Search results for: optical whitening agent

Authors: Brahim Mazian, Anne Bergeret, Jean-Charles Benezet, Sandrine Bayle, Luc Malhautier

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Hemp fibers are increasingly used as reinforcements in polymer matrix composites due to their competitive performance (low density, mechanical properties and biodegradability) compared to conventional fibres such as glass fibers. However, the huge variation of their biochemical, physical and mechanical properties limits the use of these natural fibres in structural applications when high consistency and homogeneity are required. In the hemp industry, traditional processes termed field retting are commonly used to facilitate the extraction and separation of stem fibers. This retting treatment consists to spread out the stems on the ground for a duration ranging from a few days to several weeks. Microorganisms (fungi and bacteria) grow on the stem surface and produce enzymes that degrade pectinolytic substances in the middle lamellae surrounding the fibers. This operation depends on the weather conditions and is currently carried out very empirically in the fields so that a large variability in the hemp fibers quality (mechanical properties, color, morphology, chemical composition…) is resulting. Nonetheless, if controlled, retting might be favorable for good properties of hemp fibers and then of hemp fibers reinforced composites. Therefore, the present study aims to investigate the influence of controlled retting within a designed environmental chamber (lab-scale pilot unit) on the quality of the hemp fibres harvested at the seed maturity growth stage. Various assessments were applied directly on fibers: color observations, morphological (optical microscope), surface (ESEM), biochemical (gravimetry) analysis, spectrocolorimetric measurements (pectins content), thermogravimetric analysis (TGA) and tensile testing. The results reveal that controlled retting leads to a rapid change of color from yellow to dark grey due to development of microbial communities (fungi and bacteria) at the stem surface. An increase of thermal stability of fibres due to the removal of non-cellulosic components along retting is also observed. A separation of bast fibers to elementary fibers occurred with an evolution of chemical composition (degradation of pectins) and a rapid decrease in tensile properties (380MPa to 170MPa after 3 weeks) due to accelerated retting process. The influence of controlled retting on the biocomposite material (PP / hemp fibers) properties is under investigation.

Keywords: controlled retting, hemp fibre, mechanical properties, thermal stability

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Authors: Sara Assi, Berthe Hayar, Claudio Pisano, Nadine Darwiche, Walid Saad

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Nanomedicine, the application of nanotechnology to medicine, is an emerging discipline that has gained significant attention in recent years. Current breakthroughs in nanomedicine have paved the way to develop effective drug delivery systems that can be used to target cancer. The use of nanotechnology provides effective drug delivery, enhanced stability, bioavailability, and permeability, thereby minimizing drug dosage and toxicity. As such, the use of nanoparticle (NP) formulations in drug delivery has been applied in various cancer models and have shown to improve the ability of drugs to reach specific targeted sites in a controlled manner. Cancer is one of the major causes of death worldwide; in particular, colorectal cancer (CRC) is the third most common type of cancer diagnosed amongst men and women and the second leading cause of cancer related deaths, highlighting the need for novel therapies. Retinoids, consisting of natural and synthetic derivatives, are a class of chemical compounds that have shown promise in preclinical and clinical cancer settings. However, retinoids are limited by their toxicity and resistance to treatment. To overcome this resistance, various synthetic retinoids have been developed, including the adamantyl retinoid ST1926, which is a potent anti-cancer agent. However, due to its limited bioavailability, the development of ST1926 has been restricted in phase I clinical trials. We have previously investigated the preclinical efficacy of ST1926 in CRC models. ST1926 displayed potent inhibitory and apoptotic effects in CRC cell lines by inducing early DNA damage and apoptosis. ST1926 significantly reduced the tumor doubling time and tumor burden in a xenograft CRC model. Therefore, we developed ST1926-NPs and assessed their efficacy in CRC models. ST1926-NPs were produced using Flash NanoPrecipitation with the amphiphilic diblock copolymer polystyrene-b-ethylene oxide and cholesterol as a co-stabilizer. ST1926 was formulated into NPs with a drug to polymer mass ratio of 1:2, providing a stable formulation for one week. The contin ST1926-NP diameter was 100 nm, with a polydispersity index of 0.245. Using the MTT cell viability assay, ST1926-NP exhibited potent anti-growth activities as naked ST1926 in HCT116 cells, at pharmacologically achievable concentrations. Future studies will be performed to study the anti-tumor activities and mechanism of action of ST1926-NPs in a xenograft mouse model and to detect the compound and its glucuroconjugated form in the plasma of mice. Ultimately, our studies will support the use of ST1926-NP formulations in enhancing the stability and bioavailability of ST1926 in CRC.

Keywords: nanoparticles, drug delivery, colorectal cancer, retinoids

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Authors: M. Aboalazayem, Essam A. Gouda, Ahmed M. Moussa, Amr E. Flifl

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Field data collecting is considered one of the most difficult work due to the difficulty of accessing large zones such as large lakes. Also, it is well known that the cost of obtaining field data is very expensive. Remotely monitoring of lake water quality (WQ) provides an economically feasible approach comparing to field data collection. Researchers have shown that lake WQ can be properly monitored via Remote sensing (RS) analyses. Using satellite images as a method of WQ detection provides a realistic technique to measure quality parameters across huge areas. Landsat (LS) data provides full free access to often occurring and repeating satellite photos. This enables researchers to undertake large-scale temporal comparisons of parameters related to lake WQ. Satellite measurements have been extensively utilized to develop algorithms for predicting critical water quality parameters (WQPs). The goal of this paper is to use RS to derive WQ indicators in Aswan High Dam Reservoir (AHDR), which is considered Egypt's primary and strategic reservoir of freshwater. This study focuses on using Landsat8 (L-8) band surface reflectance (SR) observations to predict water-quality characteristics which are limited to Turbidity (TUR), total suspended solids (TSS), and chlorophyll-a (Chl-a). ArcGIS pro is used to retrieve L-8 SR data for the study region. Multiple linear regression analysis was used to derive new correlations between observed optical water-quality indicators in April and L-8 SR which were atmospherically corrected by values of various bands, band ratios, and or combinations. Field measurements taken in the month of May were used to validate WQP obtained from SR data of L-8 Operational Land Imager (OLI) satellite. The findings demonstrate a strong correlation between indicators of WQ and L-8 .For TUR, the best validation correlation with OLI SR bands blue, green, and red, were derived with high values of Coefficient of correlation (R2) and Root Mean Square Error (RMSE) equal 0.96 and 3.1 NTU, respectively. For TSS, Two equations were strongly correlated and verified with band ratios and combinations. A logarithm of the ratio of blue and green SR was determined to be the best performing model with values of R2 and RMSE equal to 0.9861 and 1.84 mg/l, respectively. For Chl-a, eight methods were presented for calculating its value within the study area. A mix of blue, red, shortwave infrared 1(SWR1) and panchromatic SR yielded the greatest validation results with values of R2 and RMSE equal 0.98 and 1.4 mg/l, respectively.

Keywords: remote sensing, landsat 8, nasser lake, water quality

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Authors: Muhammad Qamar Masood, Syed Abbas Raza, Umar Yousaf Raja, Imran Hassan, Bilal Afzal, Muhammad Aleem Zahir, Atika Shaheer

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Background: Cardiovascular disease (CVD) is a major burden among people with type 2 diabetes (T2D) with 1 in 3 reported to have CVD. Therefore, understanding real-world clinical characteristics and prescribing patterns could help in better care. Objective: The CONVERGE (Cardiovascular Outcomes and Value in the Real world with GLP-1RAs) study characterized demographics and medication usage patterns in T2D intensified (add-on to metformin) overall population. The data were further divided into subgroups {dipeptidyl peptidase-4 inhibitors (DPP-4is), sulfonylureas (SUs), insulins, glucagon-like peptide-1 receptor agonists (GLP-1 RAs) and sodium-glucose cotransporter-2 inhibitors (SGLT-2is)}, according to the latest prescribed antidiabetic agent (ADA) in India/Pakistan/Thailand. Here, we report findings from Pakistan. Methods: A multi-center retrospective study utilized data from medical records between 13-Sep-2008 (post-market approval of GLP-1RAs) and 31-Dec-2017 in adults (≥18-year-old). The data for this study were collected from 05 centers / institutes located in major cities of Pakistan, including Karachi, Lahore, Islamabad, and Multan. These centers included National Hospital, Aga Khan University Hospital, Diabetes Endocrine Clinic Lahore, Shifa International Hospital, Mukhtar A Sheikh Hospital Multan. Data were collected at start of medical record and at 6 or 12-months prior to baseline based on variable type; analyzed descriptively. Results: Overall, 1,010 patients were eligible. At baseline, overall mean age (SD) was 51.6 (11.3) years, T2D duration was 2.4 (2.6) years, HbA1c was 8.3% (1.9) and 35% received ≥1CVD medications in the past 1-year (before baseline). Most frequently prescribed ADAs post-metformin were DPP-4is and SUs (~63%). Only 6.5% received GLP-1RAs and SGLT-2is were not available in Pakistan during the study period. Overall, it took a mean of 4.4 years and 5 years to initiate GLP-1RAs and SGLT-2is, respectively. In comparison to other subgroups, more patients from GLP-1RAs received ≥3 types of ADA (58%), ≥1 CVD medication (64%) and had higher body mass index (37kg/m2). Conclusions: Utilization of GLP-1RAs and SGLT-2is was low, took longer time to initiate and not before trying multiple ADAs. This may be due to lack of evidence for CV benefits for these agents during the study period. The planned phase 2 of the CONVERGE study can provide more insights into utilization and barriers to prescribe GLP-1RAs and SGLT-2is post 2018 in Pakistan.

