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1.
Wang F  Li Y  Chen L  Chen D  Wu X  Wang H 《Biomicrofluidics》2012,6(1):14120-1412012
Hyperthermia can be used as an adjunctive method of chemotherapy, radiotherapy, and gene therapy to improve cancer treatment. In this study, we investigate the hyperthermic cell death of cervix cancer CaSki cells in a microchannel integrated with a directional heating scheme. Heat was applied from the inner end to the outer end of the channel and a temperature distribution from 60 °C to 30 °C was established. A three dimensional (3D) numerical model was conducted for the heat transfer simulation, based on which a simple fitting method was proposed to easily estimate the temperature distribution along the channel. Cell death along the channel was mapped 22 h after the heating treatment by dual fluorescent labeling and phase-contrast microscopy imaging. Upstream, where the temperature is higher than 42 °C, we observe necrotic death, late-stage and early stage apoptotic death in sequence along the channel. Downstream and in the middle of the channel, where the temperature is lower than 42 °C, significant cell detachment was noted. Vigorous detachment was observed even in the non-hyperthermic zone (temperature lower than 37 °C), which we believe is due to the direct effect of the hyperthermic zones (higher than 37 °C). The present work not only gives a vivid map of cell responses under a temperature gradient, but also reveals the potential interactions of the heated tumor cells and non-heated tumor cells, which are seldom investigated in conventional petri-dish experiments.  相似文献   

2.
Quality control (QC) materials are crucial for internal quality control (IQC) and external quality assessment scheme (EQAS). However, many developing countries are disadvantaged by unavailability and high cost of commercial control material. Therefore, preparing home-made lyophilized human serum will be cost effective for used as a QC material in Bhutan. We prepared lyophilized QC material using serum collected from Bhutanese volunteers. The stability of lyophilized serum was studied at 3 selected conditions by analyzing at certain intervals. The results were statistically compared with initial values. The significant p values (<0.05) were seen in glucose, BUN, ALT, total bilirubin and protein at 2–8 °C but no significant difference were observed at −20 °C at the end of 9 months. We concluded that, home-made lyophilized human serum prepared without stabilizers could be used at least up to 9 months if stored at −20 °C and 7 months at 2–8 °C. Stabilizers and additives are necessary if the materials are to be used longer than 7–9 months.  相似文献   

3.
Thalassemia has been recognized by the World Health Organization as important inherited disorders principally impacting on the populations of low income countries. In this report, the prevalence of common β-thalassemia mutations in India was defined in 126 β-thalassemia carrier subjects in a western Indian population mainly from the south-western Maharashtra. The six most common β-thalassemia mutations were detected, which included IVS I-5 (G–C), IVS I-1 (G–T), codon 8–9 (+G), codon 41/42 (–TCTT), Codon 15 (G–A), and 619 bp deletion at 3′ end of β-globin gene. These mutations accounted for 93.66 % in 126 β-thalassemia carrier subjects and 6.34 % remained uncharacterized. Out of 126, 82 (65.07 %) showed the most common (prevalent) type of mutation, IVS I-5 (G–C), followed by IVS I-1 (G–T) showed by 12 (9.52 %) subjects. Three (2.38 %) subjects showed 619 bp deletion, codon 8/9 (+G) and codon 15 (G–A) mutations were present in eight subjects each (6.34 %). Only five (3.96 %) subjects showed codon 41/42 (–TCTT). There were eight (6.34 %) subjects where mutation was not any of the six mutations studied. This study provides the pattern of β thalassemia mutations from south-western Maharashtra, which will help to prevent β-thalassemia using prenatal diagnosis and proper counseling.

Electronic supplementary material

The online version of this article (doi:10.1007/s12291-012-0230-y) contains supplementary material, which is available to authorized users.  相似文献   

