Search results for: calcination and methylene blue

Authors: Aruma Baduge Kithma De Silva

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Haskap (Lonicera caerulea L.), also known as blue honeysuckle, is a newly commercialized berry crop in Canada. Haskap berries are rich in polyphenols, including, anthocyanins, which are known for potential health-promoting properties. Cyanidin-3-O-glucoside (C3G) is the most abundant anthocyanin of haskap berries. The compound C3G has the ability to reduce the risk of type 2 diabetes (T2D), which has become an increasingly common health issue around the world. The T2D is characterized as a metabolic disorder of hyperglycemia and insulin resistance. It has been demonstrated that C3G has anti-diabetic effects through several ways, including inhibition of dipeptidyl peptidase-4 (DPP-4), reduction of gluconeogenesis, improvement in insulin sensitivity, and inhibition of activities of carbohydrate hydrolyzing enzymes, including α-amylase and α-glucosidase. The goal of this study was to investigate the influence of variety and harvests maturity of haskap on C3G, other fruit quality characteristics and anti-diabetic activities of haskap berries using in vitro studies. The polyphenols present in four commercially grown haskap cultivars, Aurora, Rebecca, Larissa, and Evie harvested at five harvesting dates (H1-H5) apart from 2-3 days, were extracted separately. High-performance liquid chromatography electrospray ionization mass spectrometry (HPLC-ESI-MS) analyzes of polyphenols revealed that haskap berries contain predominantly anthocyanins, flavonols, flavan-3-ols, and phenolic acids. The compound C3G was the most prominent anthocyanin, which is available in approximately 79% of total anthocyanin in four cultivars. The Larissa at H5 contained the highest C3G content. The antioxidant capacity of Evie at H5 was greater than other cultivars. Furthermore, Larissa H5 showed the greatest inhibition of carbohydrate hydrolyzing enzymes including alpha-glucosidase and alpha-amylase. In conclusion, the haskap variety and harvesting date influenced the polyphenol composition and biological properties. The variety Larissa, at H5 harvesting date, contained the highest polyphenol content and the ability of inhibition of the carbohydrate hydrolyzing enzyme as well as DPP4 enzyme in order to reduce type 2 diabetes.

Keywords: anthocyanin, Haskap, type 2 diabetes, polyphenol

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Authors: Girija Jha

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This paper investigates the need of cotton to integrate sustainability. The methodology used in the paper is to do secondary research to find out the various environmental implications of cotton as textile material across its life cycle and try to look at ways and possibilities of minimizing its ecological footprint. Cotton is called ‘The Fabric of Our Lives’. History is replete with examples where this fabric used to be more than a fabric of lives. It used to be a miracle fabric, a symbol India’s pride and social Movement of Swaraj, Gandhijee’s clarion call to self reliance. Cotton is grown in more than 90 countries across the globe on 2.5 percent of the world's arable land in countries like China, India, United States, etc. accounting for almost three fourth of global production. But cotton as a raw material has come under the scanner of sustainability experts because of myriad reasons a few have been discussed here. It may take more than 20,000 liters of water to produce 1kg of cotton. Cotton harvest is primarily done from irrigated land which leads to Salinization and depletion of local water reservoirs, e.g., Drying up of Aral Sea. Cotton is cultivated on 2.4% of total world’s crop land but accounts for 24% usage of insecticide and shares the blame of 11% usage of pesticides leading to health hazards and having an alarmingly dangerous impact on the ecosystem. One of the possible solutions to these problems as proposed was GM, Genetically Modified cotton crop. However, use of GM cotton is still debatable and has many ethical issues. The practice of mass production and increasing consumerism and especially fast fashion has been major culprits to disrupt this delicate balance. Disposable fashion or fast fashion is on the rise and cotton being one of the major choices adds on to the problem. Denims – made of cotton and have a strong fashion statement and the washes being an integral part of their creation they share a lot of blame. These are just a few problems listed. Today Sustainability is the need of the hour and it is inevitable to incorporate have major changes in the way we cultivate and process cotton to make it a sustainable choice. The answer lies in adopting minimalism and boycotting fast fashion, in using Khadi, in saying no to washed denims and using selvedge denims or using better methods of finishing the washed out fabric so that the environment does not bleed blue. Truly, the answer lies in integrating state of art technology with age old sustainable practices so that the synergy of the two may help us come out of the vicious circle.

