Search results for: colorimetric gold nanoparticle assay

Authors: Chen-Chang Lee, Po-Chou Chen, Jo-Chi Jao, Chun-Chung Lui, Leung-Chit Tsang, Lain-Chyr Hwang

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Fetal Magnetic Resonance Imaging (MRI) is a challenge task because the fetal movements could cause motion artifact in MR images. The remedy to overcome this problem is to use fast scanning pulse sequences. The Single-Shot Fast Spin-Echo (SSFSE) T2-weighted imaging technique is routinely performed and often used as a gold standard in clinical examinations. Fast spoiled gradient-echo (FSPGR) T1-Weighted Imaging (T1WI) is often used to identify fat, calcification and hemorrhage. Fast Imaging Employing Steady-State Acquisition (FIESTA) is commonly used to identify fetal structures as well as the heart and vessels. The contrast of FIESTA image is related to T1/T2 and is different from that of SSFSE. The advantages and disadvantages of these two scanning sequences for fetal imaging have not been clearly demonstrated yet. This study aimed to compare these three rapid MRI techniques (SSFSE, FIESTA, and FSPGR) for fetal MRI examinations. The image qualities and influencing factors among these three techniques were explored. A 1.5T GE Discovery 450 clinical MR scanner with an eight-channel high-resolution abdominal coil was used in this study. Twenty-five pregnant women were recruited to enroll fetal MRI examination with SSFSE, FIESTA and FSPGR scanning. Multi-oriented and multi-slice images were acquired. Afterwards, MR images were interpreted and scored by two senior radiologists. The results showed that both SSFSE and T2W-FIESTA can provide good image quality among these three rapid imaging techniques. Vessel signals on FIESTA images are higher than those on SSFSE images. The Specific Absorption Rate (SAR) of FIESTA is lower than that of the others two techniques, but it is prone to cause banding artifacts. FSPGR-T1WI renders lower Signal-to-Noise Ratio (SNR) because it severely suffers from the impact of maternal and fetal movements. The scan times for these three scanning sequences were 25 sec (T2W-SSFSE), 20 sec (FIESTA) and 18 sec (FSPGR). In conclusion, all these three rapid MR scanning sequences can produce high contrast and high spatial resolution images. The scan time can be shortened by incorporating parallel imaging techniques so that the motion artifacts caused by fetal movements can be reduced. Having good understanding of the characteristics of these three rapid MRI techniques is helpful for technologists to obtain reproducible fetal anatomy images with high quality for prenatal diagnosis.

Keywords: fetal MRI, FIESTA, FSPGR, motion artifact, SSFSE

Procedia PDF Downloads 508

Authors: Lukman Ahmad Jamil, Heather M. Wallace

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Introduction: Numerous epidemiological studies have shown a positive as well as negative association and no association in some cases between chronic use of calcium channel blockers and the increased risk of developing cancer. However, these associations were enmeshed with controversies in the absence of laboratory based studies to back up those claims. Aim: The aim of this study was to determine in mechanistic terms the association between the long-term administration of nifedipine and diltiazem and increased risk of developing cancer using the human embryonic kidney (HEK293) cell line. Methods: Cell counting using the Trypan blue dye exclusion and 3-4, 5-Dimethylthiazol-2-yl-2, 5-diphenyl-tetrazolium bromide (MTT) assays were used to investigate the effect of nifedipine and diltiazem on the growth pattern of HEK293 cells. Protein assay using modified Lowry method and analysis of intracellular polyamines concentration using Liquid Chromatography – Tandem Mass Spectrometry (LC-MS) were performed to ascertain the mechanism through which chronic use of nifedipine increases the risk of developing cancer. Results: Both nifedipine and diltiazem significantly increased the proliferation of HEK293 cells dose and time dependently. This proliferative effect after 24, 48 and 72-hour incubation period was observed at 0.78, 1.56 and 25 µM for nifedipine and 0.39, 1.56 and 25 µM for diltiazem, respectively. The increased proliferation of the cells was found to be statistically significantly (p<0.05). Furthermore, the increased proliferation of the cells induced by nifedipine was associated with the increase in the protein content and elevated intracellular polyamines concentration level. Conclusion: The chronic use of nifedipine is associated with increased proliferation of cells with concomitant elevation of polyamines concentration and elevated polyamine levels have been implicated in many malignant transformations and hence, these provide a possible explanation on the link between long term use of nifedipine and development of some human cancers. Further studies are needed to evaluate the cause of this association.

Keywords: cancer, nifedipine, polyamine, proliferation

Procedia PDF Downloads 182

Authors: Aliro Villacorta, Laura Rubio, Mohamed Alaraby, Montserrat López Mesas, Victor Fuentes-Cebrian, Oscar H. Moriones, Ricard Marcos, Alba Hernández.

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Micro and nano plastics (MNPLs) are emergent environmental pollutants requiring urgent information on their potential risks to human health. One of the problems associated with the evaluation of their undesirable effects is the lack of real samples matching those resulting from the environmental degradation of plastic wastes. To such end, we propose an easy method to obtain polyethylene terephthalate nano plastics from water plastic bottles (PET-NPLs) but, in principle, applicable to any other plastic goods sources. An extensive characterization indicates that the proposed process produces uniform samples of PET-NPLs of around 100 nm, as determined by using a multi-angle and dynamic light scattering methodology. An important point to be highlighted is that to avoid the metal contamination resulting from methods using metal blades/burrs for milling, trituration, or sanding, we propose to use diamond burrs to produce metal-free samples. To visualize the toxicological profile of the produced PET-NPLs, we have evaluated their ability to be internalized by cells, their cytotoxicity, and their ability to induce oxidative stress and induce DNA damage. In this preliminary approach, we have detected their cellular uptake, but without the induction of significant biological effects. Thus, no relevant increases in toxicity, reactive oxygen species (ROS) induction, or DNA damage -as detected with the comet assay- have been observed. The use of real samples, as produced in this study, will generate relevant data in the discussion about the potential health risks associated with MNPLs exposures.

Keywords: nanoplastics, polyethylene terephthalate, physicochemical characterization, cell uptake, cytotoxicity

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Authors: Younus Mohammad, Anita B. Fallah, Ben J. Boyd, Shakila B. Rizwan

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N-acylethanolamines (NAEs) are endogenous lipids, which have neuromodulatory properties. NAEs have shown neuroprotective properties in various neurodegenerative diseases including Alzheimer's disease, Parkinson's disease and ischemic stroke. However, NAEs are eliminated rapidly in vivo by enzymatic hydrolysis. We propose to encapsulate NAEs in liquid crystalline nanoparticles (cubosomes) to increase their biological half-life and explore their therapeutic potential. Recently, we have reported the co-formulation and nanostructural characterization of cubosomes containing the NAE, oleoylethanolamide and a synthetic cubosome forming lipid phytantriol. Here, we report on the formulation of cubosomes with the NAE, linoleoylethanolamide (LEA) as the core cubosome forming lipid. LEA-cubosomes were formulated in the presence of three different steric stabilisers: two brain targeting ligands, Tween 80 and Pluronic P188 and a control, Pluronic F127. Size, morphology and internal structure of formulations were characterized by dynamic light scattering (DLS), cryogenic transmission electron microscopy (Cryo–TEM) and small angle X–ray scattering (SAXS), respectively. Chemical stability of LEA in formulations was investigated using high-performance liquid chromatography (HPLC). Cytotoxicity of formulations towards human cerebral microvascular endothelial cell line (hCMEC/D3) was also investigated using an MTT (3-[4, 5- dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide) assay. All cubosome formulations had mean particle size of less than 250 nm and were uniformly distributed with polydispersity indices less than 0.2. Cubosomes produced had a bicontinuous cubic internal structure with an Im3m space group but different lattice parameters, indicating the different modes of interaction between the stabilisers and LEA. LEA in formulations was found to be chemically stable. At concentrations of up to 20 µg/mL LEA in the presence of all the stabilisers, greater than 80% cell viability was observed.