4.
Malondialdehyde (MDA) is widely used as oxidative stress biomarker in biomedical research. Plasma is stored in deep freezers generally till analysis. Effect of such storage on MDA values, which may be variable and prolong, was incidentally observed in the ongoing study which is to estimate oxidative stress with oral iron. Plasma from blood samples of pregnant women (20–30 years age) in third trimester of singleton pregnancy (n = 139), consuming oral iron tablets was stored at −20 °C with intention of MDA estimation, as soon as possible. However logistic problems led this storage for prolonged and variable period (1–708 days). When values of MDA estimated using “Ohkawa” 79 method and readings were plotted against time to check the temporal effect, it showed a hyperbolic curve. Standard deviation (SD) was lowest when samples were tested within 3 weeks time. The samples analyzed within 3 weeks had mean ± SD value of 31.59 ± 26.11 μmol/L, while 123.7 ± 93.97 and 366.5 ± 189.8 μmol/L for samples stored for 1–3 and 4 months to 1 year respectively. Mean ± SD were 539.9 ± 196.8 in the samples store for more than a year. Rate of change in values was also lowest (0.0433 μmol/L/day) in the samples tested within first 3 weeks, which rose to 1.2 μmol/L/day during 3 month’s storage. This rate peaked at storage of 120 days (1.87 μmol/L/day) and fell to 0.502 μmol/L/day in the second year of storage. It is concluded that at −20 °C, only 3 weeks of storage time should be considered valid for fairly acceptable stability in MDA values.  相似文献   

5.
The present report describes the molecular study of HbD Iran (beta) 22 Glu → Gln associated with β-Thalassemia IVS1–5 (G > C) found in India, and the first case in which mutation has been identified using mass spectrometry. Given the apparent ethnic origin and the mobility of the variant hemoglobin at alkaline pH, hemoglobin D-Punjab would be suspected, but HPLC excluded this possibility. Further characterization of hemoglobinopathy was made by using nondenaturing gel electrophoresis and matrix assisted laser desorption ionization mass spectrometry and IVS1–5 being validated by reverse dot blot hybridization followed by sequencing of the β-globin gene.  相似文献   

6.
It is clear that Haptoglobin binds to Hemoglobin strongly and irreversibly. This binding, protects body tissues against heme-mediated oxidative tissue damages via peroxidase activity of Haptoglobin–Hemoglobin complex. Peroxidase activity of Haptoglobin(2-2)–Hemoglobin complex was determined via measurement of following increase in absorption of produced tetraguaiacol as the second substrate of Haptoglobin–Hemoglobin complex by UV–Vis spectrophotometer at 470 nm and 42°C. The results are showing that peroxidase activity of Haptoglobin(2-2)–Hemoglobin complex is modulated by homotropic effect of hydrogen peroxide as the allosteric substrate. On the other hand, antioxidant activity of Haptoglobin(2-2)–Hemoglobin is increased via heterotropic effect of two antibiotics (especially ampicillin) on the peroxidase activity of the complex. The condition of pathologic temperature along with the administration of ampicillin and/or coamoxiclav is in favor of amplification in antioxidant activity of Haptoglobin(2-2)–Hemoglobin and combating against free radicals in individuals with Hp2-2 phenotype. Therefore, oxidative stress effects have been diminished in the population with this phenotype.  相似文献   

7.
The objective of this study is to induce experimental diabetes mellitus by streptozotocin in normal adult Wistar rats via comparison of changes in body weight, consumption of food, volume of water, urine and levels of glucose, insulin and C-peptide in serum, between normal and diabetic rats. Intra-venous injection of 60 mg/kg dose of streptozotocin in 250–300 g (75–90 days) adult Wistar rats makes pancreas swell and causes degeneration in Langerhans islet β-cells and induces experimental diabetes mellitus in 2–4 days. For a microscopic study of degeneration of Langerhans islet β-cells of diabetic rats, biopsy from pancreas tissue of diabetic and normal rats, staining and comparison between them, were done. In this process, after collagenase digestion of pancreas, islets were isolated, dissociated and identified by dithizone method and then with enzymatic procedure by DNase and trypsin, the islet cells changed into single cells and β-cells were identified by immune fluorescence method and then assayed by flow-cytometer. Donor tissue in each step of work was prepared from 38 adult male Wistar rats weighted 250–300 g (75–90 days). Transplantation was performed in rats after 2–4 weeks of diabetes induction. In this study, the levels of insulin, C-peptide and glucose in diabetic rats reached to normal range as compared to un-diabetic rats in 20 days after transplantation of islet cells. Transplantation was performed under the cortex of testis as immunoisolated place for islet cells transplantation.  相似文献   

