Journal articles on the topic '35.07 laboratory technique of chemistry'

Tuberculosis remains a major threat to world health. The main aim of this study was to improve the microscopic detection of AFB from sputum by Bleach sedimentation technique using household bleach. This was a cross sectional type of study conducted for a period of one year from July 2010 to June 2011 in the Department of Laboratory Services, Khwaja Yunus Ali Medical College Hospital (KYAMCH), Sirajgonj. A total of 115 clinically suspected tuberculosis patients aged between 7-85 years were included in the study. Among them 35 (30.4%) patients had TB positive by routine (Direct microscopy) method; whereas, by bleach concentration method 42 (36.5%) were found positive; diagnosing 07 additional patients. The rise of 16.7% in sputum positivity by bleach sedimentation microscopy over the direct smear microscopy was found to be statistically significant (p= <0.005). Paired samples t test analysis of sputum showed a significant correlation between this two methods (r=0.896, p <0.001). This study suggests that Bleach sedimentation method is better than direct conventional method. So it should be applied to all laboratories across the country because it is cheap early available and also safety to the laboratory staffs.KYAMC Journal Vol. 5, No.-2, Jan 2015, Page 497-502

Abstract A binary mixture of hyoscine butylbromide and ketoprofen was determined by 4 different methods. The first involved determination of hyoscine butylbromide and ketoprofen using the ratio-spectra first-derivative spectrophotometric technique at 211 and 234 nm over the concentration ranges of 2-14 and 5-45 μg/mL with mean accuracies 99.84 ± 0.92 and 99.98 ± 0.64%, respectively. The second method utilized second-derivative spectrophotometry over the concentration ranges of 2-14 and 5-35 μg/mL with mean accuracies 99.32 ± 1.06 and 99.55 ± 1.15%, respectively. The third method was based on the resolution of the 2 components by bivariate calibration depending on a simple and rapid mathematical algorithm and quantitative evaluation of the absorbances at 206 and 254 nm over concentration ranges of 2-16 and 5-35 μg/mL; mean accuracies of 100.21 ± 1.30 and 100.19 ± 1.07% were obtained for hyoscine butylbromide and ketoprofen, respectively. The fourth method was reversed-phase liquid chromatography using 0.05 M ammonium dihydrogen phosphateacetonitrilemethanol (20 + 30 + 6, v/v) as the mobile phase with ultraviolet detection at 220 nm over concentration ranges of 1-90 and 5-70 μg/mL; mean accuracies were 99.92 ± 1.02 and 99.61 ± 0.98%, respectively. The suggested procedures were checked using laboratory-prepared mixtures and were successfully applied for the analysis of pharmaceutical preparations. The methods retained their accuracy and precision when the standard addition technique was applied. The results obtained by applying the proposed methods were statistically analyzed and compared with those obtained by the manufacturer's method.

This article presents an experimental study of estimating stresses in concrete by applications of coda wave interferometry to establish an acoustoelastic modulus database. Under well-controlled laboratory conditions, uniaxial load cycles were performed on three groups of 15 × 15 × 35-cm concrete prisms, with ultrasonic signals being collected continuously. Then, the coda wave interferometry technique, together with acoustoelastic and Kaiser theories, are utilized to analyze the stress-velocity relations for the distinct ranges before and after historical maximum loads, forming an acoustoelastic modulus database. When applied to different concrete samples, their stresses are estimated with a high degree of accuracy. This study could be used to promote the development of novel nondestructive techniques that aid in structural stress monitoring.

In this study, a powder mixture consisting of TiC (titanium carbide) and WC (tungsten carbide) reinforced AlCuNiFeCr was produced using the hot pressing technique. The AlCuNiFeCr powder mixture, TiC and WC were added at a rate of 5 %, 10 % and 15 %, respectively. In order to produce and control to produce parameters have been produced having automation systems type of laboratory a hot pressing machine. Graphite moulds were used in production and sintering processes were carried out in a protective argon gas atmosphere. Composites were produced at a sintering temperature of 900?C and under a pressure 35 MPa and for 4 minutes. To understand the microstructure and mechanical properties of produced specimens, their SEM and EDS analysis, XRD analysis, fracture surface SEM images, SEM mapping of elemental distribution and EDX spectrums, hardness, three-point bending, and corrosion tests were investigated. As a result, the best sample has been identified that belong to AlCuNiFeCr-15 % WC with great features.

