Search results for: Centrifugal Partition Chromatography

Authors: F. Al-Sheikh, C. Moralejo, M. Pritzker, W. A. Anderson, A. Elkamel

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Ammonia in mining wastewater is a significant problem, and treatment can be especially difficult in cold climates where biological treatment is not feasible. An adsorption process is one of the alternative processes that can be used to reduce ammonia concentrations to acceptable limits, and therefore a LEWATIT resin strongly acidic H+ form ion exchange resin and a Bowie Chabazite Na form AZLB-Na zeolite were tested to assess their effectiveness. For these adsorption tests, two packed bed columns (a mini-column constructed from a 32-cm long x 1-cm diameter piece of glass tubing, and a 60-cm long x 2.5-cm diameter Ace Glass chromatography column) were used containing varying quantities of the adsorbents. A mining wastewater with ammonia concentrations of 22.7 mg/L was fed through the columns at controlled flowrates. In the experimental work, maximum capacities of the LEWATIT ion exchange resin were 0.438, 0.448, and 1.472 mg/g for 3, 6, and 9 g respectively in a mini column and 1.739 mg/g for 141.5 g in a larger Ace column while the capacities for the AZLB-Na zeolite were 0.424, and 0.784 mg/g for 3, and 6 g respectively in the mini column and 1.1636 mg/g for 38.5 g in the Ace column. In the theoretical work, Thomas, Adams-Bohart, and Yoon-Nelson models were constructed to describe a breakthrough curve of the adsorption process and find the constants of the above-mentioned models. In the regeneration tests, 5% hydrochloric acid, HCl (v/v) and 10% sodium hydroxide, NaOH (w/v) were used to regenerate the LEWATIT resin and AZLB-Na zeolite with 44 and 63.8% recovery, respectively. In conclusion, continuous flow adsorption using a LEWATIT ion exchange resin and an AZLB-Na zeolite is efficient when using a co-flow technique for removal of the ammonia from wastewater. Thomas, Adams-Bohart, and Yoon-Nelson models satisfactorily fit the data with R² closer to 1 in all cases.

Keywords: AZLB-Na zeolite, continuous adsorption, Lewatit resin, models, regeneration

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Authors: Dalvir Kataria, Khushminder Kaur Chahal, Amit Kumar

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The carrot seed essential oil was obtained by hydrodistillation. Hexane, dichloromethane, and methanol solvents were used for extraction of carrot seed by Soxhlet extraction methods. The major and minor compounds identified in carrot seed essential oil were carotol (52.73), daucol (5.10), daucene (5.68), (E)-β-farnesene (5.40), β-cubebene (3.19), longifolenaldehyde (3.23), β-elimene (3.23), (E)-caryophyllene (1.22), β-bisabolene (2.95) etc. The chemical composition of hexane, dichloromethane, and methanol extracts was different. Carotol was the common compound present. Major compounds isolated were from the carrot seed essential oil by column chromatography. Chemical transformations of carotol (2) with mercuric acetate/sodium borohydride, dry hydrochloric acid gas, acetonitrile/sulfuric acid, selenium dioxide/t-butyl hydrogen peroxide, N-bromosuccinimide, hydrogen iodide, and phenol were carried out. The derivatives of carotol were designed to explore the significance of some structural modifications in relation to antioxidant activities. The structures of major compounds and derivatives were confirmed on the basis of FT-IR, 1HNMR and 13CNMR spectroscopy. Antioxidant activity of carrot seed essential oil, various extracts and isolated compounds were tested by in vitro models involving 2, 2-diphenyl-1-picrylhydrazyl (DPPH•), hydroxyl (OH•), nitric oxide (NO•), superoxide radical scavenging methods and ferric reducing antioxidant power assay (FRAP). Chemical transformations of major isolated compound carotol were carried out, and antioxidant activity of all compounds was undertaken. The major sesquiterpenoidcarotol isolated from carrot seed essential oil showed the highest antioxidant activity in all the methods. The methanol extract showed higher antioxidant potential as compared to carrot seed essential oil, hexane, and dichloromethane extracts.

Keywords: antioxidant, carotol, carrot, DPPH

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Authors: Yuri A. Kalvachev, Totka D. Todorova

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Zeolite mordenite has been investigated as a transition metal support for the preparation of efficient catalysts in the oxidation of volatile organic compounds (VOCs). The highly crystalline mordenite samples were treated with hydrofluoric acid and ammonium fluoride to get hierarchical material with secondary porosity. The obtained supports by this method have a high active surface area, good diffusion properties and prevent the extraction of metal components during catalytic reactions. The active metal phases platinum and copper were loaded by impregnation on both mordenite materials (parent and acid treated counterparts). Monometalic Pt and Cu, and bimetallic Pt/Cu catalysts were obtained. The metal phases were fine dispersed as nanoparticles on the functional porous materials. The catalysts synthesized in this way were investigated in the reaction of complete oxidation of propane and benzene. Platinum, copper and platinum/copper were loaded and there catalytic activity was investigated and compared. All samples are characterized by X-ray diffraction analysis, nitrogen adsorption, scanning electron microscopy (SEM), X-ray photoelectron measurements (XPS) and temperature programed reduction (TPR). The catalytic activity of the samples obtained is investigated in the reaction of complete oxidation of propane and benzene by using of Gas Chromatography (GC). The oxidation of three organic molecules was investigated—methane, propane and benzene. The activity of metal loaded mordenite catalysts for methane oxidation is almost the same for parent and treated mordenite as a support. For bigger molecules as propane and benzene, the activity of catalysts based on treated mordenite is higher than those based on parent zeolite.

Keywords: metal loaded catalysts, mordenite, VOCs oxidation, zeolites

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Authors: Ahmed F. Azmy, Amal E. Saafan, Tamer M. Essam, Magdy A. Amin, Shaban H. Ahmed

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Bacterial strains capable of degradation of malathion from the domestic sewage were isolated by an enrichment culture technique. Three bacterial strains were screened and identified as Acinetobacter baumannii (AFA), Pseudomonas aeruginosae (PS1),andPseudomonas mendocina (PS2) based on morphological, biochemical identification and 16S rRNA sequence analysis. Acinetobacter baumannii AFA was the most efficient malathion degrading bacterium, so used for further biodegradation study. AFA was able to grow in mineral salt medium (MSM) supplemented with malathion (100 mg/l) as a sole carbon source, and within 14 days, 84% of the initial dose was degraded by the isolate measured by high performance liquid chromatography. Strain AFA could also degrade other organophosphorus compounds including diazenon, chlorpyrifos and fenitrothion. The effect of different culture conditions on the degradation of malathion like inoculum density, other carbon or nitrogen sources, temperature and shaking were examined. Degradation of malathion and bacterial cell growth were accelerated when culture media were supplemented with yeast extract, glucose and citrate. The optimum conditions for malathion degradation by strain AFA were; an inoculum density of 1.5x 1012CFU/ml at 30°C with shaking. A specific polymerase chain reaction primers were designed manually using multiple sequence alignment of the corresponding carboxylesterase enzymes of Acinetobacter species. Sequencing result of amplified PCR product and phylogenetic analysis showed low degree of homology with the other carboxylesterase enzymes of Acinetobacter strains, so we suggested that this enzyme is a novel esterase enzyme. Isolated bacterial strains may have potential role for use in bioremediation of malathion contaminated.