8.
Jena RK  Yue CY 《Biomicrofluidics》2012,6(1):12822-1282212
This report studies the surface modification of cyclic olefin copolymer (COC) by 2-methacryloyloxyethyl phosphorylcholine (MPC) monomer using photografting technique for the purpose of biointerface applications, which demonstrate resistance to both protein adsorption and cell adhesion in COC-based microfluidic devices. This is essential because the hydrophobic nature of COC can lead to adsorption of specific compounds from biological fluids in the microchannel, which can affect the results during fluidic analysis and cause clogging inside the microchannel. A correlation was found between the irradiation time and hydrophobicity of the modified substrate. Static water contact angle results show that the hydrophilicity property of the MPC-grafted substrate improves with increasing irradiation time. The contact angle of the modified surface decreased to 20 ± 5° from 88 ± 3° for the untreated substrate. The surface characterization of the modified surface was evaluated using x-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FTIR spectroscopy). Attenuated total reflection-FTIR and XPS results show the presence of the phosphate group (P-O) on modified COC substrates, indicating that the hydrophilic MPC monomer has successfully grafted on COC. Finally, it was demonstrated that cell adhesion and protein adsorption on the MPC modified COC specimen has reduced significantly.  相似文献   

9.
Ovarian cancer has been emerged as a most common and lethal gynecological malignancy in India. High serum insulin and low adiponectin have been associated with increased risk of ovarian cancer. But their role in development of ovarian cancer is conflicting and little evidence is available. We aimed to evaluate blood levels of insulin and adiponectin in epithelial ovarian cancer (EOC) patients and their association with the risk to develop EOC. The study included following three groups; Group 1: fifty cases of cytohistopathologically confirmed cases of EOC, Group 2: fifty age matched cases of benign ovarian conditions and Group 3: fifty ages matched healthy controls with no evidence of any benign or malignant ovarian pathology as ruled out by clinical examination and relevant investigations. Cytohistopathologically confirmed and newly diagnosed cases of EOC and benign ovarian cancer were included in this study. The median value of fasting serum insulin was significantly high (15.0 µlU/ml, P = 0.02) and adiponectin were significantly low (5.1 µg/ml, P < 0.001) in ovarian cancer patients compared to benign ovarian tumors and healthy controls group. A significant increase risk of ovarian cancer was found in high tertile (≥ 18.7 µlU/ml) of serum insulin level (OR = 2.7; 95% CI = 1.00–6.67, P = 0.04) and lower tertile (≤ 5.45 µg/ml) of adiponectin level (OR = 3.2; 95% CI = 1.10–9.71, P = 0.03). High serum insulin level and low adiponectin levels were significantly associated with increased risk for development of ovarian cancer.  相似文献   

10.
We have developed a method for studying cellular adhesion by using a custom-designed microfluidic device with parallel non-connected tapered channels. The design enables investigation of cellular responses to a large range of shear stress (ratio of 25) with a single input flow-rate. For each shear stress, a large number of cells are analyzed (500–1500 cells), providing statistically relevant data within a single experiment. Besides adhesion strength measurements, the microsystem presented in this paper enables in-depth analysis of cell detachment kinetics by real-time videomicroscopy. It offers the possibility to analyze adhesion-associated processes, such as migration or cell shape change, within the same experiment. To show the versatility of our device, we examined quantitatively cell adhesion by analyzing kinetics, adhesive strength and migration behaviour or cell shape modifications of the unicellular model cell organism Dictyostelium discoideum at 21 °C and of the human breast cancer cell line MDA-MB-231 at 37 °C. For both cell types, we found that the threshold stresses, which are necessary to detach the cells, follow lognormal distributions, and that the detachment process follows first order kinetics. In addition, for particular conditions’ cells are found to exhibit similar adhesion threshold stresses, but very different detachment kinetics, revealing the importance of dynamics analysis to fully describe cell adhesion. With its rapid implementation and potential for parallel sample processing, such microsystem offers a highly controllable platform for exploring cell adhesion characteristics in a large set of environmental conditions and cell types, and could have wide applications across cell biology, tissue engineering, and cell screening.  相似文献   

