Search results for: methyl blue

Authors: Barun Chakrabarti, Evangelos Kalamaras, Vladimir Yufit, Xinhua Liu, Billy Wu, Nigel Brandon, C. T. John Low

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In this work, we perform electrophoretic deposition of activated carbon on a number of substrates to prepare symmetrical coin cells for supercapacitor applications. From several recipes that involve the evaluation of a few solvents such as isopropyl alcohol, N-Methyl-2-pyrrolidone (NMP), or acetone to binders such as polyvinylidene fluoride (PVDF) and charging agents such as magnesium chloride, we display a working means for achieving supercapacitors that can achieve 100 F/g in a consistent manner. We then adapt this EPD method to deposit reduced graphene oxide on SGL 10AA carbon paper to achieve cathodic materials for testing in a hydrogen/vanadium flow battery. In addition, a self-supported hierarchical carbon nano-fibre is prepared by means of electrospray deposition of an iron phthalocyanine solution onto a temporary substrate followed by carbonisation to remove heteroatoms. This process also induces a degree of nitrogen doping on the carbon nano-fibres (CNFs), which allows its catalytic performance to improve significantly as detailed in other publications. The CNFs are then used as catalysts by attaching them to graphite felt electrodes facing the membrane inside an all-vanadium flow battery (Scribner cell using serpentine flow distribution channels) and efficiencies as high as 60% is noted at high current densities of 150 mA/cm². About 20 charge and discharge cycling show that the CNF catalysts consistently perform better than pristine graphite felt electrodes. Following this, we also test the CNF as an electro-catalyst in the hydrogen/vanadium flow battery (cathodic side as mentioned briefly in the first paragraph) facing the membrane, based upon past studies from our group. Once again, we note consistently good efficiencies of 85% and above for CNF modified graphite felt electrodes in comparison to 60% for pristine felts at low current density of 50 mA/cm² (this reports 20 charge and discharge cycles of the battery). From this preliminary investigation, we conclude that the CNFs may be used as catalysts for other systems such as vanadium/manganese, manganese/manganese and manganese/hydrogen flow batteries in the future. We are generating data for such systems at present, and further publications are expected.

Keywords: electrospinning, carbon nano-fibres, all-vanadium redox flow battery, hydrogen-vanadium fuel cell, electrocatalysis

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Authors: Abdul Ghaffar Memon, Xiao Hong Zhou, Yunpeng Xing, Ruoyu Wang, Miao He

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Due to deleterious environmental and health effects of the Hg²⁺ ions, various online, detection methods apart from the traditional analytical tools have been developed by researchers. Biosensors especially, label, label-free, colorimetric and optical sensors have advanced with sensitive detection. However, there remains a gap of ultrasensitive quantification as noise interact significantly especially in the AuNP based label-free colorimetry. This study reported an amplification strategy using Exo-III enzyme for target recycling of Hg²⁺ ions in a T-rich hairpin loop metallobase label-free colorimetric nanosensor with an improved sensitivity using unmodified gold nanoparticles (uGNPs) as an indicator. The two T-rich metallobase hairpin loop structures as 5’- CTT TCA TAC ATA GAA AAT GTA TGT TTG -3 (HgS1), and 5’- GGC TTT GAG CGC TAA GAA A TA GCG CTC TTT G -3’ (HgS2) were tested in the study. The thermodynamic properties of HgS1 and HgS2 were calculated using online tools (http://biophysics.idtdna.com/cgi-bin/meltCalculator.cgi). The lab scale synthesized uGNPs were utilized in the analysis. The DNA sequence had T-rich bases on both tails end, which in the presence of Hg²⁺ forms a T-Hg²⁺-T mismatch, promoting the formation of dsDNA. Later, the Exo-III incubation enable the enzyme to cleave stepwise mononucleotides from the 3’ end until the structure become single-stranded. These ssDNA fragments then adsorb on the surface of AuNPs in their presence and protect AuNPs from the induced salt aggregation. The visible change in color from blue (aggregation stage in the absence of Hg²⁺) and pink (dispersion state in the presence of Hg²⁺ and adsorption of ssDNA fragments) can be observed and analyzed through UV spectrometry. An ultrasensitive quantitative nanosensor employing Exo-III assisted target recycling of mercury ions through label-free colorimetry with nanomolar detection using uGNPs have been achieved and is further under the optimization to achieve picomolar range by avoiding the influence of the environmental matrix. The proposed strategy will supplement in the direction of uGNP based ultrasensitive, rapid, onsite, label-free colorimetric detection.

Keywords: colorimetric, Exo-III, gold nanoparticles, Hg²⁺ detection, label-free, signal amplification

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Authors: A. J. S. P. Nileema, S. Kulatunga , S. H. Palihawadana

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Computer aided detection (CAD) system was developed for the diagnosis of pulmonary tuberculosis using digital chest X-rays with MATLAB image processing techniques using a statistical approach. The study comprised of 200 digital chest radiographs collected from the National Hospital for Respiratory Diseases - Welisara, Sri Lanka. Pre-processing was done to remove identification details. Lung fields were segmented and then divided into four quadrants; right upper quadrant, left upper quadrant, right lower quadrant, and left lower quadrant using the image processing techniques in MATLAB. Contrast, correlation, homogeneity, energy, entropy, and maximum probability texture features were extracted using the gray level co-occurrence matrix method. Descriptive statistics and normal distribution analysis were performed using SPSS. Depending on the radiologists’ interpretation, chest radiographs were classified manually into PTB - positive (PTBP) and PTB - negative (PTBN) classes. Features with standard normal distribution were analyzed using an independent sample T-test for PTBP and PTBN chest radiographs. Among the six features tested, contrast, correlation, energy, entropy, and maximum probability features showed a statistically significant difference between the two classes at 95% confidence interval; therefore, could be used in the classification of chest radiograph for PTB diagnosis. With the resulting value ranges of the five texture features with normal distribution, a classification algorithm was then defined to recognize and classify the quadrant images; if the texture feature values of the quadrant image being tested falls within the defined region, it will be identified as a PTBP – abnormal quadrant and will be labeled as ‘Abnormal’ in red color with its border being highlighted in red color whereas if the texture feature values of the quadrant image being tested falls outside of the defined value range, it will be identified as PTBN–normal and labeled as ‘Normal’ in blue color but there will be no changes to the image outline. The developed classification algorithm has shown a high sensitivity of 92% which makes it an efficient CAD system and with a modest specificity of 70%.

Keywords: chest radiographs, computer aided detection, image processing, pulmonary tuberculosis

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

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Backgrounds: Sulforaphane, one of the most important isothiocyanates in the human diet, is known to have chemopreventive and antioxidant activities in different tissues via activation of NF-E2-related factor 2 (Nrf2)-mediated induction of antioxidant/phase II enzymes, such as heme oxygenase-1 (HO-1) and NAD(P)H quinone oxidoreductase 1 (NQO1). However, its effects on muscular dystrophy remain unknown. This work was undertaken to evaluate the effects of Sulforaphane on Duchenne muscular dystrophy (DMD). Methods: 4-week-old mdx mice were treated with SFN by gavage (2 mg/kg body weight per day) for 8 weeks. Blood was collected from eye socket every week, and tibial anterior, extensor digitorum longus, gastrocnemius, soleus, triceps brachii muscles and heart samples were collected after 8-week gavage. Force measurements and mice exercise capacity assays were detected. GSH/GSSG ratio, TBARS, CK and LDH levels were analyzed by spectrophotometric methods. H&E staining was used to analyze histological and morphometric of skeletal muscles of mdx mice, and Evas blue dye staining was made to detect sarcolemmal integrity of mdx mice. Further, the role of Sulforaphane on Nrf2/ARE signaling pathway was analyzed by ELISA, western blot and qRT-PCR. Results: Our results demonstrated that SFN treatment increased the expression and activity of muscle phase II enzymes NQO1 and HO-1 with Nrf2 dependent manner. SFN significantly increased skeletal muscle mass, muscle force (~30%), running distance (~20%) and GSH/GSSG ratio (~3.2 folds) of mdx mice, and decreased the activities of plasma creatine phosphokinase (CK) (~45%) and lactate dehydrogenase (LDH) (~40%), gastrocnemius hypertrophy (~25%), myocardial hypertrophy (~20%) and MDA levels (~60%). Further, SFN treatment also reduced the central nucleation (~40%), fiber size variability, inflammation and improved the sarcolemmal integrity of mdx mice. Conclusions: Collectively, these results show that SFN can improve muscle function, pathology and protect dystrophic muscle from oxidative damage in mdx mice through Nrf2 signaling pathway, which indicate Nrf2 may have clinical implications for the treatment of patients with muscular dystrophy.

