these fatty acids in their composition. The objective of this project is to extract these compounds from fishing wastes, to do this, heads of known species as Pinchagua (Opistonema libertate) were used. The conducted study represents a simplified alternative for obtaining and simultaneous saponification of oil through basic hydrolysis, which separates lipids from protein and saponifies sample all the same time to isolate the fatty acid accurately through salts formation. To do these different concentrations of sodium hydroxide were used, it was demonstrated at a concentration of 1 M the highest yield of saponified oil recovery corresponding a value of 3,64% was obtained. Subsequently, the saponified oil was subjected to an acid hydrolysis in which fatty acids were isolated. Different sulfuric acid concentrations and temperatures for the process were tested. Thus, it was shown that the great fatty acids variety were obtained at a 60 °C temperature and sulfuric acid concentration of 50% v/v. Among the obtained compounds the presence of acids such as palmitic, lauric, caproic and myristic are highlighted. Applications of this type of elements are varied and widely used in the nutritional supplements development. Thus, the described methodology proposes a simple mechanism in the revaluation of fishing industry wastes that allow directly generate high added value elements.

Keywords: fatty acids, hydrolysis, Pinchagua, saponification

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Authors: Harish Rajak, Preeti Patel

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HDAC inhibitors can reactivate gene expression and inhibit the growth and survival of cancer cells. The 3D-QSAR and Pharmacophore modeling studies were performed to identify important pharmacophoric features and correlate 3D-chemical structure with biological activity. The pharmacophore hypotheses were developed using e-pharmacophore script and phase module. Pharmacophore hypothesis represents the 3D arrangement of molecular features necessary for activity. A series of 55 compounds with well-assigned HDAC inhibitory activity was used for 3D-QSAR model development. Best 3D-QSAR model, which is a five PLS factor model with good statistics and predictive ability, acquired Q2 (0.7293), R2 (0.9811) and standard deviation (0.0952). Molecular docking were performed using Histone Deacetylase protein (PDB ID: 1t69) and prepared series of hydroxamic acid based HDAC inhibitors. Docking study of compound 43 show significant binding interactions Ser 276 and oxygen atom of dioxine cap region, Gly 151 and amino group and Asp 267 with carboxyl group of CONHOH, which are essential for anticancer activity. On docking, most of the compounds exhibited better glide score values between -8 to -10.5. We have established structure activity correlation using docking, energetic based pharmacophore modelling, pharmacophore and atom based 3D QSAR model. The results of these studies were further used for the design and testing of new HDAC analogs.

Keywords: Docking, e-pharmacophore, HDACIs, QSAR, Suberoylanilidehydroxamic acid.

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Authors: Liang Jiansheng, Zhu Wei

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Light and brassinosteroids are essential external and internal cues for plant survival. Although the coordination of light with phytohormone signals is crucial for plant growth and development, the molecular connection between light and brassinosteroid signaling during root soil penetration remains elusive. Here, we reveal that light-stabilized RACK1 couples a brassinosteroid signaling cascade to drive cell division in root meristems. RACK1 family scaffold proteins positively regulate light-induced the promotion of root elongation during soil penetration. Under the light condition, RACK1A interacts with both phyB and SPA1, then reinforces the phyB-SPA1 association to accumulate its abundance in roots. In response to brassinosteroid signals, RACK1A competes with BKI1 to attenuate the BRI1-BKI1 interaction, thereby leading to activating BRI1 actions in root development. Furthermore, RACK1A binds to BES1 to repress its DNA binding activity toward the target gene CYCD3;1. This ultimately allows to release the inhibition of CYCD3;1 transcription, and promotes cell division during root growth. Our study illustrates a new mechanistic model of how plants engage scaffold proteins in transducing light information to facilitate brassinosteroid signaling for root growth in the soil.

Keywords: root growth, cell division, light signaling, brassinosteroid signaling, soil penetration, scaffold protein, RACK1

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Authors: Adriano Losso, Birgit Dämon, Stefan Mayr

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In vascular plants, water flows from roots to leaves in a metastable state, and even a small perturbation of the system can lead a sudden transition from the liquid to the vapor phase, resulting in xylem embolism (cavitation). Xylem embolism, induced by drought stress and/or freezing stress is caused by the aspiration of gaseous bubbles into xylem conduits from adjacent gas-filled compartments through pit membrane pores (‘air seeding’). At water potentials less negative than the threshold for air seeding, the surface tension (γ) stabilizes the air-water interface and thus prevents air from passing the pit pores. This hold is probably also true for conifers, where this effect occurs at the edge of the sealed torus. Accordingly, it was experimentally demonstrated that γ influences air seeding, but information on the relevance of this effect under field conditions is missing. In this study, we analyzed seasonal changes in γ of the xylem sap in two conifers growing at the alpine timberline (Picea abies and Pinus mugo). In addition, cut branches were perfused (40 min perfusion at 0.004 MPa) with different γ solutions (i.e. distilled and degassed water, 2, 5 and 15% (v/v) ethanol-water solution corresponding to a γ of 74, 65, 55 and 45 mN m-1, respectively) and their vulnerability to drought-induced embolism analyzed via the centrifuge technique (Cavitron). In both species, xylem sap γ changed considerably (ca. 53-67 and ca. 50-68 mN m-1 in P. abies and P. cembra, respectively) over the season. Branches perfused with low γ solutions showed reduced resistance against drought-induced embolism in both species. A significant linear relationship (P < 0.001) between P12, P50 and P88 (i.e. water potential at 12, 50 and 88% of the loss of conductivity) and xylem sap γ was found. Based on this correlation, a variation in P50 between -3.10 and -3.83 MPa (P. abies) and between -3.21 and -4.11 MPa (P. mugo) over the season could be estimated. Results demonstrate that changes in γ of the xylem sap can considerably influence a tree´s resistance to drought-induced embolism. They indicate that vulnerability analyses, normally conducted at a γ near that of pure water, might often underestimate vulnerabilities under field conditions. For studied timberline conifers, seasonal changes in γ might be especially relevant in winter, when frost drought and freezing stress can lead to an excessive embolism.