Keywords: type 2 diabetes, GLP-1RA, treatment intensification, cardiovascular disease

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Authors: Jannis Kountouras, Nikolaos Kapetanakis, Stergios A. Polyzos, Apostolis Papaeftymiou, Panagiotis Katsinelos, Ioannis Venizelos, Christina Nikolaidou, Christos Zavos, Iordanis Romiopoulos, Elena Tsiaousi, Evangelos Kazakos, Michael Doulberis

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Background & Aims: Helicobacter pylori infection (Hp-I) has been recognized as a substantial risk agent involved in gastrointestinal (GI) tract oncogenesis by stimulating cancer stem cells (CSCs), oncogenes, immune surveillance processes, and triggering GI microbiota dysbiosis. We aimed to investigate the possible involvement of active Hp-I in the sequence: chronic inflammation–adenoma–colorectal cancer (CRC) development. Methods: Four pillars were investigated: (i) endoscopic and conventional histological examinations of patients with CRC, colorectal adenomas (CRA) versus controls to detect the presence of active Hp-I; (ii) immunohistochemical determination of the presence of Hp; expression of CD44, an indicator of CSCs and/or bone marrow-derived stem cells (BMDSCs); expressions of oncogene Ki67 and anti-apoptotic Bcl-2 protein; (iii) expression of CD45, indicator of immune surveillance locally (assessing mainly T and B lymphocytes locally); and (iv) correlation of the studied parameters with the presence or absence of Hp-I. Results: Among 50 patients with CRC, 25 with CRA, and 10 controls, a significantly higher presence of Hp-I in the CRA (68%) and CRC group (84%) were found compared with controls (30%). The presence of Hp-I with accompanying immunohistochemical expression of CD44 in biopsy specimens was revealed in a high proportion of patients with CRA associated with moderate/severe dysplasia (88%) and CRC patients with moderate/severe degree of malignancy (91%). Comparable results were also obtained for Ki67, Bcl-2, and CD45 immunohistochemical expressions. Concluding Remarks: Hp-I seems to be involved in the sequence: CRA – dysplasia – CRC, similarly to the upper GI tract oncogenesis, by several pathways such as the following: Beyond Hp-I associated insulin resistance, the major underlying mechanism responsible for the metabolic syndrome (MetS) that increase the risk of colorectal neoplasms, as implied by other Hp-I related MetS pathologies, such as non-alcoholic fatty liver disease and upper GI cancer, the disturbance of the normal GI microbiota (i.e., dysbiosis) and the formation of an irritative biofilm could contribute to a perpetual inflammatory upper GIT and colon mucosal damage, stimulating CSCs or recruiting BMDSCs and affecting oncogenes and immune surveillance processes. Further large-scale relative studies with a pathophysiological perspective are necessary to demonstrate in-depth this relationship.

Keywords: Helicobacter pylori, colorectal cancer, colorectal adenomas, gastrointestinal oncogenesis

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Authors: Jalil ur Rehman, Ramesh C. Tailor, Muhammad Isa Khan, Jahnzeeb Ashraf, Muhammad Afzal, Geofferry S. Ibbott

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Purpose: The purpose of this analysis was to investigate dose distribution of different techniques (3D-CRT, IMRT and VMAT) of head and neck cancer using 3-dimensional dosimeter called PRESAGETM Dosimeter. Materials and Methods: Computer tomography (CT) scans of radiological physics center (RPC) head and neck anthropomorphic phantom with both RPC standard insert and PRESAGETM insert were acquired separated with Philipp’s CT scanner and both CT scans were exported via DICOM to the Pinnacle version 9.4 treatment planning system (TPS). Each plan was delivered twice to the RPC phantom first containing the RPC standard insert having TLD and film dosimeters and then again containing the Presage insert having 3-D dosimeter (PRESAGETM) by using a Varian True Beam linear accelerator. After irradiation, the standard insert including point dose measurements (TLD) and planar Gafchromic® EBT film measurement were read using RPC standard procedure. The 3D dose distribution from PRESAGETM was read out with the Duke Midsized optical scanner dedicated to RPC (DMOS-RPC). Dose volume histogram (DVH), mean and maximal doses for organs at risk were calculated and compared among each head and neck technique. The prescription dose was same for all head and neck radiotherapy techniques which was 6.60 Gy/friction. Beam profile comparison and gamma analysis were used to quantify agreements among film measurement, PRESAGETM measurement and calculated dose distribution. Quality assurances of all plans were performed by using ArcCHECK method. Results: VMAT delivered the lowest mean and maximum doses to organ at risk (spinal cord, parotid) than IMRT and 3DCRT. Such dose distribution was verified by absolute dose distribution using thermoluminescent dosimeter (TLD) system. The central axial, sagittal and coronal planes were evaluated using 2D gamma map criteria(± 5%/3 mm) and results were 99.82% (axial), 99.78% (sagital), 98.38% (coronal) for VMAT plan and found the agreement between PRESAGE and pinnacle was better than IMRT and 3D-CRT plan excludes a 7 mm rim at the edge of the dosimeter. Profile showed good agreement for all plans between film, PRESAGE and pinnacle and 3D gamma was performed for PTV and OARs, VMAT and 3DCRT endow with better agreement than IMRT. Conclusion: VMAT delivered lowered mean and maximal doses to organs at risk and better PTV coverage during head and neck radiotherapy. TLD, EBT film and PRESAGETM dosimeters suggest that VMAT was better for the treatment of head and neck cancer than IMRT and 3D-CRT.

Keywords: RPC, 3DCRT, IMRT, VMAT, EBT2 film, TLD, PRESAGETM

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Authors: Dimitrios Bakavos, Dimitrios Tsivoulas, Chaowalit Limmaneevichitr

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Aluminium cast alloys, of the Al-Si system, are widely used for shape castings. Their microstructures can be further improved on one hand, by alloying modification and on the other, by optimised artificial ageing. In this project four hypoeutectic Al-alloys, the A356, A356+ Ag, A356+Sc, and A356+Ag+Sc have been studied. The interactions of Ag and Sc during solidification and artificial ageing at 170°C to a T6 temper have been investigated in details. The evolution of the eutectic microstructure is studied by thermal analysis and interrupted solidification. The ageing kinetics of the alloys has been identified by hardness measurements. The precipitate phases, number density, and chemical composition has been analysed by means of transmission electron microscopy (TEM) and EDS analysis. Furthermore, the SHT effect onto the Si eutectic particles for the four alloys has been investigated by means of optical microscopy, image analysis, and the UTS strength has been compared with the UTS of the alloys after casting. The results suggest that the Ag additions, significantly enhance the ageing kinetics of the A356 alloy. The formation of β” precipitates were kinetically accelerated and an increase of 8% and 5% in peak hardness strength has been observed compared to the base A356 and A356-Sc alloy. The EDS analysis demonstrates that Ag is present on the β” precipitate composition. After prolonged ageing 100 hours at 170°C, the A356-Ag exhibits 17% higher hardness strength compared to the other three alloys. During solidification, Sc additions change the macroscopic eutectic growth mode to the propagation of a defined eutectic front from the mold walls opposite to the heat flux direction. In contrast, Ag has no significance effect on the solidification mode revealing a macroscopic eutectic growth similar to A356 base alloy. However, the mechanical strength of the as cast A356-Ag, A356-Sc, and A356+Ag+Sc additions has increased by 5, 30, and 35 MPa, respectively. The outcome is a tribute to the refining of the eutectic Si that takes place which it is strong in the A356-Sc alloy and more profound when silver and scandium has been combined. Moreover after SHT the Al alloy with the highest mechanical strength, is the one with Ag additions, in contrast to the as-cast condition where the Sc and Sc+Ag alloy was the strongest. The increase of strength is mainly attributed to the dissolution of grain boundary precipitates the increase of the solute content into the matrix, the spherodisation, and coarsening of the eutectic Si. Therefore, we could safely conclude for an A356 hypoeutectic alloy additions of: Ag exhibits a refining effect on the Si eutectic which is improved when is combined with Sc. In addition Ag enhance, the ageing kinetics increases the hardness and retains its strength at prolonged artificial ageing in a Al-7Si 0.3Mg hypoeutectic alloy. Finally the addition of Sc is beneficial due to the refinement of the α-Al grain and modification-refinement of the eutectic Si increasing the strength of the as-cast product.

Keywords: ageing, casting, mechanical strength, precipitates

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Authors: Takaaki Nemoto

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Case presentation: A 69-year-old Japanese man presented with a low-grade fever and fatigue that had persisted for one month. The patient had an aortic dissection on the aortic arch 13 years prior, an abdominal aortic aneurysm seven years prior, and an aortic dissection on the distal aortic arch one year prior, which were all treated with artificial blood-vessel replacement surgery. Laboratory tests revealed an inflammatory response (CRP 7.61 mg/dl), high serum creatinine (Cr 1.4 mg/dL), and elevated transaminase (AST 47 IU/L, ALT 45 IU/L). The patient was admitted to our hospital on suspicion of prosthetic vascular graft infection. Following further workups on the inflammatory response, an enhanced chest computed tomography (CT) and a non-enhanced chest DWI (MRI) were performed. The patient was diagnosed with a pulmonary fistula and a prosthetic vascular graft infection on the distal aortic arch. After admission, the patient was administered Ceftriaxion and Vancomycine for 10 days, but his fever and inflammatory response did not improve. On day 13 of hospitalization, a lung fistula repair surgery and an omental filling operation were performed, and Meropenem and Vancomycine were administered. The fever and inflammatory response continued, and therefore we took repeated blood cultures. M. fortuitum was detected in a blood culture on day 16 of hospitalization. As a result, we changed the treatment regimen to Amikacin (400 mg/day), Meropenem (2 g/day), and Cefmetazole (4 g/day), and the fever and inflammatory response began to decrease gradually. We performed a test of sensitivity for Mycobacterium fortuitum, and found that the MIC was low for fluoroquinolone antibacterial agent. The clinical course was good, and the patient was discharged after a total of 8 weeks of intravenous drug administration. At discharge, we changed the treatment regimen to Levofloxacin (500 mg/day) and Clarithromycin (800 mg/day), and prescribed these two drugs as a long life suppressive therapy. Discussion: There are few cases of prosthetic vascular graft infection caused by mycobacteria, and a standard therapy remains to be established. For prosthetic vascular graft infections, it is ideal to provide surgical and medical treatment in parallel, but in this case, surgical treatment was difficult and, therefore, a conservative treatment was chosen. We attempted to increase the treatment success rate of this refractory disease by conducting a susceptibility test for mycobacteria and treating with different combinations of antimicrobial agents, which was ultimately effective. With our treatment approach, a good clinical course was obtained and continues at the present stage. Conclusion: Although prosthetic vascular graft infection resulting from mycobacteria is a refractory infectious disease, it may be curative to administer appropriate antibiotics based on the susceptibility test in addition to surgical treatment.