Dosage regimens of drugs that are cleared mainly by glomerular filtration as well as fluid management in preterm infants should be based on the glomerular filtration rate (GFR) of the individual patient. However, GFR measurements and collection of urine in newborns are difficult to perform. The 24 to 48 h continuous inulin infusion technique does not require the collection of urine and is considered the most reliable indicator of GFR.1,2 This method is invasive, time-consuming, and expensive. In contrast, serum creatinine measurements can be obtained easily and determined quickly in the clinical chemistry laboratory. Most laboratories use an automated kinetic Jaffé method, which is subject to negative interference by plasma hemoglobin above 0.06 mmol/L, and to negative interference by bilirubin (about 35 µmol/L by a serum bilirubin of about 100 µmol/L).

Abstract Abstract 747 Introduction: RUNX1 (runt-related transcription factor 1) deregulations constitute a disease-defining aberration in AML. RUNX1 mutations were proposed as clinically useful biomarkers to follow disease progression from MDS to s-AML, as well as to monitor minimal residual disease (MRD). Study design: First, a next-generation amplicon deep-sequencing (NGS) assay was developed and a validation study was performed on genomic DNA obtained from mononuclear cells on a longitudinal series of 116 retrospective samples obtained from 25 patients. These samples were collected between 11/2005 and 6/2010 and were characterized for RUNX1 mutations by DHPLC and Sanger sequencing (conventional methods). In median, 3,346 reads per amplicon were generated and in all cases NGS analyses concordantly detected the mutations known from conventional methods. Furthermore, in 2/25 (8%) cases, NGS detected additional low-level mutations with 0.9% and 3.2% of reads mutated that were not observed by standard Sanger technique. Concerning MRD monitoring, in 7/25 (28%) cases an increasing clone size, i.e. mutations as low as 0.2% - 7.0%, was detectable up to 9 months earlier than by conventional methods. This established assay then was applied to characterize an unselected prospectively collected cohort during the subsequent 12-months routine diagnostics period starting 07/2010. Results: In total, 2,705 NGS RUNX1 mutation analyses were performed on a variety of hematological malignancies. We report on analyses on 460 AML cases at diagnosis including 369 de novo AML, 57 s-AML, and 34 t-AML cases (median age: 68 years; females: 204; males 256). 51% of cases presented with a normal karyotype, 38% harbored non-complex cytogenetic alterations, 10% carried a complex aberrant karyotype, and 1% of patients were characterized by favorable cytogenetics. Overall, 141 RUNX1 mutations were observed in 24.3% (112/460) of cases. At diagnosis, the clone size ranged from 2% to 95% (median: 40%). 82% (92/112) of mutated patients carried one, whereas 18% (20/112) harbored two (n=17) or more (n=3) mutations. The 141 mutations were characterized as follows: 43% (60/141) frame-shift mutations, 34% (49/141) missense, 15% (21/141) nonsense, 5% (7/141) exon-skipping, and 3% (4/141) in-frame insertion/deletion alterations, respectively. The mutations were predominantly located in the RHD domain (54%) or TAD domain (20%). In subsequent serial NGS analyses 31/112 evaluable RUNX1 mutated cases were studied and in 88 individual samples the alterations detected at diagnosis were specifically investigated with high coverage. With a median sampling interval of 50 days for the NGS analyses between 2 and 9 samples per patient were analyzed during the first year of treatment. In this cohort, three categories of patients were detectable: (i) 55% (17/31) of patients responded to therapy and were characterized by a total clearance of the mutated clone at the first time point of follow-up (804-fold median sequencing coverage; sensitivity ∼1:800). (ii) A second group consisted of 10% (3/31) of patients with refractory disease that stayed mutated, but were excluded from further analyses since they underwent transplantation. (iii) The third group comprised 35% (11/31) of patients: None of these patients demonstrated a clone size reduction below 0.7% of reads at the first follow-up analysis (reduction to a median of 21% mutated reads; range 0.7% - 41%). Also, at the second time point (in median 108 days after initial diagnosis), mutated clones were still detectable (reduction to a median of 8% mutated reads; range 4% - 15%). Most of these cases (10/11) had refractory disease as assessed by cytomorphology or molecular analyses. 10/11 cases did harbor a normal karyotype; n=1 with del(7q). Further, 6 of these 11 patients with refractory disease, as defined using NGS, were found to carry RUNX1 double mutations. Finally, in all (3/3) cases with double mutations in the same domain and refractory disease a changing antiparallel distribution of the clone size from initial diagnosis to first follow-up was observed. Conclusions: NGS accurately detects and quantifies RUNX1 mutations in AML with high sensitivity. The technique of deep-sequencing was observed to be superior to current routine methods, in particular during follow-up and in detecting MRD and thus has the potential to enable an individualized monitoring of disease progression and treatment efficacy. Disclosures: Kohlmann: MLL Munich Leukemia Laboratory: Employment; Roche Diagnostics: Honoraria. Grossmann:MLL Munich Leukemia Laboratory: Employment. Harbich:MLL Munich Leukemia Laboratory: Employment. Dicker:MLL Munich Leukemia Laboratory: Employment. Alpermann:MLL Munich Leukemia Laboratory: Employment. Nadarajah:MLL Munich Leukemia Laboratory: Employment. Kern:MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Haferlach:MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Haferlach:MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Schnittger:MLL Munich Leukemia Laboratory: Employment, Equity Ownership.