Keywords: cotton, sustainability, denim, Khadi

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Authors: M. Mutoti, J. Gumbo, A. Jideani

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Cyanobacteria or blue green algae have been part of the human diet for thousands of years. Cyanobacteria can multiply quickly in surface waters and form blooms when favorable conditions prevail, such as high temperature, intense light, high pH, and increased availability of nutrients, especially phosphorous and nitrogen, artificially released by anthropogenic activities. Consumption of edible cyanotoxins such as Spirulina may reduce risks of cataracts and age related macular degeneration. Sulfate polysaccharides exhibit antitumor, anticoagulant, anti-mutagenic, anti-inflammatory, antimicrobial, and even antiviral activity against HIV, herpes, and hepatitis. In humans, exposure to cyanotoxins can occur in various ways; however, the oral route is the most important. This is mainly through drinking water, or by eating contaminated foods; it may even involve ingesting water during recreational activities. This paper seeks to present a review on cyanobacteria/cyanotoxin contamination of water and food and implications for human health. In particular, examining the water quality used during maize seed that passes through mill grinding processes. In order to fulfil the objective, this paper starts with the theoretical framework on cyanobacteria contamination of food that will guide review of the present paper. A number of methods for decontaminating cyanotoxins in food is currently available. Therefore, physical, chemical, and biological methods for treating cyanotoxins are reviewed and compared. Furthermore, methods that are utilized for detecting and identifying cyanobacteria present in water and food were also informed in this review. This review has indicated various routes through which humans can be exposed to cyanotoxins. Accumulation of cyanotoxins, mainly microcystins, in food has raised an awareness of the importance of food as microcystins exposure route to human body. Therefore, this review demonstrates the importance of expanding research on cyanobacteria/cyanotoxin contamination of water and food for water treatment and water supply management, with focus on examining water for domestic use. This will help providing information regarding the prevention or minimization of contamination of water and food, and also reduction or removal of contamination through treatment processes and prevention of recontamination in the distribution system.

Keywords: biofilm, cyanobacteria, cyanotoxin, food contamination

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Authors: F. Portet-Koltalo, Y. Tian, I. Berger, C. Boulanger-Lecomte, A. Benamar, N. Machour

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Sediments are complex environments which can accumulate a great variety of persistent toxic contaminants such as polychlorobiphenyles (PCBs), polycyclic aromatic hydrocarbons (PAHs) and some of their more toxic degradation metabolites such as hydroxylated PAHs (OH-PAHs). Owing to their composition, fine clayey sediments can be more difficult to extract than soils using conventional solvent extraction processes. So this study aimed to compare the potential of MSPD (matrix solid phase dispersive extraction) to extract PCBs, PAHs and OH-PAHs, in comparison with microwave assisted extraction (MAE). Methodologies: MAE extraction with various solvent mixtures was used to extract PCBs, PAHs and OH-PAHs from sediments in two runs, followed by two GC-MS analyses. MSPD consisted in crushing the dried sediment with dispersive agents, introducing the mixture in cartridges and eluting the target compounds with an appropriate volume of selected solvents. So MSPD combined with cartridges containing MIPs (molecularly imprinted polymers) designed for OH-PAHs was used to extract the three families of target compounds in only one run, followed by parallel analyses in GC-MS for PAHs/PCBs and HPLC-FLD for OH-PAHs. Results: MAE extraction was optimized to extract from clayey sediments, in two runs, PAHs/PCBs in one hand and OH-PAHs in the other hand. Indeed, the best conditions of extractions (mixtures of extracting solvents, temperature) were different if we consider the polarity and the thermodegradability of the different families of target contaminants: PAHs/PCBs were better extracted using an acetone/toluene 50/50 mixture at 130°C whereas OH-PAHs were better extracted using an acetonitrile/toluene 90/10 mixture at 100°C. Moreover, the two consecutive GC-MS analyses contributed to double the total analysis time. A matrix solid phase dispersive (MSPD) extraction procedure was also optimized, with the first objective of increasing the extraction recovery yields of PAHs and PCBs from fine-grained sediment. The crushing time (2-10 min), the nature of the dispersing agents added for purifying and increasing the extraction yields (Florisil, octadecylsilane, 3-chloropropyle, 4-benzylchloride), the nature and the volume of eluting solvents (methylene chloride, hexane, hexane/acetone…) were studied. It appeared that in the best conditions, MSPD was a better extraction method than MAE for PAHs and PCBs, with respectively, mean increases of 8.2% and 71%. This method was also faster, easier and less expensive. But the other advantage of MSPD was that it allowed to introduce easily, just after the first elution process of PAHs/PCBs, a step permitting the selective recovery of OH-PAHs. A cartridge containing MIPs designed for phenols was coupled to the cartridge containing the dispersed sediment, and various eluting solvents, different from those used for PAHs and PCBs, were tested to selectively concentrate and extract OH-PAHs. Thereafter OH-PAHs could be analyzed at the same time than PAHs and PCBs: the OH-PAH extract could be analyzed with HPLC-FLD, whereas the PAHs/PCBs extract was analyzed with GC-MS, adding only few minutes more to the total duration of the analytical process. Conclusion: MSPD associated to MIPs appeared to be an easy, fast and low expensive method, able to extract in one run a complex mixture of toxic apolar and more polar contaminants present in clayey fine-grained sediments, an environmental matrix which is generally difficult to analyze.