Keywords: conifers, Picea abies, Pinus mugo, timberline

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Authors: Farzad Doostishoar, Abbas Pardakhty, Abdolreza Hassanzadeh, Sudeh salarpour, Elham Sharif

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Background: Sericin is a protein extracted from silk and has antioxidant, antimicrobial, antineoplastic, wound healing and moisturizing properties. Different cosmetic formulation of sericin is available in different countries such as Japan and the other south-eastern Asian countries. We formulated and evaluated the sunscreen properties of topical formulations of sericin by an in vitro method. Method: Niosomes composed of sorbitan palmitate (Span 40), polysorbate 40 (Tween 40) and cholesterol (300 µmol, 3.5:3.5:3 molar ratio) were prepared by film hydration technique. Sericin was dissolved in normal saline and the lipid hydration was carried out at 60°C and the niosomes were incorporated in a Carbomer gel base. A W/O cream was also prepared and the release of sericin was evaluated by using Franz diffusion cell. Particle size analysis, sericin encapsulation efficiency measurement, morphological studies and stability evaluation were done in niosomal formulations. SPF was calculated by using Transpore tape in vitro method for both formulations. Results: Niosomes had high stability during 6 months storage at 4-8°C. The mean volume diameter of niosomes was less than 7 µm which is ideal for sustained release of drugs in topical formulations. The SPF of niosomal gel was 25 and higher than sericin cream with a diffusion based release pattern of active material. Conclusion: Sericin can be successfully entrapped in niosomes with sustained release pattern and relatively high SPF.

Keywords: sericin, niosomes, sun protection factor, cream, gel

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Authors: S. Roy, R. Rekha, K. Sriram, G. Subhadra, R. Johana

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Cardiovascular disease remains a leading cause of death in most industrialized countries. Many chemical drugs are available in the market which targets different receptor proteins related to cardiovascular diseases. Of late the traditional herbal drugs are safer when compared to chemical drugs because of its side effects. However, many herbal remedies used in treating cardiovascular diseases have not undergone scientific assessment to prove its pharmacological activities. There are many natural compounds, nature derived and Natural product mimic compounds are available which are in the market as approved drug. In the most of the cases drug activity at the molecular level are not known. Here we have categorized those compounds with our experimental compounds in different classes based on the structural similarity and physicochemical properties, using a tool, Chemmine and has attempted to understand the mechanism of the action of a experimental compound, which are clustered with Simvastatin, Lovastatin, Mevastatin and Pravastatin. Target protein molecule for Simvastatin, Lovastatin, Mevastatin and Pravastatin is HMG-CoA reductase, so we concluded that the experimental compound may be able to bind to the same target. Molecular docking and atomic interaction studies with simvastatin and our experimental compound were compared. A pharmacophore modeling was done based on the experimental compound and HMG-CoA reductase inhibitor.

Keywords: molecular docking, physicochemical properties, pharmacophore modeling structural similarity, pravastatin

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Authors: Siddharth Soni, Gourav Kumar Pandey, Sneha Kumari, Dev Mani Pandey, Koel Mukherjee

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The silkworm Bombyx mori is a commercially important insect, but a major roadblock in silk production are silkworm diseases. Flacherie is one of the diseases of the silkworm, that affects the midgut of the 4th and 5th instar larvae and eventually makes them lethargic, stop feeding and finally result in their death. The concerned disease is a result of bacterial and viral infection and in some instances a combination of both. The present study aims to identify and study the expression level of genes in the flacherie condition. For the said work, total RNA was isolated from the infected larvae at their most probable infectious instar and cDNA was synthesized using Reverse Transcriptase PCR (RT-PCR). This cDNA was then used to amplify disease relalted genes whose expression levels were checked using quantitaive PCR (qPCR) using the double delta Ct method. Cry toxin receptors like APN and BtR-175, ROS mediator Dual Oxidase are few proteins whose genes were overexpressed. Interestingly, pattern recognition receptors (PRRs) C-type lectins' genes were found to be downregulated. The results explain about the strong expression of genes that can distinguish the concerned protein in the midgut of diseased silkworm and thereby aiding knowledge in the field of inhibitor designing research.