Keywords: blood-brain barrier, cubosomes, linoleoyl ethanolamide, N-acylethanolamines (NAEs)

Procedia PDF Downloads 193

Authors: Lin Feng, Ling-Ling E., Hong-Chen Liu

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The development of chemoresistance in patients represents a major challenge in cancer treatment. Lactate dehydrogenase‑A (LDHA) is one of the principle isoforms of LDH that is expressed in breast tissue, controlling the conversion of pyruvate to lactate and also playing a significant role in the metabolism of glucose. The aim of this study was to identify whether LDHA was involved in oral cancer cell resistance to Taxol and whether the downregulation of LDHA, as a result of cisplatin treatment, may overcome Taxol resistance in human oral squamous cells. The OECM‑1 oral epidermal carcinoma cell line was used, which has been widely used as a model of oral cancer in previous studies. The role of LDHA in Taxol and cisplatin resistance was investigated and the synergistic cytotoxicity of cisplatin and/or Taxol in oral squamous cells was analyzed. Cell viability was analyzed by MTT assay, LDHA expression was analyzed by western blot analysis and siRNA transfection was performed to knock down LDHA expression. The present study results showed that decreased levels of LDHA were responsible for the resistance of oral cancer cells to cisplatin (CDDP). CDDP treatments downregulated LDHA expression and lower levels of LDHA were detected in the CDDP‑resistant oral cancer cells compared with the CDDP‑sensitive cells. By contrast, the Taxol‑resistant cancer cells showed elevated LDHA expression levels. In addition, small interfering RNA‑knockdown of LDHA sensitized the cells to Taxol but desensitized them to CDDP treatment while exogenous expression of LDHA sensitized the cells to CDDP, but desensitized them to Taxol. The present study also revealed the synergistic cytotoxicity of CDDP and Taxol for killing oral cancer cells through the inhibition of LDHA. This study highlights LDHA as a novel therapeutic target for overcoming Taxol resistance in oral cancer patients using the combined treatments of Taxol and CDDP.

Keywords: cisplatin, Taxol, carcinoma, oral squamous cells

Procedia PDF Downloads 397

Authors: Khayra Zerrouki, Noureddine Djebli, Esra Eroglu, Afife Mat, Ozhan Gul

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Neurodegenerative diseases of the human brain comprise a variety of disorders that affect an increasing percentage of the population. Alzheimer’s disease (AD) is a complex, multifactorial, heterogeneous mental illness, which is characterized by an age-dependent loss of memory and an impairment of multiple cognitive functions, but this 10 last years it concerns the population most and most young. Hypericum perforatum has traditionally been used as an external anti-inflammatory and healing remedy for the treatment of swellings, wounds and burns, diseases of the alimentary tract and psychological disorders. It is currently of great interest due to new and important therapeutic applications. In this study, the chemical composition of methanolic extract of Hypericum perforatum (HPM) was analysed by using high performance liquid chromatography – diode array detector (HPLC-DAD). The in vitro antioxidant activity of HPM was evaluated by using several antioxidant tests. HSM exhibits inhibitory capacity against posphatidylcholine liposome peroxidation, induced with iron and ascorbic acid, scavenge DPPH and superoxide radicals and act as reductants. The cytotoxic activity of HSM was also determined by using MTT cell viability assay on HeLa and NRK-52E cell lines. The in vivo activity studies in Swiss mice were determined by using behavioral, memory tests and histological study. According to tests results HPM that may be relevant to the treatment of cognitive disorders. The results of chemical analysis showed a hight level of hyperforin and quercitin that had an important antioxidant activity proved in vitro with the DPPH, anti LPO and SOD; this antioxidant activity was confirmed in vivo after the non-toxic results by means of improvement in behavioral and memory than the reducing shrunken in pyramidal cells of mice brains.

Keywords: AlCl3, alzheimer, mice, neuroprotective, neurotoxicity, phytotherapy

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Authors: Jacob Schwartz, Maxim Durach, Aniruddha Mitra, Abbas Rashidi, Glen Sage, Atin Adhikari

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NIOSH (National Institute for Occupational Safety and Health) approved N95 respirators are commonly used by workers in construction sites where there is a large amount of dust being produced from sawing, grinding, blasting, welding, etc., both electrostatically charged and not. A significant portion of airborne particles in construction sites could be nanoparticles created beside coarse particles. The penetration of the particles through the masks may differ depending on the size and charge of the individual particle. In field experiments relevant to this current study, we found that nanoparticles of medium size ranges are penetrating more frequently than nanoparticles of smaller and larger sizes. For example, penetration percentages of nanoparticles of 11.5 – 27.4 nm into a sealed N95 respirator on a manikin head ranged from 0.59 to 6.59%, whereas nanoparticles of 36.5 – 86.6 nm ranged from 7.34 to 16.04%. The possible causes behind this increased penetration of mid-size nanoparticles through mask filters are not yet explored. The objective of this study is to identify causes behind this unusual behavior of mid-size nanoparticles. We have considered such physical factors as Boltzmann distribution of the particles in thermal equilibrium with the air, kinetic energy of the particles at impact on the mask, Stoke’s drag force, and electrostatic forces in the mask stopping the particles. When the particles collide with the mask, only the particles that have enough kinetic energy to overcome the energy loss due to the electrostatic forces and the Stokes’ drag in the mask can pass through the mask. To understand this process, the following assumptions were made: (1) the effect of Stoke’s drag depends on the particles’ velocity at entry into the mask; (2) the electrostatic force is proportional to the charge on the particles, which in turn is proportional to the surface area of the particles; (3) the general dependence on electrostatic charge and thickness means that for stronger electrostatic resistance in the masks and thicker the masks’ fiber layers the penetration of particles is reduced, which is a sensible conclusion. In sampling situations where one mask was soaked in alcohol eliminating electrostatic interaction the penetration was much larger in the mid-range than the same mask with electrostatic interaction. The smaller nanoparticles showed almost zero penetration most likely because of the small kinetic energy, while the larger sized nanoparticles showed almost negligible penetration most likely due to the interaction of the particle with its own drag force. If there is no electrostatic force the fraction for larger particles grows. But if the electrostatic force is added the fraction for larger particles goes down, so diminished penetration for larger particles should be due to increased electrostatic repulsion, may be due to increased surface area and therefore larger charge on average. We have also explored the effect of ambient temperature on nanoparticle penetrations and determined that the dependence of the penetration of particles on the temperature is weak in the range of temperatures in the measurements 37-42°C, since the factor changes in the range from 3.17 10-3K-1 to 3.22 10-3K-1.