Keywords: Acinetobacter baumannii, biodegradation, malathion, organophosphate pesticides

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Authors: Norah Al Hokbany, Ibrahim Al Jammaz, Basem Al Otaibi, Yousif Al Malki, Subhani M. Okarvi

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Objective: The folate receptor is over-expressed in a wide variety of human tumors. Conjugates of folate have been shown to be selectively taken up by tumor cells via the folate receptor. In an attempt to develop new folate radiotracers with favorable biochemical properties for detecting folate receptor-positive cancers. Methods: we synthesized ⁶⁴Cu-NOTA- and ⁶⁴Cu-NOTAM-folate conjugates using a straightforward and simple one-step reaction. Radiochemical yields were greater than 95% (decay-corrected) with a total synthesis time of less than 20 min. Results: Radiochemical purities were always greater than 98% without high-performance liquid chromatography (HPLC) purification. These synthetic approaches hold considerable promise as a rapid and simple method for ⁶⁴Cu-folate conjugate preparation with high radiochemical yield in a short synthesis time. In vitro tests on the KB cell line showed that significant amounts of the radio conjugates were associated with cell fractions. Bio-distribution studies in nude mice bearing human KB xenografts demonstrated a significant tumor uptake and favorable bio-distribution profile for ⁶⁴Cu-NOTA- and ⁶⁴Cu-NOTAM-folate conjugate. The uptake in the tumors was blocked by the excess injection of folic acid, suggesting a receptor-mediated process. Conclusion: These results demonstrate that the ⁶⁴Cu-NOTAM-folate conjugate may be useful as a molecular probe for the detection and staging of folate receptor-positive cancers, such as ovarian cancer and their metastasis, as well as monitoring tumor response to treatment.

Keywords: folate, receptor, tumor imaging, ⁶⁴Cu-NOTA-folate, PET

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Authors: L. Correia-Sá, S. Norberto, Conceição Calhau, C. Delerue-Matos, V. F. Domingues

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Endocrine disruptor chemicals (EDCs) are synthetic compounds that even though being initially designed for a specific function are now being linked with a wide range of side effects. The list of possible EDCs is growing and includes phthalates and bisphenol A (BPA). Phthalates are one of the most widely used plasticizers to improve the extensibility, elasticity and workability of polyvinyl chloride (PVC), polyvinyl acetates, etc. Considered non-toxic and harmless additives for polymers, they were used unrestrainedly all over the world for several decades. However, recent studies have indicated that some phthalates and their metabolic products are reproductive and developmental toxicants in animals and suspected endocrine disruptors in humans. BPA (2,2-bis(4-hydroxyphenyl)propane) is a high production volume chemical mainly used in the production of polycarbonate plastics and epoxy resins. Although BPA was initially considered to be a weak environmental estrogen, nowadays it is known that this compound can stimulate several cellular responses at very low levels of concentrations. The aim of this study was to develop a method based on tandem SPE to evaluate the presence of phthalates, metabolites and BPA in human urine samples. The analyzed compounds included: dibutyl phthalate (DBP) and di-2-ethylhexyl phthalate (DEHP), BPA, mono-isobutyl phthalate (MiBP), monobutyl phthalate (MBP) and. mono-(2-ethyl-5-oxohexyl) (MEOHP). Two SPE cartridges were applied both from Phenomenex, the strata X polymeric reversed phase and the strata X A (Strong anion). Chromatographic analyses were carried out in a Thermo GC ULTRA GC-MS/MS. Good recoveries and linear calibration curves were obtained. After validation, the methodology was applied to human urine samples for phthalates, metabolites and BPA evaluation.

Keywords: Bisphenol A (BPA), gas chromatography, metabolites, phtalates, SPE, tandem mode

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Authors: Chi-Ching Lee, Po-Jung Huang, Kuo-Yang Huang, Petrus Tang

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Background: Recent advances in high-throughput research technologies such as new-generation sequencing and multi-dimensional liquid chromatography makes it possible to dissect the complete transcriptome and proteome in a single run for the first time. However, it is almost impossible for many laboratories to handle and analysis these “BIG” data without the support from a bioinformatics team. We aimed to provide a web-based analysis platform for users with only limited knowledge on bio-computing to study the functional genomics and proteomics. Method: We use NCI-60 as an example dataset to demonstrate the power of the web-based analysis platform and data delivering system: C-eXpress takes a simple text file that contain the standard NCBI gene or protein ID and expression levels (rpkm or fold) as input file to generate a distribution map of gene/protein expression levels in a heatmap diagram organized by color gradients. The diagram is hyper-linked to a dynamic html table that allows the users to filter the datasets based on various gene features. A dynamic summary chart is generated automatically after each filtering process. Results: We implemented an integrated database that contain pre-defined annotations such as gene/protein properties (ID, name, length, MW, pI); pathways based on KEGG and GO biological process; subcellular localization based on GO cellular component; functional classification based on GO molecular function, kinase, peptidase and transporter. Multiple ways of sorting of column and rows is also provided for comparative analysis and visualization of multiple samples.

Keywords: cancer, visualization, database, functional annotation

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Authors: Neil Jolly, Louisa Beukes, Santiago Benito-SaEz

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Kei-apple is a tree found on the African continent. Limited information exists on the effect of alcoholic and acetous fermentation on the phytochemicals. The fruit has increased L-malic, ascorbic, and phenolic acids. Juice was co-inoculated with Schizosaccharomyces pombe and Saccharomyces cerevisiae to induce alcoholic fermentation and acetous fermentation using acetic acid bacteria. Saccharomyces cerevisiae+S. pombe wines and vinegars had highest pH. Total acidity, soluble solids and L-malic acid decreased during alcoholic and acetous fermentation with highest in S. cerevisiae wines and vinegars. Volatile acidity was highest in S. pombe vinegars but not different from S. cerevisiae and S. cerevisiae+S. pombe. Gallic acid was highest in S. pombe wines and vinegars. Syringic acid was highest in S. cerevisiae wines and vinegars. S. cerevisiae+S. pombe wines were highest in caffeic, p-coumaric and protocatechuic acids. Schizosaccharomyces pombe vinegars were highest in caffeic and p-coumaric acids. Ferulic and sinapic acids were highest in S. pombe and S. cerevisiae wines, respectively. Chlorogenic acid was most abundant in both wines and vinegars. Saccharomyces cerevisiae+S. pombe and S. cerevisiae had a positive effect on most phenolic acids. Saccharomyces cerevisiae +acetic acid bacteria had an increased effect on syringic and chlorogenic acids. Schizosaccharomyces pombe+acetic acid bacteria resulted in an increase in gallic, caffeic and p-coumaric acids. Acetic acid bacteria had minimal performance with respect to volatile acidity production in comparison to commercial vinegars. Acetic acid bacteria selection should therefore be reconsidered and the decrease of certain phenolic acids during acetous fermentation needs to be investigated.