Keywords: sulforaphane, duchenne muscular dystrophy, Nrf2, oxidative stress

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Authors: Nada R. Abuknesha, John P. Morgan, Andrew J. Searle

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Information regarding post-mortem interval (PMI) in criminal investigations is vital to establish a time frame when reconstructing events. PMI is defined as the time period that has elapsed between the occurrence of death and the discovery of the corpse. Adipocere, commonly referred to as ‘grave-wax’, is formed when post-mortem adipose tissue is converted into a solid material that is heavily comprised of fatty acids. Adipocere is of interest to forensic anthropologists, as its formation is able to slow down the decomposition process. Therefore, analysing the changes in the patterns of fatty acids during the early decomposition process may be able to estimate the period of burial, and hence the PMI. The current study concerned the investigation of the fatty acid composition and patterns in buried pig fat tissue. This was in an attempt to determine whether particular patterns of fatty acid composition can be shown to be associated with the duration of the burial, and hence may be used to estimate PMI. The use of adipose tissue from the abdominal region of domestic pigs (Sus-scrofa), was used to model the human decomposition process. 17 x 20cm piece of pork belly was buried in a shallow artificial grave, and weekly samples (n=3) from the buried pig fat tissue were collected over an 11-week period. Marker fatty acids: palmitic (C16:0), oleic (C18:1n-9) and linoleic (C18:2n-6) acid were extracted from the buried pig fat tissue and analysed as fatty acid methyl esters using the gas chromatography system. Levels of the marker fatty acids were quantified from their respective standards. The concentrations of C16:0 (69.2 mg/mL) and C18:1n-9 (44.3 mg/mL) from time zero exhibited significant fluctuations during the burial period. Levels rose (116 and 60.2 mg/mL, respectively) and fell starting from the second week to reach 19.3 and 18.3 mg/mL, respectively at week 6. Levels showed another increase at week 9 (66.3 and 44.1 mg/mL, respectively) followed by gradual decrease at week 10 (20.4 and 18.5 mg/mL, respectively). A sharp increase was observed in the final week (131.2 and 61.1 mg/mL, respectively). Conversely, the levels of C18:2n-6 remained more or less constant throughout the study. In addition to fluctuations in the concentrations, several new fatty acids appeared in the latter weeks. Other fatty acids which were detectable in the time zero sample, were lost in the latter weeks. There are several probable opportunities to utilise fatty acid analysis as a basic technique for approximating PMI: the quantification of marker fatty acids and the detection of selected fatty acids that either disappear or appear during the burial period. This pilot study indicates that this may be a potential semi-quantitative methodology for determining the PMI. Ideally, the analysis of particular fatty acid patterns in the early stages of decomposition could be an additional tool to the already available techniques or methods in improving the overall processes in estimating PMI of a corpse.

Keywords: adipocere, fatty acids, gas chromatography, post-mortem interval

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Authors: D. O. Miranda, L. R. Azevedo, J. F. C. Cordeiro, A. H. Dos Santos, S. F. Lisboa, F. S. Guimarães, G. S. Bisson

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Background: The Colorectal cancer (CRC) is one of the most common cancers and the fourth leading cause of cancer death in the world. The prevalence of psychiatric-disorders among CRC patients, mainly depression, is high, resulting in impaired quality of life and side effects of primary treatment. High levels of proinflammatory cytokines at tumor microenvironment is a feature of CRC and the literature suggests that those mediators could contribute to the development of psychiatric disorders. Nevertheless, the ability of tumor-associated biological processes to affect the central nervous system (CNS) has only recently been explored in the context of symptoms of depression and is still not well understood. Therefore, the aim of the present study was to test the hypothesis that depressive-like behavior in an experimental model of CCR induced by N-methyl-N-nitro-N-nitrosoguanidine (MNNG) was correlated to proinflammatory profile in the periphery and in the brain. Methods: Colorectal carcinogenesis was induced in adult C57BL/6 mice (n=12) by administration of MNNG (5mg/kg, 0.1ml/intrarectal instillation) 2 times a week, for 2 week. Control group (n=12) received saline (0.1ml/intrarectal instillation). Eight weeks after beginning of MNNG administration animals were submitted to the forced swim test (FST) and the sucrose preference test for evaluation, respectively, of depressive- and anhedonia-like behaviors. After behavioral evaluation, the colon was collected and brain regions dissected (cortex-C, striatum-ST and hippocampus-HIP) for posterior evaluation of cytokine levels (IL-1β, IL-10, IL-17, and CX3CL1) by ELISA. Results: MNNG induced depressive-like behavior, represented by increased immobility time in the FST (Student t test, p < 0.05) and lower sucrose preference (Student t test, p < 0.05). Moreover, there were increased levels of IL-1β, IL-17 and CX3CL1 in the colonic tissue (Student t test, p < 0.05) and in the brain (IL-1 β in the ST and HIP, Student t test, p < 0.05; IL-17 and CX3CL1 in the C and HIP, p < 0.05). IL-10 levels, in contrast, were decreased in both the colon (p < 0.05) and the brain (C and HIP, p < 0.05). Conclusions: The results obtained in the present work support the notion that tumor growth induces neuroinflammation in stress-related brain regions and depressive-like behavior, which could be related to the high incidence of depression in colorectal carcinogenesis. This work have important clinical and research implications, taken into account that cytokine levels may be a marker promissory for the developing depression in CRC patients. New therapeutic strategies to assist in alleviating mental suffering in cancer patients might result from a better understanding of the role of cytokines in the pathophysiology of depression in these subjects.

Keywords: cytokines, brain, depression, colorectal cancer

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Authors: Chandragouda R. Patil, K. S. Jagadeesh, S. D. Kalolgi

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Biofertilizers containing live microbial cells can mobilize one or more nutrients to plants when applied to either seed or rhizosphere. They form an integral part of nutrient management strategies for sustainable production of agricultural crops. Annually, about 22 tons of lignite-based biofertilizers are being produced and supplied to farmers at the Institute of Organic Farming, University of Agricultural Sciences, Dharwad, Karnataka state India. Although carrier based biofertilizers are common, they have shorter shelf life, poor quality, high contamination, unpredictable field performance and high cost of solid carriers. Hence, liquid formulations are being developed to increase their efficacy and broaden field applicability. An attempt was made to develop liquid formulation of strains of Rhizobium NC-92 (Groundnut), Azospirillum ACD15 both nitrogen-fixing biofertilizers and Pseudomonas striata an efficient P-solubilizing bacteria (PSB). Different concentration of amendments such as additives (glycerol and polyethylene glycol), adjuvants (carboxyl methyl cellulose), gum arabica (GA), surfactant (polysorbate) and trehalose specifically for Azospirillum were found essential. Combinations of formulations of Rhizobium and PSB for groundnut and Azospirillum and PSB for maize were evaluated under field conditions to determine the optimum level of inoculum required. Each biofertilizer strain was inoculated at the rate of 2, 4, 8 ml per kg of seeds and the efficacy of each formulation both individually and in combinations was evaluated against the lignite-based formulation at the rate of 20 g each per kg seeds and a un-inoculated set was included to compare the inoculation effect. The field experiment had 17 treatments in three replicates and the best level of inoculum was decided based on net returns and cost: benefit ratio. In peanut, the combination of 4 ml of Rhizobium and 2 ml of PSB resulted in the highest net returns and higher cost to benefit ratio of 1:2.98 followed by treatment with a combination of 2 ml per kg each of Rhizobium and PSB with a B;C ratio of 1:2.84. The benefits in terms of net returns were to the extent of 16 percent due to inoculation with lignite based formulations while it was up to 48 percent due to the best combination of liquid biofertilizers. In maize combination of liquid formulations consisting of 4 ml of Azospirillum and 2 ml of PSB resulted in the highest net returns; about 53 percent higher than the un-inoculated control and 20 percent higher than the treatment with lignite based formulation. In both the crops inoculation with lignite based formulations significantly increased the net returns over un-inoculated control while levels higher or lesser than 4 ml of Rhizobium and Azospirillum and higher or lesser than 2 ml of PSB were not economical and hence not optimal for these two crops.