Keywords: Bombyx mori, flacherie disease, inhibitor designing, up and down regulation

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Authors: Olivier de Backer, Alexis Khelfi, Olivier Svensek, Axelle Nolmans, Dominique Desnoeck

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The SMC5-SMC6 complex helps replication and repair of DNA double-strand breaks. Nse1, Nse3 and Nse4 are non-SMC components of the complex in which Nse3 stimulates the E3 ubiquitin ligase activity of Nse1 and is required for recruiting the complex on DNA. In most eukaryotes, Nse3 is a single protein, but in eutherians (placental mammals), it belongs to a large family of proteins called MAGE (Melanoma antigen) that share a conserved domain of about 200 aa known as MHD (Mage homology domain). MAGE assembles specific RING and HECT ubiquitin ligases and determines new substrates for ubiquitination. The MHD is required for the interaction with the cognate E3 ligase. Some MAGEs (referred to as Type I) are exclusively expressed in germ cells of the testis but are often expressed ectopically in cancer cells as the result of epigenetic modifications. The 12 MAGE-A genes belong to this category. Serval MAGE-A proteins could promote tumorigenesis by targeting tumor suppressor proteins (including p53) for ubiquitination and degradation. We showed that depletion of MAGE-A proteins in melanoma cells results in impaired DNA damage response and increased double-strand breaks after exposure to camptothecin. Moreover, it was shown that other actors of the DNA Damage Response were impacted when cells were depleted of MAGEA proteins.

Keywords: DNA damage response, melanoma, camptothecin, new role, MAGEA

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Authors: Turker Yardan, Bahattin Avci, S. Sirri Bilge, Ayhan Bozkurt

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Sinapic acid (SA) is a phenylpropanoid compound with anti-inflammatory, antioxidant, and neuroprotective activities. The purpose of this study was to characterize the possible protective effect of sinapic acid on chlorpyrifos (CPF), a common organophosphorus pesticide used worldwide, induced toxicity in rats. Forty male and female rats (240-270 g) were used in this study. Each group was composed of 5 male and 5 female rats. Sinapic acid (20 mg/kg or 40 mg/kg) or vehicle (olive oil, 1 ml ⁄ rat) were given orally for 5 days. CPF (279 mg/kg) or vehicle (peanut oil, 2 ml ⁄ kg, s.c.) was administered on the sixth day, immediately after the recording of the body weight of the animals. Twenty four hours following CPF administration body weight, body temperature and locomotor activity values were recorded before decapitation of the animals. Trunk blood, brain, and liver samples were collected for biochemical examinations. Chlorpyrifos administration decreased butyrylcholinesterase activity in blood, brain, and liver, while it increased malondialdehyde (MDA) levels and advanced oxidation protein products (AOPPs) (p < 0.01 - 0.001). Additionally, CPF administration reduced the body weight, body temperature, and locomotor activity values of the animals (p < 0.01 - 0.001). All these physiological and biochemical changes induced by CPF were reduced with the 40 mg/kg dose of SA (p < 0.05 - 0.001). Our results suggest that SA administration ameliorates CPF induced toxicity in rats, possibly by supporting the antioxidant mechanism.

Keywords: antioxidant, Chlorpyrifos, poisoning, sinapic acid

Procedia PDF Downloads 177

Authors: Cuili Yang, Chengcao Sun, Ruilin Xue, Yongyong Xi, Liang Wang, Dejia Li

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Backgrounds: Previous studies showed that Sirt1 plays an important role in C2C12 myoblast cell proliferation, but the mechanism(s) involved in this process remains unclear. This work was undertaken to determine if Myostatin participates in the regulation of C2C12 proliferation by Sirt1. Methods: We administrated the Sirt1 activator resveratrol, inhibitor Nicotinamide (NAM) and Myostatin inhibitor SB431542 on C2C12 myoblast cells. Cell viability was evaluated by CCK8 assay. The expression of Sirt1 and MyoD were detected by qRT-PCR. Utilizing western blot to determinate the expression of myostatin, P107 and p-P107. Results: Our results showed that resveratrol promoted the proliferation of C2C12 myoblast cells, while NAM suppressed the proliferation of C2C12 myoblast cells; SB431542 promoted the proliferation of C2C12 myoblast cells and attenuated the inhibition effect of NAM on C2C12 myoblast cells proliferation; Resveratrol can significantly increase the expression of Sirt1 and MyoD, decrease the expression of Myostatin, while NAM can significantly down-regulate the expression of Sirt1, MyoD and the phosphorylation of P107(p-P107), but up-regulate the expression of Myostatin and the protein P107; SB431542 can significantly mitigate the effect of NAM on the expression of MyoD, P107, and p-P107. Conclusions: Taken together, these results indicate that Sirt1 promotes the proliferation of C2C12 myoblast cells via Myostatin signaling pathway.

Keywords: Sirt1, C2C12 cells, proliferation, myostatin signaling pathway

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Authors: Priya Bajaj, Baby Tabassum

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Arsenic is a poisonous metalloid, naturally occurring in soil, air, rocks and ground water. This dreadful metalloid commonly exists as inorganic compound, arsenic trioxide. WHO permitted maximum limit for arsenic in water is 0.01 mg/L, but some affected areas show ground water level of arsenic up to 3 mg/L even. Ground water arsenic pollution has created a number of health problems, viz. keratosis, melanosis, lesions and even skin cancers. The key objective of our nested study was to characterize arsenic induced hepatotoxicity and to find out some herbal protection against it. For the purpose, we selected albino rat (Rattus norvegicus) as model for arsenic induced liver injury and wild turmeric (Curcuma aromatica) leaf extract as remedy for it. The study was performed at acute (1 day) and subacute (7, 14 & 21 days) levels. The LD50 estimated for arsenic trioxide was 14.98 mg/kg body weight. In our investigation, we observed a significant restoration of altered hepatic lipid, cholesterol, protein and glycogen contents as well as liver weight, body-weight and hepato-somatic index by Curcuma aromatica leaf extract before arsenic intoxication. The results reveal excellent protective efficacy of Curcuma aromatica leaf extract that further can be exploited in remediation programme in heavy metal affected areas.