Keywords: acetic acid bacteria, liquid chromatography, phenolics, saccharomyces cerevisiae, schizosaccharomyces pombe

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Authors: Daniella L. Pereira, Emeliana G. Imperial, Ingrid Webster, Ian Wiid, Hans Strijdom, Nireshni Chellan, Sanet H. Kotzé

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Rooibos (Aspalathus linearis) is a shrub-like bush native to the Western Cape of South Africa and commonly consumed as a herbal tea. Interest on the anti-oxidant capabilities of the tea have risen based on anecdotal evidence. Rooibos contains polyphenols that contribute to the overall antioxidant capacity of the tea. These polyphenols have been reported to attenuate the effects of oxidative stress in biological systems. The bioavailability of these compounds is compromised when exposed to light, pH fluctuations, and oxidation. It is crucial to evaluate whether the polyphenols in a typical rooibos solution remain constant over time when administered to rats in a research environment. This study aimed to determine the effects of various storage scenarios on the phenolic composition of rooibos tea commonly administered to rodents in experimental studies. A standardised aqueous solution of rooibos tea was filtered and divided into three samples namely fresh, refrigerated, and frozen. Samples were stored in air tight, light excluding bottles. Refrigerated samples were stored at 4°C for seven days. Frozen samples were stored for fourteen days at -20°C. Each sample consisted of two subgroups labeled day 1 and day 7. Teas marked day 7 of each group were kept in air tight, light protected bottles at room temperature for an additional week. All samples (n=6) were freeze-dried and underwent polyphenol characterization using liquid chromatography-mass spectrometry. The phenolic composition remained constant throughout all groups. This indicates that rooibos tea can be safely stored at the above conditions without compromising the phenolic viability of the tea typically used for research purposes.

Keywords: Aspalathus linearis, experimental studies, polyphenols, storage

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Authors: Samuele Viaro, Pierre Ricco

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Görtler instabilities generate in boundary layers from an unbalance between pressure and centrifugal forces caused by concave surfaces. Their spatial streamwise evolution influences transition to turbulence. It is therefore important to understand even the early stages where perturbations, still small, grow linearly and could be controlled more easily. This work presents a rigorous theoretical framework for compressible flows using the linearized unsteady boundary region equations, where only the streamwise pressure gradient and streamwise diffusion terms are neglected from the full governing equations of fluid motion. Boundary and initial conditions are imposed through an asymptotic analysis in order to account for the interaction of the boundary layer with free-stream turbulence. The resulting parabolic system is discretize with a second-order finite difference scheme. Realistic flow parameters are chosen from wind tunnel studies performed at supersonic and subsonic conditions. The Mach number ranges from 0.5 to 8, with two different radii of curvature, 5 m and 10 m, frequencies up to 2000 Hz, and vortex spanwise wavelengths from 5 mm to 20 mm. The evolution of the perturbation flow is shown through velocity, temperature, pressure profiles relatively close to the leading edge, where non-linear effects can still be neglected, and growth rate. Results show that a global stabilizing effect exists with the increase of Mach number, frequency, spanwise wavenumber and radius of curvature. In particular, at high Mach numbers curvature effects are less pronounced and thermal streaks become stronger than velocity streaks. This increase of temperature perturbations saturates at approximately Mach 4 flows, and is limited in the early stage of growth, near the leading edge. In general, Görtler vortices evolve closer to the surface with respect to a flat plate scenario but their location shifts toward the edge of the boundary layer as the Mach number increases. In fact, a jet-like behavior appears for steady vortices having small spanwise wavelengths (less than 10 mm) at Mach 8, creating a region of unperturbed flow close to the wall. A similar response is also found at the highest frequency considered for a Mach 3 flow. Larger vortices are found to have a higher growth rate but are less influenced by the Mach number. An eigenvalue approach is also employed to study the amplification of the perturbations sufficiently downstream from the leading edge. These eigenvalue results are compared with the ones obtained through the initial value approach with inhomogeneous free-stream boundary conditions. All of the parameters here studied have a significant influence on the evolution of the instabilities for the Görtler problem which is indeed highly dependent on initial conditions.

Keywords: compressible boundary layers, Görtler instabilities, receptivity, turbulence transition

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Authors: Miranda I. Dosunmu, Orok E. Oyo-Ita, Inyang O. Oyo-Ita

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Polycyclic aromatic hydrocarbons (PAHs) are a class of priority listed organic pollutants due to their carcinogenicity, mutagenity, acute toxicity and persistency in the environment. The distribution and historical changes of PAHs contamination in recent sediment cores from the Imo River were investigated using gas chromatography coupled with mass spectrometer. The concentrations of total PAHs (TPAHs) ranging from 402.37 ng/g dry weight (dw) at the surface layer of the Estuary zone (ESC6; 0-5 cm) to 92,388.59 ng/g dw at the near surface layer of the Afam zone (ASC5; 5-10 cm) indicate that PAHs contamination was localized not only between sample sites but also within the same cores. Sediment-depth profiles for the four (Afam, Mangrove, Estuary and illegal Petroleum refinery) cores revealed irregular distribution patterns in the TPAH concentrations except the fact that these levels became maximized at the near surface layers (5-10 cm) corresponding to a geological time-frame of about 1996-2004. This time scale coincided with the period of intensive bunkering and oil pipeline vandalization by the Niger Delta militant groups. Also a general slight decline was found in the TPAHs levels from near the surface layers (5-10 cm) to the most recent top layers (0-5 cm) of the cores, attributable to the recent effort by the Nigerian government in clamping down the illegal activity of the economic saboteurs. Therefore, the recent amnesty period granted to the militant groups should be extended. Although mechanism of perylene formation still remains enigmatic, examination of its distributions down cores indicates natural biogenic, pyrogenic and petrogenic origins for the compound at different zones. Thus, the characteristic features of the Imo River environment provide a means of tracing diverse origins for perylene.