Spacecraft formation flying (SFF) in highly elliptical orbit (HEO) has attracted a great deal of attention in many space exploration applications, while precise guidance, navigation, and control (GNC) technology—especially precise ranging—are the basis of success for such SFF missions. In this paper, we introduce a novel K-band microwave ranging (MWR) equipment for the on-orbit verification of submillimeter-level precise ranging technology in future HEO SFF missions. The ranging technique is a synchronous dual one-way ranging (DOWR) microwave phase accumulation system, which achieved a ranging accuracy of tens of microns in the laboratory environment. The detailed design and development process of the MWR equipment are provided, ranging error sources are analyzed, and relative orbit dynamic models for HEO formation scenes are given with real perturbations considered. Moreover, an adaptive Kalman filter algorithm is introduced for SFF relative navigation design, incorporating process noise uncertainty. The performance of SFF relative navigation while using MWR is tested in a hardware-in-the-loop (HIL) simulation system within a high-precision six degrees of freedom (6-DOF) moving platform. The final range estimation errors from MWR using the adaptive filter were less than 35 μm and 8.5 μm/s for range rate, demonstrating the promising accuracy for future HEO formation mission applications.

Abstract Four accurate, sensitive, and reproducible stability-indicating methods for the determination of erdosteine in the presence of its acid degradation products are presented. The first method involves processing the spectra by using a first-derivative method at 229 nm in a concentration range of 10–70 μg/mL. The mean percentage recovery was 100.43 ± 0.977. The second method is based on ratio-spectra first derivative spectrophotometry at 227.4 and 255 nm over a concentration range of 10–70 μg/mL. The mean percentage recovery was 99.65 ± 1.122% and 100.02 ± 1.306% at 227.4 and 255 nm, respectively. The third method utilizes quantitative densitometric evaluation of the TLC of erdosteine in the presence of its acid degradation products, and uses methanol–chloroform–ammonia (7 + 3 + 0.01, v/v/v) asthe mobile phase. TLC chromatograms were scanned at235 nm. This method analyzes erdosteine in a concentration range of 2.4–5.6 μg/spot, with a mean percentage recovery of 100.03 ± 1.015%. The fourth method is HPLC for the simultaneousdetermination of erdosteine in the presence of its acid degradation products. The mobile phase consists of water–methanol (65 + 35, v/v). The standard curve of erdosteine showed good linearity overa concentration range of 10–80 μg/mL,with a mean percentage recovery of 99.90 ± 1.207%. These methods were successfully applied to the determination of erdosteine in bulk powder, laboratory-prepared mixtures containing different percentages of the degradation products, and pharmaceutical dosage forms. The validity of results was assessed by applying the standard addition technique. The results obtained agreed statistically with those obtained by a reported method, showing no significant differences with respect to accuracy and precision.

In recent years, the use of hop extracts in industrial and home brewing processes as an alternative to hop cones or pellets usually added to wort during boiling has become increasingly popular. These extracts represent concentrated sources of bitter compounds, i.e., α- and β-acids, which are involved in some of the main reactions that take place in the wort and are responsible for the bitterness and the final quality of beer. This work aims at proposing a novel extraction technique, using a hydrofluorocarbon solvent in subcritical conditions; this process provided an extraction yield of 19% and an α-acid recovery of approximately 49% in 120 min of process. The α-acid isomerization kinetics of thermally treated hop extracts were studied and compared with those of both hop pellets and a CO2 extract. Laboratory scale tests showed that shorter boiling times were needed using hydrofluorocarbon and CO2 extracts (approximately 25 min and 34 min, respectively) to reach the same isomerization efficiency of 16.73%, achieved in 50 min of boiling with pellets. Moreover, the process was scaled up and the possibility of considerably reducing the conventional treatment times using hydrofluorocarbon extracts was confirmed: the same isomerization yield (9.1%) obtained after 50 min using the traditional procedure with hop pellets was reached in a shorter time of approximately 35 min in a pilot apparatus.

