Search results for: beta-barrel membrane proteins

Authors: S. Essa, H. Safar, R. Raghupathy

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Human cytomegalovirus (CMV) continues to be a source of severe complications to immunologically immature and immune-compromised hosts. Effective CMV vaccine that diminishes CMV disease in transplant patients and avoids congenital infection remains of high importance as no approved vaccines exist. Though the exact links of defense mechanisms are unidentified, viral-specific antibodies and Th1/Th2 cytokine responses have been involved in controlling viral infections. CMV envelope glycoprotein B (UL55/gB), the matrix proteins (UL83/pp65, UL99/pp28, UL32/pp150), and the assembly protein UL80a/pp38 are known to be targets of antiviral immune responses. In this study, mice were immunized with five HCMV antigens (UL32/pp150, UL80a/pp38, UL99/pp28, and UL83/pp65), and serum samples were collected and evaluated for eliciting viral-specific antibody responses. Moreover, Splenocytes were collected, stimulated, and assessed for cytokine responses. The results demonstrated a CMV-antigen-specific antibody response to pp38 and pp65 (E/C >2.0). The highest titers were detected with pp38 (average E/C 16.275) followed by pp65 (average E/C 7.72). Compared to control cells, splenocytes from PP38 antigen immunized mice gave a significantly higher concentration of GM-CSF, IFN-γ, IL-2 IL-4, IL-5, and IL-17A (P<0.05). Also, splenocytes from pp65 antigen immunized mice resulted in a significantly higher concentration of GM-CSF, IFN-γ, IL-2 IL-4, IL-10, IL-12, IL-17A, and TNF- α. The designation of target CMV peptides by identifying viral-specific antibodies and cytokine responses is vital for understanding the protective immune mechanisms during CMV infection and identifying appropriate viral antigens to develop novel vaccines.

Keywords: hepatitis C virus, peripheral blood mononuclear cells, neutrophils, cytokines

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Authors: Farhan Sohail, Gul Shahnaz Irshad Hussain, Shoaib Sarwar, Ibrahim Javed, Zajif Hussain, Akhtar Nadhman

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The present study was aimed to develop a hybrid nanocarrier (NC) system with enhanced membrane permeability, bioavailability and targeted delivery of Docetaxel (DTX) in breast cancer. Hybrid NC’s based on folic acid (FA) grafted thiolated chitosan (TCS) enveloped liposomes were prepared with DTX and evaluated in-vitro and in-vivo for their enhanced permeability and bioavailability. Physicochemical characterization of NC’s including particle size, morphology, zeta potential, FTIR, DSC, PXRD, encapsulation efficiency and drug release from NC’s was determined in vitro. Permeation enhancement and p-gp inhibition were performed through everted sac method on freshly excised rat intestine which indicated that permeation was enhanced 5 times as compared to pure DTX and the hybrid NC’s were strongly able to inhibit the p-gp activity as well. In-vitro cytotoxicity and tumor targeting was done using MDA-MB-231 cell line. The stability study of the formulations performed for 3 months showed the improved stability of FA-TCS enveloped liposomes in terms of its particles size, zeta potential and encapsulation efficiency as compared to TCS NP’s and liposomes. The pharmacokinetic study was performed in vivo using rabbits. The oral bioavailability and AUC0-96 was increased 10.07 folds with hybrid NC’s as compared to positive control. Half-life (t1/2) was increased 4 times (58.76 hrs) as compared to positive control (17.72 hrs). Conclusively, it is suggested that FA-TCS enveloped liposomes have strong potential to enhance permeability and bioavailability of hydrophobic drugs after oral administration and tumor targeting.

Keywords: docetaxel, coated liposome, permeation enhancement, oral bioavailability

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Authors: Magdalena B. Kaziuk, Waldemar Kosiba

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Background: In the XXI century we have to deal with the epidemic of obesity which is important risk factor for the cardiovascular and kidney diseases. Lipo proteins are directly involved in the atherosclerotic process. Non-high-density lipo protein (non-HDL) began following widespread recognition of its superiority over LDL as a measurement of vascular event risk. Non-HDL includes residual risk which persists in patients after achieved recommended level of LDL. Materials and Methods: The study covered 111 patients (52 females, 59 males, age 51,91±14 years), hospitalized on the intern department. Body composition was assessed using the bioimpendance method and anthropometric measurements. Physical activity data were collected during the interview. The nutritional status and the obesity type were determined with the Waist to Height Ratio and the Waist to Hip Ratio. A function of the kidney was evaluated by calculating the estimated glomerular filtration rate (eGFR) using MDRD formula. Non-HDL was calculated as a difference between concentration of the Total and HDL cholesterol. Results: 10% of patients were found to be underweight; 23.9 % had correct body weight; 15,08 % had overweight, while the remaining group had obesity: 51,02 %. People with the android shape have higher non-HDL cholesterol versus with the gynoid shape (p=0.003). The higher was non-HDL, the lower eGFR had studied subjects (p < 0.001). Significant correlation was found between high non-HDL and incorrect dietary habits in patients avoiding eating vegetables, fruits and having low physical activity (p < 0.005). Conclusions: Android type of figure raises the residual risk of the heart disease associated with higher levels of non-HDL. Increasing physical activity in these patients reduces the level of non-HDL. Non-HDL seems to be the best predictor among all cholesterol measures for the cardiovascular events and worsening eGFR.

Keywords: obesity, non-HDL cholesterol, glomerular filtration rate, lifestyle

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Authors: Yahia Massinissa, Benhouda Afaf, Benbia Souhila, Meddour Noura, Takellalet Karima, Zeroual Amina

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Introduction: chenopodiaceae family species are known for their important biological activity, in which Atriplex halimus belongs . However, the inflammatory effect of this plant leaves has not been studied. This work aimed at assessing the anti- inflammatory and hemostatic activities of the methanolic extract AHMeOH of Atriplex halimus’s leaves. Methods: The extract was obtained using sonication of leaves powder in 80 % methanol. The analysis of phenolic compounds was carried out using thin-layer chromatography (TLC).The anti-inflammatory activity was determined by studying the plasmical membrane stabilization and albumin denaturation inhibition, the hemostatic activity was evaluated by measuring the plasma in the blood. Results: Quantitative determination of total flavonoids reveals that AHMeOH is rich in flavonoids (16 ± 0.88 μg Q / mg extract) and polyphenols (20 ± 0.20 μg AG / mg extract). about anti-inflammatory activity, the tests show that AHMeOH has a significant effect (P≤0.05) of inhibiting hypotonic-induced hemolysis with concentrations (100 and 200 μg / ml) with 77.55 and 90% respectively, and heat-induced hemolysis with percentages 81.75% and 87.44% respectively with significant difference (P ≤0.05). The obtained results with this plant reveal that the inhibition of denaturation of albumin is dose dependent. The concentration of 400 μg / ml gives denaturation inhibition of 81.00 ± 17.70% and the concentration 600 μg / ml gives an effect of 82.95 ± 17.40%. Regarding the haemostatic activity our extract with the doses 10 mg / ml, 20 mg / ml and 30 mg / ml confer a decrease of the plasma recalcification time in the tube, these concentrations could prolong the time of coagulation significantly compared to the control (P≤0.001). This result is an interesting indication in favor of haemostatic activity of AHMeOH. Conclusion: Atriplex Halimus has a strong anti-inflammatory activity and constitutes a potential source for the development of new treatments.