Keywords: prosthetic vascular graft infection, lung fistula, Mycobacterium fortuitum, conservative treatment

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Authors: Justine Garraud, Hervé Capiaux, Cécile Le Guern, Pierre Gaudin, Clémentine Lapie, Samuel Chaffron, Erwan Delage, Thierry Lebeau

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The repeated use of Bordeaux mixture (copper sulphate) and other chemical forms of copper (Cu) has led to its accumulation in wine-growing soils for more than a century, to the point of modifying the ecosystem of these soils. Phytoextraction of copper could progressively reduce the Cu load in these soils, and even to recycle copper (e.g. as a micronutrient in animal nutrition) by cultivating the extracting plants in the inter-row of the vineyards. Soil cleaning up usually requires several years because the chemical speciation of Cu in solution is mainly based on forms complexed with dissolved organic matter (DOM) that are not phytoavailable, unlike the "free" forms (Cu2+). Indeed, more than 98% of Cu in the solution is bound to DOM. The selection and inoculation of invineyardsoils in vineyard soils ofbacteria(bioaugmentation) able to degrade Cu-DOM complexes could increase the phytoavailable pool of Cu2+ in the soil solution (in addition to bacteria which first mobilize Cu in solution from the soil bearing phases) in order to increase phytoextraction performance. In this study, sevenCu-accumulating plants potentially usable in inter-row were tested for their Cu phytoextraction capacity in hydroponics (ray-grass, brown mustard, buckwheat, hemp, sunflower, oats, and chicory). Also, a bacterial consortium was tested: Pseudomonas sp. previously studied for its ability to mobilize Cu through the pyoverdine siderophore (complexing agent) and potentially to degrade Cu-DOM complexes, and a second bacterium (to be selected) able to promote the survival of Pseudomonas sp. following its inoculation in soil. Interaction network method was used based on the notions of co-occurrence and, therefore, of bacterial abundance found in the same soils. Bacteria from the EcoVitiSol project (Alsace, France) were targeted. The final step consisted of incoupling the bacterial consortium with the chosen plant in soil pots. The degradation of Cu-DOMcomplexes is measured on the basis of the absorption index at 254nm, which gives insight on the aromaticity of the DOM. The“free” Cu in solution (from the mobilization of Cu and/or the degradation of Cu-MOD complexes) is assessed by measuring pCu. Eventually, Cu accumulation in plants is measured by ICP-AES. The selection of the plant is currently being finalized. The interaction network method targeted the best positive interactions ofFlavobacterium sp. with Pseudomonassp. These bacteria are both PGPR (plant growth promoting rhizobacteria) with the ability to improve the plant growth and to mobilize Cu from the soil bearing phases (siderophores). Also, these bacteria are known to degrade phenolic groups, which are highly present in DOM. They could therefore contribute to the degradation of DOM-Cu. The results of the upcoming bacteria-plant coupling tests in pots will be also presented.

Keywords: complexes Cu-DOM, bioaugmentation, phytoavailability, phytoextraction

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Authors: Rangrong Yoksan, Amporn Sane, Nattaporn Khanoonkon, Chanakorn Yokesahachart, Narumol Noivoil, Khanh Minh Dang

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Polylactic acid (PLA) is the most commercially available bio-based and biodegradable plastic at present. PLA has been used in plastic related industries including single-used containers, disposable and environmentally friendly packaging owing to its renewability, compostability, biodegradability, and safety. Although PLA demonstrates reasonably good optical, physical, mechanical, and barrier properties comparable to the existing petroleum-based plastics, its brittleness and mold shrinkage as well as its price are the points to be concerned for the production of rigid and semi-rigid packaging. Blending PLA with other bio-based polymers including thermoplastic starch (TPS) is an alternative not only to achieve a complete bio-based plastic, but also to reduce the brittleness, shrinkage during molding and production cost of the PLA-based products. TPS is a material produced mainly from starch which is cheap, renewable, biodegradable, compostable, and non-toxic. It is commonly prepared by a plasticization of starch under applying heat and shear force. Although glycerol has been reported as one of the most plasticizers used for preparing TPS, its migration caused the surface stickiness of the TPS products. In some cases, mixed plasticizers or natural fibers have been applied to impede the retrogradation of starch or reduce the migration of glycerol. The introduction of fibers into TPS-based materials could reinforce the polymer matrix as well. Therefore, the objective of the present research is to study the effect of starch type (i.e. native starch and phosphate starch), plasticizer type (i.e. glycerol and xylitol with a weight ratio of glycerol to xylitol of 100:0, 75:25, 50:50, 25:75, and 0:100), and fiber content (i.e. in the range of 1-25 % wt) on properties of PLA/TPS blend and composite. PLA/TPS blends and composites were prepared using a twin-screw extruder and then converted into dumbbell-shaped specimens using an injection molding machine. The PLA/TPS blends prepared by using phosphate starch showed higher tensile strength and stiffness than the blends prepared by using the native one. In contrast, the blends from native starch exhibited higher extensibility and heat distortion temperature (HDT) than those from the modified starch. Increasing xylitol content resulted in enhanced tensile strength, stiffness, and water resistance, but decreased extensibility and HDT of the PLA/TPS blend. Tensile properties and hydrophobicity of the blend could be improved by incorporating silane treated-jute fibers.

Keywords: polylactic acid, thermoplastic starch, Jute fiber, composite, blend

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Authors: Ana M. Antolín, Sandra Contreras, Francesc Medina, Didier Tichit

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Introduction: High levels of nitrates (> 50 ppm NO3-) in drinking water are potentially risky to human health. In the recent years, the trend of nitrate concentration in groundwater is rising in the EU and other countries. Conventional catalytic nitrate reduction processes into N2 and H2O lead to some toxic intermediates and by-products, such as NO2-, NH4+, and NOx gases. Alternatively, photocatalytic nitrate removal using solar irradiation and heterogeneous catalysts is a very promising and ecofriendly technique. It has been scarcely performed and more research on highly efficient catalysts is still needed. In this work, different nanocatalysts supported on Aeroxide Titania P25 (P25) have been prepared varying: 0.5-4 % wt. Ag); Pt (2, 4 % wt.); Pt precursor (H2PtCl6/K2PtCl6); and impregnation order of both metals. Pt was chosen in order to increase the selectivity to N2 and decrease that to NO2-. Catalysts were characterized by nitrogen physisorption, X-Ray diffraction, UV-visible spectroscopy, TEM and X Ray-Photoelectron Spectroscopy. The aim was to determine the influence of the composition and the preparation method of the catalysts on the conversion and selectivity in the nitrate reduction, as well as going through an overall and better understanding of the process. Nanocatalysts synthesis: For the mono and bimetallic catalysts preparation, wise-drop wetness impregnation of the precursors (AgNO3, H2PtCl6, K2PtCl6) followed by a reduction step (NaBH4) was used to obtain the metal colloids. Results and conclusions: Denitration experiments were performed in a 350 mL PTFE batch reactor under inert standard operational conditions, ultraviolet irradiations (λ=254 nm (UV-C); λ=365 nm (UV-A)), and presence/absence of hydrogen gas as a reducing agent, contrary to most studies using oxalic or formic acid. Samples were analyzed by Ionic Chromatography. Blank experiments using respectively P25 (dark conditions), hydrogen only and UV irradiations without hydrogen demonstrated a clear influence of the presence of hydrogen on nitrate reduction. Also, they demonstrated that UV irradiation increased the selectivity to N2. Interestingly, the best activity was obtained under ultraviolet lamps, especially at a closer wavelength to visible light irradiation (λ = 365 nm) and H2. 2% Ag/P25 leaded to the highest NO3- conversion among the monometallic catalysts. However, nitrite quantities have to be diminished. On the other hand, practically no nitrate conversion was observed with the monometallics based on Pt/P25. Therefore, the amount of 2% Ag was chosen for the bimetallic catalysts. Regarding the bimetallic catalysts, it is observed that the metal impregnation order, amount and Pt precursor highly affects the results. Higher selectivity to the desirable N2 gas is obtained when Pt was firstly added, especially with K2PtCl6 as Pt precursor. This suggests that when Pt is secondly added, it covers the Ag particles, which are the most active in this reaction. It could be concluded that Ag allows the nitrate reduction step to nitrite, and Pt the nitrite reduction step toward the desirable N2 gas.