Keywords: Rhizobium, Azospirillum, phosphate solubilizing bacteria, liquid formulation, benefit-cost ratio

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

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

Keywords: hazardous organic compounds, irradiation, nitroaromatics, photocatalysis

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Authors: Chinedu F. Anochie

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The presence of gangue particles in recovered metal concentrates has been a serious challenge to ore dressing engineers. Middlings lower the quality of concentrates, and in most cases, drastically affect the smelter terms, owing to exorbitant amounts paid by Mineral Processing industries as treatment charge. Models which encourage optimization of liberation operations have been utilized in most ore beneficiation industries to reduce the presence of locked particles in valuable concentrates. Moreover, methods such as incorporation of regrind mills, scavenger, rougher and cleaner cells, to the milling and flotation plants has been widely employed to tackle these concerns, and to optimize the grade–recovery relationship of metal concentrates. This work compared the crushing and grinding method of liberation, to the mine to mill route, by evaluating the proportion of middlings present in selectively processed complex Pb-Zn ore samples. To establish the effect of size reduction operations on the percentage of locked particles present in recovered concentrates, two similar samples of complex Pb- Zn ores were processed. Following blasting operation, the first ore sample was ground directly in a ball mill (Mine to Mill Route of Comminution), while the other sample was manually crushed, and subsequently ground in the ball mill (Crushing and Grinding Route of Comminution). The two samples were separately sieved in a mesh to obtain the desired representative particle sizes. An equal amount of each sample that would be processed in the flotation circuit was then obtained with the aid of a weighing balance. These weighed fine particles were simultaneously processed in the flotation circuit using the selective flotation technique. Sodium cyanide, Methyl isobutyl carbinol, Sodium ethyl xanthate, Copper sulphate, Sodium hydroxide, Lime and Isopropyl xanthate, were the reagents used to effect differential flotation of the two ore samples. Analysis and calculations showed that the degree of liberation obtained for the ore sample which went through the conventional crushing and grinding route of comminution, was higher than that of the directly milled run off mine (ROM) ore. Similarly, the proportion of middlings obtained from the separated galena (PbS) and sphalerite (ZnS) concentrates, were lower for the crushed and ground ore sample. A concise data which proved that the mine to mill method of size reduction is not the most ideal technique for the recovery of quality metal concentrates has been established.

Keywords: comminution, degree of liberation, middlings, mine to mill

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Authors: Nandhakumar Elumalai, Purushothaman Ayyakannu, Sachidanandam T. Panchanatham

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Background: Understanding the basic mechanisms and factors underlying the tumor growth and invasion has gained attention in recent times. The processes of angiogenesis and apoptosis are known to play a vital role in various stages of cancer. The vascular endothelial growth factor (VEGF) is well established as one of the key regulators of tumor angiogenesis while MMPs are known for their exclusive ability to degrade ECM. Objective: The present study was designed to evaluate the pro apoptotic and anti angiogenic activity of the herbal formulation Shemamruthaa. The anticancer activity of Shemamruthaa was tested in breast cancer cell line (MCF-7). Results of MTT, trypan blue and flow cytometric analysis of apoptotis suggested that Shemamruthaa can induce cytotoxicity in cancer cells, in a concentration- and time dependent manner and induce apoptosis. With these results, we further evaluated the antiangiogenic and pro-apoptotic activities of Shemamruthaa in DMBA induced mammary carcinoma in Sprague Dawley rats. Flavono tumour was induced in 8-week-old Sprague-Dawley rats by gastric intubation of 25 mg DMBA in 1ml olive oil. After 90 days of induction period, the rats were orally administered with Shemamruthaa (400 mg/kg body wt) for 45 days. Treatment with the drug SM significantly modulated the expression of p53, MMP-2, MMP-3, MMP-9 and VEGF by means of its anti angiogenic and protease inhibiting activity. Conclusion: Based on these results, it might be concluded that the formulation, Shemamruthaa, constituted of dried flowers of Hibiscus rosa-sinensis, fruits of Emblica officinalis, and honey has been found to exhibit pronounced antiproliferative and apoptotic effects. This enhanced anticancer effect of Shemamruthaa might be attributed to the synergistic action of polyphenols such as flavonoids, tannins, alkaloids, glycosides, saponins, steroids, terpenoids, vitamin C, niacin, pyrogallol, hydroxymethylfurfural, trilinolein, and other compounds present in the formulation. Collectively, these results demonstrate that Shemamruthaa holds potential to be developed as a potent chemotherapeutic agent against mammary carcinoma.

Keywords: Shemamruthaa, flavonoids, MCF-7 cell line, mammary cancer

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Authors: Harsha B. R., K. S. Anil Kumar

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Land characterization, classification, and soil suitability evaluation of grapes-growing pedons were assessed at fifteen taluks covering four agro climatic zones of Karnataka. Study on problems and potentials of grapes cultivation in selected agro-climatic zones was carried out along with the plant sample analysis. Twenty soil profiles were excavated as study site based on the dominance of area falling under grapes production and existing spatial variability of soils. The detailed information of profiles and horizon wise soil samples were collected to study the morphological, physical, chemical, and fertility characteristics. Climatic analysis and water retention characteristics of soils of major grapes-growing areas were also done. Based on the characterisation and classification study, it was revealed that soils of Doddaballapur (Bangalore Blue and Wine grapes), Bangalore North (GKVK Farm, Rajankunte, and IIHR Farm), Devanahalli, Magadi, Hoskote, Chikkaballapur (Dilkush and Red globe), Yelaburga, Hagari Bommanahalli, Bagalkot (UHS farm) and Indi fall under the soil order Alfisol. Vijaypur pedon of northern dry zone was keyed out as Vertisols whereas, Jamkhandi and Athani as Inceptisols. Properties of Aridisols were observed in B. Bagewadi (Manikchaman and Thompson Seedless) and Afzalpur. Soil fertility status and its mapping using GIS technique revealed that all the nutrients were found to be in adequate range except nitrogen, potassium, zinc, iron, and boron, which indicated the need for application along with organic matter to improve the SOC status. Varieties differed among themselves in yield and plant nutrient composition depending on their age, climatic, soil, and management requirements. Bangalore North (GKVK farm) and Jamkhandi are having medium soil organic carbon stocks of 6.21 and 6.55 kg m⁻³, respectively. Soils of Bangalore North (Rajankunte) were highly suitable (S1) for grapes cultivation. Under northern Karnataka, Vijayapura, B. Bagewadi, Indi, and Afzalpur vineyards were good performers despite the limitations of fertility and free lime content.

Keywords: land characterization, suitability, soil orders, soil organic carbon stock

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Authors: Nirali Sanghavi, Julia Ash, Amy Wasserman

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Introduction: To the best of our knowledge, there is only one case report that found an association between leuprolide and scleroderma renal crisis (SRC). Leuprolide has been noted to cause acute renal failure in some patients. Given the close timing of the leuprolide injection and the worsening renal function in our patient, leuprolide likely caused exacerbation of lupus nephritis and SRC. Interestingly, our patient on long-term hydroxychloroquine (HCQ) with normal baseline cardiac function was found to have HCQ cardiomyopathy highlighting the need for close monitoring of HCQ toxicity. We know that some of the risk factors that are involved in HCQ induced cardiomyopathy are older age, females, increased dose and >10 years of HCQ use, and pre-existing cardiac and renal insufficiency. Case presentation: A 34-year-old African American woman with a history of overlap of systemic lupus erythematosus (SLE) and scleroderma features and class III lupus nephritis presented with severe headaches, elevated blood pressure (180/120 mmHg) and worsening creatinine levels (2.07 mg/dL). The headaches started 1 month ago after she started leuprolide injections for fibroids. She was being treated with mycophenolate mofetil 1 gm twice a day, belimumab weekly, HCQ 200mg, and prednisone 5 mg daily. She has been on HCQ since her teenage years. The examination was unremarkable except for proximal interphalangeal joint contractures in the right hand and sclerodactyly of bilateral hands, unchanged from baseline. Laboratory findings include urinalysis, which showed 3+ protein, 1+ blood, 6 red blood cells, and 14 white blood cells ruling out thrombotic microangiopathy. C3 was 32 mg/dL, C4 <5 mg/dL, and +dsDNA increased >1000. She was started on captopril and discharged once creatinine and blood pressure was controlled. She was readmitted with hypertension, hyperkalemia, worsening creatinine, nephrotic range proteinuria, complaints of chest pressure, and shortness of breath with pleuritic chest pain. Physical examination and lab findings were unchanged. She was treated with pulse dose methyl prednisone followed by taper and multiple anti-hypertensive agents, including captopril, for presumed lupus nephritis flare versus SRC. Renal biopsy was consistent with SRC and class IV lupus nephritis and was started on cyclophosphamide. While cardiac biopsy showed borderline myocarditis without necrosis and cytoplasmic vacuolization consistent with HCQ cardiomyopathy, hence HCQ was discontinued. Summary: It highlights a rare association of leuprolide causing exacerbation of lupus nephritis or SRC. Although rare, the current case reinforces the importance of close monitoring for HCQ toxicity in patients with renal insufficiency.