Keywords: perylene, historical trend, distribution, origin, Imo River

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Authors: Lopamudra Ray, Malla Padma, Dibya Bhol, Samir Ranjan Mishra, A. N. Panda, Gurdeep Rastogi, T. K. Adhya, Ajit Kumar Pattnaik, Mrutyunjay Suar, Vishakha Raina

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Chilika Lake is the largest coastal estuarine brackish water lagoon in Asia situated on the east coast of India and is a designated Ramsar site. In the current study, several chitinolytic microorganisms were isolated and screened by appearance of clearance zone on 0.5% colloidal chitin agar plate. A strain designated as RC 1830 displayed maximum colloidal chitin degradation by release of 112 μmol/ml/min of N-acetyl D-glucosamine (GlcNAc) in 48h. The strain was taxonomically identified by polyphasic approach based on a range of phenotypic and genotypic properties and was found to be a novel species named Streptomyces chilikensis RC1830. The organism was halophilic (12% NaCl w/v), alkalophilic (pH10) and was capable of hydrolyzing chitin, starch, cellulose, gelatin, casein, tributyrin and tween 80. The partial purification of chitinase enzymes from RC1830 was performed by DEAE Sephacel anion exchange chromatography which revealed the presence of a very low molecular weight chitinase(10.5kD) which may be a probable chitobiosidase enzyme. The study reports the presence of a low MW chitinase (10.5kD) and a chitin decaetylase from a novel Streptomyces strain RC1830 isolated from Chilika Lake. Previously chitinases less than 20.5kD have not been reported from any other Streptomyces species. The enzymes was characterized with respect to optimum pH, temperature, and substrate specificity and temperature stability.

Keywords: chitinases, chitobiosidase, Chilika Lake, India

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Authors: Bruna Goncalves, Jennifer Henck, Romulo Uliana, Eliana Kamimura, Carlos Oliveira, Carlos Corassin

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Studies have shown evidence of human exposure to aflatoxin M1 due to the consumption of contaminated milk and dairy products (mainly cheeses). This poses a great risk to public health, since milk and milk products are frequently consumed by a portion of the population considered immunosuppressed, children and the elderly. Knowledge of the negative impacts of aflatoxins on health and economics has led to investigations of strategies to prevent their formation in food, as well as to eliminate, inactivate or reduce the bioavailability of these toxins in contaminated products This study evaluated the effect of microbiological methods using lactic acid bacteria on aflatoxin M1 (AFM1) reduction in Minas Frescal cheese (typical Brazilian product, being among the most consumed cheeses in Brazil) spiked with 1 µg/L AFM1. Inactivated lactic acid bacteria (0,5%, v/v de L. rhamnosus e L. lactis) were added during the cheese production process. Nine cheeses were produced, divided into three treatments: negative controls (without AFM1 or lactic acid bacteria), positive controls (AFM1 only), and lactic acid bacteria + AFM1. Samples of cheese were collected on days 2, 10, 20 and 30 after the date of production and submitted to composition analyses and determination of AFM1 by high-performance liquid chromatography. The reductions of AFM1 in cheese by lactic acid bacteria at the end of the trial indicate a potential application of inactivated lactic acid bacteria in reducing the bioavailability of AFM1 in Minas frescal cheese without physical-chemical and microbiological modifications during the 30-day experimental period. The authors would like to thank São Paulo Research Foundation – FAPESP (grants #2017/20081-6 and #2017/19683-1).

Keywords: aflatoxin, milk, minas frescal cheese, decontamination

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Authors: Pengzhou Lu, Liming Xin, Payam Tavakoli, Zhonghua Lin, Roberto V. P. Ribeiro, Mitesh V. Badiwala

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Ex vivo Heart Perfusion (EVHP) has been developed as an alternative strategy to expand cardiac donation by enabling resuscitation and functional assessment of hearts donated from marginal donors, which were previously not accepted. EVHP parameters, such as perfusion flow (PF) and perfusion pressure (PP) are crucial for optimal organ preservation. However, with the heart’s constant physiological changes during EVHP, such as coronary vascular resistance, manual control of these parameters is rendered imprecise and cumbersome for the operator. Additionally, low control precision and the long adjusting time may lead to irreversible damage to the myocardial tissue. To solve this problem, an automatic heart perfusion system was developed by applying a Human-Machine Interface (HMI) and a Programmable-Logic-Controller (PLC)-based circuit to control PF and PP. The PLC-based control system collects the data of PF and PP through flow probes and pressure transducers. It has two control modes: the RPM-flow mode and the pressure mode. The RPM-flow control mode is an open-loop system. It influences PF through providing and maintaining the desired speed inputted through the HMI to the centrifugal pump with a maximum error of 20 rpm. The pressure control mode is a closed-loop system where the operator selects a target Mean Arterial Pressure (MAP) to control PP. The inputs of the pressure control mode are the target MAP, received through the HMI, and the real MAP, received from the pressure transducer. A PID algorithm is applied to maintain the real MAP at the target value with a maximum error of 1mmHg. The precision and control speed of the RPM-flow control mode were examined by comparing the PLC-based system to an experienced operator (EO) across seven RPM adjustment ranges (500, 1000, 2000 and random RPM changes; 8 trials per range) tested in a random order. System’s PID algorithm performance in pressure control was assessed during 10 EVHP experiments using porcine hearts. Precision was examined through monitoring the steady-state pressure error throughout perfusion period, and stabilizing speed was tested by performing two MAP adjustment changes (4 trials per change) of 15 and 20mmHg. A total of 56 trials were performed to validate the RPM-flow control mode. Overall, the PLC-based system demonstrated the significantly faster speed than the EO in all trials (PLC 1.21±0.03, EO 3.69±0.23 seconds; p < 0.001) and greater precision to reach the desired RPM (PLC 10±0.7, EO 33±2.7 mean RPM error; p < 0.001). Regarding pressure control, the PLC-based system has the median precision of ±1mmHg error and the median stabilizing times in changing 15 and 20mmHg of MAP are 15 and 19.5 seconds respectively. The novel PLC-based control system was 3 times faster with 60% less error than the EO for RPM-flow control. In pressure control mode, it demonstrates a high precision and fast stabilizing speed. In summary, this novel system successfully controlled perfusion flow and pressure with high precision, stability and a fast response time through a user-friendly interface. This design may provide a viable technique for future development of novel heart preservation and assessment strategies during EVHP.

Keywords: automatic control system, biomedical engineering, ex-vivo heart perfusion, human-machine interface, programmable logic controller

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Authors: Ozlem Bahadir Acikara, Mert Ilhan, Ekin Kurtul, Karel Smejkal, Esra Kupeli Akkol

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Present study is aimed to investigate in vitro inhibitory effects of the extracts prepared from the aerial parts of Podospermum canum (Asteraceae) on hyaluronidase, collagenase, and elastase enzymes using a bioassay-guided fractionation. Inhibitory effects of the extract, sub-extracts, fractions obtained by column chromatography, and isolated compounds on collagenase, elastase, and hyaluronidase were performed by using in vitro enzyme inhibitory assays based on spectrophotometric evaluation. The ethyl acetate and remaining water extracts prepared from the plant displayed significant inhibitory activities on collagenase and elastase, while petroleum ether and chloroform extracts did not show any inhibitory activity. Eleven known compounds: arbutin, 6'-O-caffeoylarbutin, cichoriin, 3,5-dicaffeoylquinic acid methyl ester, apigenin-7-O-β-glucoside, luteolin-7-O-β-glucoside, apigenin-7-O-β-rutinoside, isoorientin, orientin, vitexin, procatechuic acid, and compound 4-hydroxy-benzoic acid 4-(6-O-α-rhamnopyranosyl-β-glucopyranosyl) benzyl ester have been obtained from ethyl acetate sub-extract of the plant through bioassay-guided fractionation and isolation. Results of the present study have revealed that among the isolated compounds, apigenin-7-O-β-glucoside, luteolin-7-O-β-glucoside, apigenin-7-O-β-rutinoside and isoorientin showed potent enzyme inhibitory activities. However, methanolic extract of P. canum displayed a greater inhibitory activity than fractions and isolated compounds both on collagenase and elastase.