Keywords: albumin, atriplex halimus, hemostatic activity, methanolic extract

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Authors: Hai-Ying Liu, Yi-Hao Chen, Shu-Yin Pang, Feng-Hua Wang, Xiao-Fang Peng, Li-Yuan Yang, Zheng-Rong Chen, Yi Chen, Bing Zhu

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Objective: Biliary atresia (BA) is characterized by progressive liver fibrosis. Epithelial-mesenchymal transition (EMT) has been implicated as a key mechanism in the pathogenesis of organ fibrosis. MiR-200a has been shown to repress EMT. We aim to explore the role of miR-200a in the fibrogenesis of BA. Methods: We obtained the plasma samples and liver samples from patients with BA or controls to examine the role of miR-200a. Histological liver fibrosis was assessed using the Ishak fibrosis scores. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was performed to detect the expression of miR-200a in plasma. We also evaluated the expression of miR-200a in liver tissues using tyramide signal amplification fluorescence in situ hybridization (TSA-FISH). The expression of EMT related proteins zinc finger E-box-binding homeobox 1 (ZEB1), E-cadherin and α-smooth muscle actin (α-SMA) in the liver sections were detected by immunohistochemical staining. Results: We found that the expression of miR-200a was both elevated in the plasma and liver tissues from BA patients compared with the controls. The hepatic expression of ZEB1 and α-SMA were markedly increased in the liver sections from BA patients compared to the controls, whereas E-cadherin was downregulated in the BA group. Simultaneously, we noted that the hepatic expression of miR-200a, E-cadherin and α-SMA were upregulated with the progression of liver fibrosis in the BA group, while ZEB1 was downregulated with the progression of liver fibrosis in BA patients. Conclusion: These findings suggest EMT has a critical effect on the fibrotic process of BA, and the interaction between miR-200a and ZEB1 may regulate EMT and eventually influence liver fibrogenesis of BA.

Keywords: biliary atresia, liver fibrosis, MicroRNA, epithelial-mesenchymal transition, zinc finger E-box-binding homeobox 1

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Authors: Ridhish Kumar, Sudeep Nadukkandy, Anirban Roy

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The development of reverse osmosis happened in 1960. Along the years this technique has been widely accepted all over the world for varied applications ranging from seawater desalination to municipal water treatment. Forward osmosis (FO) is one of the foremost technologies for low energy consuming solutions for water purification. In this study, we have carried out a detailed analysis on selection, design, and pricing for a prototype of potable water system for purifying water in emergency situations. The portable and light purification system is envisaged to be driven by FO. This pouch will help to serve as an emergency water filtration device. The current effort employs a model to understand the interplay of permeability and area on the rate of purification of water from any impure source/brackish water. The draw solution for the FO pouch is considered to be a combination of salt and sugar such that dilution of the same would result in an oral rehydration solution (ORS) which is a boon for dehydrated patients. However, the effort takes an extra step to actually estimate the cost and pricing of designing such a prototype. While the mathematical model yields the best membrane (compositions are taken from literature) combination in terms of permeability and area, the pricing takes into account the feasibility of such a solution to be made available as a retail item. The product is envisaged to be a market competitor for packaged drinking water and ORS combination (costing around $0.5 combined) and thus, to be feasible has to be priced around the same range with greater margins in order to have a better distribution. Thus a proper business plan and production of the same has been formulated in order to be a feasible solution for unprecedented calamities and emergency situations.

Keywords: forward osmosis, water treatment, oral rehydration solution, prototype

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Authors: Sanja O. Podunavac-Kuzmanović, Strahinja Z. Kovačević, Lidija R. Jevrić

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Prion is a protein substance whose certain form is considered as infectious agent. It is presumed to be the cause of the transmissible spongiform encephalopathies (TSEs). The protein it is composed of, called PrP, can fold in structurally distinct ways. At least one of those 3D structures is transmissible to other prion proteins. Prions can be found in brain tissue of healthy people and have certain biological role. The structure of prions naturally occurring in healthy organisms is marked as PrPc, and the structure of infectious prion is labeled as PrPSc. PrPc may play a role in synaptic plasticity and neuronal development. Also, it may be required for neuronal myelin sheath maintenance, including a role in iron uptake and iron homeostasis. PrPSc can be considered as an environmental pollutant. The main aim of this study was to carry out the molecular modeling and calculation of molecular descriptors (lipophilicity, physico-chemical and topological descriptors) of structurally diverse compounds which can be considered as anti-prion agents. Molecular modeling was conducted applying ChemBio3D Ultra version 12.0 software. The obtained 3D models were subjected to energy minimization using molecular mechanics force field method (MM2). The cutoff for structure optimization was set at a gradient of 0.1 kcal/Åmol. The Austin Model 1 (AM-1) was used for full geometry optimization of all structures. The obtained set of molecular descriptors is applied in analysis of similarities and dissimilarities among the tested compounds. This study is an important step in further development of quantitative structure-activity relationship (QSAR) models, which can be used for prediction of anti-prion activity of newly synthesized compounds.

Keywords: chemometrics, molecular modeling, molecular descriptors, prions, QSAR

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Authors: Sofia G. Meirinho, Luis G. Dias, António M. Peres, Lígia R. Rodrigues

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The emerging development of electrochemical aptasen sors has enabled the easy and fast detection of protein biomarkers in standard and real samples. Biomarkers are produced by body organs or tumours and provide a measure of antigens on cell surfaces. When detected in high amounts in blood, they can be suggestive of tumour activity. These biomarkers are more often used to evaluate treatment effects or to assess the potential for metastatic disease in patients with established disease. Osteopontin (OPN) is a protein found in all body fluids and constitutes a possible biomarker because its overexpression has been related with breast cancer evolution and metastasis. Currently, biomarkers are commonly used for the development of diagnostic methods, allowing the detection of the disease in its initial stages. A previously described RNA aptamer was used in the current work to develop a simple and sensitive electrochemical aptasensor with high affinity for human OPN. The RNA aptamer was biotinylated and immobilized on a gold electrode by avidin-biotin interaction. The electrochemical signal generated from the aptamer–target molecule interaction was monitored electrochemically using cyclic voltammetry in the presence of [Fe (CN) 6]−3/− as a redox probe. The signal observed showed a current decrease due to the binding of OPN. The preliminary results showed that this aptasensor enables the detection of OPN in standard solutions, showing good selectivity towards the target in the presence of others interfering proteins such as bovine OPN and bovine serum albumin. The results gathered in the current work suggest that the proposed electrochemical aptasensor is a simple and sensitive detection tool for human OPN and so, may have future applications in cancer disease monitoring.