Keywords: arsenic, Curcuma aromatica, glycogen, lipids

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Authors: Sawale Pravin Digambar, G. R. Patil, Shaik Abdul Hussain

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Terminalia arjuna (TA), an important medicinal plant in Indian System of Medicine, is specifically recognized for its recuperative effect on heart ailments. Alcoholic extract of TA (both free and encapsulated) was incorporated into milk to obtain functional dairy beverages. The respective beverages were appropriately flavored and optimized using response surface methodology to improve the sensory appeal. The beverages were evaluated for their compositional, anti-oxidative and various other physico-chemical aspects. Addition of herb (0.3%) extract to flavoured dairy drink (Drink 1) resulted in significantly lowered (p>0.05) HMF content and increased antioxidant activity, total phenol content as compared with control (Control 1). Subsequently, a significant (p>0.05) increase in acidity and sedimentation was also observed. Encapsulated herb (1.8%) incorporated drink (Drink 2) had significantly (P>0.05) enhanced HMF value and decreased antioxidant activity, phenol content as compared to herb added vanilla chocolate dairy drink (Drink 1). It can be concluded that addition of encapsulated TA extract and non-encapsulated TA extract to chocolate dairy drink at 0.3% concentration altered the functional properties vanilla chocolate dairy drink which could be related to the interaction of herb components such as polyphenol with milk protein or maltodextrin/ gum Arabic matrix.

Keywords: Terminalia arjuna, encapsulate, antioxidant activity, physicochemical study

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Authors: Namsoo Kim, So-Hee Son, Jin-Soo Maeng, Yong-Jin Cho, Chong-Tai Kim

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It has been reported that some enzymes such as trypsin and Alcalase 2.4L are tolerant to a medium high pressure of 300 MPa and preparation of protein hydrolyzates under 300 MPa was advantageous with regard to hydrolysis rate and thus production yield compared with the counterpart under ambient pressure.1,2) In this study, nine fish comprising halibut, soft shell clam and carp were hydrolyzed using Flavourzyme 500MG only, and the combination of Flavourzyme 500 mg, Alcalase 2.4 L, Marugoto E, and Protamex under 300 MPa. Then, the effects of single and multiple protease treatments were determined with respect to contents of soluble solid (SS) and soluble nitrogen, sensory attributes, electrophoretic profiles, and HPLC peak patterns of the fish hydrolyzates (FHs) from various species. The contents of SS of the FHs were quite species-specific and the hydrolyzates of halibut showed the highest SS contents. At this point, multiple protease treatment increased SS content conspicuously in all fish tested. The contents of total soluble nitrogen and TCA-soluble nitrogen were well correlated with those of SS irrespective of fish species and methods of enzyme treatment. Also, it was noticed that multiple protease treatment improved sensory attributes of the FHs considerably. Electropherograms of the FHs showed fast migrating peptide bands that had the molecular masses mostly lower than 1 kDa and this was confirmed by peptide patterns from HPLC analysis for some FHs that had good sensory quality.

Keywords: production, fish hydrolyzates, protease treatments, high pressure

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Authors: Etika Sukma Adiyanti, H. N. Nurul Huda Putu Ekapraja, Gugun Gunawan

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Global warming causes climate change and sea level rise. This is a threat for coastal regions, especially for coastal settlements with activities that are influenced by this natural phenomenon. Consequences are damage of houses, humid house environment, sustainability of the houses, obstructed economic activities and domestic works, disruption of sanitation facilities, lack of electricity, failure of transport system, psychological issues and other. Icons Tambak Lorok as 'Fisherman Village' is not something familiar to residents of the city of Semarang. Especially for the housewife who every day have to buy the ingredients high in protein and omega fish auction which is adjacent to the main street market in the village of Tambak Lorok. However, there are major problems that are being experienced by this small neighborhood. In fact, this issue includes seven infrastructure that should spoil the fishermen in activity with marine life. With this research, we will investigate water urbanism and climate change resiliency in Semarang, specifically the traditional fisher community of Tambak Lorok. We intend to find out how the local people in the fisher settlement Tambak Lorok deal with water urbanism, proverty and living with floods. So, we have a good solution for this problem, Floating Stage. We think that Tambak Lorok needs a new design for the common future. With this, One Building at A Time for Tambak Lorok, will be a good solution.