Keywords: heterogeneous catalysis, hydrogenation, nanocatalyst, nitrate removal, photocatalysis

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Authors: Robin Anderson, Elizabeth Latham, David Nisbet

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Propagation and dissemination of antimicrobial resistant and pathogenic microbes from spoiled silages and composts represents a serious public health threat to humans and animals. In the present study, the antimicrobial activity of the short chain nitro-compound, 2-nitro-1-propanol (9 mM) as well as the medium chain fatty acid, lauric acid, and its glycerol monoester, monolaurin, (each at 25 and 17 µmol/mL, respectfully) were investigated against select pathogenic and multi-drug resistant antimicrobial resistant Gram-positive bacteria common to spoiled silages and composts. In an initial study, we found that growth rates of a multi-resistant Enterococcus faecalis (expressing resistance against erythromycin, quinupristin/dalfopristin and tetracycline) and Staphylococcus aureus strain 12600 (expressing resistance against erythromycin, linezolid, penicillin, quinupristin/dalfopristin and vancomycin) were more than 78% slower (P < 0.05) by 2-nitro-1-propanol treatment during culture (n = 3/treatment) in anaerobically prepared ½ strength Brain Heart Infusion broth at 37oC when compared to untreated controls (0.332 ± 0.04 and 0.108 ± 0.03 h-1, respectively). The growth rate of 2-nitro-1-propanol-treated Listeria monocytogenes was also decreased by 96% (P < 0.05) when compared to untreated controls cultured similarly (0.171 ± 0.01 h-1). Maximum optical densities measured at 600 nm were lower (P < 0.05) in 2-nitro-1-propanol-treated cultures (0.053 ± 0.01, 0.205 ± 0.02 and 0.041 ± 0.01, respectively) than in untreated controls (0.483 ± 0.02, 0.523 ± 0.01 and 0.427 ± 0.01, respectively) for E. faecalis, S. aureus and L. monocytogenes, respectively. When tested against mixed microbial populations during anaerobic 24 h incubation of spoiled silage, significant effects of treatment with 1 mg 2-nitro-1-propanol (approximately 9.5 µmol/g) or 5 mg lauric acid/g (approximately 25 µmol/g) on populations of wildtype Enterococcus and Listeria were not observed. Mixed populations treated with 5 mg monolaurin/g (approximately 17 µmol/g) had lower (P < 0.05) viable cell counts of wildtype enterococci than untreated controls after 6 h incubation (2.87 ± 1.03 versus 5.20 ± 0.25 log10 colony forming units/g, respectively) but otherwise significant effects of monolaurin were not observed. These results reveal differential susceptibility of multi-drug resistant enterococci and staphylococci as well as L. monocytogenes to the inhibitory activity of 2-nitro-1-propanol and the medium chain fatty acid, lauric acid and its glycerol monoester, monolaurin. Ultimately, these results may lead to improved treatment technologies to preserve the microbiological safety of silages and composts.

Keywords: 2-nitro-1-propanol, lauric acid, monolaurin, gram positive bacteria

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Authors: Nothando Gwazani, K. R. Marembo

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An increase in the atmospheric concentration of carbon dioxide (CO₂) from fossil fuel and land use change necessitates identification of strategies for mitigating threats associated with global warming. Oceans are insufficient to offset the accelerating rate of carbon emission. However, the challenges of oceans as a source of reducing carbon footprint can be effectively overcome by the storage of carbon in terrestrial carbon sinks. The gases with special optical properties that are responsible for climate warming include carbon dioxide (CO₂), water vapors, methane (CH₄), nitrous oxide (N₂O), nitrogen oxides (NOₓ), stratospheric ozone (O₃), carbon monoxide (CO) and chlorofluorocarbons (CFC’s). Amongst these, CO₂ plays a crucial role as it contributes to 50% of the total greenhouse effect and has been linked to climate change. Because plants act as carbon sinks, interest in terrestrial carbon sequestration has increased in an effort to explore opportunities for climate change mitigation. Removal of carbon from the atmosphere is a topical issue that addresses one important aspect of an overall strategy for carbon management namely to help mitigate the increasing emissions of CO₂. Thus, terrestrial ecosystems have gained importance for their potential to sequester carbon and reduce carbon sink in oceans, which have a substantial impact on the ocean species. Field data and electromagnetic spectrum bands were analyzed using ArcGIS 10.2, QGIS 2.8 and ERDAS IMAGINE 2015 to examine the vegetation distribution. Satellite remote sensing data coupled with Normalized Difference Vegetation Index (NDVI) was employed to assess future potential changes in vegetation distributions in Eastern Cape Province of South Africa. The observed 5-year interval analysis examines the amount of carbon absorbed using vegetation distribution. In 2015, the numerical results showed low vegetation distribution, therefore increased the acidity of the oceans and gravely affected fish species and corals. The outcomes suggest that the study area could be effectively utilized for carbon sequestration so as to mitigate ocean acidification. The vegetation changes measured through this investigation suggest an environmental shift and reduced vegetation carbon sink, and that threatens biodiversity and ecosystem. In order to sustain the amount of carbon in the terrestrial ecosystems, the identified ecological factors should be enhanced through the application of good land and forest management practices. This will increase the carbon stock of terrestrial ecosystems thereby reducing direct loss to the atmosphere.

Keywords: remote sensing, vegetation dynamics, carbon sequestration, terrestrial carbon sink

Procedia PDF Downloads 128

Authors: Vasiliki Markantoni, Eftychia Platsidaki, Georgios Chaidemenos, Georgios Kontochristopoulos

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Introduction: Bromhidrosis, also known as osmidrosis, is a common distressing condition with a significant negative effect on patient’s quality of life. Its etiology is multifactorial. It usually affects axilla, genital skin, breasts and soles, areas where apocrine glands are mostly distributed. Therapeutic treatments include topical antibacterial agents, antiperspirants and neuromuscular blocker agents-toxins. In this study, we aimed to evaluate the efficacy and possible complications of topical glycopyrrolate, an anticholinergic agent, for treatment of bromhidrosis. Glycopyrrolate, applied topically as a cream, solution or spray at concentrations between 0,5% and 4%, has been successfully used to treat different forms of focal hyperhidrosis. Materials and Methods: Twenty patients, six males and fourteen females, meeting the criteria for bromhidrosis were treated with topical glycopyrrolate for two months. The average age was 36. Eleven patients had bromhidrosis located to the axillae, four to the soles, four to both axillae and soles and one to the genital folds. Glycopyrrolate was applied topically as a cream at concentration 2%, formulated in Fitalite. During the first month, patients were using the cream every night and thereafter twice daily. The degree of malodor was assessed subjectively by patients and scaled averagely as ‘none’, ‘mild’, ‘moderate’, and ‘severe’ with corresponding scores of 0, 1, 2, and 3, respectively. The modified Dermatology Life Quality Index (DLQI) was used to assess the quality of life. The clinical efficacy was graded by the patient scale of excellent, good, fair and poor. In the end, patients were given the power to evaluate whether they were totally satisfied with, partially satisfied or unsatisfied and possible side effects during the treatment were recorded. Results: All patients were satisfied at the end of the treatment. No patient defined the response as no improvement. The subjectively assessed score level of bromhidrosis was remarkably improved after the first month of treatment and improved slightly more after the second month. DLQI score was also improved to all patients. Adverse effects were reported in 2 patients. In the first case, topical irritation was reported. This was classed as mild (erythema and desquamation), appeared during the second month of treatment and was treated with low-potency topical corticosteroids. In the second case, mydriasis was reported, that recovered without specific treatment, as soon as we insisted to the importance of careful hygiene after cream application so as not to contaminate the periocular skin or ocular surface. Conclusions: Dermatologists often encounter patients with bromhidrosis, therefore should be aware of treatment options. To the best of our knowledge, this is the first study to evaluate the use of topical glycopyrrolate as a therapeutic approach for bromhidrosis. Our findings suggest that topical glycopyrrolate has an excellent safety profile and demonstrate encouraging results for the management of this distressful condition.

Keywords: Bromhidrosis, glycopyrrolate, topical treatment, osmidrosis

Procedia PDF Downloads 144

Authors: Maria Botero-Uribe, Melissa Fitzgerald, Robert Gilbert, Kim Bryceson, Jocelyn Midgley

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French fry manufacturing involves a series of processes in which structural properties of potatoes are modified to produce crispy french fries which consumers enjoy. In addition to the traditional french fry manufacturing process, the industry is applying a relatively new process called pulsed electrical field (PEF) to the whole potatoes. There is a wealth of information on the technical treatment conditions of PEF, however, there is a lack of information about its effect on the structural properties that affect texture and its synergistic interactions with the other manufacturing steps of french fry production. The effect of PEF on starch gelatinisation properties of Russet Burbank potato was measured using a Differential Scanning Calorimeter. Cation content (K+, Ca2+ and Mg2+) was determined by inductively coupled plasma optical emission spectrophotometry. Firmness, and toughness of raw and blanched potatoes were determined in an uniaxial compression test. Moisture content was determined in a vacuum oven and oil content was measured using the soxhlet system with hexane. The final texture of the french fries – crispness - was determined using a three bend point test. Triangle tests were conducted to determine if consumers were able to perceive sensory differences between French fries that were PEF treated and those without treatment. The concentration of K+, Ca2+ and Mg2+ decreased significantly in the raw potatoes after the PEF treatment. The PEF treatment significantly increased modulus of elasticity, compression strain, compression force and toughness in the raw potato. The PEF-treated raw potato were firmer and stiffer, and its structure integrity held together longer, resisted higher force before fracture and stretched further than the untreated ones. The strain stress relationship exhibited by the PEF-treated raw potato could be due to an increase in the permeability of the plasmalema and tonoplasm allowing Ca2+ and Mg2+ cations to reach the cell wall and middle lamella, and be available for cross linking with the pectin molecule. The PEF-treated raw potato exhibited a slightly higher onset gelatinisation temperatures, similar peak temperatures and lower gelatinisation ranges than the untreated raw potatoes. The final moisture content of the french fries was not significantly affected by the PEF treatment. Oil content in the PEF- treated potatoes was lower than the untreated french fries, however, not statistically significant at 5 %. The PEF treatment did not have an overall significant effect on french fry crispness (modulus of elasticity), flexure stress or strain. The triangle tests show that most consumers could not detect a difference between French fries that received a PEF treatment from those that did not.