Keywords: contaminated fine-grained sediments, matrix solid phase dispersive extraction, microwave assisted extraction, molecularly imprinted polymers, multi-pollutant analysis

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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: Claudio Clemente, Valentina Gargiulo, Alessio Occhicone, Giovanni Piero Pepe, Giovanni Ausanio, Michela Alfè

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Volatile organic compound (VOC) emissions represent a serious risk to human health and the integrity of the ecosystems, especially at high concentrations. For this reason, it is very important to continuously monitor environmental quality and develop fast and reliable portable sensors to allow analysis on site. Chemiresistors have become promising candidates for VOC sensing as their ease of fabrication, variety of suitable sensitive materials, and simple sensing data. A chemoresistive gas sensor is a transducer that allows to measure the concentration of an analyte in the gas phase because the changes in resistance are proportional to the amount of the analyte present. The selection of the sensitive material, which interacts with the target analyte, is very important for the sensor performance. The most used VOC detection materials are metal oxides (MOx) for their rapid recovery, high sensitivity to various gas molecules, easy fabrication. Their sensing performance can be improved in terms of operating temperature, selectivity, and detection limit. Metal-organic frameworks (MOFs) have attracted a lot of attention also in the field of gas sensing due to their high porosity, high surface area, tunable morphologies, structural variety. MOFs are generated by the self-assembly of multidentate organic ligands connecting with adjacent multivalent metal nodes via strong coordination interactions, producing stable and highly ordered crystalline porous materials with well-designed structures. However, most MOFs intrinsically exhibit low electrical conductivity. To improve this property, MOFs can be combined with organic and inorganic materials in a hybrid fashion to produce composite materials or can be transformed into more stable structures. MOFs, indeed, can be employed as the precursors of metal oxides with well-designed architectures via the calcination method. The MOF-derived MOx partially preserved the original structure with high surface area and intrinsic open pores, which act as trapping centers for gas molecules, and showed a higher electrical conductivity. Core-shell heterostructures, in which the surface of a metal oxide core is completely coated by a MOF shell, forming a junction at the core-shell heterointerface, can also be synthesized. Also, nanocomposite in which MOF structures are intercalated with graphene related materials can also be produced, and the conductivity increases thanks to the high mobility of electrons of carbon materials. As MOF structures, zinc-based MOFs belonging to the ZIF family were selected in this work. Several Zn-based materials based and/or derived from MOFs were produced, structurally characterized, and arranged in a chemo resistive architecture, also exploring the potentiality of different approaches of sensing layer deposition based on PLD (pulsed laser deposition) and, in case of thermally labile materials, MAPLE (Matrix Assisted Pulsed Laser Evaporation) to enhance the adhesion to the support. The sensors were tested in a controlled humidity chamber, allowing for the possibility of varying the concentration of ethanol, a typical analyte chosen among the VOCs for a first survey. The effect of heating the chemiresistor to improve sensing performances was also explored. Future research will focus on exploring new manufacturing processes for MOF-based gas sensors with the aim to improve sensitivity, selectivity and reduce operating temperatures.