Keywords: leuprolide, lupus nephritis, scleroderma, SLE

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Authors: Varsha Salian, Shama Rao, N. Narendra, B. Mohana Kumar

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Evolving evidence proposes the existence of a highly tumorigenic subpopulation of undifferentiated, self-renewing cancer stem cells, responsible for exhibiting resistance to conventional anti-cancer therapy, recurrence, metastasis and heterogeneous tumor formation. Importantly, the mechanisms exploited by cancer stem cells to resist chemotherapy are very less understood. Oral squamous cell carcinoma (OSCC) is one of the most regularly diagnosed cancer types in India and is associated commonly with alcohol and tobacco use. Therefore, the isolation and in vitro characterization of cancer stem-like cells from patients with OSCC is a critical step to advance the understanding of the chemoresistance processes and for designing therapeutic strategies. With this, the present study aimed to establish and characterize cancer stem-like cells in vitro from OSCC. The primary cultures of cancer stem-like cell lines were established from the tissue biopsies of patients with clinical evidence of an ulceroproliferative lesion and histopathological confirmation of OSCC. The viability of cells assessed by trypan blue exclusion assay showed more than 95% at passage 1 (P1), P2 and P3. Replication rate was performed by plating cells in 12-well plate and counting them at various time points of culture. Cells had a more marked proliferative activity and the average doubling time was less than 20 hrs. After being cultured for 10 to 14 days, cancer stem-like cells gradually aggregated and formed sphere-like bodies. More spheroid bodies were observed when cultured in DMEM/F-12 under low serum conditions. Interestingly, cells with higher proliferative activity had a tendency to form more sphere-like bodies. Expression of specific markers, including membrane proteins or cell enzymes, such as CD24, CD29, CD44, CD133, and aldehyde dehydrogenase 1 (ALDH1) is being explored for further characterization of cancer stem-like cells. To summarize the findings, the establishment of OSCC derived cancer stem-like cells may provide scope for better understanding the cause for recurrence and metastasis in oral epithelial malignancies. Particularly, identification and characterization studies on cancer stem-like cells in Indian population seem to be lacking thus provoking the need for such studies in a population where alcohol consumption and tobacco chewing are major risk habits.

Keywords: cancer stem-like cells, characterization, in vitro, oral squamous cell carcinoma

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Authors: Sakeena Naikwadi, K. Jagaluraiah Sannapapamma

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Vetiveria zizanioides is an aromatic grass and traditionally been used in aromatherapy and ayurvedic medicine. Vetiver root is multi-functional biopolymer and has highly aromatic, antimicrobial, UV blocking, antioxidant properties suitable for textile finishing. The vetiver root (Gulabi) powder of different concentration (2, 4, 6,8 percent) were extracted by aqueous and solvent methods subjected to bioassay for antimicrobial efficiency and GCMS spectral analysis. The organic cotton fabric was finished with vetiver root extract (8 percent) by exhaust and pad dry cure methods. The finished fabric was assessed for functional properties viz., UV protective factor, antimicrobial efficiency and aroma intensity. The results revealed that Ethanol extraction showed a greater zone of inhibition compared to aqueous extract in root powder. Among the concentrations, 8 percent root extract in ethanol showed a greater zone of inhibition against gram-positive organism S. aureus and gram-negative organism E. coli. The major compounds present in vetiver root extracts were diethyl pathalate with greater percentage (87.73 %) followed by 7- Isopropyl dimethyl carboxylic acid (4.05 %), 2-butanone 4-trimethyle cyclohexen (1.21 %), phenanthrene carboxylic acid (1.03 %), naphthalene pentanoic acid (0.99 %), 1-phenanthrene carboxylic acid and 1 cyclohexenone 2-methyl oxobuty (0.89 %). The sample finished by pad dry cure method exhibited better UV protection even after 10th wash as compared to exhaust method. Vetiver extract treated samples exhibited maximum zone of inhibition against S. aureus than the E. coli organism. The vetiver root extract treated organic cotton fabric through pad dry cure method possessed good antimicrobial activity against S. aureus and E. coli even after 20th washes compared to vetiver root extract treated by exhaust method. The olfactory analysis was carried out by 30 panels of members and opined that vetiver root extract treated fabric has very good and pleasant aroma with better tactile properties that provide cooling, soothing effect and enhances the mood of the wearer. Vetiver root extract finished organic cotton fabric possessed aroma, antimicrobial and UV properties which are aptly suitable for medical and healthcare textiles viz., wound dressing, bandage gauze, surgical cloths, baby diapers and sanitary napkins. It can be used as after finishing agent for variegated garments and made-ups and can be replaced with commercial after finishing agents.

Keywords: antimicrobial, olfactory analysis, UV protection factor, vetiver root extract

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Authors: Aayushi Pandey, Manoj Kumar Pandey, Ashutosh Tiwari, Kireet Kumar

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Glaciers play an important role in climate change and are sensitive phenomena of global climate change scenario. Glaciers in Himalayas are unique as they are predominantly valley type and are located in tropical, high altitude regions. These glaciers are often covered with debris which greatly affects ablation rate of glaciers and work as a sensitive indicator of glacier health. The aim of this study is to map high altitude Glacier surface with a focus on glacial lake and debris estimation using different techniques in Nagling glacier of dhauliganga basin in Himalayan region. Different Image Classification techniques i.e. thresholding on different band ratios and supervised classification using maximum likelihood classifier (MLC) have been used on high resolution sentinel 2A level 1c satellite imagery of 14 October 2017.Here Near Infrared (NIR)/Shortwave Infrared (SWIR) ratio image was used to extract the glaciated classes (Snow, Ice, Ice Mixed Debris) from other non-glaciated terrain classes. SWIR/BLUE Ratio Image was used to map valley rock and Debris while Green/NIR ratio image was found most suitable for mapping Glacial Lake. Accuracy assessment was performed using high resolution (3 meters) Planetscope Imagery using 60 stratified random points. The overall accuracy of MLC was 85 % while the accuracy of Band Ratios was 96.66 %. According to Band Ratio technique total areal extent of glaciated classes (Snow, Ice ,IMD) in Nagling glacier was 10.70 km2 nearly 38.07% of study area comprising of 30.87 % Snow covered area, 3.93% Ice and 3.27 % IMD covered area. Non-glaciated classes (vegetation, glacial lake, debris and valley rock) covered 61.93 % of the total area out of which valley rock is dominant with 33.83% coverage followed by debris covering 27.7 % of the area in nagling glacier. Glacial lake and Debris were accurately mapped using Band ratio technique Hence, Band Ratio approach appears to be useful for the mapping of debris covered glacier in Himalayan Region.

Keywords: band ratio, Dhauliganga basin, glacier mapping, Himalayan region, maximum likelihood classifier (MLC), Sentinel-2 satellite image

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Authors: T. Ferreira, A. Kulkarni, C. Bretscher, K. Richter, M. Ehrlich, A. Marchini

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H-1 protoparvovirus (H-1PV) is a virus with inherent oncolytic and oncosuppressive activities while remaining non-pathogenic in humans. H-1PV was the first oncolytic parvovirus to undergo clinical testing. Results from trials in patients with glioblastoma or pancreatic carcinoma showed an excellent safety profile and first signs of efficacy. H-1PV infection is vastly dependent on cellular factors, from cell attachment and entry to viral replication and egress. Hence, we believe that the characterisation of the parvovirus life cycle would ultimately help further improve H-1PV clinical outcome. In the present study, we explored the entry pathway of H-1PV in cervical HeLa and glioma NCH125 cancer cell lines. Electron and confocal microscopy showed viral particles associated with clathrin-coated pits and vesicles, providing the first evidence that H-1PV cell entry occurs through clathrin-mediated endocytosis. Accordingly, we observed that by blocking clathrin-mediated endocytosis with hypertonic sucrose, chlorpromazine, or pitstop 2, H-1PV transduction was markedly decreased. Accordingly, siRNA-mediated knockdown of AP2M1, which retains a crucial role in clathrin-mediated endocytosis, verified the reliance of H-1PV on this route to enter HeLa and NCH125 cancer cells. By contrast, we found no evidence of viral entry through caveolae-mediated endocytosis. Indeed, pre-treatment of cells with nystatin or methyl-β-cyclodextrin, both inhibitors of caveolae-mediated endocytosis, did not affect viral transduction levels. Unexpectedly, siRNA-mediated knockdown of caveolin-1, the main driver of caveolae-mediated endocytosis, increased H-1PV transduction, suggesting caveolin-1 is a negative modulator of H-1PV infection. We also show that H-1PV entry is dependent on dynamin, a protein responsible for mediating the scission of vesicle neck and promoting further internalisation. Furthermore, since dynamin inhibition almost completely abolished H-1PV infection, makes it unlikely that H-1PV uses macropinocytosis as an alternative pathway to enter cells. After viral internalisation, H-1PV passes through early to late endosomes as observed by confocal microscopy. Inside these endocytic compartments, the acidic environment proved to be crucial for a productive infection. Inhibition of acidification of pH dramatically reduced H-1PV transduction. Besides, a fraction of H-1PV particles was observed inside LAMP1-positive lysosomes, most likely following a non-infectious route. To the author's best knowledge, this is the first study to characterise the cell entry pathways of H-1PV. Along these lines, this work will further contribute to understand H-1PV oncolytic properties as well as to improve its clinical potential in cancer virotherapy.