Background: The criminal investigation starts at crime scene and ends in the laboratory. The most vital of which is mortuary where the postmortem examination of the deceased is carried out to answer the questions of interest in that investigatory menu. Aim: Unaided eye estimation of age from the lambdoid suture of skull on autopsy. Methods: The subjects under study were taken from mortuary of the department of Forensic Medicine and Toxicology in the King Edward Medical University Lahore during the year 2016. The targeted population for study was the draining are of the mortuary of King Edward Medical College having designated police station. A pretested questionnaire was used to collect the calculated sample among the research population as a research method. A non-probability convenient consecutive sampling technique was applied to collect the data. The comparative descriptive study design was utilized to analyze the results. Results: The study revealed that among the targeted population a higher percentage more than third of the sample size were adolescents of age between 21 – 30 years being 35% followed by elderlies of age between 41 – 50 years amount to 20%. The research depicted that age can be determined from degree of closure of lambdoid suture in the dead body on the autopsy table with closure taking place earlier in males as compared to that of females. Conclusion: This study concludes that it is possible to estimate age from the degree of lambdoid suture closure of the deceased skull vault during the postmortem examination. Keywords: Age, Lambdoid, Suture, Autopsy, Cranial, Post-mortem Examination, Skull

Abstract The MC-Media Pad ACplus™ is a dry, rehydratable film medium for the enumeration of aerobic bacterial colonies. The performance of the method in a variety of foods was compared to that of U.S. reference methods: U.S. Department of Agriculture (USDA) Food Safety and Inspection Service (FSIS) Microbiology Laboratory Guidebook (MLG) Chapter 3.02 “Quantitative Analysis of Bacteria in Foods as Sanitary Indicators” (USDA/FSIS MLG 3.02); Standard Methods for the Examination of Dairy Products (SMEDP) Chapter 6 “Microbiological Count Methods, Standard Plate Count Method” (SMEDP 6); AOAC Official MethodSM966.23Microbiological Methods; and ISO 4833-1:2013 “Microbiology of the food chain—Horizontal method for the enumeration of microorganisms—Part 1: Colony count at 30 degrees C by the pour plate technique.” The validated matrixes included raw chicken breast and raw ground pork for USDA/FSIS MLG 3.02; cream cheese and yogurt drink for SMEDP 6; parsley, vegetable juice, prawns, tuna pate, sandwiches, and pasta salad for AOAC Method 966.23, and raw chicken breast, raw ground pork, cream cheese, yogurt drink, parsley, vegetable juice, prawns, tuna pate, sandwiches, and pasta salad for ISO 4833-1:2013. In each matrix study, five replicates at each of three contamination levels were tested as paired test portions. All 10 matrixes were compared to the appropriate U.S. reference methods under MC-Media Pad ACplus standard-usage conditions (35 ± 1°C for 48 ± 2 h). Across all matrixes, the difference of mean log10 values ranged from –0.43 to 0.44, within the acceptable range of –0.50 to 0.50. The candidate method repeatability SD (sr) varied from 0.03 to 0.23 log10 CFU/g, comparing favorably to the reference method SD, which ranged from 0.06 to 0.30 log10 CFU/g. Seven matrixes were compared to the appropriate U.S. reference methods under MC-Media Pad ACplus rapid-usage conditions (35 ± 1°C for 24 ± 2 h). Of the 21 matrix/concentration combinations, only three instances of difference of mean &gt;0.5 log were observed. The ranges of sr values of the rapid-usage candidate method (0.023–0.324) and the reference method (0.013–0.236) were similar for the seven matrixes tested. All 10 matrixes were compared to the International Organization for Standardization (ISO) reference method under MC-Media Pad ACplus alternate-method conditions (30 ± 1°C for 72 ± 3 h). All 10 matrixes yielded a mean difference between methods of &lt;0.5 log, and the ranges of sr values were similar between the candidate alternate method (0.037–0.378) and the ISO reference method (0.037–0.437). The product consistency study demonstrated no significant difference between lots of product and supported the 2-year shelf life. Robustness testing yielded no significant differences when small variations were made in sample volume, incubation temperature, and incubation time. Thus, the data show equivalent or better performance of the MC-Media Pad ACplus method compared to the relevant reference methods in support of AOAC Performance Tested MethodSM certification.