Keywords: osteopontin, aptamer, aptasensor, screen-printed electrode, cyclic voltammetry

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Authors: Thanusha D. Abeywickrama, Inoka C. Perera, Genji Kurisu

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Tuberculosis, considered being as the ninth leading cause of death worldwide, cause from a single infectious agent M. tuberculosis and the drug resistance nature of this bacterium is a continuing threat to the world. Therefore TB preventing treatment is expanding, where this study designed to analyze the regulatory mechanism of GntR transcriptional regulator gene Rv0792c, which lie between several genes codes for some hypothetical proteins, a monooxygenase and an oxidoreductase. The gene encoding Rv0792c was cloned into pET28a and expressed protein was purified to near homogeneity by Nickel affinity chromatography. It was previously reported that the protein binds within the intergenic region (BS region) between Rv0792c gene and monooxygenase (Rv0793). This resulted in binding of three protein molecules with the BS region suggesting tight control of monooxygenase as well as its own gene. Since monooxygenase plays a key role in metabolism, this gene may have a global regulatory role. The natural ligand for this regulator is still under investigation. In relation to the Rv0792 protein structure, a Circular Dichroism (CD) spectrum was carried out to determine its secondary structure elements. Percentage-wise, 17.4% Helix, 21.8% Antiparallel, 5.1% Parallel, 12.3% turn and 43.5% other were revealed from CD spectrum data under room temperature. Differential Scanning Calorimetry (DSC) was conducted to assess the thermal stability of Rv0792, which the melting temperature of protein is 57.2 ± 0.6 °C. The graph of heat capacity (Cp) versus temperature for the best fit was obtained for non-two-state model, which concludes the folding of Rv0792 protein occurs through stable intermediates. Peak area (∆HCal ) and Peak shape (∆HVant ) was calculated from the graph and ∆HCal / ∆HVant was close to 0.5, suggesting dimeric nature of the protein.

Keywords: CD spectrum, DSC analysis, GntR transcriptional regulator, protein structure

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Authors: Mehmet Bulent Topkaya, Mustafa Yildirim

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Wastewater treatment systems are designed and used to minimize adverse impacts of the wastewater on the environment before discharging. Various treatment options for wastewater treatment have been developed, and each of them has different performance characteristics and environmental impacts (e.g. material and land usage, energy consumption, greenhouse gas emission, water and soil emission) during construction, operation or maintenance phases. Assessing the environmental impacts during these phases are essential for the overall evaluation of the treatment systems. In this study, wastewater treatment options, such as vegetated land treatment, constructed wetland, rotating biological contactor, conventional activated sludge treatment, membrane bioreactor, extended aeration and stabilization pond are evaluated. The comparison of the environmental impacts is conducted under the assumption that the effluents will be discharged to sensitive and less sensitive areas respectively. The environmental impacts of each alternative are evaluated by life cycle assessment (LCA) approach. For this purpose, data related to energy usage, land requirement, raw material consumption, and released emissions from the life phases were collected with inventory studies based on field studies and literature. The environmental impacts were assessed by using SimaPro 7.1 LCA software. As the scale of the LCA results is global, an MS-Excel based decision support tool that includes the LCA result is developed in order to meet also the local demands. Using this tool, it is possible to assign weight factors on the LCA results according to local conditions by using Analytical Hierarchy Process and finally the most environmentally appropriate treatment option can be selected.

Keywords: analytical hierarchy process, decision support system, life cycle assessment, wastewater treatment

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Authors: Kai Pei Huang, Ting Chung Hung, Li Ching Wu

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Amyloidosis refers to a variety of conditions wherein amyloid proteins are abnormally deposited in organ or tissues and cause harm. Among the several forms of amyloidosis, the principal types of that in inpatient medical services are the AL amyloidosis (primary) and AA amyloidois (secondary). AL Amyloidois is due to deposition of protein derived from overproduction of immunoglobulin light chain in plasma cell myeloma. Furthermore, it is a systemic disorder that can present with a variety of symptoms, including heavy proteinemia and edema, heptosplenomegaly, otherwise unexplained heart failure. We reported a 78-year-old female presenting dysuria, oliguria and leg edema for several months. Laboratory data showed proteinuria (UPCR:1679.8), leukocytosis (WBC:16.2 x 10^3/uL), results of serum urea nitrogen (39mg/dL), creatinine (0.76 mg/dL), IgG (748 mg/dL.), IgA (635 mg/dL), IgM (63 mg/dL), kappa light chain(18.8 mg/dL), lambda light chain (110.0 mg/dL) and kappa/lambda ratio (0.17). Renal biopsy found amyloid fibrils in glomerular mesangial area, and Congo red stain highlights amyloid deposition in glomeruli. Additional lab studies included serum protein electrophoresis, which shows a major monoclonal peak in β region and minor small peak in gamma region, and the immunotyping studies for serum showed two IgA/λ type. We treated sample with beta-mercaptoethanol which reducing the polymerized immunoglobulin to clarify two IgA/λ are secreted from the same plasma cell clone in bone marrow. Later examination confirmed it existed plasma cell infiltration in bone marrow, and the immunohistochemical staining showed monotypic for λ light chain and are positive for IgA. All findings mentioned above reveal it is a case of plasma cell myeloma with λ Light Chain Amyloidosis.