Keywords: respiratory protection, industrial hygiene, aerosol, electrostatic force

Procedia PDF Downloads 182

Authors: Emine Aydin, Duygu Gocmen

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In this study, pumpkin flours (PFs) were used to replace wheat flour in cookie formulation at three different levels (10%, 20% and 30% w/w). For this purpose PFs produced by two different applications (with or without metabisulfite pre-treatment) and then dried in freeze dryer. Control sample included no PFs. The total phenolic contents of the cookies supplemented with PFs were higher than that of control and gradually increased in total phenolic contents of cookies with increasing PF supplementation levels. Phenolic content makes also significant contribution on nutritional excellence of the developed cookies. Pre-treatment with metabisulfite (MS) had a positive effect on free, bound and total phenolics of cookies which are supplemented with various levels of MS-PF. This is due to a protective effect of metabisulfite pretreatment for phenolic compounds in the pumpkin flour. Phenolic antioxidants may act and absorb in a different way in humans and thus their antioxidant and health effects will be changed accordingly. In the present study phenolics’ bioavailability of cookies was investigated in order to assess PF as sources of accessible phenolics. The content of bioaccessible phenolics and phenolic bioaccessibility of cookies supplemented with PFs had higher than those of control sample. Cookies enriched with 30% MS-PF had the highest bioaccessible phenolics (597.86 mg GAE 100g-1) and phenolic bioaccessibility (41.71%). MS application in PF production caused a significant increase in phenolic bioaccessibility of cookies. According to all assay (ABTS, CUPRAC, FRAP and DPPH), antioxidant activities of cookies with PFs higher than that of control cookie. It was also observed that the cookies supplemented with MS-PF had significantly higher antioxidant activities than those of cookies including PF. In presented study, antioxidative bioaccessibilities of cookies were also determined. The cookies with PFs had significantly (p ≤ 0.05) higher antioxidative bioaccessibilities than control ones. Increasing PFs levels enhanced antioxidative bioaccessibilities of cookies. As a result, PFs addition improved the nutritional and functional properties of cookie by causing increase in antioxidant activity, total phenolic content, bioaccessible phenolics and phenolic bioaccessibilities.

Keywords: phenolic compounds, antioxidant activity, dietary fiber, pumpkin, freeze drying, cookie

Procedia PDF Downloads 246

Authors: Godwin Ojochogu Adejo, Evelyn UnekwuOjo Adejo, Sunday UnenwOjo Adejo

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The mainstream contemporary approach to controlling the impact of diseases among poultry birds rely largely on curative measures through the administration of drugs to infected birds. Most times as observed in the deep liter poultry farming system, entire flocks including uninfected birds receive the treatment they do not need. As such, unguarded use of chemical drugs and antibiotics has led to wastage and accumulation of chemical residues in poultry products with associated health hazards to humans. However, wanton and frequent drug usage in poultry is avoidable if feeding and drinking equipment are designed to curb infection transmission among birds. Using toxicological assays as guide and with efficiency and simplicity in view, two newly field-tested and recently patented equipments called 'healthy liquid drinking trough (HDT)' and 'healthy feeding trough (HFT)' that systematically eliminate contamination of the feeding and drinking channels, thereby, curbing wide-spread infection and transmission of diseases in the (intensive) deep litter poultry farming system were designed. Upon combined usage, they automatically and drastically reduced both the amount and frequency of antibiotics use in poultry by over > 50%. Additionally, they conferred optimization of feed and water utilization/elimination of wastage by > 80%, reduced labour by > 70%, reduced production cost by about 15%, and reduced chemical residues in poultry meat or eggs by > 85%. These new and cheap technologies which require no energy input are likely to elevate safety of poultry products for consumers' health, increase marketability locally and for export, and increase output and profit especially among poultry farmers and poor people who keep poultry or inevitably utilize poultry products in developing countries.

Keywords: healthy, trough, toxicological, assay-guided, poultry

Procedia PDF Downloads 138

Authors: Jayeola A. O., Ayodele O. S., Olususi J. I.

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From field studies, six (6) lithological units were identified to be common around the study area, which includes quartzites, granites, granite gneiss, porphyritic granites, amphibolite and pegmatites. Petrographical analysis was done to establish the major mineral assemblages and accessory minerals present in selected rock samples, which represents the major rock types in the area. For the purpose of this study, twenty (20) pulverized rock samples were taken to the laboratory for geochemical analysis with their results used in the classification, as well as suggest the geochemical attributes of the rocks. Results from petrographical studies of the rocks under both plane and cross polarized lights revealed the major minerals identified under thin sections to include quartz, feldspar, biotite, hornblende, plagioclase and muscovite with opaque other accessory minerals, which include actinolite, spinel and myrmekite. Geochemical results obtained and interpreted using various geochemical plots or discrimination plots all classified the rocks in the area as belonging to both the peralkaline metaluminous and peraluminous types. Results for the major oxides ratios produced for Na₂O/K₂O, Al₂O₃/Na₂O + CaO + K₂O and Na₂O + CaO + K₂O/Al₂O₃ show the excess of alumina, Al₂O₃ over the alkaline Na₂O +CaO +K₂O thus suggesting peraluminous rocks. While the excess of the alkali over the alumina suggests the peralkaline metaluminous rock type. The results of correlation coefficient show a perfect strong positive correlation, which shows that they are of same geogenic sources, while negative correlation coefficient values indicate a perfect weak negative correlation, suggesting that they are of heterogeneous geogenic sources. From factor analysis, five component groups were identified as Group 1 consists of Ag-Cr-Ni elemental associations suggesting Ag, Cr, and Ni mineralization, predicting the possibility of sulphide mineralization. in the study area. Group ll and lll consist of As-Ni-Hg-Fe-Sn-Co-Pb-Hg element association, which are pathfinder elements to the mineralization of gold. Group 1V and V consist of Cd-Cu-Ag-Co-Zn, which concentrations are significant to elemental associations and mineralization. In conclusion, from the potassium radiometric anomaly map produced, the eastern section (northeastern and southeastern) is observed to be the hot spot and mineralization zone for the study area.

Keywords: petrography, Ila Orangun, petrochemistry, pegmatites, peraluminous

Procedia PDF Downloads 43

Authors: Gary Pattemore, Michael Edgar, Andrew Job, Marina Auad, Kathryn Job

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As critical commodities become more scarce, significant time and resources have been used to better understand complicated ore bodies and extract their full potential. These challenging ore bodies provide several pain points for geologists and engineers to overcome, poor handling of these issues flows downs stream to the processing plant affecting throughput rates and recovery. Many open cut mines utilise blast hole drilling to extract additional information to feed back into the modelling process. This method requires samples to be collected during or after blast hole drilling. Samples are then sent for assay with turnaround times varying from 1 to 12 days. This method is time consuming, costly, requires human exposure on the bench and collects elemental data only. To address this challenge, research has been undertaken to utilise hyperspectral imaging across a broad spectrum to scan samples, collars or take down hole measurements for minerals and moisture content and grade abundances. Automation of this process using unmanned vehicles and on-board processing reduces human in pit exposure to ensure ongoing safety. On-board processing allows data to be integrated into modelling workflows with immediacy. The preliminary results demonstrate numerous direct and indirect benefits from this new technology, including rapid and accurate grade estimates, moisture content and mineralogy. These benefits allow for faster geo modelling updates, better informed mine scheduling and improved downstream blending and processing practices. The paper presents recommendations for implementation of the technology in open cut mining environments.