Keywords: french fries, mechanical properties, PEF, potatoes

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Authors: Klaudia Pluta, Katarzyna Bialik-Wąs, Dagmara Malina, Mateusz Barczewski

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Hydrogel networks, due to their unique properties, are highly attractive materials for wound dressing. The three-dimensional structure of hydrogels provides tissues with optimal moisture, which supports the wound healing process. Moreover, a characteristic feature of hydrogels is their absorption properties which allow for the absorption of wound exudates. For the fabrication of biomedical hydrogels, a combination of natural polymers ensuring biocompatibility and synthetic ones that provide adequate mechanical strength are often used. Sodium alginate (SA) is one of the polymers widely used in wound dressing materials because it exhibits excellent biocompatibility and biodegradability. However, due to poor strength properties, often alginate-based hydrogel materials are enhanced by the addition of another polymer such as poly(vinyl alcohol) (PVA). This paper is concentrated on the preparation methods of sodium alginate/polyvinyl alcohol hydrogel system incorporating Aloe vera extract and glycerin for wound healing material with particular focus on the role of their composition on structure, thermal properties, and stability. Briefly, the hydrogel preparation is based on the chemical cross-linking method using poly(ethylene glycol) diacrylate (PEGDA, Mn = 700 g/mol) as a crosslinking agent and ammonium persulfate as an initiator. In vitro degradation tests of SA/PVA/AV hydrogels were carried out in Phosphate-Buffered Saline (pH – 7.4) as well as in distilled water. Hydrogel samples were firstly cut into half-gram pieces (in triplicate) and immersed in immersion fluid. Then, all specimens were incubated at 37°C and then the pH and conductivity values were measurements at time intervals. The post-incubation fluids were analyzed using SEC/GPC to check the content of oligomers. The separation was carried out at 35°C on a poly(hydroxy methacrylate) column (dimensions 300 x 8 mm). 0.1M NaCl solution, whose flow rate was 0.65 ml/min, was used as the mobile phase. Three injections with a volume of 50 µl were made for each sample. The thermogravimetric data of the prepared hydrogels were collected using a Netzsch TG 209 F1 Libra apparatus. The samples with masses of about 10 mg were weighed separately in Al2O3 crucibles and then were heated from 30°C to 900°C with a scanning rate of 10 °C∙min−1 under a nitrogen atmosphere. Based on the conducted research, a fast and simple method was developed to produce potential wound dressing material containing sodium alginate, poly(vinyl alcohol) and Aloe vera extract. As a result, transparent and flexible SA/PVA/AV hydrogels were obtained. The degradation experiments indicated that most of the samples immersed in PBS as well as in distilled water were not degraded throughout the whole incubation time.

Keywords: hydrogels, wound dressings, sodium alginate, poly(vinyl alcohol)

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Authors: P. Barreto, A. Guevara, V. Ibujes

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In this study, the removal of arsenic from synthetic solutions and natural water from Papallacta Lagoon was evaluated, by using the rhizofiltration method with terrestrial and aquatic plant species. Ecuador is a country of high volcanic activity, that is why most of water sources come from volcanic glaciers. Therefore, it is necessary to find new, affordable and effective methods for treating water. The water from Papallacta Lagoon shows levels from 327 µg/L to 803 µg/L of arsenic. The evaluation for the removal of arsenic began with the selection of 16 different species of terrestrial and aquatic plants. These plants were immersed to solutions of 4500 µg/L arsenic concentration, for 48 hours. Subsequently, 3 terrestrial species and 2 aquatic species were selected based on the highest amount of absorbed arsenic they showed, analyzed by plasma optical emission spectrometry (ICP-OES), and their best capacity for adaptation into the arsenic solution. The chosen terrestrial species were cultivated from their seed with hydroponics methods, using coconut fiber and polyurethane foam as substrates. Afterwards, the species that best adapted to hydroponic environment were selected. Additionally, a control of the development for the selected aquatic species was carried out using a basic nutrient solution to provide the nutrients that the plants required. Following this procedure, 30 plants from the 3 types of species selected were exposed to a synthetic solution with levels of arsenic concentration of 154, 375 and 874 µg/L, for 15 days. Finally, the plant that showed the highest level of arsenic absorption was placed in 3 L of natural water, with arsenic levels of 803 µg/L. The plant laid in the water until it reached the desired level of arsenic of 10 µg/L. This experiment was carried out in a total of 30 days, in which the capacity of arsenic absorption of the plant was measured. As a result, the five species initially selected to be used in the last part of the evaluation were: sunflower (Helianthus annuus), clover (Trifolium), blue grass (Poa pratensis), water hyacinth (Eichhornia crassipes) and miniature aquatic fern (Azolla). The best result of arsenic removal was showed by the water hyacinth with a 53,7% of absorption, followed by the blue grass with 31,3% of absorption. On the other hand, the blue grass was the plant that best responded to the hydroponic cultivation, by obtaining a germination percentage of 97% and achieving its full growth in two months. Thus, it was the only terrestrial species selected. In summary, the final selected species were blue grass, water hyacinth and miniature aquatic fern. These three species were evaluated by immersing them in synthetic solutions with three different arsenic concentrations (154, 375 and 874 µg/L). Out of the three plants, the water hyacinth was the one that showed the highest percentages of arsenic removal with 98, 58 and 64%, for each one of the arsenic solutions. Finally, 12 plants of water hyacinth were chosen to reach an arsenic level up to 10 µg/L in natural water. This significant arsenic concentration reduction was obtained in 5 days. In conclusion, it was found that water hyacinth is the best plant to reduce arsenic levels in natural water.

Keywords: arsenic, natural water, plant species, rhizofiltration, synthetic solutions

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Authors: I. Nitoi, P. Oancea, M. Raileanu, M. Crisan, L. Constantin, I. Cristea

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Hazardous organic compounds like nitroaromatics are frequently found in chemical and petroleum industries discharged effluents. Due to their bio-refractory character and high chemical stability cannot be efficiently removed by classical biological or physical-chemical treatment processes. In the past decades, semiconductor photocatalysis has been frequently applied for the advanced degradation of toxic pollutants. Among various semiconductors titania was a widely studied photocatalyst, due to its chemical inertness, low cost, photostability and nontoxicity. In order to improve optical absorption and photocatalytic activity of TiO2 many attempts have been made, one feasible approach consists of doping oxide semiconductor with metal. The degradation of dinitrobenzene (DNB) and dinitrotoluene (DNT) from aqueous solution under UVA-VIS irradiation using heavy metal (0.5% Fe, 1%Co, 1%Ni ) doped titania was investigated. The photodegradation experiments were carried out using a Heraeus laboratory scale UV-VIS reactor equipped with a medium-pressure mercury lamp which emits in the range: 320-500 nm. Solutions with (0.34-3.14) x 10-4 M pollutant content were photo-oxidized in the following working conditions: pH = 5-9; photocatalyst dose = 200 mg/L; irradiation time = 30 – 240 minutes. Prior to irradiation, the photocatalyst powder was added to the samples, and solutions were bubbled with air (50 L/hour), in the dark, for 30 min. Dopant type, pH, structure and initial pollutant concentration influence on the degradation efficiency were evaluated in order to set up the optimal working conditions which assure substrate advanced degradation. The kinetics of nitroaromatics degradation and organic nitrogen mineralization was assessed and pseudo-first order rate constants were calculated. Fe doped photocatalyst with lowest metal content (0.5 wt.%) showed a considerable better behaviour in respect to pollutant degradation than Co and Ni (1wt.%) doped titania catalysts. For the same working conditions, degradation efficiency was higher for DNT than DNB in accordance with their calculated adsobance constants (Kad), taking into account that degradation process occurs on catalyst surface following a Langmuir-Hinshalwood model. The presence of methyl group in the structure of DNT allows its degradation by oxidative and reductive pathways, while DNB is converted only by reductive route, which also explain the highest DNT degradation efficiency. For highest pollutant concentration tested (3 x 10-4 M), optimum working conditions (0.5 wt.% Fe doped –TiO2 loading of 200 mg/L, pH=7 and 240 min. irradiation time) assures advanced nitroaromatics degradation (ηDNB=89%, ηDNT=94%) and organic nitrogen mineralization (ηDNB=44%, ηDNT=47%).

Keywords: hazardous organic compounds, irradiation, nitroaromatics, photocatalysis

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Authors: Yudo Prasetyo, Hana Sugiastu Firdaus, Diyanah Diyanah

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The phenomenon of the lack of clean water supply in several big cities in Indonesia is a major problem in the development of urban areas. Moreover, in the city of Semarang, the population density and growth of physical development is very high. Continuous and large amounts of underground water (aquifer) exposure can result in a drastically aquifer supply declining in year by year. Especially, the intensity of aquifer use in the fulfilment of household needs and industrial activities. This is worsening by the land subsidence phenomenon in some areas in the Semarang city. Therefore, special research is needed to know the spatial correlation of the impact of decreasing aquifer capacity on the land subsidence phenomenon. This is necessary to give approve that the occurrence of land subsidence can be caused by loss of balance of pressure on below the land surface. One method to observe the correlation pattern between the two phenomena is the application of remote sensing technology based on radar and optical satellites. Implementation of Differential Interferometric Synthetic Aperture Radar (DINSAR) or Small Baseline Area Subset (SBAS) method in SENTINEL-1A satellite image acquisition in 2014-2017 period will give a proper pattern of land subsidence. These results will be spatially correlated with the aquifer-declining pattern in the same time period. Utilization of survey results to 8 monitoring wells with depth in above 100 m to observe the multi-temporal pattern of aquifer change capacity. In addition, the pattern of aquifer capacity will be validated with 2 underground water cavity maps from observation of ministries of energy and natural resources (ESDM) in Semarang city. Spatial correlation studies will be conducted on the pattern of land subsidence and aquifer capacity using overlapping and statistical methods. The results of this correlation will show how big the correlation of decrease in underground water capacity in influencing the distribution and intensity of land subsidence in Semarang city. In addition, the results of this study will also be analyzed based on geological aspects related to hydrogeological parameters, soil types, aquifer species and geological structures. The results of this study will be a correlation map of the aquifer capacity on the decrease in the face of the land in the city of Semarang within the period 2014-2017. So hopefully the results can help the authorities in spatial planning and the city of Semarang in the future.