Keywords: chemiresistors, gas sensors, graphene related materials, laser deposition, MAPLE, metal-organic frameworks, metal oxides, nanocomposites, sensing performance, transduction mechanism, volatile organic compounds

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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: 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: 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: Gloria M. Ntuba-Jua, Afui M. Mih, Eneke E. T. Bechem

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Prunus africana (Hook. F.) Kalkman, commonly known as Pygeum or African cherry belongs to the Rosaceae family. It is a medium to large, evergreen tree with a spreading crown of 10 to 20 m. It is used by the traditional medical practitioners for the treatment of over 45ailments in Cameroon and sub-Sahara Africa. In modern medicine, it is used in the treatment of benign prostrate hyperplasia (BPH), prostate gland hypertrophy (enlarged prostate glands). This is possible because of its ability to produce some secondary metabolites which are believed to have bioactivity against these ailments. The ready international market for the sale of Prunus bark, uncontrolled exploitation, illegal harvesting using inappropriate techniques and poor timing of harvesting have contributed enormously to making the plant endangered. It is known to harbor a large number of endophytic fungi with the potential to produce similar secondary metabolites as the parent plant. Alternative sourcing of medicinal principles through endophytic fungi requires succinct knowledge of the endophytic fungi. This will serve as a conservation measure for Prunus africana by reducing dependence on Prunus bark for such metabolites. This work thus sought to compare the production of some major secondary metabolites produced by P. africana and some of its associated endophytic fungi. The leaves and stem bark of the plant from different provenances were soaked in methanol for 72 hrs to yield the methanolic crude extract. The phytochemical screening of the methanolic crude extracts using different standard procedures revealed the presence of tannins, flavonoids, terpenoids, saponins, phenolics and steroids. Pure cultures of some predominantly isolated endophyte species from the difference Prunus provenances such as Curvularia sp, and Morphospecies P001 were also grown in Potato Dextrose Broth (PDB) for 21 days and later extracted with Methylene dichloride (MDC) solvent after 24hrs to produce crude culture extracts. Qualitative assessment of crude culture extracts showed the presence of tannins, terpenoids, phenolics and steroids particularly β-Sitosterol, (a major bioactive metabolite) as did the plant tissues. Qualitative analysis by thin layer chromatography (TLC) was done to confirm and compare the production of β-Sitosterol (as marker compounds) in the crude extracts of the plant and endophyte. Samples were loaded on TLC silica gel aluminium barked plate (Kieselgel 60 F254, 0.2 mm, Merck) using acetone/hexane, (3.0:7.0) solvent system. They were visualized under an ultra violet lamp (UV254 and UV360). TLC revealed that leaves had a higher concentration of β-sitosterol in terms of band intensity than stem barks from the different provenances. The intensity of β-sitosterol bands in the culture extracts of endophytes was comparable to the plant extracts except for Curvularia sp (very minute) whose band was very faint. The ability of these fungi to make β-sitosterol was confirmed by TLC analysis with the compound having chromatographic properties (retention factor) similar to those of β-sitosterol standard. The ability of these major endophytes to produce secondary metabolites similar to the host has therefore been demonstrated. There is, therefore, the potential of developing the in vitro production system of Prunus secondary metabolites thereby enhancing its conservation.

Keywords: Caneroon, endophytic fungi, Prunus africana, secondary metabolite

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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 145

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: 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

Procedia PDF Downloads 362

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: Saeed Rezaeian, M. Rezaee Boroon

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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: Kangana Sengar, Sanjay Deb, Ramesh Dawar

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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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Authors: Jovana Vukcevic, Sanja Pekovic, Djurdjica Perovic, Tatjana Stanovcic