Keywords: Asteraceae, collagenase, elastase, hyaluronidase, Podospermum canum

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Authors: Amr Amer

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Cheeses are among those high-protein-containing foodstuffs in which enzymatic and microbial activities cause the formation of biogenic amines from amino acids decarboxylation. The amount of biogenic amines in cheese may act as a useful indicator of the hygienic quality of the product. In other words, their presence in cheese is related to its spoilage and safety. Formation of biogenic amines during Ras cheese (Egyptian hard cheese) ripening was investigated for 4 months. Three batches of Ras cheese were manufactured using Egyptian traditional method. From each batch, Samples were collected at 1, 7, 15, 30, 60, 90 and 120 days after cheese manufacture. The concentrations of biogenic amines (Tyramine, Histamine, Cadaverine and Tryptamine) were analyzed by high performance liquid chromatography (HPLC). There was a significant increased (P<0.05) in Tyramine levels from 4.34± 0.07 mg|100g in the first day of storage till reached 88.77± 0.14 mg|100g at a 120-day of storage. Also, Histamine and Cadaverine levels had the same increased pattern of Tyramine reaching 64.94± 0.10 and 28.28± 0.08 mg|100g in a 120- day of storage, respectively. While, there was a fluctuation in the concentration of Tryptamine level during ripening period as it decreased from 3.24± 0.06 to 2.66± 0.11 mg|100g at 60-day of storage then reached 5.38±0.08 mg|100g in a 120- day of storage. Biogenic amines can be formed in cheese during production and storage: many variables, as pH, salt concentration, bacterial activity as well as moisture, storage temperature and ripening time, play a relevant role in their formation. Comparing the obtained results with the recommended standard by Food and Drug Administration "FDA" (2001), High levels of biogenic amines in various Ras cheeses consumed in Egypt exceeded the permissible value (10 mg%) which seemed to pose a threat to public health. In this study, presence of high concentrations of biogenic amines (Tyramine, Histamine, cadaverine and Tryptamine) in Egyptian Ras cheeses reflects the bad hygienic conditions under which they produced and stored. Accordingly, the levels of biogenic amines in different cheeses should be come in accordance with the safe permissible limit recommended by FDA to ensure human safety.

Keywords: Ras cheese, biogenic amines, tyramine, histamine, cadaverine

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Authors: Boukhatem Mohamed Nadjib, Ferhat Mohamed Amine

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Essential Oils (EO) produced by medicinal plants have been traditionally used for respiratory tract infections and are used nowadays as ethical medicines for colds. The aim of this study was to test the efficacy of the Algerian EGEO against some respiratory tract pathogens by disc diffusion and vapour diffusion methods at different concentrations. The chemical composition of the EGEO was analysed by Gas Chromatography-Mass Spectrometry. Fresh leaves of E. globulus on steam distillation yielded 0.96% (v/w) of essential oil whereas the analysis resulted in the identification of a total of 11 constituents, 1.8 cineole (85.8%), α-pinene (7.2%) and β-myrcene (1.5%) being the main components. By disc diffusion method, EGEO showed potent antimicrobial activity against Gram-positive more than Gram-negative bacteria. The Diameter of Inhibition Zone (DIZ) varied from 69 mm to 75 mm for Staphylococcus aureus and Bacillus subtilis (Gram +) and from 13 to 42 mm for Enterobacter sp and Escherichia coli (Gram-), respectively. However, the results obtained by both agar diffusion and vapour diffusion methods were different. Significantly higher antibacterial activity was observed in the vapour phase at lower concentrations. A. baumanii and Klebsiella pneumoniae were the most susceptible strains to the oil vapour with DIZ varied from 38 to 42 mm. Therefore, smaller doses of EO in the vapour phase can be inhibitory to pathogenic bacteria. Else, the DIZ increased with increase in the concentration of the oil. There is growing evidence that EGEO in the vapour phase are effective antibacterial systems and appears worthy to be considered for practical uses in the treatment or prevention of patients with respiratory tract infections or as air decontaminants in the hospital. The present study indicates that EGEO has considerable antimicrobial activity, deserving further investigation for clinical applications.

Keywords: eucalyptus globulus, essential oils, respiratory tract pathogens, antimicrobial activity, vapour phase

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Authors: Belhaj Mohamed, M. Saidi, I. Bouazizi, M. Soussi, M. Ben Jrad

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The Late Cenomanian-Early Turonian laminated, black, organic-rich limestones were described in Central Tunisia and attributed to the Bahloul Formation. It covers central and northern Tunisia, and the northern part of the Gulf of Gabes. The Bahloul Formation is considered as one of the main source rocks in Tunisia and is composed of outer-shelf to slop-laminated and dark-gray to black-colored limestones and marls. This formation had been deposited in a relatively deep-marine, calm, and anoxic environment. Rock-Eval analysis and vitrinite reflectance (Ro) measurements were performed on the basis of the organic carbon content. Several samples were chosen for molecular organic geochemistry. Saturate and aromatic hydrocarbons were analyzed by gas chromatography (GC) and GC–mass spectrometry. Geochemical data of the Bahloul Formation in northern and central Tunisia show this level to be a good potential source rock as indicated by the high content of type II organic matter. This formation exhibits high total organic carbon contents (as much as 14%), with an average value of 2% and a good to excellent petroleum potential, ranging between 2 and 50 kg of hydrocarbon/ton of rock. The extracts from the Bahloul Formation are characterized by Pr/Ph ratios ranging between 1.5 and 3, a moderate diasterane content, a C27 sterane approximately equal to C29 sterane, a high C28/C29 ratio, low gammacerane index, a C35/C34 homohopane ratio less than 1 and carbon isotope compositions between -24 and -26‰. The thermal maturity is relatively low, corresponding to the beginning of the oil window in the western area near the Algerian border, in the oil window in the eastern area (Sahel basin) and late mature in northern part.