Keywords: fisher community, environment, climate change, settlement

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Authors: Mohanapriya Subramanian, V. Raj

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Direct methanol fuel cells (DMFCs) are considered to be one of the most promising candidates for portable and stationary applications in the view of their advantages such as high energy density, easy manipulation, high efficiency and they operate with liquid fuel which could be used without requiring any fuel-processing units. Electrolyte membrane of DMFC plays a key role as a proton conductor as well as a separator between electrodes. Increasing concern over environmental protection, biopolymers gain tremendous interest owing to their eco-friendly bio-degradable nature. Pectin is a natural anionic polysaccharide which plays an essential part in regulating mechanical behavior of plant cell wall and it is extracted from outer cells of most of the plants. The aim of this study is to develop and demonstrate pectin based polymer composite membranes as methanol impermeable polymer electrolyte membranes for DMFCs. Pectin based nanocomposites membranes are prepared by solution-casting technique wherein pectin is blended with chitosan followed by the addition of optimal amount of sulphonic acid modified Titanium dioxide nanoparticle (S-TiO2). Nanocomposite membranes are characterized by Fourier Transform-Infra Red spectroscopy, Scanning electron microscopy, and Energy dispersive spectroscopy analyses. Proton conductivity and methanol permeability are determined into order to evaluate their suitability for DMFC application. Pectin-chitosan blends endow with a flexible polymeric network which is appropriate to disperse rigid S-TiO2 nanoparticles. Resulting nanocomposite membranes possess adequate thermo-mechanical stabilities as well as high charge-density per unit volume. Pectin-chitosan natural polymeric nanocomposite comprising optimal S-TiO2 exhibits good electrochemical selectivity and therefore desirable for DMFC application.

Keywords: biopolymers, fuel cells, nanocomposite, methanol crossover

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Authors: Neena Kanojia, R. K. Kanojia

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Background: Osteoarthritis OA of the knee is one of the leading causes of disability characterized by degeneration of hyaline cartilage combined with reparative processes. Its strong association with metabolic syndrome is postulated to be due to both mechanical and biochemical factors. Our study aims to study differential effect of metabolic risk factors on cartilage degeneration and regeneration at biomarker level. Design: After screening 281 patients presenting with knee pain, 41 patients who met the selection criteria were included and were divided into metabolic MetS OA and non-metabolic Non-MetS OA phenotypes using National Cholesterol Education Programme-Adult Treatment Panel-III NCEP ATP III criteria for metabolic syndrome. Serum Cartilage Oligomeric Matrix Protein COMP and Procollagen type IIA N terminal Propeptide PIIANP levels were used as tools to assess cartilage degeneration and regeneration, respectively. Results: 22 among 41 patients 53.66% had metabolic syndrome. Covariates like age, gender, Kellgren Lawrence KL grades were comparable in both groups. MetS OA group showed significant increase in serum COMP levels (p 0.03 with no significant effect on serum PIIANP levels (p 0.46. Hypertriglyceridemia showed independent association with both cartilage anabolism (p 0.03 and catabolism (p 0.03. Conclusion: Metabolic syndrome, though has no effect on cartilage regeneration tends to shift cartilage homeostasis towards degeneration with hypertriglyceridemia showing significant independent effect on cartilage metabolism.

Keywords: metabolic, syndrome, cartilage, degernation

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Authors: Seung-Hwa Baek, In-Jung Nam, Sang-Han Lee

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The development of anti-melanogenic agents is important for the prevention of serious esthetic problem like a melasma, freckle, age spots, and chloasma. The aim of this study was to investigate the anti-melanogenic effect of sesamol, an active lignan isolated from sesame seed, by mushroom and cellular tyrosinase assay, melanin content and the analysis of melanogensis-related mRNA expressions in melana cells. Sesamol showed strong inhibitory activity against the mushroom tyrosinase in a dose-dependent manner. Intracellular tyrosinase inhibition activity was also confirmed by zymography. At a concentration of 50 μM, sesamol inhibited melanin production in melan-a cells with no cytoxicity while those of phenylthiourea (PTU) as a positive control were the same condition. Sesamol significantly inhibited the expression of melanogensis-related genes, such as tyrosinase, tyrosinase-related protein-1 (TRP-1), dopachrome tautomerase (Dct), microphthalmia-associated transcription factor (MITF) and melanocortin 1 receptor (MC1R). These findings indicate that sesamol could reduce melanin biosynthesis via the downregulation of tyrosinase activity and melanin production via subsequent gene expression of melanogenesis-related proteins. Together, these results suggest that the sesamol have strong potential in inhibiting melanin biosynthesis, in that the substance may be used as a new skin-whitening agent of cosmetic materials.

Keywords: sesamol, sesame seed, melanin biosynthesis, melanogenesis-related gene, skin-whitening agent

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Authors: Bhumika Wadhwa, Fayaz Malik

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The serine/threonine protein kinase B (PKB) also known as Akt, is one of the multifaceted kinase in human kinome, existing in three isoforms. Akt plays a vital role in phosphoinositide 3-kinase (PI3K) mediated oncogenesis in various malignancies and is one of the attractive targets for cancer drug discovery. The functional significance of an individual isoform of Akt is not redundant in cancer cell proliferation and metastasis instead Akt isoforms play distinct roles during metastasis; thereby regulating EMT. This study aims to determine isoform specific functions of Akt in cancer. The results obtained suggest that Akt1 restrict tumor invasion, whereas Akt2 promotes cell migration and invasion by various techniques like MTT, wound healing and invasion assay. Similarly, qRT-PCR also revealed that Akt3 has shown promising results in promoting cancer cell migration. Contrary to pro-oncogenic properties attributed to Akt, it is to be understood how various isoforms of Akt compensates each other in the regulation of common pathways during cancer progression and drug resistance. In conclusion, this study aims to target selective isoforms which is essential to inhibit cancer. However, the question now is whether, and how much, Akt inhibition will be tolerated in the clinic remains to be answered and the experiments will have to address the question of which combinations of newly devised Akt isoform specific inhibitors exert a favourable therapeutic effect in in vivo models of cancer to provide the therapeutic window with minimal toxicity.