Keywords: clathrin-mediated endocytosis, H-1 parvovirus, oncolytic virus, virus entry

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Authors: Hamada Ahmed, Mervat A. Abdel-Latif, Alaa. A. Ghoraba, Samah A. Ganna

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An experiment was conducted to determine the effect of microbial phytase (Quantum Blue®) supplementation on layer chickens fed diets with required or low phosphorous level in corn-soybean based diets. One hundred and sixteen 23-week-old Lohman brown laying hens were used in 8-week feeding trial. Hens were randomly allotted into four treatments where the group (1) (control group) was fed basal diet without phytase, group (2) fed basal diet supplemented with phytase, group (3) fed diet supplemented with phytase as a replacement of 25% of monocalcium phosphate and group (4) fed diet supplemented with phytase as a replacement of 50% of monocalcium phosphate. Records on daily egg production, egg mass, egg weight and body weight of hens at the end of experimental period were recorded. Results revealed no significant (p ≥ 0.05) differences were observed among the other dietary treatments in BW, egg production, egg mass, feed intake or feed conversion when these parameters were evaluated over the duration of the experiment while egg weight showed significant (p < 0.05) increase in all phytase supplemented groups. There was no significant (p ≥ 0.05) differences in egg quality including egg length, egg width, egg shape index, yolk height, yolk width, yolk index, yolk weight and yolk albumin ratio while egg albumin was significantly increased (p < 0.05) in group (2) and group (3). Egg shell weight increased significantly (p < 0.05) in all phytase supplemented groups when compared with the control group also shell thickness increased significantly (p < 0.05) in both group (2 &3). No significant (P ≥ 0.05) difference was observed in serum Ca, P level while alkaline phosphatase was significantly (P ˂ 0.05) increased in group (3). Egg shell analysis showed increase in egg shell ash% in all phytase supplemented groups when compared with the control group, egg shell calcium % was higher in group (3) and group (4) than the control group while group (2) showed lower egg shell calcium% than the other experimental groups, egg shell phosphorous% was higher in all phytase supplemented groups than the control group. Phosphorous digestability was significantly (P ˂ 0.05) increased in all phytase supplemented groups than the control group and the highest p digestability was in group (4). Calcium digestability showed significant (P ˂ 0.05) increase in all phytase supplemented groups when compared with the control group and the highest digetability was in group (4).

Keywords: layers, microbial phytase, Ca and P availability, egg production, egg characteristics

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Authors: Qiaoyue Kuang, Yang Li, Mizuki Endo, Takeaki Ozawa

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G protein-coupled receptors (GPCR) are the largest family of receptor proteins that detect molecules outside the cell and activate cellular responses. Of the GPCRs, dopamine receptors, which recognize extracellular dopamine, are essential to mammals due to their roles in numerous physiological events, including autonomic movement, hormonal regulation, emotions, and the reward system in the brain. To precisely understand the physiological roles of dopamine receptors, it is important to spatiotemporally control the signaling mediated by dopamine receptors, which is strongly dependent on their surface expression. Conventionally, chemical-induced interactions were applied to trigger the endocytosis of cell surface receptors. However, these methods were subjected to diffusion and therefore lacked temporal and special precision. To further understand the receptor-mediated signaling and to control the plasma membrane expression of receptors, an optogenetic tool called E-fragment was developed. The C-terminus of a light-sensitive photosensory protein cyptochrome2 (CRY2) was attached to β-Arrestin, and the E-fragment was generated by fusing the C-terminal peptide of vasopressin receptor (V2R) to CRY2’s binding partner protein CIB. The CRY2-CIB heterodimerization triggered by blue light stimulation brings β-Arrestin to the vicinity of membrane receptors and results in receptor endocytosis. In this study, the E-fragment system was applied to dopamine receptors 1 and 2 (DRD1 and DRD2) to control dopamine signaling. First, confocal fluorescence microscope observation qualitatively confirmed the light-induced endocytosis of E-fragment fused receptors. Second, NanoBiT bioluminescence assay verified quantitatively that the surface amount of E-fragment labeled receptors decreased after light treatment. Finally, GloSensor bioluminescence assay results suggested that the E-fragment-dependent receptor light-induced endocytosis decreased cAMP production in DRD1 signaling and attenuated the inhibition effect of DRD2 on cAMP production. The developed optogenetic tool was able to induce receptor endocytosis by external light, providing opportunities to further understand numerous physiological activities by controlling receptor-mediated signaling spatiotemporally.

Keywords: dopamine receptors, endocytosis, G protein-coupled receptors, optogenetics

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Authors: Jorge Martínez-Rodríguez, Esther Pérez-Carrillo, Diana Laura Anchondo Álvarez, Julia Lucía Leal Villarreal, Mariana Juárez Dominguez, Luisa Fernanda Torres Hernández, Daniela Salinas Morales, Erick Heredia-Olea

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Maize tortillas are a staple food in Mexico which are mostly made by nixtamalization, which includes the cooking and steeping of maize kernels in alkaline conditions. The cooking step in nixtamalization demands a lot of energy and also generates nejayote, a water pollutant, at the end of the process. The aim of this study was to reduce the cooking time by adding a maize soaking step before nixtamalization while maintaining the quality properties of masa and tortillas. Maize kernels were soaked for 36 h to increase moisture up to 36%. Then, the effect of different cooking times (0, 5, 10, 15, 20, 20, 25, 30, 35, 45-control and 50 minutes) was evaluated on viscosity profile (RVA) of masa to select the treatments with a profile similar or equal to control. All treatments were left steeping overnight and had the same milling conditions. Treatments selected were 20- and 25-min cooking times which had similar values for pasting temperature (79.23°C and 80.23°C), Maximum Viscosity (105.88 Cp and 96.25 Cp) and Final Viscosity (188.5 Cp and 174 Cp) to those of 45 min-control (77.65 °C, 110.08 Cp, and 186.70 Cp, respectively). Afterward, tortillas were produced with the chosen treatments (20 and 25 min) and for control, then were analyzed for texture, damage starch, colorimetry, thickness, and average diameter. Colorimetric analysis of tortillas only showed significant differences for yellow/blue coordinates (b* parameter) at 20 min (0.885), unlike the 25-minute treatment (1.122). Luminosity (L*) and red/green coordinates (a*) showed no significant differences from treatments with respect control (69.912 and 1.072, respectively); however, 25 minutes was closer in both parameters (73.390 and 1.122) than 20 minutes (74.08 and 0.884). For the color difference, (E), the 25 min value (3.84) was the most similar to the control. However, for tortilla thickness and diameter, the 20-minute with 1.57 mm and 13.12 cm respectively was closer to those of the control (1.69 mm and 13.86 cm) although smaller to it. On the other hand, the 25 min treatment tortilla was smaller than both 20 min and control with 1.51 mm thickness and 13.590 cm diameter. According to texture analyses, there was no difference in terms of stretchability (8.803-10.308 gf) and distance for the break (95.70-126.46 mm) among all treatments. However, for the breaking point, all treatments (317.1 gf and 276.5 gf for 25 and 20- min treatment, respectively) were significantly different from the control tortilla (392.2 gf). Results suggest that by adding a soaking step and reducing cooking time by 25 minutes, masa and tortillas obtained had similar functional and textural properties to the traditional nixtamalization process.

Keywords: tortilla, nixtamalization, corn, lime cooking, RVA, colorimetry, texture, masa rheology

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Authors: G. Ghamrawi, Y. Abunnasr, M. Fawaz, S. Yazigi

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Coastal cities are constantly exposed to environmental degradation and economic regression fueled by rapid and uncontrolled urban growth as well as continuous resource depletion. This is the case of the City of Mina in Tripoli (Lebanon), where lack of awareness to preserve social, ecological, and historical assets, coupled with the increasing development pressures, are threatening the socioeconomic status of the city residents, the quality of life and accessibility to the coast. To address these challenges, a holistic coastal urban design and planning approach was developed to analyze the environmental, political, legal, and socioeconomic context of the city. This approach aims to investigate the potential of balancing urban development with the protection and enhancement of cultural, ecological, and environmental assets under an integrated coastal zone management approach (ICZM). The analysis of Mina's different sectors adopted several tools that include direct field observation, interviews with stakeholders, analysis of available data, historical maps, and previously proposed projects. The findings from the analysis were mapped and graphically represented, allowing the recognition of character zones that become the design intervention units. Consequently, the thesis proposes an urban, city-scale intervention that identifies 6 different character zones (the historical fishing port, Abdul Wahab island, the abandoned Port Said, Hammam el Makloub, the sand beach, and the new developable area) and proposes context-specific design interventions that capitalize on the main characteristics of each zone. Moreover, the intervention builds on the institutional framework of ICZM as well as other studies previously conducted for the coast and adopts nature-based solutions with hybrid systems for providing better environmental design solutions for developing the coast. This enables the realization of an all-inclusive, well-connected shoreline with easy and free access towards the sea; a developed shoreline with an active local economy, and an improved urban environment.