Keywords: biomarkers, organic geochemistry, source rock, Tunisia

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Authors: Miguel Pereira, Lígia Pimentel, Manuela Pintado

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Animal blood is a major by-product of slaughterhouses and still represents a cost and environmental problem in some countries. To be eliminated, blood should be stabilised by cooking and afterwards the slaughterhouses must have to pay for its incineration. In order to reduce the elimination costs and valorise the high protein content the aim of this study was the optimization of hydrolysis conditions, in terms of enzyme ratio and time, in order to obtain hydrolysates with biological activity. Two enzymes were tested in this assay: pepsin and proteases from Cynara cardunculus (cardosins). The latter has the advantage to be largely used in the Portuguese Dairy Industry and has a low price. The screening assays were carried out in a range of time between 0 and 10 h and using a ratio of enzyme/reaction volume between 0 and 5%. The assays were performed at the optimal conditions of pH and temperature for each enzyme: 55 °C at pH 5.2 for cardosins and 37 °C at pH 2.0 for pepsin. After reaction, the hydrolysates were evaluated by FPLC (Fast Protein Liquid Chromatography) and tested for their antioxidant activity by ABTS method. FPLC chromatograms showed different profiles when comparing the enzymatic reactions with the control (no enzyme added). The chromatogram exhibited new peaks with lower MW that were not present in control samples, demonstrating the hydrolysis by both enzymes. Regarding to the antioxidant activity, the best results for both enzymes were obtained using a ratio enzyme/reactional volume of 5% during 5 h of hydrolysis. However, the extension of reaction did not affect significantly the antioxidant activity. This has an industrial relevant aspect in what concerns to the process cost. In conclusion, the enzymatic blood hydrolysis can be a better alternative to the current elimination process allowing to the industry the reuse of an ingredient with biological properties and economic value.

Keywords: antioxidant activity, blood, by-products, enzymatic hydrolysis

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Authors: Mthokozisi Simelane

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Background: Malaria remains a life-threatening disease in tropical regions despite the advances in the treatment of this disease, it still remains a significant burden as some parasites have become resistant to the currently available drugs. This has created a necessity for the development of alternative, more efficient antimalarial drugs. Warburgia salutaris is a traditional medicinal plant used in malaria treatment by Zulu traditional healers. Materials and methods: The W. salutaris stem-bark was extracted with dichloromethane and the compound was isolated through column chromatography. The compound was identified and characterized by spectroscopic analysis (1H NMR, 13C NMR, IR and MS) and the structure was also confirmed by x-ray crystallography. The anti-plasmodial activity (in vitro) was studied on NF54 Plasmodium falciparum strain (CQS). Cytotoxicity was measured using the MTT assay on HEK239 and HEPG2 cell lines. Docking of Mukaadial acetate was conducted in AutoDock Vina. Structural modifications were conducted in UCSF Chimera and molecular interactions examined in LigPlot. Results: The compound, Mukaadial Acetate showed appreciable inhibition (IC50 0.44±0.10 µg/ml) of the parasite growth and cytotoxicity activity of 0.124±0.109 and 0.199±0.083 (µg/ml) on HEK293 and HEPG2 cells respectively. Molecular docking revealed that Mukaadial Acetate binds to the purine, pyrophosphate and ribose binding sites of the PfHGXPRT with an optimum binding conformation and forms hydrogen bond, steric and hydrophobic interactions with the residues inhabiting the respective binding sites. Conclusion: It is apparent that W. salutaris contains components (including Mukaadial Acetate) that exhibit antimalarial activity. This study scientifically validates the use of this plant in folk medicine.

Keywords: plasmodium falciparum, molecular docking, antimalarial activity, PfHGXPRT, Warburgia salutaris, mukaadial acetate

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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: Majid Farsadrouh Rashti, Parisa Jahani, Amir Shafiee, Mehrdad Mofidi

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The dihydroxybenzene isomers, hydroquinone (HQ), catechol (CC) and resorcinol (RS) have been widely recognized as important environmental pollutants due to their toxicity and low degradability in the ecological environment. Speciation of HQ, CC and RS is very important for environmental analysis because they co-exist of these isomers in environmental samples and are too difficult to degrade as an environmental contaminant with high toxicity. There are many analytical methods have been reported for detecting these isomers, such as spectrophotometry, fluorescence, High-performance liquid chromatography (HPLC) and electrochemical methods. These methods have attractive advantages such as simple and fast response, low maintenance costs, wide linear analysis range, high efficiency, excellent selectivity and high sensitivity. A novel modified glassy carbon electrode (GCE) with Pd@ Cu/CNTs core−shell nanowires for the simultaneous determination of hydroquinone (HQ), catechol (CC) and resorcinol (RS) is described. A detailed investigation by field emission scanning electron microscopy and electrochemistry was performed in order to elucidate the preparation process and properties of the GCE/ Pd/CuNWs-CNTs. The electrochemical response characteristic of the modified GPE/LFOR toward HQ, CC and RS were investigated by cyclic voltammetry, differential pulse voltammetry (DPV) and Chronoamperometry. Under optimum conditions, the calibrations curves were linear up to 228 µM for each with detection limits of 0.4, 0.6 and 0.8 µM for HQ, CC and RS, respectively. The diffusion coefficient for the oxidation of HQ, CC and RS at the modified electrode was calculated as 6.5×10⁻⁵, 1.6 ×10⁻⁵ and 8.5 ×10⁻⁵ cm² s⁻¹, respectively. DPV was used for the simultaneous determination of HQ, CC and RS at the modified electrode and the relative standard deviations were 2.1%, 1.9% and 1.7% for HQ, CC and RS, respectively. Moreover, GCE/Pd/CuNWs-CNTs was successfully used for determination of HQ, CC and RS in real samples.

Keywords: dihydroxybenzene isomers, galvanized copper nanowires, electrochemical sensor, Palladium, speciation

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Authors: Samuel K. Degife, Kamal K. Pant, Sapna Jain

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The world´s population and industrialization of countries continued to grow in an alarming rate irrespective of the security for food, energy supply, and pure water availability. As a result, the global energy consumption is observed to increase significantly. Fossil energy resources that mainly comprised of crude oil, coal, and natural gas have been used by mankind as the main energy source for almost two centuries. However, sufficient evidences are revealing that the consumption of fossil resource as transportation fuel emits environmental pollutants such as CO2, NOx, and SOx. These resources are dwindling rapidly besides enormous amount of problems associated such as fluctuation of oil price and instability of oil-rich regions. Biomass is a promising renewable energy candidate to replace fossil-based transportation fuel and chemical production. The present study aims at valorization of hexose sugars (glucose and fructose) using zeolite based catalysts in imidazolium based ionic liquid (1-butyl-3-methylimidazolium chloride, [BMIM] Cl) reaction media. The catalytic effect chromium impregnated H-ZSM-5 (Cr/H-ZSM-5) was studied for dehydration of hexose sugars. The wet impregnation method was used to prepare Cr/H-ZSM-5 catalyst. The characterization of the prepared catalyst was performed using techniques such as Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction analysis (XRD), Temperature-programmed desorption of ammonia (NH3-TPD) and BET-surface area analysis. The dehydration product, 5-hydroxymethylfurfural (5-HMF), was analyzed using high-performance liquid chromatography (HPLC). Cr/H-ZSM-5 was effective in dehydrating fructose with 87% conversion and 55% yield 5-HMF at 180 oC for 30 min of reaction time compared with H-ZSM-5 catalyst which yielded only 31% of 5-HMF at identical reaction condition.