Keywords: amyloidosis, immunoglobulin light chain, plasma cell myeloma, serum protein electrophoresis

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Authors: Ouchemoukh Salim, Amessis-Ouchemoukh Nadia, Guenaoui Nawel, Moumeni Lynda, Zaidi Hicham, Otmani Amar, Sadou Dyhia

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Honey is a hive food rich in carbohydrates and water and it also has a lot of nutrients (enzymes, minerals, organic acids, phytochemicals...). It is used in different nutritional and therapeutic fields. Algerian honey was studied for its physicochemical parameters, nutritional values (moisture, brix, pH, electrical conductivity, and amounts of HMF, proteins, proline, total phenolic compounds and flavonoids) and some biological activities (antioxidant, anti-inflammatory and enzymatic anti-browning). The antioxidant activities of the samples were estimated using different methods (ABTS, DPPH free radicals scavenging, reducing power, and chelating ferrous activity). All honeys were acidic (3.45≤pH≤4.65). The color varied from mimosa yellow to dark brown. The specific rotation was levorotatory in most honey samples, and the electrical conductivity, hydroxymethylfurfural, and proline values agreed with the international honey requirements. For anti-inflammatory activity, the results showed that the inhibiting capacity of the denaturation of the BSA of the honey analyzed varied from 15 to 75 % with a maximum of activity at the concentration of 0,5 mg/ml. All honey exhibited enzymatic anti-browning on different slices of fruits. In fact, the results showed that the controls have the greatest browning unit compared to the honeys studied and PPO and POD enzymes had the lowest enzyme activity. High significant correlations were found between the color of honey, its antioxidant content and its biological activities (antioxidant, anti-inflammatory and enzymatic anti-browning). The dark color of honey is a good indicator of the best biological properties, therefore, the best nutritional and therapeutic values.

Keywords: honey, physico-chemical parameters, bioactive compounds, biological properties

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Authors: Jasna Čanadanović-Brunet, Gordana Ćetković, Sonja Djilas, Vesna Tumbas-Šaponjac, Jelena Vulić, Sladjana Stajčić

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Honey, produced by the honeybee, is a natural saturated sugar solution, which is mainly composed of a complex mixture of carbohydrates. Besides this, it also contains certain minor constituents, proteins, enzymes, amino and organic acids, lipids, vitamins, phenolic acids, flavonoids and carotenoids. Honey serves as a source of natural antioxidants, which are effective in reducing the risk of heart disease, cancer, immune-system decline, cataracts, and different inflammatory processes. Honey is consumed in its natural form alone, but also in combination with nuts and various kinds of dried fruits (plums, figs, cranberries, apricots etc.). The aim of this research was to investigate the contribution of dried apricot addition to polyphenols and flavonoids contents and antioxidant activities of honey. Some individual phenolic compounds in Serbian polyfloral honey (PH), linden honey (LH) and also in their mixtures with dried apricot, in 40% mass concentrations (PH40; LH40), were identified and quantified by HPLC. The most dominant phenolic compound was: gallic acid in LH (11.14 mg/100g), LH40 (42.65 mg/100g), PH (7.24 mg/100g) and catehin in PH40 (11.83 mg/100g). The antioxidant activity of PH, LH, PH40 and LH40 was tested by measuring their ability to scavenge hydroxyl radicals (OH) by electron spin resonance spectroscopy (ESR). Honey samples with 40% dried apricot exhibited better antioxidant activity measured by hydroxyl radical scavenging activity. The EC50 values, the amount of antioxidant necessary to decrease the initial concentration of OH radicals by 50%, were: EC50PH=3.36 mg/ml, EC50LH=13.36 mg/ml, EC50PH40=2.29 mg/ml, EC50 LH40=7.78 mg/ml. Our results indicate that supplementation of polyfloral honey and linden honey with dried apricots improves antioxidant activity of honey by enriching the phenolic composition.

Keywords: honey, dried apricot, HPLC, hydroxyl radical

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Authors: M. Kashfi, K. Jahani

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Sound attenuation in absorptive silencers has been analyzed in this paper. The structure of such devices is as follows. When the rigid duct of an expansion chamber has been lined by a packed absorptive material under a perforated membrane, incident sound waves will be dissipated by the absorptive liners. This kind of silencer, usually are applicable for medium to high frequency ranges. Several conditions for different absorptive materials, variety in their thicknesses, and different shapes of the expansion chambers have been studied in this paper. Also, graphs of sound attenuation have been compared between empty expansion chamber and duct of silencer with applying liner. Plane waves have been assumed in inlet and outlet regions of the silencer. Presented results that have been achieved by applying finite element method (FEM), have shown the dependence of the sound attenuation spectrum to flow resistivity and the thicknesses of the absorptive materials, and geometries of the cross section (configuration of the silencer). As flow resistivity and thickness of absorptive materials increase, sound attenuation improves. In this paper, diagrams of the transmission loss (TL) for absorptive silencers in five different cross sections (rectangle, circle, ellipse, square, and rounded rectangle as the main geometry) have been presented. Also, TL graphs for silencers using different absorptive material (glass wool, wood fiber, and kind of spongy materials) as liner with three different thicknesses of 5 mm, 15 mm, and 30 mm for glass wool liner have been exhibited. At first, the effect of substances of the absorptive materials with the specific flow resistivity and densities on the TL spectrum, then the effect of the thicknesses of the glass wool, and at last the efficacy of the shape of the cross section of the silencer have been investigated.

Keywords: transmission loss, absorptive material, flow resistivity, thickness, frequency

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Authors: J. P. Chica Cano, G. Cabot, S. de Persis, F. Foucher

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Among all possibilities to combat global warming, CO₂ capture and sequestration (CCS) is presented as a great alternative to reduce greenhouse gas (GHG) emission. Several strategies for CCS from industrial and power plants are being considered. The concept of combined oxy-fuel combustion has been the most alternative solution. Nevertheless, due to the high cost of pure O₂ production, additional ways recently emerged. In this paper, an innovative combustion process for a gas turbine cycle was studied: it was composed of methane combustion with oxygen enhanced air (OEA), exhaust gas recirculation (EGR) and H₂O issuing from STIG (Steam Injection Gas Turbine), and the CO₂ capture was realized by membrane separator. The effect on this combustion process was emphasized, and it was shown that a study of the influence of H₂O dilution on the combustion parameters by experimental and numerical approaches had to be carried out. As a consequence, the laminar burning velocities measurements were performed in a stainless steel spherical combustion from atmospheric pressure to high pressure (up to 0.5 MPa), at 473 K for an equivalence ratio at 1. These experimental results were satisfactorily compared with Chemical Workbench v.4.1 package in conjunction with GRIMech 3.0 reaction mechanism. The good correlations so obtained between experimental and calculated flame speed velocities showed the validity of the GRIMech 3.0 mechanism in this domain of combustion: high H₂O dilution, low N₂, medium pressure. Finally, good estimations of flame speed and pollutant emissions were determined in other conditions compatible with real gas turbine. In particular, mixtures (composed of CH₄/O₂/N₂/H₂O/ or CO₂) leading to the same adiabatic temperature were investigated. Influences of oxygen enrichment and H₂O dilution (compared to CO₂) were disused.