Keywords: blue green infrastructure, coastal cities, hybrid solutions, integrated coastal zone management, sustainable development, urban planning

Procedia PDF Downloads 143

Authors: Aurea C. L. Lacerda, Paulo R. H. Moreno, Bruna M. P. Vianna, Cristina H. R. Serra, Airton Martin, André R. Baby, Vladi O. Consiglieri, Telma M. Kaneko

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The modified Franz cell is the most widely used model for in vitro permeation studies, however it still presents some disadvantages. Thus, some alternative methods have been developed such as Skin PAMPA, which is a bio- artificial membrane that has been applied for skin penetration estimation of xenobiotics based on HT permeability model consisting. Skin PAMPA greatest advantage is to carry out more tests, in a fast and inexpensive way. The membrane system mimics the stratum corneum characteristics, which is the primary skin barrier. The barrier properties are given by corneocytes embedded in a multilamellar lipid matrix. This layer is the main penetration route through the paracellular permeation pathway and it consists of a mixture of cholesterol, ceramides, and fatty acids as the dominant components. However, there is no consensus on the membrane composition. The objective of this work was to compare the performance among different bio-artificial membranes for studying the permeation in skin PAMPA system. Material and methods: In order to mimetize the lipid composition`s present in the human stratum corneum six membranes were developed. The membrane composition was equimolar mixture of cholesterol, ceramides 1-O-C18:1, C22, and C20, plus fatty acids C20 and C24. The membrane integrity assay was based on the transport of Brilliant Cresyl Blue, which has a low permeability; and Lucifer Yellow with very poor permeability and should effectively be completely rejected. The membrane characterization was performed using Confocal Laser Raman Spectroscopy, using stabilized laser at 785 nm with 10 second integration time and 2 accumulations. The membrane behaviour results on the PAMPA system were statistically evaluated and all of the compositions have shown integrity and permeability. The confocal Raman spectra were obtained in the region of 800-1200 cm-1 that is associated with the C-C stretches of the carbon scaffold from the stratum corneum lipids showed similar pattern for all the membranes. The ceramides, long chain fatty acids and cholesterol in equimolar ratio permitted to obtain lipid mixtures with self-organization capability, similar to that occurring into the stratum corneum. Conclusion: The artificial biological membranes studied for Skin PAMPA showed to be similar and with comparable properties to the stratum corneum.

Keywords: bio-artificial membranes, comparison, confocal Raman, skin PAMPA

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Authors: Sadao Araki, Daisuke Gondo, Satoshi Imasaka, Hideki Yamamoto

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The separation of organic compounds from aqueous solutions is a key technology for recycling valuable organic compounds and for the treatment of wastewater. The wastewater from chemical plants often contains organic compounds such as ethyl acetate (EA), methylethyl ketone (MEK) and isopropyl alcohol (IPA). In this study, we prepared hydrophobic silica membranes by a sol-gel method. We used phenyltrimethoxysilane (PhTMS), ethyltrimethoxysilan (ETMS), Propyltrimethoxysilane (PrTMS), N-butyltrimethoxysilane (BTMS), N-Hexyltrimethoxysilane (HTMS) as silica sources to introduce each functional groups on the membrane surface. Cetyltrimethyl ammonium bromide (CTAB) was used as a molecular template to create suitable pore that enable the permeation of organic compounds. These membranes with five different functional groups were characterized by SEM, FT-IR, and permporometry. Thicknesses and pore diameters of silica layer for all membrane were about 1.0 μm and about 1 nm, respectively. In other words, functional groups had an insignificant effect on the membrane thicknesses and the formation of the pore by CTAB. We confirmed the effect of functional groups on the flux and separation factor for ethyl acetate (EA), methyl ethyl ketone, acetone and 1-butanol (1-BtOH) /water mixtures. All membranes showed a high flux for ethyl acetate compared with other compounds. In particular, the hydrophobic silica membrane prepared by using BTMS showed 0.75 kg m-2 h-1 of flux for EA. For all membranes, the fluxes of organic compounds showed the large values in the order corresponding to EA > MEK > acetone > 1-BtOH. On the other hand, carbon chain length of functional groups among ETMS, PrTMS, BTMS, PrTMS and HTMS did not have a major effect on the organic flux. Although we confirmed the relationship between organic fluxes and organic molecular diameters or fugacity of organic compounds, these factors had a low correlation with organic fluxes. It is considered that these factors affect the diffusivity. Generally, permeation through membranes is based on the diffusivity and solubility. Therefore, it is deemed that organic fluxes through these hydrophobic membranes are strongly influenced by solubility. We tried to estimate the organic fluxes by Hansen solubility parameter (HSP). HSP, which is based on the cohesion energy per molar volume and is composed of dispersion forces (δd), intermolecular dipole interactions (δp), and hydrogen-bonding interactions (δh), has recently attracted attention as a means for evaluating the resolution and aggregation behavior. Evaluation of solubility for two substances can be represented by using the Ra [(MPa)1/2] value, meaning the distance of HSPs for both of substances. A smaller Ra value means a higher solubility for each substance. On the other hand, it can be estimated that the substances with large Ra value show low solubility. We established the correlation equation, which was based on Ra, of organic flux at low concentrations of organic compounds and at 295-325 K.

Keywords: hydrophobic, membrane, Hansen solubility parameter, functional group

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Authors: Komal Khan, Hafsa Zaneb, Saima Masood, Muhammad Younus, Sanan Raza

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Cigarette smoke induces many physiological and pathological changes in respiratory tract like goblet cell hyperplasia and regional distention of airspaces. It is also associated with elevation of inflammatory profiles in different airway compartments. As probiotics are generally known to promote mucosal tolerance, it was postulated that prophylactic use of probiotics can be helpful in reduction of respiratory damage induced by cigarette smoke exposure. Twenty-four adult mice were randomly divided into three groups (cigarette-smoke (CS) group, cigarette-smoke+ Lactobacillus (CS+ P) group, control (Cn) group), each having 8 mice. They were exposed to cigarette smoke for 28 days (6 cigarettes/ day for 6 days/week). Wright-Giemsa staining of bronchoalveolar lavage fluid (BALF) was performed in three mice per group. Tissue samples of trachea and lungs of 7 mice from each group were processed by paraffin embedding technique for haematoxylin & eosin (H & E) and alcian blue- periodic acid-Schiff (AB-PAS) staining. Then trachea (goblet cell number, ratio and loss of cilia) and lungs (airspace distention) were studied. The results showed that the number of goblet cells was increased in CS group as a result of defensive mechanism of the respiratory system against irritating substances. This study also revealed that the cells of CS group having acidic glycoprotein were found to be higher in quantity as compared to those containing neutral glycoprotein. However, CS + P group showed a decrease in goblet cell index due to enhanced immunity by prophylactically used probiotics. Moreover, H & E stained tracheas showed significant loss of cilia in CS group due to propelling of mucous but little loss in CS + P group because of having good protective tracheal epithelium. In lungs, protection of airspaces was also much more evident in CS+ P group as compared to CS group having distended airspaces, especially at 150um distance from terminal bronchiole. In addition, a comprehensive analysis of inflammatory cells population of BALF showed neutrophilia and eosinophilia was significantly reduced in CS+ P group. This study proved that probiotics are found to be useful for reduction of changes in micro-architecture of the respiratory system. Thus, dietary supplementation of probiotic as prophylactic measure can be useful in achieving immunomodulatory effects.