Keywords: aquifer, differential interferometric synthetic aperture radar (DINSAR), land subsidence, small baseline area subset (SBAS)

Procedia PDF Downloads 165

Authors: Sulalit Bandyopadhyay, Muhammad Awais Ashfaq Alvi, Anuvansh Sharma, Wilhelm R. Glomm

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Release of drugs from nanogels and nanogel-based systems can occur under the influence of external stimuli like temperature, pH, magnetic fields and so on. pNIPAm-AAc nanogels respond to the combined action of both temperature and pH, the former being mostly determined by hydrophilic-to-hydrophobic transitions above the volume phase transition temperature (VPTT), while the latter is controlled by the degree of protonation of the carboxylic acid groups. These nanogels based systems are promising candidates in the field of drug delivery. Combining nanogels with magneto-plasmonic nanoparticles (NPs) introduce imaging and targeting modalities along with stimuli-response in one hybrid system, thereby incorporating multifunctionality. Fe@Au core-shell NPs possess optical signature in the visible spectrum owing to localized surface plasmon resonance (LSPR) of the Au shell, and superparamagnetic properties stemming from the Fe core. Although there exist several synthesis methods to control the size and physico-chemical properties of pNIPAm-AAc nanogels, yet, there is no comprehensive study that highlights the dependence of incorporation of one or more layers of NPs to these nanogels. In addition, effective determination of volume phase transition temperature (VPTT) of the nanogels is a challenge which complicates their uses in biological applications. Here, we have modified the swelling-collapse properties of pNIPAm-AAc nanogels, by combining with Fe@Au NPs using different solution based methods. The hydrophilic-hydrophobic transition of the nanogels above the VPTT has been confirmed to be reversible. Further, an analytical method has been developed to deduce the average VPTT which is found to be 37.3°C for the nanogels and 39.3°C for nanogel coated Fe@Au NPs. An opposite swelling –collapse behaviour is observed for the latter where the Fe@Au NPs act as bridge molecules pulling together the gelling units. Thereafter, Cyt C, a model protein drug and L-Dopa, a drug used in the clinical treatment of Parkinson’s disease were loaded separately into the nanogels and nanogel coated Fe@Au NPs, using a modified breathing-in mechanism. This gave high loading and encapsulation efficiencies (L Dopa: ~9% and 70µg/mg of nanogels, Cyt C: ~30% and 10µg/mg of nanogels respectively for both the drugs. The release kinetics of L-Dopa, monitored using UV-vis spectrophotometry was observed to be rather slow (over several hours) with highest release happening under a combination of high temperature (above VPTT) and acidic conditions. However, the release of L-Dopa from nanogel coated Fe@Au NPs was the fastest, accounting for release of almost 87% of the initially loaded drug in ~30 hours. The chemical structure of the drug, drug incorporation method, location of the drug and presence of Fe@Au NPs largely alter the drug release mechanism and the kinetics of these nanogels and Fe@Au NPs coated with nanogels.

Keywords: controlled release, nanogels, volume phase transition temperature, l-dopa

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Authors: Kanduru Lokesh, Venkitasamy Kesavan

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The unique structural features of spiro-oxindoles with diverse biological activities have made them privileged structures in new drug discovery. The key structural characteristic of these compounds is the spiro ring fused at the C-3 position of the oxindole core with varied heterocyclic motifs. Structural diversification of heterocyclic scaffolds to synthesize new chemical entities as pharmaceuticals and agrochemicals is one of the important goals of synthetic organic chemists. Nitrogen and oxygen containing heterocycles are by far the most widely occurring privileged structures in medicinal chemistry. The structural complexity and distinct three-dimensional arrangement of functional groups of these privileged structures are generally responsible for their specificity against biological targets. Structurally diverse compound libraries have proved to be valuable assets for drug discovery against challenging biological targets. Thus, identifying a new combination of substituents at C-3 position on oxindole moiety is of great importance in drug discovery to improve the efficiency and efficacy of the drugs. The development of suitable methodology for the synthesis of spiro-oxindole compounds has attracted much interest often in response to the significant biological activity displayed by the both natural and synthetic compounds. So creating structural diversity of oxindole scaffolds is need of the decade and formidable challenge. A general way to improve synthetic efficiency and also to access diversified molecules is through the annulation reactions. Annulation reactions allow the formation of complex compounds starting from simple substrates in a single transformation consisting of several steps in an ecologically and economically favorable way. These observations motivated us to develop the annulation reaction protocol to enable the synthesis of a new class of spiro-oxindole motifs which in turn would enable the enhancement of molecular diversity. As part of our enduring interest in the development of novel, efficient synthetic strategies to enable the synthesis of biologically important oxindole fused spirocarbocyclic systems, We have developed an efficient methodology for the construction of highly functionalized spirocarbocyclic oxindoles through [4+2] annulation of phthalides via Hauser annulation. functionalized spirocarbocyclic oxindoles was accomplished for the first time in the literature using Hauser annulation strategy. The reaction between methyleneindolinones and arylsulfonylphthalides catalyzed by cesium carbonate led to the access of new class of biologically important spiro[indoline-3,2'-naphthalene] derivatives in very good yields. The synthetic utility of the annulated product was further demonstrated by fluorination Using NFSI as a fluorinating agent to furnish corresponding fluorinated product.

Keywords: Hauser-Kraus annulation, spiro carbocyclic oxindoles, oxindole-ester, fluoridation

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Authors: Evrim Colak, Andriy E. Serebryannikov, Pavel V. Usik, Ekmel Ozbay

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Working with a dielectric photonic crystal (PC) structure which does not include surface corrugations, unidirectional transmission and dual-beam splitting are observed under normal incidence as a result of the strong diffractions caused by the embedded defect layer. The defect layer has twice the period of the regular PC segments which sandwich the defect layer. Although the PC has even number of rows, the structural symmetry is broken due to the asymmetric placement of the defect layer with respect to the symmetry axis of the regular PC. The simulations verify that efficient splitting and occurrence of strong diffractions are related to the dispersion properties of the Floquet-Bloch modes of the photonic crystal. Unidirectional and bi-directional splitting, which are associated with asymmetric transmission, arise due to the dominant contribution of the first positive and first negative diffraction orders. The effect of the depth of the defect layer is examined by placing single defect layer in varying rows, preserving the asymmetry of PC. Even for deeply buried defect layer, asymmetric transmission is still valid even if the zeroth order is not coupled. This transmission is due to evanescent waves which reach to the deeply embedded defect layer and couple to higher order modes. In an additional selected performance, whichever surface is illuminated, i.e., in both upper and lower surface illumination cases, incident beam is split into two beams of equal intensity at the output surface where the intensity of the out-going beams are equal for both illumination cases. That is, although the structure is asymmetric, symmetric bidirectional transmission with equal transmission values is demonstrated and the structure mimics the behavior of symmetric structures. Finally, simulation studies including the examination of a coupled-cavity defect for two different permittivity values (close to the permittivity values of GaAs or Si and alumina) reveal unidirectional splitting for a wider band of operation in comparison to the bandwidth obtained in the case of a single embedded defect layer. Since the dielectric materials that are utilized are low-loss and weakly dispersive in a wide frequency range including microwave and optical frequencies, the studied structures should be scalable to the mentioned ranges.

Keywords: asymmetric transmission, beam deflection, blazing, bi-directional splitting, defect layer, dual beam splitting, Floquet-Bloch modes, isofrequency contours, line defect, oblique incidence, photonic crystal, unidirectionality

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Authors: Surekha Barkur, Aseefhali Bankapur, Santhosh Chidangil

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Raman Tweezers technique has become prevalent in single cell studies. This technique combines Raman spectroscopy which gives information about molecular vibrations, with optical tweezers which use a tightly focused laser beam for trapping the single cells. Thus Raman Tweezers enabled researchers analyze single cells and explore different applications. The applications of Raman Tweezers include studying blood cells, monitoring blood-related disorders, silver nanoparticle-induced stress, etc. There is increased interest in the toxic effect of nanoparticles with an increase in the various applications of nanoparticles. The interaction of these nanoparticles with the cells may vary with their size. We have studied the effect of silver nanoparticles of sizes 10nm, 40nm, and 100nm on erythrocytes using Raman Tweezers technique. Our aim was to investigate the size dependence of the nanoparticle effect on RBCs. We used 785nm laser (Starbright Diode Laser, Torsana Laser Tech, Denmark) for both trapping and Raman spectroscopic studies. 100 x oil immersion objectives with high numerical aperture (NA 1.3) is used to focus the laser beam into a sample cell. The back-scattered light is collected using the same microscope objective and focused into the spectrometer (Horiba Jobin Vyon iHR320 with 1200grooves/mm grating blazed at 750nm). Liquid nitrogen cooled CCD (Symphony CCD-1024x256-OPEN-1LS) was used for signal detection. Blood was drawn from healthy volunteers in vacutainer tubes and centrifuged to separate the blood components. 1.5 ml of silver nanoparticles was washed twice with distilled water leaving 0.1 ml silver nanoparticles in the bottom of the vial. The concentration of silver nanoparticles is 0.02mg/ml so the 0.03mg of nanoparticles will be present in the 0.1 ml nanoparticles obtained. The 25 ul of RBCs were diluted in 2 ml of PBS solution and then treated with 50 ul (0.015mg) of nanoparticles and incubated in CO2 incubator. Raman spectroscopic measurements were done after 24 hours and 48 hours of incubation. All the spectra were recorded with 10mW laser power (785nm diode laser), 60s of accumulation time and 2 accumulations. Major changes were observed in the peaks 565 cm-1, 1211 cm-1, 1224 cm-1, 1371 cm-1, 1638 cm-1. A decrease in intensity of 565 cm-1, increase in 1211 cm-1 with a reduction in 1224 cm-1, increase in intensity of 1371 cm-1 also peak disappearing at 1635 cm-1 indicates deoxygenation of hemoglobin. Nanoparticles with higher size were showing maximum spectral changes. Lesser changes observed in case of 10nm nanoparticle-treated erythrocyte spectra.