Keywords: CO₂ capture, oxygen enrichment, water dilution, laminar burning velocity, pollutants emissions

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Authors: Ivan Zupan, Tomislav Saric, Suzana Tkalcic

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IMUNO-2865® is a commercially available, β–glucan based, natural hemicellulose compound with proven immunostimulative properties in people, domestic and some aquatic animals. During the experimental feeding trial with IMUNO-2865® in juvenile wild-caught chub under laboratory conditions, supplementation resulted in overall higher growth performance for all experimental groups regardless of the concentration of the added compound. The maximum, 5% concentration of the supplement, resulted in highest weight gain and calculated specific growth rate. In sea bream, as economically most important species in the Mediterranean aquaculture, significant increases in numbers of monocytes and heterophils were observed in the group supplemented with 2.5 % of IMUNO-2865® in the feed. An overall increase of erythrocytes was noted by the end of the experiment, although with variable distribution among groups. Blood Ca++ levels, total proteins, and total NH₃ were significantly higher after 60 days of feeding in all treatment groups compared to the control and remained elevated in the treated group following the secession of supplementation. Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and serum paraoxonase PON1 (U/L) showed similar trends. All these parameters are playing a significant role in either oxygen supplementation of tissues, or anabolic and catabolic processes that on molecular levels contribute to the overall health and immune-building capacity of cells and tissues. The complete lack of mortality in sea bream and presented increases in cellular, biochemical and oxidative stress parameters in the blood suggest that the IMUNO-2865® represents a safe dietary supplement for in aquaculture, with an overall positive and potentially immunostimulative effect on farmed fish.

Keywords: IMUNO-2865®, β–glucans, Mediterranean aquaculture, fish imunnostimulans

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Authors: Boudjenah-Haroun Saliha, Isselnane Souad, Nouani Abdelwahab, Baaissa Babelhadj, Mati Abderrahmane

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Algeria is experiencing significant development of the dairy sector, where consumption of milk and milk products increased by 2.7 million tons in 2008 to 4,400,000 tons in 2013, and cheese production has reached 1640 tons in the year 2014 with average consumption of 0.7 kg/person/year. Although rennet is still the most used coagulating enzyme in cheese, its production has been growing worldwide shortage. This shortage is primarily due to a growing increase in the production and consumption of cheese, and the inability to increase in parallel the production of rennet. This shortage has caused very large fluctuations in its price). To overcome these obstacles, much research has been undertaken to find effective and competitive substitutes used industrially. For this, the selection of a local production of rennet substitute is desirable. It would allow a permanent supply with limited dependence on imports and price fluctuations. Investigations conducted by our research team showed that extracts coagulants from the stomachs of older camels are characterized by a coagulating power than those from younger camels. The objective of this work is to study the possibility of substituting commercial rennet coagulant by gastric enzymes from adult camels for coagulation bovine milk. Excerpts from the raw camel coagulants obtained are characterized through their teneures proteins and clotting and proteolytic activities. Milk clotting conditions by the action of these extracts were optimized. Milk clotting time all treated with enzyme preparations and under different conditions was calculated. Bovine rennet has been used for comparison. The results show that crude extracts from gastric adult camel can be good substituting bovine rennet.

Keywords: Algeria, camel, cheese, coagulation, gastric extracts, milk

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Authors: Javad Mohiti-Ardekani, Shabodin Asadii, Ali Moradi

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Introduction: Curcumin, the yellow pigment in turmeric, has been shown as an anti-diabetic agent for centuries but only in recent few years, its mechanism of action has been under investigation. Some studies showed that curcumin might exert its anti-diabetic effect via increasing glucose transporter isotype-4 (GLUT4) gene and glycoprotein contents in cells. To investigate this possibility, we investigate the effects of extract and commercial curcumin with and without insulin on GLUT4 translocation from intracellular compartments of nuclear or endoplasmic reticulum membranes (N/ER) into the cytoplasmic membrane (CM). Methods and Material: C2C12 myoblastic cell line were seeded in DMEM plus 20 % FBS and differentiated to myotubes using 2 % horse serum. After myotubes formation, 40 µmolar Extract and Commercial curcumin, with or without insulin as intervention, and as control 1 % DMSO were added for 3 h. Cells were washed and homogenized followed by ultracentrifuge fractionation, protein separation by SDS-PAGE and GLUT4 detection using semi-quantitative Western blotting. Data analysis was done by two independent samples t-test for comparison of mean ± SD of GLUT4 percent in categories. GLUT4 contents were higher in CM groups curcumin and curcumin with insulin in comparison to 1 % DMSO-treated myotubes control group. Results: As our results have shown extract and commercial curcumin induces GLUT4 translocation from intra-cell into cell surface. The results have also shown synergic effect of curcumin on translocation of GLUT4 from intra-cell into cell surface in the presence of 100 nm insulin. Discussion: We conclude that curcumin may be a choice of type-2 diabetes mellitus treatment because its extract and commercial enhances GLUT4 contents in CM where it facilitates glucose entrance into the cell. However, it is necessary to trace the signaling pathways which are activated by curcumin.

Keywords: Curcumin, insulin, Diabetes type-2, GLUT4

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Authors: Mohammad K. Parvez, Mohammed S. Al-Dosari

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Hepatitis E virus (HEV) is an emerging RNA virus that causes acute and chronic liver disease with a global mortality rate of about 2%. Despite milestone developments in understanding of HEV biology, there is still lack of a robust culture system or animal model. Therefore, in a novel approach, two recombinant-baculoviruses (vBac-ORF2 and vBac-ORF3) that could overexpress HEV ORF2 (structural/capsid) and ORF3 (nonstructural/regulatory) proteins, respectively were constructed. The established HEV-SAR55 (genotype 1) replicon that contained GFP gene, in place of ORF2/ORF3 sequences was in vitro transcribed, and GFP production in RNA transfected S10-3 cells was scored by FACS. Enhanced infectivity, if any, of nascent virions produced by exogenously-supplied ORF2 and viral RNA by co-expression of ORF3 was tested on naïve HepG2 cells. Co-transduction with vBac-ORF2/vBac-ORF3 (108 pfu/microL) produced high amounts of native ORF2/ORF3 in approximately 60% of S10-3 cells, determined by immunofluorescence microscopy and Western analysis. FACS analysis showed about 9% GFP positivity of S10-3 cells on day6 post-transfection (i.e, day5 post-transduction). Further, FACS scoring indicated that lysates from S10-3 cultures receiving the RNA plus vBac-ORF2 were capable of producing HEV particles with about 4% infectivity in HepG2 cells. However, lysates of cultures co-transduced with vBac-ORF3, were found to further enhance virion infectivity by approximately 17%. This supported a previously proposed role of ORF3 as a minor-structural protein in HEV virion assembly and infectivity. In conclusion, the present model for efficient genomic RNA packaging and production of infectious virions could be a valuable tool to study various aspects of HEV molecular biology, in vitro.