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This paper presents a critical inquiry into the role of uncomfortable heritage in nation branding with the particular focus on the specificities of the politics of memory, forgetting and revisionism in the post-communist post-Yugoslavia. It addresses legacies of unwanted, ambivalent or unacknowledged past and different strategies employed by the former-Yugoslav states and private actors in “rebranding” their heritage, ensuring its preservation, but re-contextualizing the narrative of the past through contemporary tourism practices. It questions the interplay between nostalgia, heritage and market, and the role of heritage in polishing the history of totalitarian and authoritarian regimes in the Balkans. It argues that in post-socialist Yugoslavia, the necessity to limit correlations with former ideology and the use of the commercial brush in shaping a marketable version of the past instigated the emergence of the profit-oriented heritage practices. Building on that argument, the paper addresses these issues as “commodification” and “disneyfication” of Balkans’ ambivalent heritage, contributing to the analysis of changing forms of memorialisation and heritagization practices in Europe. It questions the process of ‘coming to terms with the past’ through marketable forms of heritage tourism, fetching the boundary between market-driven nostalgia and state-imposed heritage policies. In order to analyse plurality of ways of dealing with controversial, ambivalent and unwanted heritage of dictatorships in the Balkans, the paper considers two prominent examples of heritage commodification in Serbia and Montenegro, and the re-appropriations of those narratives for the nation branding purposes. The first one is the story of the Tito’s Blue Train, the landmark of the socialist past and the symbol of Yugoslavia which has nowadays being used for birthday parties and marriage celebrations, while the second emphasises the unusual business arrangement turning the fortress Mamula, former concentration camp through the Second World War, into a luxurious Mediterranean resort. Questioning how the ‘uneasy’ past was acknowledged and embedded into the official heritage institutions and tourism practices, study examines the changing relation towards the legacies of dictatorships, inviting us to rethink the economic models of the things past. Analysis of these processes should contribute to better understanding of the new mnemonics strategies and (converging?) ways of ‘doing’ past in Europe.

Keywords: commodification, heritage tourism, totalitarianism, Serbia, Montenegro

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Authors: Bahaa Eldin A. Fouda, Khaled N. Yossef, Mohamed Elhosseny, Ahmed Lotfy, Mohamed Salama, Mohamed Sobh

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Developmental neurotoxicity (DNT) entails the toxic effects imparted by various chemicals on the brain during the early childhood period. As human brains are vulnerable during this period, various chemicals would have their maximum effects on brains during early childhood. Some toxicants have been confirmed to induce developmental toxic effects on CNS e.g. lead, however; most of the agents cannot be identified with certainty due the defective nature of predictive toxicology models used. A novel alternative method that can overcome most of the limitations of conventional techniques is the use of 3D neurospheres system. This in-vitro system can recapitulate most of the changes during the period of brain development making it an ideal model for predicting neurotoxic effects. In the present study, we verified the possible DNT of epoxomicin which is a naturally occurring selective proteasome inhibitor with anti-inflammatory activity. Rat neural progenitor cells were isolated from rat embryos (E14) extracted from placental tissue. The cortices were aseptically dissected out from the brains of the fetuses and the tissues were triturated by repeated passage through a fire-polished constricted Pasteur pipette. The dispersed tissues were allowed to settle for 3 min. The supernatant was, then, transferred to a fresh tube and centrifuged at 1,000 g for 5 min. The pellet was placed in Hank’s balanced salt solution cultured as free-floating neurospheres in proliferation medium. Two doses of epoxomicin (1µM and 10µM) were used in cultured neuropsheres for a period of 14 days. For proliferation analysis, spheres were cultured in proliferation medium. After 0, 4, 5, 11, and 14 days, sphere size was determined by software analyses. The diameter of each neurosphere was measured and exported to excel file further to statistical analysis. For viability analysis, trypsin-EDTA solution were added to neurospheres for 3 min to dissociate them into single cells suspension, then viability evaluated by the Trypan Blue exclusion test. Epoxomicin was found to affect proliferation and viability of neuropsheres, these effects were positively correlated to doses and progress of time. This study confirms the DNT effects of epoxomicin on 3D neurospheres model. The effects on proliferation suggest possible gross morphologic changes while the decrease in viability propose possible focal lesion on exposure to epoxomicin during early childhood.