Keywords: chromium, hexose, ionic liquid, , zeolite

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Authors: Abiola A. Ojesanmi, Eric O. Amonsou

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The formation of soluble complexes is usually within a narrow pH range characterized by weak interactions. Moreover, the rigid conformation of globular proteins restricts the number of charged groups capable of interacting with polysaccharides, thereby limiting food applications. Hence, this study investigated the impact of tryptic-limited hydrolysis on the formation of Bambara protein-gum arabic soluble complexes formation. The electrostatic interactions were monitored through turbidimetry analysis. The Bambara protein hydrolysates at a specified degree of hydrolysis, and DHs (2, 5, and 7.5) were characterized using size exclusion chromatography, zeta potential, surface hydrophobicity, and intrinsic fluorescence. The stability of the complexes was investigated using differential scanning calorimetry and rheometry. The limited tryptic hydrolysis significantly widened the pH range of the formation of soluble complexes, with DH 5 having a wider range (pH 7.0 - 4.3) compared to DH 2 and DH 7.5, while there was no notable difference in the optimum complexation pH of the insoluble complexes. Larger peptides (140, 118 kDa) were detected in DH 2 relative to 144, 70, and 61 kDa in DH 5, which were larger than 140, 118, 48, and 32 kDa in DH 7. 5. An increase in net negative charge (- 30 Mv for DH 7.5) and a slight shift in the net neutrality (from pH 4.9 to 4.3) of the hydrolysates were observed which consequently impacted the electrostatic interaction with gum arabic. There was exposure of the hydrophobic amino acids up to 4-fold in comparison with the isolate and a red shift in maximum fluorescence wavelength in DH dependent manner following the hydrolysis. The denaturation temperature of the soluble complex from the hydrolysates shifted to higher values, having DH 5 with the maximum temperature (94.24 °C). A highly interconnected gel-like soluble complex network was formed having DH 5 with a better structure relative to DH 2 and 7.5. The study showed the use of limited tryptic hydrolysis at DH 5 as an effective approach to modify Bambara protein and provided a more stable and wider pH range of formation for soluble complex, thereby enhancing the food application.

Keywords: Bambara groundnut, gum arabic, interaction, soluble complex

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Authors: Jasneet Kaur, Swapandeep Singh Chimni

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The unfavorable use of metal-based catalysts that are often extortionate and toxic can be overcome by using small organic molecules known as organocatalysts. A variety of small organic molecules, including Brønsted/Lewis bases and acids, based on sulfonic acids, phosphoric acids, amines, phosphines or carbenes, Cinchona alkaloids, have been used as organocatalysts. One of the key reasons for using organocatalysis is their ability to be effectively removed from the final product in comparison to the metallic counterparts, which are exceedingly difficult to remove. The present investigation seeks to explore the catalytic nature of Cinchona alkaloids as an organocatalyst for enantioselective synthesis of 3-substituted-3-aminooxindole, which is known to exhibit a variety of biological activities and pharmacological activities. In this context, an organocatalytic asymmetric route for the synthesis of 3-aminooxindoles via reaction of isatin imine with α-acetoxy-β-ketoesters has been developed. The bifunctional Cinchona derived thiourea catalyzed the reaction of α-acetoxy-β-ketoesters derivatives with isatin imine to afford 3-substituted-aminooxindole derivatives in up to 93% yield, 95% enantiomeric excess and >20:1 diastereomeric ratio. The reaction was performed at room temperature for two hours using 10 mol% of catalyst, in the presence of 4Å molecular sieves in tetrahydrofuran as a solvent at ambient temperature. After the completion of the reaction, the pure product could be easily separated by using column chromatography using hexane and ethyl acetate as solvents. In conclusion, the catalytic potential of Cinchona derived chiral thiourea-tertiary amine catalyst was explored for an organocatalytic enantioselective Mannich reaction of β-ketoester derivatives with various isatin imine derivatives under mild conditions.

Keywords: asymmetric synthesis, aminooxindoles, enantioselective, isatin imine

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Authors: Thavasimuthu Citarasu, Mariavincent Michaelbabu, Vikram Vakharia

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The rhizome of Java grass, Cyperus rotundus was extracted different organic polar and non-polar solvents and performed the in vitro antiviral and immunostimulant activities against White Spot Syndrome Virus (WSSV) and Vibrio harveyi respectively. Based on the initial screening the ethyl acetate extract of C. rotundus was strong activities and further it was purified through silica column chromatography and the fractions were screened again for antiviral and immunostimulant activity. Among the different fractions screened against the WSSV and V. harveyi, the fractions, F-III to FV had strong activities. In order to study the in vivo influence of C. rotundus, the fractions (F-III to FV) were pooled and delivered to the F. indicus through artificial feed for 30 days. After the feeding trail the experimental and control diet fed F. indicus were challenged with virulent WSSV and studied the survival, molecular diagnosis, biochemical, haematological and immunological parameters. Surprisingly, the pooled fractions (F-III to FV) incorporated diets helped to significantly (P < 0.01) suppressed viral multiplication, showed significant (P < 0.01) differences in protein and glucose levels, improved total haemocyte count (THC), coagulase activity, significantly increased (P < =0.001) prophenol oxidase and intracellular superoxide anion production compared to the control shrimps. Based on the results, C. rotundus extracts effectively suppressed WSSV multiplication and improve the immune system in F. indicus against WSSV infection and this knowledge will helps to develop novel drugs from C. rotundus against WSSV.

Keywords: antiviral drugs, cyperus rotundus, fenneropenaeus indicus, WSSV

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Authors: Ogbuagu Josephat Okechukwu, Okeke Abuchi Princewill, Arinze Rosemary Uche, Tabugbo Ifeyinwa Blessing, Ogbuagu Adaora Stellamaris

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Phthalates or Phthalate esters (PAEs), categorized as an endocrine disruptor and persistent organic pollutants, are known for their environmental contamination and toxicological effects. In this study, the concentration of selected phthalates was determined across the sampling site to investigate their occurrence and the ecological and health risk assessment they pose to the environment. Water and sediment samples were collected following standard procedures. Solid phase and ultrasonic methods were used to extract seven different PAEs, which were analyzed by Gas Chromatography with Mass Detector (GCMS). The analytical average recovery was found to be within the range of 83.4% ± 2.3%. The results showed that PAEs were detected in six out of seven samples with a high percentage of detection rate in water. Di-n-butyl phthalate (DPB) and disobutyl phthalates (DiBP) showed a greater detection rate compared to other PAE monomers. The concentration of PEs was found to be higher in sediment samples compared to water samples due to the fact that sediments serve as a sink for most persistent organic pollutants. The concentrations of PAEs in water samples and sediments ranged from 0.00 to 0.23 mg/kg and 0.00 to 0.028 mg/l, respectively. Ecological risk assessment using the risk quotient method (RQ) reveals that the estimated environmental risk caused by phthalates lies within the moderate level as RQ ranges from 0.1 to 1.0, whereas the health risk assessment caused by phthalates on estimating the average daily dose reveals that the ingestion of phthalates was found to be approaching permissible limit which can cause serious carcinogenic occurrence in the human system with time due to excess accumulation.