Keywords: chronic liver disease, hepatitis E virus, ORF2, ORF3, replicon

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Authors: Chen Jun-Hong, He Pu, Tao Wen-Quan

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Proton exchange membrane fuel cell (PEMFC) is the most promising future energy source owing to its low operating temperature, high energy efficiency, high power density, and environmental friendliness. In this paper, a comprehensive PEMFC system control-oriented model is developed in the Matlab/Simulink environment, which includes the hydrogen supply subsystem, air supply subsystem, and thermal management subsystem. Besides, Improved Artificial Bee Colony (IABC) is used in the parameter identification of PEMFC semi-empirical equations, making the maximum relative error between simulation data and the experimental data less than 0.4%. Operation temperature is essential for PEMFC, both high and low temperatures are disadvantageous. In the thermal management subsystem, water pump and fan are both controlled with the PID controller to maintain the appreciate operation temperature of PEMFC for the requirements of safe and efficient operation. To improve the control effect further, fuzzy control is introduced to optimize the PID controller of the pump, and the Radial Basis Function (RBF) neural network is introduced to optimize the PID controller of the fan. The results demonstrate that Fuzzy-PID and RBF-PID can achieve a better control effect with 22.66% decrease in Integral Absolute Error Criterion (IAE) of T_st (Temperature of PEMFC) and 77.56% decrease in IAE of T_in (Temperature of inlet cooling water) compared with traditional PID. In the end, a novel thermal management structure is proposed, which uses the cooling air passing through the main radiator to continue cooling the secondary radiator. In this thermal management structure, the parasitic power dissipation can be reduced by 69.94%, and the control effect can be improved with a 52.88% decrease in IAE of T_in under the same controller.

Keywords: PEMFC system, parameter identification, temperature control, Fuzzy-PID, RBF-PID, parasitic power

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Authors: Szu-Chieh Yu, Pei-Chin Chiand, Chih-Yi Lin, Yi-Wen Chien

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UV radiation from sunlight cause numbers of acute and chronic skin damage which can result in inflammation, immune changes, physical changes and DNA damage that facilitates skin aging and the development of skin carcinogenesis. Reactive oxygen species (ROS) are generated by excessive solar UV radiation, resulting in oxidative damage to cellar components, proteins, lipids, and nucleic acids. Thus, antioxidation plays an important role that protects skin against ROS-induced injury. Safflower (Carthamus tinctorius L.) is an important Chinese medicine contained abundance flavones and hydroxysafflor yellow A (HSYA) which is main active ingredient. HSYA is part of quinochalcone and has unique structures of hydroxy groups that provided the antioxidant effect. In this study, the aim was to investigate the protective role of HYSA in human keratinocytes (HaCaT) against UVA-induced oxidative damage and the possible mechanism. The HaCaT cells were UVA-irradiated and the effects of HYSA on cell viability, reactive oxygen species generation, DNA fragmentation and lipid peroxidation were measured. The mRNA expression of matrix metalloproteinase Ι (MMP Ι), cyclooxygenase-2 (COX-2) were determined by RT-PCR. In this study, UVA exposure lead to decrease in cell viability and increase in reactive oxygen species generation in HaCaT cells. HYSA could effectively increase the viability of HaCaT cells after UVA exposure and protect them from UVA-induced oxidative stress. Moreover, HYSA can reduce inflammation through inhibition the mRNA expression of MMP Ι and COX-2. Our results suggest that HSYA can act as a free radical scavenger while keratinocytes were photodamaged. HYSA could be a useful natural medicine for the protection of epidermal cells from UVA-induced damage and will be developed into products for skin care.

Keywords: HaCaT keratinocytes, hydroxysafflor yellow A (HSYA), MMP Ι, oxidative stress

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Authors: Geno Paul S. Cumla, Jan Mariel M. Gentiles, M. Brenda Gajelan-Samson

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Eutrophication is a process where in there is a surplus of nutrients present in a lake or river. Harmful cyanobacteria, hypoxia, and primarily algae, which contain toxins, grow because of the excess nutrients. Algal blooms can cause fish kills, limiting the light penetration which reduces growth of aquatic organisms, causing die-offs of plants and produce conditions that are dangerous to aquatic and human life. The main cause for eutrophication is the presence of excessive amounts of phosphorus (P) and nitrogen (N). Nitrogen is necessary for the production of the plant tissues and is usually used to synthesize proteins. Nitrate is a compound that contains nitrogen, and at elevated levels it can cause harmful effects. Excessive amounts of phosphorus, displaced through human activity, is the major cause of algae growth and as well as degraded water quality. To accomplish this study the Assessment of Soluble inorganic nitrogen (SIN), Assessment of Soluble reactive phosphate (SRP), Determination of Chlorophyll a (Chl-a) concentration, and Determination of Dominating Taxa were done. The study addresses the high probability of algal blooms in Maqueda Bay by assessing the biotic and abiotic factors of Antiao and Jiabong rivers. The data predicts the overgrowth of algae and to create awareness to prevent the event from taking place. The study assesses the adverse effects that could be prevented by understanding and controlling algae. This should predict future cases of algal blooms and allow government agencies which require data to create programs to prevent and assess these issues.

Keywords: eutrophication, chlorophyll a, nitrogen, phosphorus, red tide, Kjeldahl method, spectrophotometer, assessment of soluble inorganic nitrogen, SIN, assessment of soluble reactive phosphate, SRP

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Authors: Devinder Kaur Sugga, Ekamdeep Kaur, Jaspreet Kaur, C. Rajesh, Preeti Rajesh, Harsimran Kaur

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Withanolides are steroidal lactones and are highly oxygenated phytoconstituents that can be developed as potential anti-carcinogenic agents. The two main withanolides, namely Withaferin A and Withanolides D, have been extensively studied for their pharmacological activities. Both these withanolides are present in the Withania somnifera (WS) leaves belonging to the family Solanaceae, also known as “Indian ginseng .”In this study effects of WS leaf extract on the MCF7 breast cancer cell line were investigated by performing a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay to evaluate the cytotoxic effects and in vitro wound-healing assay to study the effect on cancer cell migration. Our data suggest WS extracts have cytotoxic effects and are effective anti-migrating agents and thus can be a source of potential candidates for the development of potential agents against metastasis. Thus, it can be a source of potential candidates for the development of potential agents against metastasis. Insight into these results, the in-silico approach to identify the possible protein targets interacting with withanolides was taken. Protein kinase C alpha (PKCα) was among the selected 5 top-ranked target proteins identified by the Swiss Target Prediction tool. PKCα is known to promote the growth and invasion of cancer cells and is being evaluated as a prognostic biomarker and therapeutic target in clinically aggressive tumors. Molecular docking of Withaferin A and Withanolides D was performed using AutoDock Vina. Both the bioactive compounds interacted with PKCα. The targets predicted using this approach will serve as leads for the possible therapeutic potential of withanolides, the bioactive ingredients of WS extracts, as anti-cancer drugs.