Keywords: cigarette smoke, probiotics, goblet cells, airspace enlargement, BALF

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Authors: Melanie Ostermann, Eivind Birkeland, Ying Xue, Alexander Sauter, Mihaela R. Cimpan

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Background: New reliable and robust techniques to assess biological effects of nanomaterials (NMs) in vitro are needed to speed up safety analysis and to identify key physicochemical parameters of NMs, which are responsible for their acute cytotoxicity. The central aim of this study was to validate and evaluate the applicability and reliability of an impedance-based flow cytometry (IFC) technique for the high-throughput screening of NMs. Methods: Eight inorganic NMs from the European Commission Joint Research Centre Repository were used: NM-302 and NM-300k (Ag: 200 nm rods and 16.7 nm spheres, respectively), NM-200 and NM- 203 (SiO₂: 18.3 nm and 24.7 nm amorphous, respectively), NM-100 and NM-101 (TiO₂: 100 nm and 6 nm anatase, respectively), and NM-110 and NM-111 (ZnO: 147 nm and 141 nm, respectively). The aim was to assess the biological effects of these materials on human monoblastoid (U937) cells. Dispersions of NMs were prepared as described in the NANOGENOTOX dispersion protocol and cells were exposed to NMs at relevant concentrations (2, 10, 20, 50, and 100 µg/mL) for 24 hrs. The change in electrical impedance was measured at 0.5, 2, 6, and 12 MHz using the IFC AmphaZ30 (Amphasys AG, Switzerland). A traditional toxicity assay, Trypan Blue Dye Exclusion assay, and dark-field microscopy were used to validate the IFC method. Results: Spherical Ag particles (NM-300K) showed the highest toxic effect on U937 cells followed by ZnO (NM-111 ≥ NM-110) particles. Silica particles were moderate to non-toxic at all used concentrations under these conditions. A higher toxic effect was seen with smaller sized TiO2 particles (NM-101) compared to their larger analogues (NM-100). No interferences between the IFC and the used NMs were seen. Uptake and internalization of NMs were observed after 24 hours exposure, confirming actual NM-cell interactions. Conclusion: Results collected with the IFC demonstrate the applicability of this method for rapid nanotoxicity assessment, which proved to be less prone to nano-related interference issues compared to some traditional toxicity assays. Furthermore, this label-free and novel technique shows good potential for up-scaling in directions of an automated high-throughput screening and for future NM toxicity assessment. This work was supported by the EC FP7 NANoREG (Grant Agreement NMP4-LA-2013-310584), the Research Council of Norway, project NorNANoREG (239199/O70), the EuroNanoMed II 'GEMN' project (246672), and the UH-Nett Vest project.

Keywords: cytotoxicity, high-throughput, impedance, nanomaterials

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Authors: Manami Masuda, Mariko Era, Takayoshi Kawahara, Takahide Kanyama, Hiroshi Morita

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Objectives: Fatty acid salts are a type of anionic surfactant and are produced from fatty acids and alkali. Moreover, fatty acid salts are known to have potent antibacterial activities. Acanthamoeba is ubiquitously distributed in the environment including sea water, fresh water, soil and even from the air. Although generally free-living, Acanthamoeba can be an opportunistic pathogen, which could cause a potentially blinding corneal infection known as Acanthamoeba keratitis. So, in this study, we evaluated the anti-amoeba activity of fatty acid salts and fatty acids to Acanthamoeba castellanii ATCC 30010. Materials and Methods: The antibacterial activity of 9 fatty acid salts (potassium butyrate (C4K), caproate (C6K), caprylate (C8K), caprate (C10K), laurate (C12K), myristate (C14K), oleate (C18:1K), linoleate (C18:2K), linolenate (C18:3K)) tested on cells of Acanthamoeba castellanii ATCC 30010. Fatty acid salts (concentration of 175 mM and pH 10.5) were prepared by mixing the fatty acid with the appropriate amount of KOH. The amoeba suspension mixed with KOH with a pH adjusted solution was used as the control. Fatty acids (concentration of 175 mM) were prepared by mixing the fatty acid with Tween 80 (20 %). The amoeba suspension mixed with Tween 80 (20 %) was used as the control. The anti-amoeba method, the amoeba suspension (3.0 × 104 cells/ml trophozoites) was mixed with the sample of fatty acid potassium (final concentration of 175 mM). Samples were incubated at 30°C, for 10 min, 60 min, and 180 min and then the viability of A. castellanii was evaluated using plankton counting chamber and trypan blue stainings. The minimum inhibitory concentration (MIC) against Acanthamoeba was determined using the two-fold dilution method. The MIC was defined as the minimal anti-amoeba concentration that inhibited visible amoeba growth following incubation (180 min). Results: C8K, C10K, and C12K were the anti-amoeba effect of 4 log-unit (99.99 % growth suppression of A. castellanii) incubated time for 180 min against A. castellanii at 175mM. After the amoeba, the suspension was mixed with C10K or C12K, destroying the cell membrane had been observed. Whereas, the pH adjusted control solution did not exhibit any effect even after 180 min of incubation with A. castellanii. Moreover, C6, C8, and C18:3 were the anti-amoeba effect of 4 log-unit incubated time for 60 min. C4 and C18:2 exhibited a 4-log reduction after 180 min incubation. Furthermore, the minimum inhibitory concentration (MIC) was determined. The MIC of C10K, C12K and C4 were 2.7 mM. These results indicate that C10K, C12K and C4 have high anti-amoeba activity against A. castellanii and suggest C10K, C12K and C4 have great potential for antimi-amoeba agents.

Keywords: Fatty acid salts, anti-amoeba activities, Acanthamoeba, fatty acids

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Authors: Kristina Zuzek, Xuan Xu, Awais Ikram, Richard Sheridan, Allan Walton, Saso Sturm

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Recycling of end-of-life REEs based Nd-Fe-B magnets is an important strategy for reducing the environmental dangers associated with rare-earth mining and overcoming the well-documented supply risks related to the REEs. However, challenges on their reprocessing still remain. We report on the possibility of direct electrochemical recycling and reprocessing of Nd-Fe(B)-based magnets. In this investigation, we were able first to electrochemically leach the end-of-life NdFeB magnet and to electrodeposit Nd–Fe using a 1-ethyl-3-methyl imidazolium dicyanamide ([EMIM][DCA]) ionic liquid-based electrolyte. We observed that Nd(III) could not be reduced independently. However, it can be co-deposited on a substrate with the addition of Fe(II). Using advanced TEM techniques of electron-energy-loss spectroscopy (EELS) it was shown that Nd(III) is reduced to Nd(0) during the electrodeposition process. This gave a new insight into determining the Nd oxidation state, as X-ray photoelectron spectroscopy (XPS) has certain limitations. This is because the binding energies of metallic Nd (Nd0) and neodymium oxide (Nd₂O₃) are very close, i. e., 980.5-981.5 eV and 981.7-982.3 eV, respectively, making it almost impossible to differentiate between the two states. These new insights into the electrodeposition process represent an important step closer to efficient recycling of rare piles of earth in metallic form at mild temperatures, thus providing an alternative to high-temperature molten-salt electrolysis and a step closer to deposit Nd-Fe-based magnetic materials. Further, we propose a new concept of recycling the sintered Nd-Fe-B magnets by direct recovering the 2:14:1 matrix phase. Via an electrochemical etching method, we are able to recover pure individual 2:14:1 grains that can be re-used for new types of magnet production. In the frame of physical reprocessing, we have successfully synthesized new magnets out of hydrogen (HDDR)-recycled stocks with a contemporary technique of pulsed electric current sintering (PECS). The optimal PECS conditions yielded fully dense Nd-Fe-B magnets with the coercivity Hc = 1060 kA/m, which was boosted to 1160 kA/m after the post-PECS thermal treatment. The Br and Hc were tackled further and increased applied pressures of 100 – 150 MPa resulted in Br = 1.01 T. We showed that with a fine tune of the PECS and post-annealing it is possible to revitalize the Nd-Fe-B end-of-life magnets. By applying advanced TEM, i.e. atomic-scale Z-contrast STEM combined with EDXS and EELS, the resulting magnetic properties were critically assessed against various types of structural and compositional discontinuities down to atomic-scale, which we believe control the microstructure evolution during the PECS processing route.