Keywords: Akt isoforms, cancer, drug resistance, epithelial mesenchymal transition

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Authors: Özlem Ateş Sönmezoğlu, Begüm Terzi, Ahmet Yıldırım, Ramazan Özbey

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The basic criteria which determine durum wheat quality is its suitability for pasta processing that is pasta making quality. Bright yellow color is a desired property in pasta products. Durum wheat pasta making quality is affected by grain pigment content and oxidative enzymes which affect adversely bright yellow color. Of the oxidative enzymes, lipoxygenase LOX is the most effective one on oxidative bleaching of yellow pigments in durum wheat products. Thus, wheat cultivars that are high in yellow pigments but low in LOX enzyme activity should be preferred for the production of pasta with high color quality. The aim of this study was to reduce lipoxygenase activities of the backcross durum wheat lines that were previously improved for their protein quality. For this purpose, two advanced lines with different parents (TMB2 and TMB3) were used recurrent parents. Also, Gediz-75 wheat with low LOX enzyme activity was used as the gene source. In all of the generations, backcrossed plants carrying the targeted gene region (Lpx-B1.1) were selected using SSR markers by marker assisted selection method. As a result, the study will be completed in three years instead of six years required in a classical backcross breeding study, leading to the development of high-quality candidate varieties. This research has been financially supported by TÜBİTAK (Project No: 112T910).

Keywords: durum wheat, lipoxygenase, LOX, Lpx-B1.1, MAS, Triticum durum

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Authors: Yanghui Xu, Qin Ou, Xintu Wang, Feng Hou, Peng Li, Jan Peter van der Hoek, Gang Liu

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The level and removal of microplastics (MPs) in wastewater treatment plants (WWTPs) has been well evaluated by the particle number, while the mass concentration of MPs and especially nanoplastics (NPs) remains unclear. In this study, microfiltration, ultrafiltration and hydrogen peroxide digestion were used to extract MPs and NPs with different size ranges (0.01−1, 1−50, and 50−1000 μm) across the whole treatment schemes in two WWTPs. By identifying specific pyrolysis products, pyrolysis gas chromatography−mass spectrometry were used to quantify their mass concentrations of selected six types of polymers (i.e., polymethyl methacrylate (PMMA), polypropylene (PP), polystyrene (PS), polyethylene (PE), polyethylene terephthalate (PET), and polyamide (PA)). The mass concentrations of total MPs and NPs decreased from 26.23 and 11.28 μg/L in the influent to 1.75 and 0.71 μg/L in the effluent, with removal rates of 93.3 and 93.7% in plants A and B, respectively. Among them, PP, PET and PE were the dominant polymer types in wastewater, while PMMA, PS and PA only accounted for a small part. The mass concentrations of NPs (0.01−1 μm) were much lower than those of MPs (>1 μm), accounting for 12.0−17.9 and 5.6− 19.5% of the total MPs and NPs, respectively. Notably, the removal efficiency differed with the polymer type and size range. The low-density MPs (e.g., PP and PE) had lower removal efficiency than high-density PET in both plants. Since particles with smaller size could pass the tertiary sand filter or membrane filter more easily, the removal efficiency of NPs was lower than that of MPs with larger particle size. Based on annual wastewater effluent discharge, it is estimated that about 0.321 and 0.052 tons of MPs and NPs were released into the river each year. Overall, this study investigated the mass concentration of MPs and NPs with a wide size range of 0.01−1000 μm in wastewater, which provided valuable information regarding the pollution level and distribution characteristics of MPs, especially NPs, in WWTPs. However, there are limitations and uncertainties in the current study, especially regarding the sample collection and MP/NP detection. The used plastic items (e.g., sampling buckets, ultrafiltration membranes, centrifugal tubes, and pipette tips) may introduce potential contamination. Additionally, the proposed method caused loss of MPs, especially NPs, which can lead to underestimation of MPs/NPs. Further studies are recommended to address these challenges about MPs/NPs in wastewater.