Keywords: grade control, hyperspectral scanning, artificial intelligence, autonomous mining, machine learning

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Authors: Maria Manzoor, Iram Gul, Muhammad Arshad

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Bacterial strains were isolated from contaminated soil collected from Rawalpindi and Islamabad. The strains were investigated for lead resistance and their effect on Pb solubility and PGPR activity. Incubation experiments were carried for inoculated and unoculated soil containing different levels of Pb. Results revealed that few stains (BTM-4, BTM-11, BTM-14) were able to tolerate Pb up to 600 mg L-1, whereas five strains (BTM-3, BTM-6, BTM-10, BTM-21 and BTM-24) showed significant increase in solubility of Pb when compared to all other strains and control. The CaCl2 extractable Pb was increased by 13.6, 6.8, 4.4 and 2.4 folds compared to un-inoculated control soil at increased soil Pb concentration (500, 1000, 1500 and 200 mg kg-1, respectively). The selected bacterial strains (11) were further investigated for plant growth promotion activity through PGPR assays including. Germination and root elongation assays were also conducted under elevated metal concentration in controlled conditions to elucidate the effects of microbial strains upon plant growth and development. The results showed that all the strains tested in this study, produced significantly varying concentrations of IAA, siderophores and gibberellic acid along with ability to phosphorus solubilization index (PSI). The results of germination and root elongation assay further confirmed the beneficial role of the microbial strains in elevating metal stress through PGPR activity. Among all tested strains, BTM-10 significantly improved plant growth. 1.3 and 2.7 folds increase in root and shoot length was observed when compared to control. Which may be attributed to presence of important plant growth promoting enzymes (IAA 74.6 μg/ml; GA 19.23 μg/ml; Sidrophore units 49% and PSI 1.3 cm). The outcome of this study indicates that these Pb tolerant and solubilizing strains may have the potential for plant growth promotion under metal stress and can be used as mediator when coupled with heavy metal hyperaccumulator plants for phytoremediation of Pb contaminated soil.

Keywords: Pb resistant bacteria, Pb mobilizing bacteria, Phytoextraction of Pb, PGPR activity of bacteria

Procedia PDF Downloads 199

Authors: A. Khan, I. Khan, M. Younus, S. E. Haque, U. Waheed, H. Neubauer, A. A. Anjum, S. A. Muhammad, A. Idrees T. Abbas, S. Raza, M. A. Ali, M. Farooq, M. Mahmood, A. Hussain, H. Danish, U. Tayyab, M. Zafar, M. Aslam.

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Brucellosis is one of the major problems of milk producing animals in our country which deteriorate the health of livestock. It is a disease of zoonotic significance which is capable of producing disease in humans leading to infertility, orchitis, abortions, and synovitis. In this particular study, milk and serum samples of cattle and buffalo (n=402) were collected from different districts of Punjab including Narowal, Gujranwala and Gujrat. Milk samples were analyzed by Milk Ring Test (MRT), while serum samples were tested through Rose Bengal Plate agglutination Test (RBPT) and Indirect Enzyme Linked Immunosorbant Assay (i-ELISA). The sample tested with MRT were 9.5% positive, including cattle 9.6% and buffalo 9.3%. While using the RBPT test for the detection of serum samples and for screening purpose it was observed that 16.4% animals were seropositive, cattle were 18.8% and buffalo were 13.9% seropositive. The higher prevalence of brucellosis indicates the danger of the disease to human population. The serum samples positive by RBPT were further confirmed by the use of most specific and sensitive serological test known as i-ELISA. 11.4% animals were confirmed as seropositive by i-ELISA including cattle 13.5% seropositive and buffalo 9.3%. The results indicated high seroprevalence of brucellosis in cattle as compared to buffalos. Different risk factors were also studied to know the association between disease and their spread. Advanced age, larger herds, history of abortion and pregnancy of the animals is considered to be the important factors for the prevalence and spread of the hazardous zoonotic disease. It is a core issue of developing countries like Pakistan and has major public health impact.

Keywords: humans, bovines, infertility, orchitis, abortions, seroprevalence, brucellosis

Procedia PDF Downloads 468

Authors: L. Hoade, O. O. Onafowokan, K. Anderson, G. E. Bartlett, E. D. Fern, M. R. Norton, R. G. Middleton

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Aims: Femoroacetabular impingement (FAI) was first recognised as a potential driver for hip pain at the turn of the last millennium. While there is an increasing trend towards surgical management of FAI by arthroscopic means, open surgical hip dislocation and debridement (SHD) remains the Gold Standard of care in terms of reported outcome measures. (1) Long-term functional and survivorship outcomes of SHD as a treatment for FAI are yet to be sufficiently reported in the literature. This study sets out to help address this imbalance. Methods: We undertook a retrospective review of our institutional database for all patients who underwent SHD for FAI between January 2003 and December 2008. A total of 223 patients (241 hips) were identified and underwent a ten year review with a standardised radiograph and patient-reported outcome measures questionnaire. The primary outcome measure of interest was survivorship, defined as progression to total hip arthroplasty (THA). Negative predictive factors were analysed. Secondary outcome measures of interest were survivorship to further (non-arthroplasty) surgery, functional outcomes as reflected by patient reported outcome measure scores (PROMS) scores, and whether a learning curve could be identified. Results: The final cohort consisted of 131 females and 110 males, with a mean age of 34 years. There was an overall native hip joint survival rate of 85.4% at ten years. Those who underwent a THA were significantly older at initial surgery, had radiographic evidence of preoperative osteoarthritis and pre- and post-operative acetabular undercoverage. In those whom had not progressed to THA, the average Non-arthritic Hip Score and Oxford Hip Score at ten year follow-up were 72.3% and 36/48, respectively, and 84% still deemed their surgery worthwhile. A learning curve was found to exist that was predicated on case selection rather than surgical technique. Conclusion: This is only the second study to evaluate the long-term outcomes (beyond ten years) of SHD for FAI and the first outside the originating centre. Our results suggest that, with correct patient selection, this remains an operation with worthwhile outcomes at ten years. How the results of open surgery compared to those of arthroscopy remains to be answered. While these results precede the advent of collison software modelling tools, this data helps set a benchmark for future comparison of other techniques effectiveness at the ten year mark.

Keywords: femoroacetabular impingement, hip pain, surgical hip dislocation, hip debridement

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Authors: Tatjana Jurić, Bojana Blagojević, Denis Uka, Ružica Ždero Pavlović, Boris M. Popović

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Natural deep eutectic solvents (NADES) represent a class of modern systems that have been developed as a green alternative to toxic organic solvents, which are commonly used as extraction media. It has been considered that hydrogen bonding is the main interaction leading to the formation of NADES. The aim of this study was phytochemical characterization and determination of the antioxidant and antibacterial activity of Mentha piperita leaf extracts obtained by six choline chloride-based NADES. NADES were prepared by mixing choline chloride with different hydrogen bond donors in 1:1 molar ratio following the addition of 30% (w/w) water. The mixtures were then heated (60 °C) and stirred (650 rpm) until the clear homogenous liquids were obtained. The Mentha piperita extracts were prepared by mixing 75 mg of peppermint leaves with 1 mL of NADES following by the heating and stirring (60 °C, 650 rpm) within 30 min. The content of six phenolics in extracts was determined using HPLC-PDA. The dominant compounds presented in peppermint leaves - rosmarinic acid and luteolin 7-O-glucoside, were extracted by NADES at a similar level as 70% ethanol. The microdilution method was applied to test the antibacterial activity of extracts. Compared with 70% ethanol, all NADES systems showed higher antibacterial activity towards Pseudomonas aeruginosa (Gram -), Staphylococcus aureus (Gram +), Escherichia coli (Gram -), and Salmonella enterica (Gram -), especially NADES containing organic acids. The majority of NADES extracts showed a better ability to neutralize DPPH radical than conventional solvent and similar ability to reduce Fe3+ to Fe2+ ions in FRAP assay. The obtained results introduce NADES systems as the novel, sustainable, and low-cost solvents with a variety of applications.