11.
Estimation of low density lipoprotein cholesterol (LDL-C) is crucial in management of coronary artery disease patients. Though a number of homogenous assays are available for estimation of LDL-C, use of calculated LDL-C by Friedewald’s formula (FF) is common in Indian laboratories for logistic reasons. Recently Anandaraja and colleagues have derived a new formula for calculating LDL-C. This formula needs to be evaluated before it is extensively applied in diagnosis. We measured LDL-C by homogenous method (D-LDL-C) in 515 fasting samples. Friedewald’s and Anandaraja’s formulas were used for calculation of LDL-C (F-LDL-C and A-LDL-C, respectively). The mean LDL-C levels were 123.3 ± 53.2, 112.4 ± 50.2 and 109.2 ± 49.8 mg/dl for D-LDL-C, F-LDL-C and A-LDL-C, respectively. There was a statistically significant difference between the results (P > 0.001) obtained by calculation formulas compared to the measured LDL-C. There was underestimation of LDL-C by 10.8 and 14 mg/dl by Friedewald’s and Anandaraja’s formulas respectively. The Pearson’s correlation between F-LDL-C and D-LDL-C was 0.931 and that between A-LDL-C and D-LDL-C was 0.930. Bland–Altman graphs showed a definite agreement between mean and differences of the calculation formulas and direct LDL-C with 95% of values lying with in ±2 SD limits. The mean percentage difference (calculated as {(Calculated LDL-C)-(D-LDL-C)}/D-LDL-C × 100) for F-LDL-C was maximum (−11.6%) at HDL-C ≥ 60 mg/dl and TG levels of 200–300 mg/dl (−10.4%) compared to D-LDL-C. A-LDL-C results gave highest mean percentage difference at total cholesterol concentrations <100 mg/dl (−37.3%) and HDL-C < 40 mg/dl (−17.1%), respectively. The results of our study showed that FF is better in agreement with D-LDL-C than Anandaraja’s formula for estimation of LDL-C by calculation though both lead to its underestimation.  相似文献   

12.
Recent studies have demonstrated the presence of prostate specific antigen (PSA) in cord blood of male as well as female babies. The placental progesterone and estradiol up-regulate the synthesis and secretion of PSA in Placenta. This PSA is presumed to play a role in intrauterine growth of fetus by virtue of its proteolytic action on several substrates including insulin-like-growth-factor-binding-protein-3, insulin chains and Interleukin-2. This study was planned with the objective of correlating the levels of PSA in cord blood to gestation at delivery, the type of delivery and gender of the fetus. Fifty-seven cord blood samples were collected from the umbilical cord during delivery or mid-trimester abortion and analyzed for PSA using ‘Active PSA DSL-9700 ultra sensitive’ kit employing two-site immuno-radiometric assay principle and having a detection limit of 0.001 ng/ml. Mean PSA levels in cord blood were found to be 0.112 ± 0.027 ng/ml. The concentration of PSA in cord blood was found to be higher in case of higher gestational age, male baby and operative delivery. 50 % of cord bloods for female babies had PSA below detection limit (range <0.001–0.460 ng/ml), while all the male samples had detectable PSA (range 0.11–0.973 ng/ml). Higher Progesterone levels found in prenatal maternal blood in case of male babies may be responsible for the higher cord blood PSA. Mean cord blood PSA was 0.150 ± 0.150 ng/ml in forceps delivery and 0.078 ± 0.012 ng/ml in normal vaginal delivery. Forceps delivery causes much more stress and strain as compared to a normal vaginal delivery, resulting in increased levels of adrenal glucocorticoids, and therefore, higher cord blood PSA.  相似文献   

13.
Chaurey V  Polanco C  Chou CF  Swami NS 《Biomicrofluidics》2012,6(1):12806-1280614
We present an electrokinetic framework for designing insulator constriction-based dielectrophoresis devices with enhanced ability to trap nanoscale biomolecules in physiological media of high conductivity, through coupling short-range dielectrophoresis forces with long-range electrothermal flow. While a 500-fold constriction enables field focusing sufficient to trap nanoscale biomolecules by dielectrophoresis, the extent of this high-field region is enhanced through coupling the constriction to an electrically floating sensor electrode at the constriction floor. However, the enhanced localized fields due to the constriction and enhanced current within saline media of high conductivity (1 S/m) cause a rise in temperature due to Joule heating, resulting in a hotspot region midway within the channel depth at the constriction center, with temperatures of ∼8°–10°K above the ambient. While the resulting vortices from electrothermal flow are directed away from the hotspot region to oppose dielectrophoretic trapping, they also cause a downward and inward flow towards the electrode edges at the constriction floor. This assists biomolecular trapping at the sensor electrode through enabling long-range fluid sampling as well as through localized stirring by fluid circulation in its vicinity.  相似文献   