Keywords: electrochemistry, Nd-Fe-B, pulsed electric current sintering, recycling, reprocessing

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Authors: Rabindranath Jana, Biswajit Maity, Keka Rana

Abstract:

The most promising clean energy source is the fuel cell, since it does not generate toxic gases and other hazardous compounds. Again the direct methanol fuel cell (DMFC) is more user-friendly as it is easy to be miniaturized and suited as energy source for automobiles as well as domestic applications and portable devices. And unlike the hydrogen used for some fuel cells, methanol is a liquid that is easy to store and transport in conventional tanks. The most important part of a fuel cell is its membrane. Till now, an overall efficiency for a methanol fuel cell is reported to be about 20 ~ 25%. The lower efficiency of the cell may be due to the critical factors, e.g. slow reaction kinetics at the anode and methanol crossover. The oxidation of methanol is composed of a series of successive reactions creating formaldehyde and formic acid as intermediates that contribute to slow reaction rates and decreased cell voltage. Currently, the investigation of new anode catalysts to improve oxidation reaction rates is an active area of research as it applies to the methanol fuel cell. Surprisingly, there are very limited reports on nanostructured membranes, which are rather simple to manufacture with different tuneable compositions and are expected to allow only the proton permeation but not the methanol due to their molecular sizing effects and affinity to the membrane surface. We have developed a nanostructured fuel cell membrane from polydimethyl siloxane rubber (PDMS), ethylene methyl co-acrylate (EMA) and multi-walled carbon nanotubes (MWNTs). The effect of incorporating different proportions of f-MWNTs in polymer membrane has been studied. The introduction of f-MWNTs in polymer matrix modified the polymer structure, and therefore the properties of the device. The proton conductivity, measured by an AC impedance technique using open-frame and two-electrode cell and methanol permeability of the membranes was found to be dependent on the f-MWNTs loading. The proton conductivity of the membranes increases with increase in concentration of f-MWNTs concentration due to increased content of conductive materials. Measured methanol permeabilities at 60oC were found to be dependant on loading of f-MWNTs. The methanol permeability decreased from 1.5 x 10-6 cm²/s for pure film to 0.8 x 10-7 cm²/s for a membrane containing 0.5wt % f-MWNTs. This is due to increasing proportion of f-MWNTs, the matrix becomes more compact. From DSC melting curves it is clear that the polymer matrix with f-MWNTs is thermally stable. FT-IR studies show good interaction between EMA and f-MWNTs. XRD analysis shows good crystalline behavior of the prepared membranes. Significant cost savings can be achieved when using the blended films which contain less expensive polymers.

Keywords: fuel cell membrane, polydimethyl siloxane rubber, carbon nanotubes, proton conductivity, methanol permeability

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Authors: Saeed Rezaeian, M. Rezaee Boroon

Abstract:

Nitrogen accumulates in soils during the process of fertilizer addition to promote the plant growth. When the organic matter decomposes, the form of available nitrogen produced is in the form of nitrate, which is highly mobile. The most significant health effect of nitrate ingestion is methemoglobinemia in infants under six months of age (blue baby syndrome). The mobile nutrients, like nitrate nitrogen, are not stored in the soil as the available forms for the long periods and in large amounts. It depends on the needs for the crops such as vegetables. On the other hand, the vegetables will compete actively for nitrate nitrogen as a mobile nutrient and water. The mobile nutrients must be shared. The fewer the plants, the larger this share is for each plant. Also, this nitrate nitrogen is poisonous for the people who use these vegetables. Nitrate is converted to nitrite by the existing bacteria in the stomach and the Gastro-Intestinal (GI) tract. When nitrite is entered into the blood cells, it converts the hemoglobin to methemoglobin, which causes the anoxemia and cyanosis. The increasing use of pesticides and chemical fertilizers, especially the fertilizers with nitrates compounds, which have been common for the increased production of agricultural crops, has caused the nitrate pollution in the (soil, water, and environment). They have caused a lot of damage to humans and animals. In this research, the nitrate accumulation in different kind of vegetables such as; green pepper, tomatoes, egg plants, watermelon, cucumber, and red pepper were observed in the suburbs of Mashhad, Neisabour, and Sabzevar cities. In some of these cities, the information forms of agronomical practices collected were such as; different vegetable crops fertilizer recommendations, varieties, pesticides, irrigation schedules, etc., which were filled out by some of our colleagues in the research areas mentioned above. Analysis of the samples was sent to the soil and water laboratory in our department in Mashhad. The final results from the chemical analysis of samples showed that the mean levels of nitrates from the samples of the fruit crops in the mentioned cities above were all lower than the critical levels. These fruit crop samples were in the order of: 35.91, 8.47, 24.81, 6.03, 46.43, 2.06 mg/kg dry matter, for the following crops such as; tomato, cucumber, eggplant, watermelon, green pepper, and red pepper. Even though, this study was conducted with limited samples and by considering the mean levels, the use of these crops from the nutritional point of view will not cause the poisoning of humans.

Keywords: environmental pollution, human health, nitrate accumulations, nitrate fertilizers

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Authors: Hamsasew Hankebo Lemago, Dóra Hessz, Zoltán Erdélyi, Imre Miklós Szilágyi

Abstract:

Metal oxide inverse opals are a unique class of photocatalysts with a hierarchical structure that mimics the natural opal gemstone. They are composed of a network of interconnected pores, which provides a large surface area and efficient pathways for the transport of light and reactants. Atomic layer deposition (ALD) is a versatile technique for the synthesis of high-precision metal oxide thin films, including inverse opals. ALD allows for precise control over the thickness, composition, and morphology of the synthesized films, making it an ideal technique for the fabrication of photocatalysts with tailored properties. In this study, we report the synthesis of TiO2, ZnO, and Al2O3 inverse opal photocatalysts using thermal or plasma-enhanced ALD. The synthesized photocatalysts were characterized using a variety of techniques, including scanning electron microscopy (SEM)-energy dispersive X-ray spectroscopy (EDX), X-ray diffraction (XRD), Raman spectroscopy, photoluminescence (PL), ellipsometry, and UV-visible spectroscopy. The results showed that the ALD-synthesized metal oxide inverse opals had a highly ordered structure and a tunable pore size. The PL spectroscopy results showed low recombination rates of photogenerated electron-hole pairs, while the ellipsometry and UV-visible spectroscopy results showed tunable optical properties and band gap energies. The photocatalytic activity of the samples was evaluated by the degradation of methylene blue under visible light irradiation. The results showed that the ALD-synthesized metal oxide inverse opals exhibited high photocatalytic activity, even under visible light irradiation. The composites photocatalysts showed even higher activity than the individual metal oxide inverse opals. The enhanced photocatalytic activity of the composites can be attributed to the synergistic effect between the different metal oxides. For example, Al2O3 can act as a charge carrier scavenger, which can reduce the recombination of photogenerated electron-hole pairs. The ALD-synthesized metal oxide inverse opals and their composites are promising photocatalysts for a variety of applications, such as wastewater treatment, air purification, and energy production. For example, they can be used to remove organic pollutants from wastewater, decompose harmful gases in the air, and produce hydrogen fuel from water.

Keywords: ALD, metal oxide inverse opals, composites, photocatalysis

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Authors: Kangana Sengar, Sanjay Deb, Ramesh Dawar

Abstract:

Introduction: Ki-67 can be a useful marker in determining proliferative activity in patients with multiple myeloma (MM). However, using Ki-67 alone results in the erroneous inclusion of non-myeloma cells leading to false high counts. We have used Dual IHC (immunohistochemistry) staining with Ki-67 and CD138 to enhance specificity in assessing proliferative activity of bone marrow plasma cells. Aims and objectives: To estimate the proportion of proliferating (Ki-67 expressing) plasma cells in patients with MM and correlation of Ki-67 with other known prognostic parameters. Materials and Methods: Fifty FFPE (formalin fixed paraffin embedded) blocks of trephine biopsies of cases diagnosed as MM from 2010 to 2015 are subjected to H & E staining and Dual IHC staining for CD 138 and Ki-67. H & E staining is done to evaluate various histological parameters like percentage of plasma cells, pattern of infiltration (nodular, interstitial, mixed and diffuse), routine parameters of marrow cellularity and hematopoiesis. Clinical data is collected from patient records from Medical Record Department. Each of CD138 expressing cells (cytoplasmic, red) are scored as proliferating plasma cells (containing a brown Ki¬67 nucleus) or non¬proliferating plasma cells (containing a blue, counter-stained, Ki-¬67 negative nucleus). Ki-67 is measured as percentage positivity with a maximum score of hundred percent and lowest of zero percent. The intensity of staining is not relevant. Results: Statistically significant correlation of Ki-67 in D-S Stage (Durie & Salmon Stage) I vs. III (p=0.026) and ISS (International Staging System) Stage I vs. III (p=0.019), β2m (p=0.029) and percentage of plasma cells (p < 0.001) is seen. No statistically significant correlation is seen between Ki-67 and hemoglobin, platelet count, total leukocyte count, total protein, albumin, S. calcium, S. creatinine, S. LDH, blood urea and pattern of infiltration. Conclusion: Ki-67 index correlated with other known prognostic parameters. However, it is not determined routinely in patients with MM due to little information available regarding its relevance and paucity of studies done to correlate with other known prognostic factors in MM patients. To the best of our knowledge, this is the first study in India using Dual IHC staining for Ki-67 and CD138 in MM patients. Routine determination of Ki-67 will help to identify patients who may benefit with more aggressive therapy. Recommendation: In this study follow up of patients is not included, and the sample size is small. Studying with larger sample size and long follow up is advocated to prognosticate Ki-67 as a marker of survival in patients with multiple myeloma.

Keywords: bone marrow, dual IHC, Ki-67, multiple myeloma

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