Keywords: antibacterial activity, antioxidant activity, green extraction, natural deep eutectic solvents, polyphenols

Procedia PDF Downloads 168

Authors: B. Makoudi, R. Ghanimi, A. Bargaz, M. Mouradi, M. Farissi, A. Kabbaj, J. J. Drevon, C. Ghoulam

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Legume species are able to establish a nitrogen fixing symbiosis with soil rhizobia that allows them, when it operates normally, to ensure their necessary nitrogen nutrition. This biological process needs high phosphorus (P) supply and consequently it is limited under low phosphorus availability. To overcome this constraint, legume-rhizobia symbiosis develops many mechanisms to increase P availability in the rhizosphere and also the efficiency of P fertilizers. The objectives of our research works are to understand the physiological and biochemical mechanisms implemented by legume-rhizobia symbiosis to increase its P use efficiency (PUE) in order to select legume genotypes-rhizobia strains combination more performing for BNF under P deficiency. Our studies were carried out on two grain legume species, common bean (Phaseolus vulgaris) and faba bean (Vicia faba) tested in farmers’ fields and in experimental station fewer than two soil phosphorus levels. Under field conditions, the P deficiency caused a significant decrease of Plant and nodule biomasses in all of the tested varieties with a difference between them. This P limitation increased the contents of available P in the rhizospheric soils that was positively correlated with the increase of phosphatases activities in the nodules and the rhizospheric soil. Some legume genotypes showed a significant increase of their P use efficiency under P deficiency. The P solubilization test showed that some rhizobia strains isolated from Haouz region presented an important capacity to grow on solid and liquid media with tricalcium phosphate as the only P source and their P solubilizing activity was confirmed by the assay of the released P in the liquid medium. Also, this P solubilizing activity was correlated with medium acidification and the excretion of acid phosphatases and phytases in the medium. Thus, we concluded that medium acidification and excretion of phosphatases in the rhizosphere are the prominent reactions for legume-rhizobia symbiosis to improve its P nutrition.

Keywords: legume, phosphorus deficiency, rhizobia, rhizospheric soil

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Authors: John Onolame Unuofin, Uchechukuw U. Nwodo, Anthony I. Okoh

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Laccase is constantly gaining status as important biocatalyst in biotechnology. The illimitable potential of its industrial applications and the corresponding aggressive need for phenomenal volumes of extracellularly secreted laccases have called for its interminable production from sources which are able to meet this demand within a relatively short period of time, preferably bacteria. In response to this call, this study was designed to source for laccase-producing bacteria from different environmental matrices. Three sampling environments were chosen such as wastewater treatment plants, University of Fort Hare vicinity and the Hogback woodland, all within the Eastern Cape, South Africa. Samples such as effluents, sediments, leaf litters, degrading wood and rock scrapings were selectively enriched with some model aromatic compounds and were further screened qualitatively and quantitatively on five phenolic substrates ABTS (2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), Guaiacol, 1-Naphthol, Potassium Ferric Cyanide and Syringaldazine). Basis for selection was their ability to elicit a colour change on at least three of the above mentioned agar based assay substrates. The choice isolates were further identified based on 16S rRNA molecular identification techniques. 33 isolates were screened out of the 40 representative distinct colonies during the qualitative plate screens, while quantitative screens selected out 11 bacterial isolates. They were, based on molecular identification, desginated as members of the genera Pseudomonas, Stenotrophomonas and Citrobacter of the gammaproteobacteria and Bordetalla and Achromobacter of the betaproteobacteria respectively. We therefore conclude based on our outcomes that we may have isolated efficient laccase-producing bacteria, which might be of beneficial significance in catalysis and biotechnology.

Keywords: beta proteobacteria, catalysis, gammaproteobacteria, laccase

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Authors: Aulanni’am Aulanni’am

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Incidence of Diabetes Mellitus (DM) progressively increasing it became a serious problem in Indonesia and it is a disease that government is priority to be addressed. The longer a person is suffering from diabetes the more likely to develop complications particularly diabetic patients who are not well maintained. Therefore, Incidence of Diabetes Mellitus needs to be done in the early diagnosis of pre-phase of the disease. In this pre-phase disease, already happening destruction of pancreatic beta cells and declining in beta cell function and the sign autoimmunity reactions associated with beta cell destruction. Type 1 DM is a multifactorial disease triggered by genetic and environmental factors, which leads to the destruction of pancreatic beta cells. Early marker of "beta cell autoreactivity" is the synthesis of autoantibodies against 65-kDa protein, which can be a molecule that can be detected early in the disease pathomechanism. The importance of early diagnosis of diabetic patients held in the phase of pre-disease is to determine the progression towards the onset of pancreatic beta cell destruction and take precautions. However, the price for this examination is very expensive ($ 150/ test), the anti-GAD65 abs examination cannot be carried out routinely in most or even in all laboratories in Indonesia. Therefore, production-based Rapid Test Recombinant Human Protein GAD65 with "Reverse Flow Immunchromatography Technique" in Indonesia is believed to reduce costs and improve the quality of care of patients with diabetes in Indonesia. Rapid Test Product innovation is very simple and suitable for screening and routine inspection of GAD65 autoantibodies. In the blood serum of patients with diabetes caused by autoimmunity, autoantibody-GAD65 is a major serologic marker to detect autoimmune reaction because their concentration level of stability.GAD65 autoantibodies can be found 10 years before clinical symptoms of diabetes. Early diagnosis is more focused to detect the presence autontibodi-GAD65 given specification and high sensitivity. Autoantibodies- GAD65 that circulates in the blood is a major indicator of the destruction of the islet cells of the pancreas. Results of research in collaboration with Biofarma has produced GAD65 autoantibodies based Rapid Test had conducted the soft launch of products and has been tested with the results of a sensitivity of 100 percent and a specificity between 90 and 96% compared with the gold standard (import product) which worked based on ELISA method.

Keywords: diabetes mellitus, GAD65 autoantibodies, rapid test, sensitivity, specificity

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Authors: Anuj Kumar, Kummara M. Rao, Sung S. Han

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Bone tissue engineering has emerged as a potentially alternative method for localized bone defects or diseases, congenital deformation, and surgical reconstruction. The designing and the fabrication of the ideal scaffold is a great challenge, in restoring of the damaged bone tissues via cell attachment, proliferation, and differentiation under three-dimensional (3D) biological micro-/nano-environment. In this case, hydrogel system composed of high hydrophilic 3D polymeric-network that is able to mimic some of the functional physical and chemical properties of the extracellular matrix (ECM) and possibly may provide a suitable 3D micro-/nano-environment (i.e., resemblance of native bone tissues). Thus, this proposed hydrogel system is highly permeable and facilitates the transport of the nutrients and metabolites. However, the use of hydrogels in bone tissue engineering is limited because of their low mechanical properties (toughness and stiffness) that continue to posing challenges in designing and fabrication of tough and stiff hydrogels along with improved bioactive properties. For this purpose, in our lab, polyacrylamide-based hybrid hydrogels were synthesized by involving sodium alginate, cellulose nanocrystals and silica-based glass using one-step free-radical polymerization. The results showed good in vitro apatite-forming ability (biomineralization) and improved mechanical properties (under compression in the form of strength and stiffness in both wet and dry conditions), and in vitro osteoblastic (MC3T3-E1 cells) cytocompatibility. For in vitro cytocompatibility assessment, both qualitative (attachment and spreading of cells using FESEM) and quantitative (cell viability and proliferation using MTT assay) analyses were performed. The obtained hybrid hydrogels may potentially be used in bone tissue engineering applications after establishment of in vivo characterization.