Keywords: neural progentor cells, epoxomicin, neurosphere, medical and health sciences

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Authors: Julie Anne Braun, Leonardo D. da Fonseca, Gerson C. Parreira, Ricardo J. E. Andrade

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Polyurethanes (PU) are multifunctional polymers used across various industries. In construction, thermosetting polyurethanes are applied as coatings for flooring, paints, and waterproofing. They are widely specified in Brazil for waterproofing concrete structures like roof slabs and parking decks. Applied to concrete, they form a fully adhered membrane, providing a protective barrier with low water absorption, high chemical resistance, impermeability to liquids, and low vapor permeability. Their mechanical properties, including tensile strength (1 to 35 MPa) and Shore A hardness (83 to 88), depend on resin molecular weight and functionality, often using Methylene diphenyl diisocyanate. PU production, reliant on fossil-derived isocyanates and polyols, contributes significantly to carbon emissions. Sustainable alternatives, such as biopolyurethanes from renewable sources, are needed. Castor oil is a viable option for synthesizing sustainable polyurethanes. As a bio-based feedstock, castor oil is extensively cultivated in Brazil, making it a feasible option for the national market and ranking third internationally. This study aims to develop and characterize castor oil-based biopolyurethane for high-performance waterproofing and coating applications. A comparative analysis between castor oil-based PU and polyether polyol-based PU was conducted. Mechanical tests (tensile strength, Shore A hardness, abrasion resistance) and surface properties (contact angle, water absorption) were evaluated. Thermal, chemical, and morphological properties were assessed using thermogravimetric analysis (TGA), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). The results demonstrated that both polyurethanes exhibited high mechanical strength. Specifically, the tensile strength for castor oil-based PU was 19.18 MPa, compared to 12.94 MPa for polyether polyol-based PU. Similarly, the elongation values were 146.90% for castor oil-based PU and 135.50% for polyether polyol-based PU. Both materials exhibited satisfactory performance in terms of abrasion resistance, with mass loss of 0.067% for castor oil PU and 0.043% for polyether polyol PU and Shore A hardness values of 89 and 86, respectively, indicating high surface hardness. The results of the water absorption and contact angle tests confirmed the hydrophilic nature of polyether polyol PU, with a contact angle of 58.73° and water absorption of 2.53%. Conversely, the castor oil-based PU exhibited hydrophobic properties, with a contact angle of 81.05° and water absorption of 0.45%. The results of the FTIR analysis indicated the absence of a peak around 2275 cm-1, which suggests that all of the NCO groups were consumed in the stoichiometric reaction. This conclusion is supported by the high mechanical test results. The TGA results indicated that polyether polyol PU demonstrated superior thermal stability, exhibiting a mass loss of 13% at the initial transition (around 310°C), in comparison to castor oil-based PU, which experienced a higher initial mass loss of 25% at 335°C. In summary, castor oil-based PU demonstrated mechanical properties comparable to polyether polyol PU, making it suitable for applications such as trafficable coatings. However, its higher hydrophobicity makes it more promising for watertightness. Increasing environmental concerns necessitate reducing reliance on non-renewable resources and mitigating the environmental impacts of polyurethane production. Castor oil is a viable option for sustainable polyurethanes, aligning with emission reduction goals and responsible use of natural resources.

Keywords: polyurethane, castor oil, sustainable, waterproofing, construction industry

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Authors: Samuel Munaf, Kevin Swingler, Franz Brülisauer, Anthony O’Hare, George Gunn, Aaron Reeves

Abstract:

Background: Avian influenza, more commonly known as Bird flu, is a viral zoonotic respiratory disease stemming from various species of poultry, including pets and migratory birds. Researchers have purported that the accessibility of health information online, in addition to the low-cost data collection methods the internet provides, has revolutionized the methods in which epidemiological and disease surveillance data is utilized. This paper examines the feasibility of using internet data sources, such as Twitter and livestock forums, for the early detection of the avian flu outbreak, through the use of text mining algorithms and social network analysis. Methods: Social media mining was conducted on Twitter between the period of 01/01/2021 to 31/12/2021 via the Twitter API in Python. The results were filtered firstly by hashtags (#avianflu, #birdflu), word occurrences (avian flu, bird flu, H5N1), and then refined further by location to include only those results from within the UK. Analysis was conducted on this text in a time-series manner to determine keyword frequencies and topic modeling to uncover insights in the text prior to a confirmed outbreak. Further analysis was performed by examining clinical signs (e.g., swollen head, blue comb, dullness) within the time series prior to the confirmed avian flu outbreak by the Animal and Plant Health Agency (APHA). Results: The increased search results in Google and avian flu-related tweets showed a correlation in time with the confirmed cases. Topic modeling uncovered clusters of word occurrences relating to livestock biosecurity, disposal of dead birds, and prevention measures. Conclusions: Text mining social media data can prove to be useful in relation to analysing discussed topics for epidemiological surveillance purposes, especially given the lack of applied research in the veterinary domain. The small sample size of tweets for certain weekly time periods makes it difficult to provide statistically plausible results, in addition to a great amount of textual noise in the data.