14.
Here, we utilize microfluidic droplet technology to generate photopolymerizeable polyethylene glycol (PEG) hydrogel microbeads incorporating a fluorescence-based glucose bioassay. A microfluidic T-junction and multiphase flow of fluorescein isothiocyanate dextran, tetramethyl rhodamine isothiocyanate concanavalin A, and PEG in water were used to generate microdroplets in a continuous stream of hexadecane. The microdroplets were photopolymerized mid-stream with ultraviolet light exposure to form PEG microbeads and were collected at the outlet for further analysis. Devices were prototyped in PDMS and generated highly monodisperse 72 ± 2 μm sized microbeads (measured after transfer into aqueous phase) at a continuous flow rate between 0.04 ml/h—0.06 ml/h. Scanning electron microscopy analysis was conducted to analyze and confirm microbead integrity and surface morphology. Glucose sensing was carried out using a Förster resonance energy transfer (FRET) based assay. A proportional fluorescence intensity increase was measured within a 1–10 mM glucose concentration range. Microfluidically synthesized microbeads encapsulating sensing biomolecules offer a quick and low cost method to generate monodisperse biosensors for a variety of applications including cell cultures systems, tissue engineering, etc.  相似文献   

15.
We present a 91 MHz surface acoustic wave resonator with integrated microfluidics that includes a flow focus, an expansion region, and a binning region in order to manipulate particle trajectories. We demonstrate the ability to change the position of the acoustic nodes by varying the electronic phase of one of the transducers relative to the other in a pseudo-static manner. The measurements were performed at room temperature with 3 μm diameter latex beads dispersed in a water-based solution. We demonstrate the dependence of nodal position on pseudo-static phase and show simultaneous control of 9 bead streams with spatial control of −0.058 μm/deg ± 0.001 μm/deg. As a consequence of changing the position of bead streams perpendicular to their flow direction, we also show that the integrated acoustic-microfluidic device can be used to change the trajectory of a bead stream towards a selected bin with an angular control of 0.008 deg/deg ± 0.000(2) deg/deg.  相似文献   

16.
A study of iron, zinc, copper and selenium concentration levels was carried out in three compartments namely, maternal serum (MS), colostrums and cord blood serum (CS) of healthy Indian mothers (n = 42) who delivered healthy normal neonates without any congenital anomalies at Bhabha Atomic Research Centre hospital, Mumbai. Fe, Zn, Cu in maternal serum, cord blood and colostrums were estimated by flame atomic absorption spectrometry while Se was determined by graphite furnace absorption spectrometry. It was seen that there was a significant difference in the level of trace elements in the three compartments. The average levels of Fe in the three compartments were 1,132 ± 519, 2,312 ± 789 and 1,183 ± 602 μg/L while Zn was 514 ± 149, 819 ± 224 and 7,148 ± 2,316 μg/L respectively. Mean Cu values were 1,614 ± 295, 301 ± 77 and 392 ± 174 μg/L respectively while Se values were 70 ± 15, 36 ± 10 and 23 ± 8 μg/L respectively. The results indicated a positive correlation of Fe and Zn concentrations in MS versus CS which were (r = 0.386), (r = 0.572) respectively and Fe levels in MS and colostrums (r = 0.235). A few inter element correlations were found within compartments. Zn and Se showed a negative correlation in both MS (r = −0.489) and colostrums (r = −0.258) while a positive inter correlation of Fe and Zn was seen in MS (r = 0.44) and in CS (r = 0.54). This study gave us an overview of the serum and colostrum values of mother and neonates in Indian population, data of which are scarce.  相似文献   