Keywords: erythrocytes, nanoparticle-induced toxicity, Raman tweezers, silver nanoparticles

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Authors: Gulmira Zhakupova, Tamara Tultabayeva, Aknur Muldasheva, Assem Sagandyk

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The development of technology and production of products with increased biological value based on the use of natural food raw materials are important tasks in the policy of the food market of the Republic of Kazakhstan. For Kazakhstan, livestock farming, in particular sheep farming, is the most ancient and developed industry and way of life. The history of the Kazakh people is largely connected with this type of agricultural production, with established traditions using dairy products from sheep's milk. Therefore, the development of new technologies from sheep’s milk remains relevant. In addition, one of the most promising areas for the development of food technology for therapeutic and prophylactic purposes is sheep milk products as a source of protein, immunoglobulins, minerals, vitamins, and other biologically active compounds. This article presents the results of research on the study of milk processing technology. The objective of the study is to study the possibilities of processing sheep milk and its role in human nutrition, as well as the results of research to improve the technology of sheep milk products. The studies were carried out on the basis of sanitary and hygienic requirements for dairy products in accordance with the following test methods. To perform microbiological analysis, we used the method for identifying Salmonella bacteria (Horizontal method for identifying, counting, and serotyping Salmonella) in a certain mass or volume of product. Nutritional value is a complex of properties of food products that meet human physiological needs for energy and basic nutrients. The protein mass fraction was determined by the Kjeldahl method. This method is based on the mineralization of a milk sample with concentrated sulfuric acid in the presence of an oxidizing agent, an inert salt - potassium sulfate, and a catalyst - copper sulfate. In this case, the amino groups of the protein are converted into ammonium sulfate dissolved in sulfuric acid. The vitamin composition was determined by HPLC. To determine the content of mineral substances in the studied samples, the method of atomic absorption spectrophotometry was used. The study identified the technological parameters of sheep milk products and determined the prospects for researching sheep milk products. Microbiological studies were used to determine the safety of the study product. According to the results of the microbiological analysis, no deviations from the norm were identified. This means high safety of the products under study. In terms of nutritional value, the resulting products are high in protein. Data on the positive content of amino acids were also obtained. The results obtained will be used in the food industry and will serve as recommendations for manufacturers.

Keywords: dairy, milk processing, nutrition, colostrum

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Authors: Rui Huang, Thomas Richard, Masood Mostofi

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The lifetime of impregnated diamond bits and their drilling efficiency are in part governed by the bit wear conditions, not only the extent of the diamonds’ wear but also their exposure or protrusion out of the matrix bonding. As much as individual diamonds wear, the bonding matrix does also wear through two-body abrasion (direct matrix-rock contact) and three-body erosion (cuttings trapped in the space between rock and matrix). Although there is some work dedicated to the study of diamond bit wear, there is still a lack of understanding on how matrix erosion and diamond exposure relate to the bit drilling response and drilling efficiency, as well as no literature on the process that governs bit sharpening a procedure commonly implemented by drillers when the extent of diamond polishing yield extremely low rate of penetration. The aim of this research is (i) to derive a correlation between the wear state of the bit and the drilling performance but also (ii) to gain a better understanding of the process associated with tool sharpening. The research effort combines specific drilling experiments and precise mapping of the tool-cutting face (impregnated diamond bits and segments). Bit wear is produced by drilling through a rock sample at a fixed rate of penetration for a given period of time. Before and after each wear test, the bit drilling response and thus efficiency is mapped out using a tailored design experimental protocol. After each drilling test, the bit or segment cutting face is scanned with an optical microscope. The test results show that, under the fixed rate of penetration, diamond exposure increases with drilling distance but at a decreasing rate, up to a threshold exposure that corresponds to the optimum drilling condition for this feed rate. The data further shows that the threshold exposure scale with the rate of penetration up to a point where exposure reaches a maximum beyond which no more matrix can be eroded under normal drilling conditions. The second phase of this research focuses on the wear process referred as bit sharpening. Drillers rely on different approaches (increase feed rate or decrease flow rate) with the aim of tearing worn diamonds away from the bit matrix, wearing out some of the matrix, and thus exposing fresh sharp diamonds and recovering a higher rate of penetration. Although a common procedure, there is no rigorous methodology to sharpen the bit and avoid excessive wear or bit damage. This paper aims to gain some insight into the mechanisms that accompany bit sharpening by carefully tracking diamond fracturing, matrix wear, and erosion and how they relate to drilling parameters recorded while sharpening the tool. The results show that there exist optimal conditions (operating parameters and duration of the procedure) for sharpening that minimize overall bit wear and that the extent of bit sharpening can be monitored in real-time.

Keywords: bit sharpening, diamond exposure, drilling response, impregnated diamond bit, matrix erosion, wear rate

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Authors: Tim J. Walsh, Lindsay E. Young, John A. Schneider

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Background: The disproportionate rate of HIV infection among Black men who have sex with men (BMSM) in the United States suggest the importance of Preexposure Prophylaxis (PrEP) interventions for this population. As such, there is an urgent need for innovative outreach strategies that extend beyond the traditional patient-provider relationship to reach at-risk populations. Training members of the BMSM community as peer change agents (PCAs) is one such strategy. An important piece of this training is goal-setting. Goal-setting not only encourages PCAs to define the parameters of the intervention according to their lived experience, it also helps them plan courses of action. Therefore, the aims of this mixed methods study are: (1) Characterize the goals that BMSM set at the end of their PrEP training and (2) Assess the relationship between goal types and PCA engagement. Methods: Between March 2016 and July 2016, preliminary data were collected from 68 BMSM, ages 18-33, in Chicago as part of an ongoing PrEP intervention. Once enrolled, PCAs participate in a half-day training in which they learn about PrEP, practice initiating conversations about PrEP, and identify strategies for supporting at-risk peers through the PrEP adoption process. Training culminates with a goal-setting exercise, whereby participants establish a goal related to their role as a PCA. Goals were coded for features that either emerged from the data itself or existed in extant goal-setting literature. The main outcomes were (1) number of PrEP conversations PCAs self-report during booster conversations two weeks following the intervention and (2) number of peers PCAs recruit into the study that completed the PrEP workshop. Results: PCA goals (N=68) were characterized in terms of four features: Specificity, target population, personalization, and purpose defined. To date, PCAs report a collective 52 PrEP conversations. 56, 25, and 6% of PrEP conversations occurred with friends, family, and sexual partners, respectively. PCAs with specific goals had more PrEP conversations with at-risk peers compared to those with vague goals (58% vs. 42%); PCAs with personalized goals had more PrEP conversations compared to those with de-personalized goals (60% vs. 53%); and PCAs with goals that defined a purpose had more PrEP conversations compared to those who did not define a purpose (75% vs. 52%). 100% of PCAs with goals that defined a purpose recruited peers into the study compared to 45 percent of PCAs with goals that did not define a purpose. Conclusion: Our preliminary analysis demonstrates that BMSM are motivated to set and work toward a diverse set of goals to support peers in PrEP adoption. PCAs with goals involving a clearly defined purpose had more PrEP conversations and greater peer recruitment than those with goals lacking a defined purpose. This may indicate that PCAs who define their purpose at the outset of their participation will be more engaged in the study than those who do not. Goal-setting may be considered as a component of future HIV prevention interventions to advance intervention goals and as an indicator of PCAs understanding of the intervention.

Keywords: HIV prevention, MSM, peer change agent, preexposure prophylaxis

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Authors: Michelle Belinda S. Weerawardana, Gobika Thiripuranathar, Priyani A. Paranagama

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Most of fresh vegetables and fruits available in the retail markets undergo a physiological disorder in its appearance and coloration, which indeed discourages consumer purchase. A loss of millions of dollars yearly to the food industry had been due to this pronounced color reaction called Enzymatic Browning which is driven due to the catalytic activity by an oxidoreductase enzyme, polyphenol oxidase (PPO). The enzyme oxidizes the phenolic compounds which are abundantly available in fruits and vegetables as substrates into quinones, which could react with proteins in its surrounding to generate black pigments, called melanins, which are highly UV-active compounds. Annona muricata (Katu anoda) and Musa acuminata (Ash plantains) is a fruit and a vegetable consumed by Sri Lankans widely due to their high nutritional values, medicinal properties and economical importance. The objective of the present study was to evaluate and determine the effective natural anti-browning inhibitors that could prevent PPO activity in the selected fruit and vegetable. Enzyme extracts from Annona muricata (Katu anoda) and Musa acuminata (Ash plantains), were prepared by homogenizing with analytical grade acetone, and pH of each enzyme extract was maintained at 7.0 using a phosphate buffer. The extracts of inhibitors were prepared using powdered ginger rhizomes and essential oil from the bark of Cinnamomum zeylanicum. Water extracts of ginger were prepared and the essential oil from Ceylon cinnamon bark was extracted using steam distillation method. Since the essential oil is not soluble in water, 0.1µl of cinnamon bark oil was mixed with 0.1µl of Triton X-100 emulsifier and 5.00 ml of water. The effect of each inhibitor on the PPO activity was investigated using catechol (0.1 mol dm-3) as the substrate and two samples of enzyme extracts prepared. The dosages of the prepared Cinnamon bark oil, and ginger (2 samples) which were used to measure the activity were 0.0035 g/ml, 0.091 g/ml and 0.087 g/ml respectively. The measurements of the inhibitory activity were obtained at a wavelength of 525 nm using the UV-visible spectrophotometer. The results evaluated thus revealed that % inhibition observed with cinnamon bark oil, and ginger for Annona muricata was 51.97%, and 60.90% respectively. The effects of cinnamon bark oil, and ginger extract on PPO activity of Musa acuminata were 49.51%, and 48.10%. The experimental findings thus revealed that Cinnamomum zeylanicum bark oil was a more effective inhibitor for PPO enzyme present in Musa acuminata and ginger was effective for PPO enzyme present in Annona muricata. Overall both the inhibitors were proven to be more effective towards the activities of PPO enzyme present in both samples. These inhibitors can thus be corroborated as effective, natural, non-toxic, anti-browning extracts, which when added to the above fruit and vegetable will increase the shelf life and also the acceptance of the product by the consumers.