Keywords: veterinary epidemiology, disease surveillance, infodemiology, infoveillance, avian influenza, social media

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Authors: Andrea M. Rivas-Castillo, Marlenne Gómez-Ramirez, Isela Rodríguez-Pozos, Norma G. Rojas-Avelizapa

Abstract:

Spent catalysts are considered as hazardous residues of major concern, mainly due to the simultaneous presence of several metals in elevated concentrations. Although hydrometallurgical, pyrometallurgical and chelating agent methods are available to remove and recover some metals contained in spent catalysts; these procedures generate potentially hazardous wastes and the emission of harmful gases. Thus, biotechnological treatments are currently gaining importance to avoid the negative impacts of chemical technologies. To this end, diverse microorganisms have been used to assess the removal of metals from spent catalysts, comprising bacteria, archaea and fungi, whose resistance and metal uptake capabilities differ depending on the microorganism tested. Acidophilic sulfur oxidizing bacteria have been used to investigate the biotreatment and extraction of valuable metals from spent catalysts, namely Acidithiobacillus thiooxidans and Acidithiobacillus ferroxidans, as they present the ability to produce leaching agents such as sulfuric acid and sulfur oxidation intermediates. In the present work, the ability of A. thiooxidans DSM 26636 for the bioleaching of metals contained in five different spent catalysts was assessed by growing the culture in modified Starkey mineral medium (with elemental sulfur at 1%, w/v), and 1% (w/v) pulp density of each residue for up to 21 days at 30 °C and 150 rpm. Sulfur-oxidizing activity was periodically evaluated by determining sulfate concentration in the supernatants according to the NMX-k-436-1977 method. The production of sulfuric acid was assessed in the supernatants as well, by a titration procedure using NaOH 0.5 M with bromothymol blue as acid-base indicator, and by measuring pH using a digital potentiometer. On the other hand, Inductively Coupled Plasma - Optical Emission Spectrometry was used to analyze metal removal from the five different spent catalysts by A. thiooxidans DSM 26636. Results obtained show that, as could be expected, sulfuric acid production is directly related to the diminish of pH, and also to highest metal removal efficiencies. It was observed that Al and Fe are recurrently removed from refinery spent catalysts regardless of their origin and previous usage, although these removals may vary from 9.5 ± 2.2 to 439 ± 3.9 mg/kg for Al, and from 7.13 ± 0.31 to 368.4 ± 47.8 mg/kg for Fe, depending on the spent catalyst proven. Besides, bioleaching of metals like Mg, Ni, and Si was also obtained from automotive spent catalysts, which removals were of up to 66 ± 2.2, 6.2±0.07, and 100±2.4, respectively. Hence, the data presented here exhibit the potential of A. thiooxidans DSM 26636 for the simultaneous bioleaching of metals contained in spent catalysts from diverse provenance.

Keywords: bioleaching, metal removal, spent catalysts, Acidithiobacillus thiooxidans

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Authors: Jia-Chao Wang

Abstract:

A model of the universe without invoking space expansion is proposed to explain the observed redshift-distance relation and the cosmic microwave background radiation (CMB). The main hypothesized feature of the model is that photons traveling in space interact with the CMB photon gas. This interaction causes the photons to gradually lose energy through dissipation and, therefore, experience redshift. The interaction also causes some of the photons to be scattered off their track toward an observer and, therefore, results in beam intensity attenuation. As observed, the CMB exists everywhere in space and its photon density is relatively high (about 410 per cm³). The small average energy of the CMB photons (about 6.3×10⁻⁴ eV) can reduce the energies of traveling photons gradually and will not alter their momenta drastically as in, for example, Compton scattering, to totally blur the images of distant objects. An object moving through a thermalized photon gas, such as the CMB, experiences a drag. The cause is that the object sees a blue shifted photon gas along the direction of motion and a redshifted one in the opposite direction. An example of this effect can be the observed CMB dipole: The earth travels at about 368 km/s (600 km/s) relative to the CMB. In the all-sky map from the COBE satellite, radiation in the Earth's direction of motion appears 0.35 mK hotter than the average temperature, 2.725 K, while radiation on the opposite side of the sky is 0.35 mK colder. The pressure of a thermalized photon gas is given by Pγ = Eγ/3 = αT⁴/3, where Eγ is the energy density of the photon gas and α is the Stefan-Boltzmann constant. The observed CMB dipole, therefore, implies a pressure difference between the two sides of the earth and results in a CMB drag on the earth. By plugging in suitable estimates of quantities involved, such as the cross section of the earth and the temperatures on the two sides, this drag can be estimated to be tiny. But for a photon traveling at the speed of light, 300,000 km/s, the drag can be significant. In the present model, for the dissipation part, it is assumed that a photon traveling from a distant object toward an observer has an effective interaction cross section pushing against the pressure of the CMB photon gas. For the attenuation part, the coefficient of the typical attenuation equation is used as a parameter. The values of these two parameters are determined by fitting the 748 µ vs. z data points compiled from 643 supernova and 105 γ-ray burst observations with z values up to 8.1. The fit is as good as that obtained from the lambda cold dark matter (ΛCDM) model using online cosmological calculators and Planck 2015 results. The model can be used to interpret Hubble's constant, Olbers' paradox, the origin and blackbody nature of the CMB radiation, the broadening of supernova light curves, and the size of the observable universe.

Keywords: CMB as the lowest energy state, model of the universe, origin of CMB in a static universe, photon-CMB photon gas interaction

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Authors: Naomi Sakamoto, Eisuke Fukuma, Mika Nashimoto, Yoshitomo Koshida

Abstract:

Background: Selective localization of the metastatic lymph node with clip and removal of clipped nodes with sentinel lymph node (SLN), known as targeted axillary dissection (TAD), reduced false-negative rates (FNR) of SLN biopsy (SLNB) after neoadjuvant chemotherapy (NAC). For the patients who achieved nodal pathologic complete response (pCR), accurate staging of axilla by TAD lead to omit axillary lymph node dissection (ALND), decreasing postoperative arm morbidity without a negative effect on overall survival. This study aimed to investigate the ultrasound (US) identification rate and success removal rate of two kinds of ultrasound-visible clips placed in metastatic lymph nodes during TAD procedure. Methods: This prospective study was conducted using patients with clinically T1-3, N1, 2, M0 breast cancer undergoing NAC followed by surgery. A US-visible clip was placed in the suspicious lymph node under US guidance before neoadjuvant chemotherapy. Before surgery, US examination was performed to evaluate the detection rate of clipped node. During the surgery, the clipped node was removed using several localization techniques, including hook-wire localization, dye-injection, or fluorescence technique, followed by a dual-technique SLNB and resection of palpable nodes if present. For the fluorescence technique, after injection of 0.1-0.2 mL of indocyanine green dye (ICG) into the clipped node, ICG fluorescent imaging was performed using the Photodynamic Eye infrared camera (Hamamatsu Photonics k. k., Shizuoka, Japan). For the dye injection method, 0.1-0.2 mL of pyoktanin blue dye was injected into the clipped node. Results: A total of 29 patients were enrolled. Hydromark™ breast biopsy site markers (Hydromark, T3 shape; Devicor Medical Japan, Tokyo, Japan) was used in 15patients, whereas a UltraCor™ Twirl™ breast marker (Twirl; C.R. Bard, Inc, NJ, USA) was placed in 14 patients. US identified the clipped node marked with the UltraCore Twirl in 100% (14/14) and with the Hydromark in 93.3% (14/15, p = ns). Success removal of clipped node marked with the UltraCore Twirl was achieved in 100% (14/14), whereas the node marked with the Hydromark was removed in 80% (12/15) (p = ns). Conclusions: The ultrasound identification rate differed between the two types of ultrasound-visible clips, which also affected the success removal rate of clipped nodes. Labelling the positive node with a US-highly-visible clip allowed successful TAD.

Keywords: breast cancer, neoadjuvant chemotherapy, targeted axillary dissection, breast tissue marker, clip

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