Keywords: bone tissue engineering, cellulose nanocrystals, hydrogels, polyacrylamide, sodium alginate

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Authors: Gordana Ćetković, Sanja Podunavac-Kuzmanović, Jasna Čanadanović-Brunet, Vesna Tumbas Šaponjac, Vanja Šeregelj, Jelena Vulić, Slađana Stajčić

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The color of food is one of the decisive factors for consumers. Potential toxicity of artificial food colorants has led to the consumers' preference for natural products over products with artificial colors. Natural pigments have many bioactive functions, such as antioxidant, provitamin and many other. Having this in mind, the acceptability of natural colorants by the consumers is much higher. Being present in all photosynthetic plant tissues carotenoids are probably most widespread pigments in nature. Carrot (Daucus carota) is a good source of functional food components. Carrot is especially rich in carotenoids, mainly α- and β-carotene and lutein. For this study, carrot was extracted using classical extraction with hexane and ethyl acetate, as well as supercritical CO₂ extraction. The extraction efficiency was evaluated by estimation of carotenoid yield determined spectrophotometrically. Classical extraction using hexane (18.27 mg β-carotene/100 g DM) was the most efficient method for isolation of carotenoids, compared to ethyl acetate classical extraction (15.73 mg β-carotene/100 g DM) and supercritical CO₂ extraction (0.19 mg β-carotene/100 g DM). Three carrot extracts were tested in terms of antioxidant activity using DPPH and reducing power assay as well. Surprisingly, ethyl acetate extract had the best antioxidant activity on DPPH radicals (AADPPH=120.07 μmol TE/100 g) while hexane extract showed the best reducing power (RP=1494.97 μmol TE/100 g). Hexane extract was chosen as the most potent source of carotenoids and was encapsulated in whey protein by freeze-drying. Carotenoid encapsulation efficiency was found to be high (89.33%). Based on our results it can be concluded that carotenoids from carrot can be efficiently extracted using hexane and classical extraction method. This extract has the potential to be applied in encapsulated form due to high encapsulation efficiency and coloring capacity. Therefore it can be used for dietary supplements development and food fortification.

Keywords: carotenoids, carrot, extraction, encapsulation

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Authors: Syed Asif Hassan, Tabrej Khan

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Multiple sclerosis (MS) is a chronic demyelinating autoimmune disorder, of the central nervous system (CNS). In the present scenario, the current therapies either do not halt the progression of the disease or have side effects which limit the usage of current Disease Modifying Therapies (DMTs) for a longer period of time. Therefore, keeping the current treatment failure schema, we are focusing on screening novel analogues of the available DMTs that specifically bind and inhibit the Sphingosine1-phosphate receptor1 (S1PR1) thereby hindering the lymphocyte propagation toward CNS. The novel drug-like analogs molecule will decrease the frequency of relapses (recurrence of the symptoms associated with MS) with higher efficacy and lower toxicity to human system. In this study, an integrated approach involving ligand-based virtual screening protocol (Ultrafast Shape Recognition with CREDO Atom Types (USRCAT)) to identify the non-toxic drug like analogs of the approved DMTs were employed. The potency of the drug-like analog molecules to cross the Blood Brain Barrier (BBB) was estimated. Besides, molecular docking and simulation using Auto Dock Vina 1.1.2 and GOLD 3.01 were performed using the X-ray crystal structure of Mtb LprG protein to calculate the affinity and specificity of the analogs with the given LprG protein. The docking results were further confirmed by DSX (DrugScore eXtented), a robust program to evaluate the binding energy of ligands bound to the ligand binding domain of the Mtb LprG lipoprotein. The ligand, which has a higher hypothetical affinity, also has greater negative value. Further, the non-specific ligands were screened out using the structural filter proposed by Baell and Holloway. Based on the USRCAT, Lipinski’s values, toxicity and BBB analysis, the drug-like analogs of fingolimod and BG-12 showed that RTL and CHEMBL1771640, respectively are non-toxic and permeable to BBB. The successful docking and DSX analysis showed that RTL and CHEMBL1771640 could bind to the binding pocket of S1PR1 receptor protein of human with greater affinity than as compared to their parent compound (Fingolimod). In this study, we also found that all the drug-like analogs of the standard MS drugs passed the Bell and Holloway filter.

Keywords: antagonist, binding affinity, chemotherapeutics, drug-like, multiple sclerosis, S1PR1 receptor protein

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Authors: Maryam Monazzah, Ronak Samadpour, Mehdi Nasr-esfahani, Fatemeh Qalavand, Marziye Motamedi

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Bipolaris sorokiniana, and Heterodera filipjevi, are important wheat diseases that lead to yield losses worldwide. Identifying novel resistant sources helps us combat these devastating diseases. In this study, we studied the role of Cre3 gene and antioxidant enzymes in the immune responses of wheat genotypes to H. filipjevi and B. sorokiniana. Therefore, real-time PCR analysis using Cre3 gene marker, a resistant gene to cereal cyst nematodes, was conducted on leaves and roots, along with changes ‎in the activity of antioxidant enzymes, peroxidase, and catalase. Enzyme activity assay was performed on roots attacked by nematode and in leaves infected with Bipolaris. Wheat accessions including “Bam” (resistant), “Parsi” (moderately-resistant), “Azar2”, “Ohadi”, “Homa” (highly-susceptible) were previously screened against both stresses under greenhouse and field conditions. Results showed that Cre3 expression against cyst nematodes was significantly higher in resistant cultivars compared to susceptible cultivars. Cre3 was used in marker-assisted selection programs to identify genotypes carrying resistant genes to cyst nematodes. Interestingly, Cre3 was also up-regulated in both tissues of resistant cultivars to B. sorokiniana. Therefore, Cre3 in wheat similarly modulates immunity against B. sorokiniana and might be one of the central components of the induced immune system in wheat. The activity of antioxidant enzymes also indicated the highest increase in resistant genotypes upon both stresses that subsequently neutralize oxidative stress in tissues and decrease damage. Further studies on these resistance components may help us gain insight into the molecular basis of resistance and shed new light on the interaction and overlap between different forms of stress.