Keywords: phthalates, endocrine disruptor, risk assessment, ecological risk, health risk

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Authors: A. Banerjee, C. Grazon, B. Nadal, T. Pons, Y. Krishnan, B. Dubertret

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Quantum Dots (QDs) have emerged as novel fluorescent probes for biomedical applications. The photophysical properties of QDs such as broad absorption, narrow emission spectrum, reduced blinking, and enhanced photostability make them advantageous over organic fluorophores. However, for some biological applications, QDs need to be first targeted to specific intracellular locations. It parallel, base pairing properties and biocompatibility of DNA has been extensively used for biosensing, targetting and intracellular delivery of numerous bioactive agents. The combination of the photophysical properties of QDs and targettability of DNA has yielded fluorescent, stable and targetable nanosensors. QD-DNA conjugates have used in drug delivery, siRNA, intracellular pH sensing and several other applications; and continue to be an active area of research. In this project, a novel method to synthesise QD-DNA conjugates and their applications in bioimaging are investigated. QDs are first solubilized in water using a thiol based amphiphilic co-polymer and, then conjugated to amine functionalized DNA using a heterobifunctional linker. The conjugates are purified by size exclusion chromatography and characterized by UV-Vis absorption and fluorescence spectroscopy, electrophoresis and microscopy. Parameters that influence the conjugation yield such as reducing agents, the excess of salt and pH have been investigated in detail. In optimized reaction conditions, up to 12 single-stranded DNA (15 mer length) can be conjugated per QD. After conjugation, the QDs retain their colloidal stability and high quantum yield; and the DNA is available for hybridization. The reaction has also been successfully tested on QDs emitting different colors and on Gold nanoparticles and therefore highly generalizable. After extensive characterization and robust synthesis of QD-DNA conjugates in vitro, the physical properties of these conjugates in cellular milieu are being invistigated. Modification of QD surface with DNA appears to remarkably alter the fate of QD inside cells and can have potential implications in therapeutic applications.

Keywords: bioimaging, cellular targeting, drug delivery, photostability

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Authors: Nan Xu

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In airlines, the crew scheduling problem is usually decomposed into two stages: crew pairing and crew rostering. In the crew pairing stage, pairings are generated such that each flight is covered by exactly one pairing and the overall cost is minimized. In the crew rostering stage, the pairings generated in the crew pairing stage are combined with off days, training and other breaks to create individual work schedules. The paper focuses on cabin crew rostering problem, which is challenging due to the extremely large size and the complex working rules involved. In our approach, the objective of rostering consists of two major components. The first is to minimize the number of unassigned pairings and the second is to ensure the fairness to crew members. There are two measures of fairness to crew members, the number of overnight duties and the total fly-hour over a given period. Pairings should be assigned to each crew member so that their actual overnight duties and fly hours are as close to the expected average as possible. Deviations from the expected average are penalized in the objective function. Since several small deviations are preferred than a large deviation, the penalization is quadratic. Our model of the airline crew rostering problem is based on column generation. The problem is decomposed into a master problem and subproblems. The mater problem is modeled as a set partition problem and exactly one roster for each crew is picked up such that the pairings are covered. The restricted linear master problem (RLMP) is considered. The current subproblem tries to find columns with negative reduced costs and add them to the RLMP for the next iteration. When no column with negative reduced cost can be found or a stop criteria is met, the procedure ends. The subproblem is to generate feasible crew rosters for each crew member. A separate acyclic weighted graph is constructed for each crew member and the subproblem is modeled as resource constrained shortest path problems in the graph. Labeling algorithm is used to solve it. Since the penalization is quadratic, a method to deal with non-additive shortest path problem using labeling algorithm is proposed and corresponding domination condition is defined. The major contribution of our model is: 1) We propose a method to deal with non-additive shortest path problem; 2) Operation to allow relaxing some soft rules is allowed in our algorithm, which can improve the coverage rate; 3) Multi-thread techniques are used to improve the efficiency of the algorithm when generating Line-of-Work for crew members. Here a column generation based algorithm for the airline cabin crew rostering problem is proposed. The objective is to assign a personalized roster to crew member which minimize the number of unassigned pairings and ensure the fairness to crew members. The algorithm we propose in this paper has been put into production in a major airline in China and numerical experiments show that it has a good performance.

Keywords: aircrew rostering, aircrew scheduling, column generation, SPPRC

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Authors: O. Ubani, H. I. Atagana, M. S. Thantsha

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This study was conducted to measure the reduction in polycyclic aromatic hydrocarbons (PAHs) content in oil sludge by co-composting the sludge with pig, cow, horse and poultry manures under laboratory conditions. Four kilograms of soil spiked with 800 g of oil sludge was co-composted differently with each manure in a ratio of 2:1 (w/w) spiked soil:manure and wood-chips in a ratio of 2:1 (w/v) spiked soil:wood-chips. Control was set up similar as the one above but without manure. Mixtures were incubated for 10 months at room temperature. Compost piles were turned weekly and moisture level was maintained at between 50% and 70%. Moisture level, pH, temperature, CO2 evolution and oxygen consumption were measured monthly and the ash content at the end of experimentation. Bacteria capable of utilizing PAHs were isolated, purified and characterized by molecular techniques using polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), amplification of the 16S rDNA gene using the specific primers (16S-P1 PCR and 16S-P2 PCR) and the amplicons were sequenced. Extent of reduction of PAHs was measured using automated soxhlet extractor with dichloromethane as the extraction solvent coupled with gas chromatography/mass spectrometry (GC/MS). Temperature did not exceed 27.5O°C in all compost heaps, pH ranged from 5.5 to 7.8 and CO2 evolution was highest in poultry manure at 18.78 µg/dwt/day. Microbial growth and activities were enhanced. Bacteria identified were Bacillus, Arthrobacter and Staphylococcus species. Results from PAH measurements showed reduction between 77 and 99%. The results from the control experiments may be because it was invaded by fungi. Co-composting of spiked soils with animal manures enhanced the reduction in PAHs. Interestingly, all bacteria isolated and identified in this study were present in all treatments, including the control.

Keywords: bioremediation, co-composting, oil refinery sludge, PAHs, bacteria spp, animal manures, molecular techniques

Procedia PDF Downloads 475