17.
Abnormal glomerular permeability is the primary step towards the glomerulosclerosis. The progression rate of glomerulosclerosis is proportionate to abundance and severity of lesions created at incipient stage, which is reflected as proteinuria even though eGFR remains in the normal range. Therefore, there is a current need to find out the association between relative risks for the factors leading to proteinuria. The relations could be more informative, if it is with respect to the macromolecules like “IgG” excretion in urine. Type 2 diabetic patients were selected for this study with eGFR > 75 ml/min/1.73 m2 and grouped into four quartiles based on UIgGCR. The markers of key factors affecting progression of proteinuria were estimated through biochemical tests. The impact of these markers on proteinuria was accessed by applying multinomial logistic regression. The adjusted odds ratio for the UGAGCR was 1.186 (95 % CI: 1.061–1.327) P < 0.003 in highest quartiles of UIgGCR, followed by odds ratio for markers of collagen catabolism 1.051 (95 % CI: 1.025–1.079) P < 0.001, and USACR 1.044 (95 % CI: 1.013–1.077) P < 0.006 respectively. The marker of glycation, i.e., glycated hemoglobin showed the highest odds ratio 5.449 (95 % CI: 1.132–26.236) P < 0.035. In addition, odds for the systolic blood pressure was observed 1.387 (95 % CI: 1.124–1.712) P < 0.002. The higher odds inform and could help to discriminate the diabetic patients with fast progressive diabetic nephropathy. The study describes critical relationship between the urinary excretion of IgG and factors leading to proteinuria in type 2 diabetic patients.  相似文献   

18.
Increasingly, invitro culture of adherent cell types utilizes three-dimensional (3D) scaffolds or aggregate culture strategies to mimic tissue-like, microenvironmental conditions. In parallel, new flow cytometry-based technologies are emerging to accurately analyze the composition and function of these microtissues (i.e., large particles) in a non-invasive and high-throughput way. Lacking, however, is an accessible platform that can be used to effectively sort or purify large particles based on analysis parameters. Here we describe a microfluidic-based, electromechanical approach to sort large particles. Specifically, sheath-less asymmetric curving channels were employed to separate and hydrodynamically focus particles to be analyzed and subsequently sorted. This design was developed and characterized based on wall shear stress, tortuosity of the flow path, vorticity of the fluid in the channel, sorting efficiency and enrichment ratio. The large particle sorting device was capable of purifying fluorescently labelled embryoid bodies (EBs) from unlabelled EBs with an efficiency of 87.3% ± 13.5%, and enrichment ratio of 12.2 ± 8.4 (n = 8), while preserving cell viability, differentiation potential, and long-term function.  相似文献   

19.
Thermotaxis has been demonstrated to be an important criterion for sperm evaluation, yet clinical assessment of thermotaxis capacity is currently lacking. In this article, the on-chip thermotaxis evaluation of human sperm is presented for the first time using an interfacial valve-facilitated microfluidic device. The temperature gradient was established and accurately controlled by an external temperature gradient control system. The temperature gradient responsive sperm population was enriched into one of the branch channels with higher temperature setting and the non-responsive ones were evenly distributed into the two branch channels. We employed air-liquid interfacial valves to ensure stable isolation of the two branches, facilitating convenient manipulation of the entrapped sperm. With this device, thermotactic responses were observed in 5.7%-10.6% of the motile sperm moving through four temperature ranges (34.0-35.3 °C, 35.0-36.3 °C, 36.0-37.3 °C, and 37.0-38.3 °C, respectively). In conclusion, we have developed a new method for high throughput clinical evaluation of sperm thermotaxis and this method may allow other researchers to derive better IVF procedure.  相似文献   

20.
Multiple myeloma is a disseminated malignancy of monoclonal plasma cells that accounts for 15 % of all hematological cancers. The present study was conducted to evaluate the role of inflammation and oxidant-antioxidant dynamics in the etiology of this disease. The study population comprised of 20 cases of multiple myeloma and 20 healthy controls. The parameters evaluated were serum malondialdehyde (MDA), superoxide dismutase (SOD) and ferritin levels. The serum MDA levels were 1.9 ± 0.96 nmol/ml in cases as compared to 0.98 ± 0.55 nmol/ml in the controls. Similarly, a statistically significant difference was noted in the SOD and ferritin levels between the cases and controls (93.2 ± 23.8 vs. 210.1 ± 190.5 U/ml and 285.8 ± 216.4 vs. 131.8 ± 30.1 ng/ml respectively). Our study highlights the imbalance in the oxidant-anti oxidant mechanism and the role of smoldering inflammation in the etiology of multiple myeloma.  相似文献   

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