Keywords: microplastics, nanoplastics, mass concentration, WWTPs, Py-GC/MS

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Authors: T. Abbas, J. McEvoy, E. Khan

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Pesticide contamination in water supply is a common environmental problem in rural agricultural communities. Advanced water treatment processes such as membrane filtration and adsorption on activated carbon only remove pesticides from water without degrading them into less toxic/easily degradable compounds leaving behind contaminated brine and activated carbon that need to be managed. Rural communities which normally cannot afford expensive water treatment technologies need an economical and sustainable filter which not only treats pesticides from water but also degrades them into benign products. In this study, iron turning waste experimented as potential point-of-use filtration media for the removal/degradation of a mixture of six chlorinated pesticides (lindane, heptachlor, endosulfan, dieldrin, endrin, and DDT) in water. As a common and traditional medium for water filtration, sand was also tested along with iron turning waste. Iron turning waste was characterized using scanning electron microscopy and energy dispersive X-Ray analyzer. Four glass columns with different filter media layer configurations were set up: (1) only sand, (2) only iron turning, (3) sand and iron turning (two separate layers), and (4) sand, iron turning and sand (three separate layers). The initial pesticide concentration and flow rate were 2 μg/L and 10 mL/min. Results indicate that sand filtration was effective only for the removal of DDT (100%) and endosulfan (94-96%). Iron turning filtration column effectively removed endosulfan, endrin, and dieldrin (85-95%) whereas the lindane and DDT removal were 79-85% and 39-56%, respectively. The removal efficiencies for heptachlor, endosulfan, endrin, dieldrin, and DDT were 90-100% when sand and iron turning waste (two separate layers) were used. However, better removal efficiencies (93-100%) for five out of six pesticides were achieved, when sand, iron turning and sand (three separate layers) were used as filtration media. Moreover, the effects of water pH, amounts of media, and minerals present in water such as magnesium, sodium, calcium, and nitrate on the removal of pesticides were examined. Results demonstrate that iron turning waste efficiently removed all the pesticides under studied parameters. Also, it completely de-chlorinated all the pesticides studied and based on the detection of by-products, the degradation mechanisms for all six pesticides were proposed.

Keywords: pesticide contamination, rural communities, iron turning waste, filtration

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Authors: Carmel Khrisna Wong Moreno, Dinah M. Espina

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The increasing price of feed ingredients has prompted farmers to seek feasible feed alternatives like the utilization of locally-grown protein-rich feedstuff which is cheaper but gives a positive result in poultry production. Trichanthera gigantea, a fodder tree which is an excellent alternative as feed ingredient in the Philippines has now gained popularity as feed supplement. This study was conducted to determine the growth and laying performance of commercial hens fed with varying levels of Trichanthera gigantea leaf meal. The incorporation of Trichanthera gigantea leaf meal at 5%, 10%, and 15% into the diet of commercial hens did not affect the growth and laying performance. Results of the study revealed that the weight gain of the birds fed with Trichanthera gigantea supplemented diets was not significantly different with the control (100% commercial layer mash). The voluntary feed intake, feed conversion ratio, weekly average egg weight and egg production of the commercial hens fed with T. gigantea leaf meal supplemented diets were not significantly different from the control. Results of the study showed that the supplementation of Trichanthera gigantea leaf meal of up to 15% into the diets of commercial hens is highly acceptable since it does not affect the growth and laying performance of the birds. In addition, it would mean a 15% savings in production cost from commercial feeds.

Keywords: egg production, growth, laying performance, trichanthera gigantea (nees)

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Authors: Strahinja Kovačević, Lidija Jevrić, Miloš Kuzmanović, Sanja Podunavac-Kuzmanović

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In this paper, various derivatives of benzimidazole have been evaluated against Gram-negative bacteria Escherichia coli. For all investigated compounds the minimum inhibitory concentration (MIC) was determined. Quantitative structure-activity relationships (QSAR) attempts to find consistent relationships between the variations in the values of molecular properties and the biological activity for a series of compounds so that these rules can be used to evaluate new chemical entities. The correlation between MIC and some absorption, distribution, metabolism and excretion (ADME) parameters was investigated, and the mathematical models for predicting the antibacterial activity of this class of compounds were developed. The quality of the multiple linear regression (MLR) models was validated by the leave-one-out (LOO) technique, as well as by the calculation of the statistical parameters for the developed models and the results are discussed on the basis of the statistical data. The results of this study indicate that ADME parameters have a significant effect on the antibacterial activity of this class of compounds. Principal component analysis (PCA) and agglomerative hierarchical clustering algorithms (HCA) confirmed that the investigated molecules can be classified into groups on the basis of the ADME parameters: Madin-Darby Canine Kidney cell permeability (MDCK), Plasma protein binding (PPB%), human intestinal absorption (HIA%) and human colon carcinoma cell permeability (Caco-2).

Keywords: benzimidazoles, QSAR, ADME, in silico

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Authors: Abderrahim Cheriguene, Fatiha Arioui

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The experimental study focuses on the extraction of pectin, whey protein and gelatin, and the study of their functional properties. Microbiological, physicochemical and sensory approach integrated has been implanted to study the effect of the incorporation of these natural food additives in the matrix of a fermented milk type set yogurt, to study the stability of the product during the periods of fermentation and post-acidification over a period of 21 days at 4°C. Pectin was extracted in hot HCl solution. Thermo-precipitation was carried out to obtain the whey proteins while the gelatin was extracted by hydrolysis of the collagen from bovine ossein. The fermented milk was prepared by varying the concentration of the incorporated additives. The measures and controls carried performed periodically on fermented milk experimental tests were carried out: pH, acidity, viscosity, the enumeration of Streptococcus thermophilus, cohesiveness, adhesiveness, taste, aftertaste, whey exudation, and odor. It appears that the acidity, viscosity, and number of Streptococcus thermophilus increased with increasing concentration of additive added in the experimental tests. Indeed, it seems clear that the quality of fermented milk and storability is more improved than the incorporation rate is high. The products showed a better test and a firmer texture limiting the whey exudation.