Keywords: withania somnifera, withaferin A, withanolides D, PKCα

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Authors: Arámbula-Villa Gerónimo, García-Lara Kenia Y., Figueroa-Cárdenas J. D., Pérez-Robles J. F., Jiménez-Sandoval S., Salazar-López R., Herrera-Corredor J. A.

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The nixtamalization process (thermal-alkaline method) improves the nutritional part of the corn grain. In this process, the using of Ca(OH)₂ is basic, although the chemical mechanisms between this alkali and the carbohydrates (starch), proteins, lipids, and fiber have not been fully identified. In this study, the native corn starch was taken as a model, and it was subjected to cooking with different concentrations of lime (nixtamalization process) and specific studies of FTIR and XRD were carried out to identify the formation of chemical compounds, and the physical, physicochemical, rheological (paste) and structural properties of material obtained were determined. The FTIR spectra showed the formation of calcium-starch complexes. The treatments with Ca(OH)₂ showed a band shift towards 1675 cm⁻¹ and a band in 1436 cm⁻¹ (COO⁻), indicating the oxidation of starch. Three bands were identified (1575, 1550, and 1540 cm⁻¹) characteristics of carboxylic acid salts for three types of coordinated structures: monodentate, pseudo-bridged, and bidentate. The XRD spectra of starch treated with Ca(OH)₂ showed a peak corresponding to CaCO₃ (29.40°). The oxidation of starch was favored with low concentrations of Ca(OH)₂, producing carboxyl and carbonyl groups and increasing the residual CaCO₃. The increased concentration of Ca(OH)₂ showed the formation of calcium carboxylates, with a decrease in relative crystallinity and residual CaCO₃. Samples with low concentrations of Ca(OH)₂ slowed the onset of gelatinization and increased the swelling of the granules and the peak viscosity. The higher concentrations of Ca(OH)₂ difficulted the water absorption and decreased the viscosity rate and peak viscosity. These results can be used to improve the quality characteristics of the dough and tortillas and to get better acceptance by consumers.

Keywords: maize starch, nixtamalization, gelatinization, calcium carboxylates

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Authors: K. Nikovics, D. Riccobono, M. Oger, H. Morin, L. Barbier, T. Poyot, X. Holy, A. Bendahmane, M. Drouet, A. L. Favier

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Muscle regeneration after injury (as irradiation) is of great importance. However, the molecular and cellular mechanisms are still unclear. Cytokines are believed to play fundamental role in the different stages of muscle regeneration. They are secreted by many cell populations, but the predominant producers are macrophages and helper T cells. On the other hand, it has been shown that adipose tissue derived stromal/stem cell (ASC) injection could improve muscle regeneration. Stem cells probably induce the coordinated modulations of gene expression in different macrophage cells. Therefore, we investigated the patterns and timing of changes in gene expression of different cytokines occurring upon stem cells loading. Muscle regeneration was studied in an irradiated muscle of minipig animal model in presence or absence of ASC treatment (irradiated and treated with ASCs, IRR+ASC; irradiated not-treated with ASCs, IRR; and non-irradiated no-IRR). We characterized macrophage populations by immunolabeling in the different conditions. In our study, we found mostly M2 and a few M1 macrophages in the IRR+ASC samples. However, only few M2b macrophages were noticed in the IRR muscles. In addition, we found intensive fibrosis in the IRR samples. With in situ hybridization and immunolabeling, we analyzed the cytokine expression of the different macrophages and we showed that M2d macrophage are the most abundant in the IRR+ASC samples. By in situ hybridization, strong expression of the transforming growth factor β (TGF-β) was observed in the IRR+ASC but very week in the IRR samples. But when we analyzed TGF-β level with immunolabeling the expression was very different: many M2 macrophages showed week expression in IRR+ASC and few cells expressing stronger level in IRR muscles. Therefore, we investigated the MMP expressions in the different muscles. Our data showed that the M2 macrophages of the IRR+ASC muscle expressed MMP2 proteins. Our working hypothesis is that MMP2 expression of the M2 macrophages can decrease fibrosis in the IRR+ASC muscle by capturing TGF-β.

Keywords: adipose tissue derived stromal/stem cell, cytokine, macrophage, muscle regeneration

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Authors: Gulyas Erzsebet

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The multimerisation of proteins is a common feature of many cellular processes; however, it could also impair protein functions and/or be associated with the occurrence of diseases. Thus, development of a research tool monitoring the appearance/presence of multimeric protein forms has great importance for a variety of research fields. Such a tool is potentially applicable in the ante-mortem diagnosis of certain conformational diseases, such as transmissible spongiform encephalopathies (TSE) and Alzheimer’s disease. These conditions are accompanied by the appearance of aggregated protein multimers, present in low concentrations in various tissues. This detection is particularly relevant for TSE where the handling of tissues derived from affected individuals and of meat products of infected animals have become an enormous health concern. Here we demonstrate the potential of such a multimer detection approach in TSE by developing a facile approach. The Biospiral-Detect system resembles a traditional sandwich ELISA, except that the capturing antibody that is attached to a solid surface and the detecting antibody is directed against the same or overlapping epitopes. As a consequence, the capturing antibody shields the epitope on the captured monomer from reacting with the detecting antibody, therefore monomers are not detected. Thus, MDS is capable of detecting only protein multimers with high specificity. We developed an alternative system as well, where RNA aptamers were employed instead of monoclonal antibodies. In order to minimize degradation, the 3' and 5' ends of the aptamer contained deoxyribonucleotides and phosphorothioate linkages. When compared the monoclonal antibodies-based system with the aptamers-based one, the former proved to be superior. Thus all subsequent experiments were conducted by employing the Biospiral -Detect modified sandwich ELISA kit. Our approach showed an order of magnitude higher sensitivity toward mulimers than monomers suggesting that this approach may become a valuable diagnostic tool for conformational diseases that are accompanied by multimerization.

Keywords: diagnosis, ELISA, Prion, TSE

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Authors: Meryem Zarri, Mohame Tahiri, Fouad Amraoui

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In the context of water resources shortage that Morocco is experiencing in recent years, the mobilization of non-conventional resources becomes a necessity. The reuse of treated water and the bioconversion of biological sewage sludge into value-added products is considered an environmentally friendly and economical approach to the management of this significant resource which represent at least 80 % of consumed fresh wate In this work, we compare the quality of treated water and sewage sludge from wastewater treatment plants in the peri-urban Casablanca by analyzing different physicochemical and bacteriological parameters. The choice was made for three wastewater plants installed in different regions and monitored either by LYDEC and Commune of Had Soualem and use different technologies. Recycling of treated water in agriculture and watering of green spaces is dependent on the compliance of the parameters with international standards (WHO, FAO, …etc.) The preliminary tests of the samples taken during the second half of the year 2021 showed that the advanced technologies put in place at the level of the Mediouna and the airport zone stations (membrane reactor and activated sludge, respectively) give water to the output of the stations more respectful of the standards required in terms of physicochemical parameters (pH, Conductivity, Tubidity, COD, BOD5, TNK, and TPK) and bacteriological (fecal germs, Escherichia Coli, streptococci, Helminthes eggs). The parameters relating to the Had Soualem natural lagoon station are generally at the tolerance’s threshold. The results of analyzes relating to the residual sludge collected at the end of the cycle are, on the whole satisfactory despite a fluctuating variability of the bacteriological parameters.