Keywords: Bipolaris sorokiniana, Heterodera filipjevi, resistant gene expression, wheat

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Authors: Duaa Mughal, Syeda Areeba Nadeem, Shakil Ahmed, Ishtiaq Ahmed Khan

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The identification of porcine DNA in Halal food items is critical to ensuring compliance with dietary restrictions and religious beliefs. In Islam, Porcine is prohibited as clearly mentioned in Quran (Surah Al-Baqrah, Ayat 173). The purpose of this study was to compare two DNA extraction procedures for detecting 0.001% of porcine DNA in processed Halal food sample mixtures containing chicken, camel, veal, turkey and goat meat using the TaqMan Real-Time PCR technology. In this research, two different commercial kit protocols were compared. The processed sample mixtures were prepared by spiking known concentration of porcine DNA to non-porcine food matrices. Afterwards, TaqMan Real-Time PCR technique was used to target a particular porcine gene from the extracted DNA samples, which was quantified after extraction. The results of the amplification were evaluated for sensitivity, specificity, and reproducibility. The results of the study demonstrated that two DNA extraction techniques can detect 0.01% of porcine DNA in mixture of Halal food samples. However, as compared to the alternative approach, Eurofins| GeneScan GeneSpin DNA Isolation kit showed more effective sensitivity and specificity. Furthermore, the commercial kit-based approach showed great repeatability with minimal variance across repeats. Quantification of DNA was done by using fluorometric assay. In conclusion, the comparison of DNA extraction methods for detecting porcine DNA in Halal food sample mixes using the TaqMan Real-Time PCR technology reveals that the commercial kit-based approach outperforms the other methods in terms of sensitivity, specificity, and repeatability. This research helps to promote the development of reliable and standardized techniques for detecting porcine DNA in Halal food items, religious conformity and assuring nutritional.

Keywords: real time PCR (qPCR), DNA extraction, porcine DNA, halal food authentication, religious conformity

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Authors: Kazam Ali, Muhammad Sagheer, Mansoor-Ul- Hasan, Abdul Rashid, Chaudhary Muhammad Shahid Hanif, Fawad Zafar Ahmad Khan, Hafiz Muhammad Aatif

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Natural extracts of two medicinal plants Nicotiana tabacum and Eucalyptus globulus were tested for their toxic and enzyme inhibition effects against three insects species of stored grains Tribolium castaneum, Trogoderma granarium and Sitophilus granarius. Responses of insects varied with exposure periods and dilution levels of acetone extracts of plants. Both plant extracts were lethal to insects but the crude leaf extract of N. tabacum evidenced strong toxic action against three tested insect species. Maximum mortality 36.30% in S. granarius, 25.96% in T. castaneum, and 21.88% in T. granarium were found at 20% dilution level, after 10 days exposure to botanical extract of N. tabacum. The impact of N. tabacum and E. globulus on the activity of esterases; acetylcholinesterase (AChE), α-carboxylesterase (α-CE), β-carboxylesterase (β-CE) and phosphatses; acid phosphatase (AcP), alkaline phosphatase (AlP) of three stored grain insect species were also studied in the survivors of toxicity assay. Whole body homogenates of insects were used for enzyme determination and consumption of high dose rate N. tabacum extract containing diet resulted in maximum 55.33% inhibition of AChE and 26.17% AlP inhibition in T. castaneum, while 44.17% of α-CE and 31.67% inhibition of β-CE activity were noted in S. granarius. Maximum inhibition 23.44% of AcP activity was found in T. granarium exposed to diet treated with the extract of E. globulus. The findings indicate that acetone extracts of N. tabacum and E. globulus are naturally occurring pesticide and facts of the enzyme inhibition relations specify that their effect changes with the insect species.

Keywords: natural extract, medicinal plant, toxic effects, enzyme inhibition, acetone extract

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Authors: Cyril Deroy, Agata Rumianek, David R. Greaves, Peter R. Cook, Edmond J. Walsh

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Various microfluidic platforms have been developed in the past couple of decades offering experimental methods for the study of cell migration; however, their implementation in the laboratory has remained limited. Some reasons cited for the lack of uptake include the technical complexity of the devices, high failure rate associated with gas-bubbles, biocompatibility concerns with the use of polydimethylsiloxane (PDMS) and equipment/time/expertise requirements for operation and manufacture. As sample handling remains challenging due to the closed format of microfluidic devices, open microfluidic systems have been developed offering versatility and simplicity of use. Rather than confining fluids by solid walls, samples can be accessed directly over the open platform, by removing at least one of the solid boundaries, such as the cover. In this paper, a method for the fabrication of open fluid-walled microfluidic circuits for cell migration studies is introduced, where only materials commonly used by the life-science community are required; tissue culture dishes and cell media. The simplicity of the method, and ability to retrieve cells of interest are two key features of the method. Both passive and active flow-devices can be created in this way. To demonstrate the versatility of the method a cell migration assay is performed, which requires fabricating circuits for establishing chemical gradients, loading cells and incubating, creating chemical gradients, real time imaging of cell migration and finally retrieval of cells. The open architecture has high fidelity as it eliminates air bubble related failures and enables the precise control of gradients. The ability to fabricate custom microfluidic designs in minutes should make this method suitable for use in a wide range of cell migration studies.

Keywords: chemotaxis, fluid walls, gradient generation, open microfluidics

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Authors: Kiran Nawaz, Waheed Anwar, Sehrish Iftikhar, Muhammad Nasir Subhani, Ahmad Ali Shahid

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Plant growth promoting rhizobacteria (PGPR) have been extensively studied and applied for the biocontrol of many soilborne diseases. These rhizobacteria are very efficient against root rot and many other foliar diseases associated with solanaceous plants. These bacteria may inhibit the growth of various pathogens through direct inhibition of target pathogens or indirectly by the initiation of systemic resistance (ISR) which is active all over the complete plant. In the present study, 20 different rhizobacterial isolates were recovered from the root zone of healthy chili plants. All soil samples were collected from various chili-growing areas in Punjab. All isolated rhizobacteria species were evaluated in vitro and in vivo against Phytophthora capsici. Different species of Bacillus and Pseudomonas were tested for the antifungal activity against P. capsici the causal organism of Root rot disease in different crops together with chili. Dual culture and distance culture bioassay were carried out to study the antifungal potential of volatile and diffusible metabolites secreted from rhizobacteria. After seven days of incubation at 22°C, growth inhibition rate was recorded. Growth inhibition rate depended greatly on the tested bacteria and screening methods used. For diffusible metabolites, inhibition rate was 35-62% and 20-45% for volatile metabolites. The screening assay for plant growth promoting and disease inhibition potential of chili associated PGPR indicated 42-100% reduction in disease severity and considerable enhancement in roots fresh weight by 55-87%, aerial parts fresh weight by 35-65% and plant height by 65-76% as compared to untreated control and pathogen-inoculated plants. Pseudomonas flourescene, B. thuringiensis, and B. subtilis were found to be the most efficient isolates in inhibiting P. capsici radial growth, increase plant growth and suppress disease severity.