Keywords: fermented milk, pectin, gelatin, whey proteins, functional properties, quality, conservation, valorization

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Authors: Dominika Pihikova, Stefan Belicky, Tomas Bertok, Roman Sokol, Petra Kubanikova, Jan Tkac

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Since aberrant glycosylation is frequently accompanied by both physiological and pathological processes in a human body (cancer, AIDS, inflammatory diseases, etc.), the analysis of tumor-associated glycan patterns have a great potential for the development of novel diagnostic approaches. Moreover, altered glycoforms may assist as a suitable tool for the specificity and sensitivity enhancement in early-stage prostate cancer diagnosis. In this paper we discuss the construction and optimization of ultrasensitive sandwich biosensor platform employing lectin as glycan-binding protein. We focus on the immunoassay development, reduction of non-specific interactions and final glycoprofiling of human serum samples including both prostate cancer (PCa) patients and healthy controls. The fabricated biosensor was measured by label-free electrochemical impedance spectroscopy (EIS) with further lectin microarray verification. Furthermore, we analyzed different biosensor interfaces with atomic force microscopy (AFM) in nanomechanical mapping mode showing a significant differences in the altitude. These preliminary results revealing an elevated content of α-2,3 linked sialic acid in PCa patients comparing with healthy controls. All these experiments are important step towards development of point-of-care devices and discovery of novel glyco-biomarkers applicable in cancer diagnosis.

Keywords: biosensor, glycan, lectin, prostate cancer

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Authors: Zhang Zhenzhen

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Valproate (VPA) is commonly used in the treatment of bipolar disorder and epilepsy. The mechanism is complicated, including its ability to inhibit histone deacetylases (HDACs). Here, we show that VPA attenuated VEGF gene expression and the morphological changes in choroidal neovascularization (CNV) induced by photocoagulation in retina. C57BL/6 mice were injected subcutaneously at 300mg/kg twice daily with VPA before insult. Vascular endothelial growth factor (VEGF)-A and VEGF-B were examined in the eyes of VPA-treated mice and in human retinal pigment epithelial cell lines (ARPE-19) exposed to VPA. In addition, CNV was induced by photocoagulation in mice injected with VPA, and the volume of CNV was compared by fluorescence-labeled choroidal flat mount. Morphological changes were analyzed on stained histological sections. Western blot analysis was used to determine protein levels of VEGF-A and VEGF-B, and acetylation of histone H3 in each group. VPA injected intraperitoneally attenuated the VEGF-A and VEGF-B expression in the retina, accompanied by the hyperacetylation of retina tissue, indicating that VPA acts directly on retina tissues through acetylation to reduce the expression of VEGF. VPA also attenuated the VEGF-A mRNA expression in the retinal pigment epithelium showed by immunohistochemistry. Moreover, the administration of VPA significantly attenuated photocoagulation-induced CNV in mice. These results demonstrate that VPA attenuated VEGF production in retina associated with choroidal neovascularization possibly via the HDAC inhibition.

Keywords: retina, acetylation, chorodial neovascularization, vascular endothelial growth factor

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Authors: Nicholas Bayfield, Liam Bibo, Kaushelandra Rathore, Lucas Sanders, Mark Newman

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INTRODUCTION: Surgical pulmonary embolectomy is an established therapy for acute pulmonary embolism causing right heart dysfunction and haemodynamic instability. Massive intraoperative airway haemorrhage is a rare complication of pulmonary embolectomy. We present our institutional experience with massive airway haemorrhage complicating pulmonary embolectomy and discuss optimal therapeutic strategies. METHODS: A retrospective review of emergent surgical pulmonary embolectomy patients was undertaken. Cases complicated by massive intra-operative airway haemorrhage were identified. Intra- and peri-operative management strategies were analysed and discussed. RESULTS: Of 76 patients undergoing emergent or salvage pulmonary embolectomy, three cases (3.9%) of massive intraoperative airway haemorrhage were identified. Haemorrhage always began on weaning from cardiopulmonary bypass. Successful management strategies involved intraoperative isolation of the side of bleeding, occluding the affected airway with an endobronchial blocker, institution of veno-arterial (VA) extracorporeal membrane oxygenation (ECMO) and reversal of anticoagulation. Running the ECMO without heparinisation allows coagulation to occur. Airway haemorrhage was controlled within 24 hours of operation in all patients, allowing re-institution of dual lung ventilation and decannulation from ECMO. One case in which positive end-expiratory airway pressure was trialled initially was complicated by air embolism. Although airway haemorrhage was controlled successfully in all cases, all patients died in-hospital for reasons unrelated to the airway haemorrhage. CONCLUSION: Massive intraoperative airway haemorrhage during pulmonary embolectomy is a rare complication with potentially catastrophic outcomes. Re-perfusion alveolar and capillary injury is the likely aetiology. With a systematic approach to management, airway haemorrhage can be well controlled intra-operatively and often resolves within 24 hours. Stopping blood flow to the pulmonary arteries and support of oxygenation by the institution of VA ECMO is important. This management has been successful in our 3 cases.

Keywords: pulmonary embolectomy, cardiopulmonary bypass, cardiac surgery, pulmonary embolism

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