Keywords: urban wastewater treatment plants, purified wastewater, sewage sludge, physicochemical parameters, bacteriological parameters, peri-urban area of ​​casablanca, morocco

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Authors: Simona Moravcova, Jiri Novotny, Zdenka Bendova

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The pineal gland of the vertebrate brain is a circumventricular organ which serves as a major neuroendocrine gland with the primary function of rhythmic secretion of neurohormone melatonin under the control of the hypothalamic suprachiasmatic nucleus (SCN). Soon after the onset of the darkness, the activity of the key rate-limiting enzyme for melatonin synthesis, arylalkylamine N-acetyltransferase (AANAT), raises due to the increased release of norepinephrine from sympathetic neurons terminating on the parenchymal cells where it binds to β-adrenergic receptors. Melatonin codes the length of the night, and it is well recognized for its anti-inflammatory effects. However, to our knowledge, less is known about the effect of the immune system on the melatonin biosynthesis and the precise role of the STAT3 in the signaling pathway leading to the expression of AANAT. Lipopolysaccharide (LPS) is the essential component in the outer surface membrane of gram-negative bacteria and acts as a strong stimulator of natural and innate immunity. STAT3 acts as an important factor in immune response. Here we investigated the effect of LPS on the components of the melatonergic synthetic pathway in the pineal gland. The experiments were performed both in vivo and in vitro. The changes in AANAT activity were determined by radioenzymatic assay. PCR analyses were carried out to detect aa-nat, icer, spi-3 and stat3 gene expression. From our results, it is apparent that the high basal level of phosphorylated forms of STAT3 can be elevated after systemic as well as in vitro administration of LPS. Our experiments have shown that LPS reduces melatonin synthesis, nevertheless, the activity of AANAT was increased. Moreover, the basal level of phosphorylated STAT3 counteracts β-adrenergic receptor-mediated aa-nat gene expression and sustains its own and spi-3 gene expression. In conclusion, LPS can affect immunomodulators such as melatonin in the pineal gland.

Keywords: AANAT, lipopolysaccharide, pineal gland, rat, STAT3

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Authors: Syeda Fahria Hoque Mimmi

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Many new and promising treatments for reducing or diminishing the adverse effects of microorganisms are being discovered day by day. On the other hand, the dairy industry is accelerating the economic wheel of Bangladesh. Considering all these facts, new thoughts were developed to isolate milk proteins by the present experiment for opening up a new era of developing natural antibiotics from milk. Lactoferrin, an iron-binding glycoprotein with multifunctional properties, is crucial to strengthening the immune system and also useful for commercial applications. The protein’s iron-binding capacity makes it undoubtedly advantageous to immune system modulation and different bacterial strains. For fulfilling the purpose, 4 of raw and 17 of commercially available milk samples were collected from different farms and stores in Bangladesh (Dhaka, Chittagong, and Cox’s Bazar). Protein quantification by nanodrop technology has confirmed that raw milk samples have better quantities of protein than the commercial ones. All the samples were tested for their antimicrobial activity against 18 pathogens, where raw milk samples showed a higher percentage of antibacterial activity. In addition to this, SDS-PAGE (Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis) was performed to identify lactoferrin in the milk samples. Lactoferrin was detected in 9 samples from which 4 were raw milk samples. Interestingly, Streptococcus pyogenes, Klebsiella pneumoniae, Bacillus cereus, Pseudomonas aeruginosa, Vibrio cholera, Staphylococcus aureus, and enterotoxigenic E. coli significantly displayed sensitivity against lactoferrin collected from raw milk. Only Bacillus cereus, Pseudomonas aeruginosa, Streptococcus pneumonia, Enterococcus faecalis, and ETEC (Enterotoxigenic Escherichia coli) were susceptible to lactoferrin obtained from a commercial one. This study suggested that lactoferrin might be used as the potential alternative of antibiotics for many diseases and also can be used to reduce microbial deterioration in the food and feed industry.

Keywords: alternative of antibiotics, commercially available milk, lactoferrin, nanodrop technology, pathogens, raw milk

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Authors: L. Piñeiro, N. Cobas, L. Gómez-Limia, S. Martínez, I. Franco

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The swordfish (Xiphias gladius) is a large oceanic fish of high commercial value, which is widely distributed in waters of the world’s oceans. They are considered to be an important source of high quality proteins, vitamins and essential fatty acids, although only half of the population follows the recommendation of nutritionists to consume fish at least twice a week. Swordfish is consumed worldwide because of its low fat content and high protein content. It is generally sold as fresh, frozen, and as pieces or slices. The aim of this study was to evaluate the effect of salting and frying on the composition of the water-soluble vitamins (B₂, B₃, B₉ and B₁₂) and fat-soluble vitamins (A, D, and E) of swordfish. Three loins of swordfish from Pacific Ocean were analyzed. All the fishes had a weight between 50 and 70 kg and were transported to the laboratory frozen (-18 ºC). Before the processing, they were defrosted at 4 ºC. Each loin was sliced and salted in brine. After cleaning the slices, they were divided into portions (10×2 cm) and fried in olive oil. The identification and quantification of vitamins were carried out by high-performance liquid chromatography (HPLC), using methanol and 0.010% trifluoroacetic acid as mobile phases at a flow-rate of 0.7 mL min^-1. The UV-Vis detector was used for the detection of the water- and fat-soluble vitamins (A and D), as well as the fluorescence detector for the detection of the vitamin E. During salting, water and fat-soluble vitamin contents remained constant, observing an evident decrease in the values of vitamin B₂. The diffusion of salt into the interior of the pieces and the loss of constitution water that occur during this stage would be related to this significant decrease. In general, after frying water-soluble and fat-soluble vitamins showed a great thermolability with high percentages of retention with values among 50–100%. Vitamin B₃ is the one that exhibited higher percentages of retention with values close to 100%. However, vitamin B₉ presented the highest losses with a percentage of retention of less than 20%.

Keywords: frying, HPLC, salting, swordfish, vitamins

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