Keywords: anti-browning agent, enzymatic browning, inhibitory activity, polyphenol oxidase

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Authors: Tahisa M. Pedroso, Hérida R. N. Salgado

Abstract:

The microbiological turbidimetric assay allows the determination of potency of the drug, by measuring the turbidity (absorbance), caused by inhibition of microorganisms by ertapenem sodium. Ertapenem sodium (ERTM), a synthetic antimicrobial agent of the class of carbapenems, shows action against Gram-negative, Gram-positive, aerobic and anaerobic microorganisms. Turbidimetric assays are described in the literature for some antibiotics, but this method is not described for ertapenem. The objective of the present study was to develop and validate a simple, sensitive, precise and accurate microbiological assay by turbidimetry to quantify ertapenem sodium injectable as an alternative to the physicochemical methods described in the literature. Several preliminary tests were performed to choose the following parameters: Staphylococcus aureus ATCC 25923, IAL 1851, 8 % of inoculum, BHI culture medium, and aqueous solution of ertapenem sodium. 10.0 mL of sterile BHI culture medium were distributed in 20 tubes. 0.2 mL of solutions (standard and test), were added in tube, respectively S1, S2 and S3, and T1, T2 and T3, 0.8 mL of culture medium inoculated were transferred to each tube, according parallel lines 3 x 3 test. The tubes were incubated in shaker Marconi MA 420 at a temperature of 35.0 °C ± 2.0 °C for 4 hours. After this period, the growth of microorganisms was inhibited by addition of 0.5 mL of 12% formaldehyde solution in each tube. The absorbance was determined in Quimis Q-798DRM spectrophotometer at a wavelength of 530 nm. An analytical curve was constructed to obtain the equation of the line by the least-squares method and the linearity and parallelism was detected by ANOVA. The specificity of the method was proven by comparing the response obtained for the standard and the finished product. The precision was checked by testing the determination of ertapenem sodium in three days. The accuracy was determined by recovery test. The robustness was determined by comparing the results obtained by varying wavelength, brand of culture medium and volume of culture medium in the tubes. Statistical analysis showed that there is no deviation from linearity in the analytical curves of standard and test samples. The correlation coefficients were 0.9996 and 0.9998 for the standard and test samples, respectively. The specificity was confirmed by comparing the absorbance of the reference substance and test samples. The values obtained for intraday, interday and between analyst precision were 1.25%; 0.26%, 0.15% respectively. The amount of ertapenem sodium present in the samples analyzed, 99.87%, is consistent. The accuracy was proven by the recovery test, with value of 98.20%. The parameters varied did not affect the analysis of ertapenem sodium, confirming the robustness of this method. The turbidimetric assay is more versatile, faster and easier to apply than agar diffusion assay. The method is simple, rapid and accurate and can be used in routine analysis of quality control of formulations containing ertapenem sodium.

Keywords: ertapenem sodium, turbidimetric assay, quality control, validation

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Authors: Nima Samie, Sekaran Muniandy, Zahurin Mohamed, M. S. Kanthimathi

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Although number of indole containing compounds have been reported to have anticancer properties in vitro but only a few of them show potential as anticancer compounds in vivo. The current study was to evaluate the mechanism of cytotoxicity of selected indolic compound in vivo and in vitro. In this context, we determined the potency of the compound in the induction of apoptosis, cell cycle arrest, and cytoskeleton rearrangement. HT-29, WiDr, CCD-18Co, human monocyte/macrophage CRL-9855, and B lymphocyte CCL-156 cell lines were used to determine the IC50 of the compound using the MTT assay. Analysis of apoptosis was carried out using immunofluorescence, acridine orange/ propidium iodide double staining, Annexin-V-FITC assay, evaluation of the translocation of NF-kB, oxygen radical antioxidant capacity, quenching of reactive oxygen species content, measurement of LDH release, caspase-3/-7, -8 and -9 assays and western blotting. The cell cycle arrest was examined using flowcytometry and gene expression was assessed using qPCR array. Results displayed a potent suppressive effect on HT-29 and WiDr after 24 h of treatment with IC50 value of 2.52±0.34 µg/ml and 2.13±0.65 µg/ml respectively. This cytotoxic effect on normal, monocyte/macrophage and B-cells was insignificant. Dipping in the mitochondrial membrane potential and increased release of cytochrome c from the mitochondria indicated induction of the intrinsic apoptosis pathway by the compound. Activation of this pathway was further evidenced by significant activation of caspase-9 and 3/7. The compound was also shown to activate the extrinsic pathways of apoptosis via activation of caspase-8 which is linked to the suppression of NF-kB translocation to the nucleus. Cell cycle arrest in the G1 phase and up-regulation of glutathione reductase, based on excessive ROS production were also observed. These findings were further investigated for inhibitory efficiency of the compound on colonic aberrant crypt foci in male rats. Rats were divided in to 5 groups: vehicle, cancer control, positive control groups and the groups treated with 25 and 50 mg/kg of compounds for 10 weeks. Administration of compound suppressed total colonic ACF formation up to 73.4%. The results also showed that treatment with the compound significantly reduced the level of malondialdehyde while increasing superoxide dismutase and catalase activities. Furthermore, the down-regulation of PCNA and Bcl2 and the up-regulation of Bax was confirmed by immunohistochemical staining. The outcome of this study suggest sthat the indolic compound is a potent anti-cancer agent against colon cancer and can be further evaluated by animal trial.

Keywords: indolic compound, chemoprevention, crypt, azoxymethane, colon cancer

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Authors: Richa Sharma, Uma Nahar, Sadhna Sharma, Indu Verma

Abstract:

Counteracting the deadly pathogen Mycobacterium tuberculosis (M. tb) effectively is still a global challenge. Scrutinizing alternative weapons like antimicrobial peptides to strengthen existing tuberculosis artillery is urgently required. Considering the antimycobacterial potential of Human Beta Defensin 1 (HBD-1) along with isoniazid, the present study was designed to explore the ability of HBD-1 to act against active and dormant M. tb. HBD-1 was screened in silico using antimicrobial peptide prediction servers to identify its short antimicrobial motif. The activity of both HBD-1 and its selected motif (Pep B) was determined at different concentrations against actively growing M. tb in vitro and ex vivo in monocyte derived macrophages (MDMs). Log phase M. tb was grown along with HBD-1 and Pep B for 7 days. M. tb infected MDMs were treated with HBD-1 and Pep B for 72 hours. Thereafter, colony forming unit (CFU) enumeration was performed to determine activity of both peptides against actively growing in vitro and intracellular M. tb. The dormant M. tb models were prepared by following two approaches and treated with different concentrations of HBD-1 and Pep B. Firstly, 20-22 days old M. tbH37Rv was grown in potassium deficient Sauton media for 35 days. The presence of dormant bacilli was confirmed by Nile red staining. Dormant bacilli were further treated with rifampicin, isoniazid, HBD-1 and its motif for 7 days. The effect of both peptides on latent bacilli was assessed by colony forming units (CFU) and most probable number (MPN) enumeration. Secondly, human PBMC granuloma model was prepared by infecting PBMCs seeded on collagen matrix with M. tb(MOI 0.1) for 10 days. Histopathology was done to confirm granuloma formation. The granuloma thus formed was incubated for 72 hours with rifampicin, HBD-1 and Pep B individually. Difference in bacillary load was determined by CFU enumeration. The minimum inhibitory concentrations of HBD-1 and Pep B restricting growth of mycobacteria in vitro were 2μg/ml and 20μg/ml respectively. The intracellular mycobacterial load was reduced significantly by HBD-1 and Pep B at 1μg/ml and 5μg/ml respectively. Nile red positive bacterial population, high MPN/ low CFU count and tolerance to isoniazid, confirmed the formation of potassium deficienybaseddormancy model. HBD-1 (8μg/ml) showed 96% and 99% killing and Pep B (40μg/ml) lowered dormant bacillary load by 68.89% and 92.49% based on CFU and MPN enumeration respectively. Further, H&E stained aggregates of macrophages and lymphocytes, acid fast bacilli surrounded by cellular aggregates and rifampicin resistance, indicated the formation of human granuloma dormancy model. HBD-1 (8μg/ml) led to 81.3% reduction in CFU whereas its motif Pep B (40μg/ml) showed only 54.66% decrease in bacterial load inside granuloma. Thus, the present study indicated that HBD-1 and its motif are effective antimicrobial players against both actively growing and dormant M. tb. They should be further explored to tap their potential to design a powerful weapon for combating tuberculosis.

Keywords: antimicrobial peptides, dormant, human beta defensin 1, tuberculosis

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