Keywords: rhizobacteria, chili, phytophthora, root rot

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Authors: Ediati Budi Cahyono, Triana Hertiani, Nauval Arrazy Asawimanda, Wahyu Puji Pratomo

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Background: Doxorubicin is widely used as a chemotherapeutic drug despite having many side effects. It may cause macrophage dysfunction and decreasing proliferation of lymphocyte. Noni (Morinda citrifolia) fruit which has rich of polysaccharide content has potential as antitumor and immunostimulant effect. The isolation of polysaccharide from Noni fruit has been optimized according to four different methods based on macrophage and lymphocyte activities. We found the highest polysaccharide content from one of the four methods isolation. A method of polysaccharide isolation which has the highest immunostimulant effect was used for further observation as co-chemotherapy. The aim of the study: was to evaluate the isolated polysaccharide from the method of choice as co-chemotherapy of doxorubicin for the growth of Vero, He-La, and T47D cell lines in vitro. The method: in vitro growth assay of Vero, He-La, and T47D cell lines was done using MTT-reduction method, and apoptosis test was done by double staining method to evaluate the induction apoptotic effect of the combination. Every group was treated with doxorubicin and isolated polysaccharide from method of choice with 4 variances of concentrations (25 µg/ml, 50 µg/ml, 100 µg/ml and 200 µg/ml) a long with negative control (doxorubicin only) and normal control (without doxorubicin or polysaccharide administration). Results: The combination of polysaccharide fraction in the concentration of 100μg/ml with 2μmol of doxorubicin against He-La and T47D cell lines influenced the highest cytotoxic effect by suppressing cell viability comparing with doxorubicin only. The combination of polysaccharide fraction in the concentration of 100μg/ml with 2μmol of doxorubicin-induced apoptotic effect the He-La cell line comparing with doxorubicin only. The result of the study: it can be concluded that the combination of polysaccharide fraction and doxorubicin effect more selective toward He-La and T47D cell lines than to Vero cell line. It can be suggested isolated polysaccharide from the method of choice has co-chemotherapy activity against doxorubicin.

Keywords: polysaccharide, noni fruit, doxorubicin, cancer cell lines, vero cell line

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Authors: Ahmed Atwa, Ashraf Hafez, Mohamed Abdelhameed, Adel Nabeeh, Mohamed Dawaba, Tamer Helmy

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Introduction and Objective: Tumour staging and grading do not usually reflect the future behavior of Wilms' tumor (WT) regarding mortality. Therefore, in this study, P53, Ki67 and cyclin A immunohistochemistry were used in a trial to predict WT cancer-specific survival (CSS). Methods: In this nonconcurrent cohort study, patients' archived data, including age at presentation, gender, history, clinical examination and radiological investigations, were retrieved then the patients were reviewed at the outpatient clinic of a tertiary care center by history-taking, clinical examination and radiological investigations to detect the oncological outcome. Cases that received preoperative chemotherapy or died due to causes other than WT were excluded. Formalin-fixed, paraffin-embedded specimens obtained from the previously preserved blocks at the pathology laboratory were taken on positively charged slides for IHC with p53, Ki67 and cyclin A. All specimens were examined by an experienced histopathologist devoted to the urological practice and blinded to the patient's clinical findings. P53 and cyclin A staining were scored as 0 (no nuclear staining),1 (<10% nuclear staining), 2 (10-50% nuclear staining) and 3 (>50% nuclear staining). Ki67 proliferation index (PI) was graded as low, borderline and high. Results: Of the 75 cases, 40 (53.3%) were males and 35 (46.7%) were females, and the median age was 36 months (2-216). With a mean follow-up of 78.6±31 months, cancer-specific mortality (CSM) occurred in 15 (20%) and 11 (14.7%) patients, respectively. Kaplan-Meier curve was used for survival analysis, and groups were compared using the Log-rank test. Multivariate logistic regression and Cox regression were not used because only one variable (cyclin A) had shown statistical significance (P=.02), whereas the other significant factor (residual tumor) had few cases. Conclusions: Cyclin A IHC should be considered as a marker for the prediction of WT CSS. Prospective studies with a larger sample size are needed.

Keywords: wilms’ tumour, nephroblastoma, urology, survival

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Authors: Erin M. Durke, Monica L. McEntee, Meilu He, Suresh Dhaniyala

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Aerosols are one of the most important and significant surfaces in the atmosphere. They can influence weather, absorption, and reflection of light, and reactivity of atmospheric constituents. A notable feature of aerosol particles is the presence of a surface charge, a characteristic imparted via the aerosolization process. The existence of charge can complicate the interrogation of aerosol particles, so many researchers remove or neutralize aerosol particles before characterization. However, the charge is present in real-world samples, and likely has an effect on the physical and chemical properties of an aerosolized material. In our studies, we aerosolized different materials in an attempt to characterize the charge imparted via the aerosolization process and determine what impact it has on the aerosolized materials’ properties. The metal oxides, TiO₂ and SiO₂, were aerosolized expulsively and then characterized, using several different techniques, in an effort to determine the surface charge imparted upon the particles via the aerosolization process. Particle charge distribution measurements were conducted via the employment of a custom scanning mobility particle sizer. The results of the charge distribution measurements indicated that expulsive generation of 0.2 µm SiO₂ particles produced aerosols with upwards of 30+ charges on the surface of the particle. Determination of the degree of surface charging led to the use of non-traditional techniques to explore the impact of additional surface charge on the overall reactivity of the metal oxides, specifically TiO₂. TiO₂ was aerosolized, again expulsively, onto a gold-coated tungsten mesh, which was then evaluated with transmission infrared spectroscopy in an ultra-high vacuum environment. The TiO₂ aerosols were exposed to O₂, H₂, and CO, respectively. Exposure to O₂ resulted in a decrease in the overall baseline of the aerosol spectrum, suggesting O₂ removed some of the surface charge imparted during aerosolization. Upon exposure to H₂, there was no observable rise in the baseline of the IR spectrum, as is typically seen for TiO₂, due to the population of electrons into the shallow trapped states and subsequent promotion of the electrons into the conduction band. This result suggests that the additional charge imparted via aerosolization fills the trapped states, therefore no rise is seen upon exposure to H₂. Dosing the TiO₂ aerosols with CO showed no adsorption of CO on the surface, even at lower temperatures (~100 K), indicating the additional charge on the aerosol surface prevents the CO molecules from adsorbing to the TiO₂ surface. The results observed during exposure suggest that the additional charge imparted via aerosolization impacts the interaction with each probe gas.

Keywords: aerosols, charge, reactivity, infrared

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Authors: Rajesh S. Mony, Vaidyaratnam Oushadhasala

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An attempt is made in this study to evolve a drug development modality based on classical Ayurvedic knowledge base as well as on modern scientific methodology. The present study involves (a) identification of a specific ailment condition, (b) the selection of a polyherbal formulation, (c) deciding suitable extraction procedure, (d) confirming the efficacy of the combination by in-vitro trials and (e) fixing up the recommended dose. The ailment segment selected is arthritic condition. The selected herbal combination is Kunturushka, Vibhitaki, Guggulu, Haridra, Maricha and Nirgundi. They were selected as per Classical Ayurvedic references, Authentified as per API (Ayurvedic Pharmacopeia of India), Extraction of each drug was done by different ratios of Hydroalcoholic menstrums, Invitro assessment of each extract after removing residual solvent for anti-Inflammatory, anti-arthritic activities (by UV-Vis. Spectrophotometer with positive control), Invitro assessment of each extract for COX enzyme inhibition (by UV-Vis. Spectrophotometer with positive control), Selection of the extracts was made having good in-vitro activity, Performed the QC testing of each selected extract including HPTLC, that is the in process QC specifications, h. Decision of the single dose with mixtures of selected extracts was made as per the level of in-vitro activity and available toxicology data, Quantification of major groups like Phenolics, Flavonoids, Alkaloids and Bitters was done with both standard Spectrophotometric and Gravimetric methods, Method for Marker assay was developed and validated by HPTLC and a good resolved HPTLC finger print was developed for the single dosage API (Active Pharmaceutical Ingredient mixture of extracts), Three batches was prepared to fix the in process and API (Active Pharmaceutical Ingredient) QC specifications.

Keywords: drug development, antiinflammatory, quality stardardisation, planar chromatography

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