Abstract Abstract 2268 Since the introduction of imatinib mesylate for treating patients with chronic myeloid leukemia (CML) data has shown that a small proportion of patients in chronic phase experience refractoriness to kinase inhibitor treatment due to mutations in the ABL1 domain. A variety of molecular methods provide a clinically feasible workflow to detect these mutations but have both strengths and weaknesses: e.g. direct sequencing of nested PCR assays amplifying the ABL1 part of BCR-ABL1 fusion transcript has a low diagnostic detection sensitivity of 10%; and more sensitive methods such as allele-specific PCRs are only available for known mutations. Here, we investigated the utility of ultra-deep next-generation sequencing (NGS) to detect and monitor the composition of ABL1 kinase mutations at diagnosis and during inhibitor treatment. We included 62 samples from 13 CML patients, diagnosed between 10/2005 - 07/2009. The chimeric BCR-ABL1 fusion transcript was amplified from cDNA. Subsequently, 2 overlapping amplicons were designed to amplify a 740 bp stretch of the ABL1 kinase domain to be processed by 454 Titanium amplicon chemistry protocol (454 Life Sciences, Branford, CT). In median, 454 sequencing data was generated for each patient across 4 time points (range 3–11) with a median time span of 11 months (range 5–35) from state of first diagnosis to the most recent investigation. For each investigation a median of 2234 reads specific for the ABL1 kinase domain was obtained. In this selected cohort, in 12/13 patients, previous Sanger sequencing had detected a spectrum of 10 different kinase domain mutations during course of inhibitor treatment. In all 12 patients deep-sequencing enabled the quantitative detection of these mutations with 100% concordance. In 1/13 cases carrying a t(2;3)(q31,q27), neither NGS nor routine methods detected any mutation over four distinct time points, although the %BCR-ABL1/ABL1 was persistently high (ratios of 108, 84, 80, and 108, respectively). According to European LeukemiaNet guidelines (Baccarani et al., JCO 2009), kinase domain mutation screening is only recommended if BCR-ABL1 transcript levels are increasing at consecutive time points. We therefore investigated at which point in time NGS would allow to detect resistant clones subsequent to inhibitor therapy even though alternative testings had not been performed. In 11/12 cases resistant clones were already early detectable after start of treatment with tyrosine kinase inhibitors, with a median time span of 3.9 months from first diagnosis (range 1.1–19.5). We here demonstrate in 6 patients, treated with Imatinib or Dasatinib that even at transcript levels of BCR-ABL1/ABL1, ranging in our cohort from 1.1%-13%, mutations were detectable (range of mutations: 4%-97% of sequencing reads). These points to a very fast selection process of resistance-defining clones. More importantly, NGS was able to quantitate in 4 patients an increasing mutational burden of resistant clones while the %BCR-ABL1/ABL1 stayed consistently low. For example, one patient harbored the L387M mutation, detected with 54% sequencing reads harboring the mutation (1.1 %BCR-ABL1/ABL1). After 8 months, the L387M mutation was detectable in 97% NGS reads, while the %BCR-ABL1/ABL1 was 3.7. Moreover, it was possible to monitor in detail the molecular composition in 3 patients harboring concomitantly more than one mutation. NGS demonstrated that one of these mutations or even a novel clone was dominant in a subsequent time point, in particular after changing the inhibitor. For example, one patient harbored concomitantly the mutations L284V (30%), Y253H (17%), and T315I (4%). The L284V and Y253H mutations disappeared during Dasatinib treatment, and T315I was thereafter persistent -within 38 days- with a high mutation burden of 98%. Importantly, 454 NGS never detected more than 3 resistant clones concomitantly per patient and thus in no case indicated the existence of a variety of coexisting low-level mutations. However, in all 12 patients at the last time point of our investigation there was only one resistance mutation detectable (in 6/12 cases: T315I). In conclusion, as investigated in this work, NGS is technically feasible for high-throughput serial monitoring of CML patients and, moreover, provides an accurate and highly-sensitive quantitative assessment of mutations leading to therapy resistance but also allows uncovering novel mutations. Disclosures: Grossmann: MLL Munich Leukemia Laboratory: Employment. Wild: MLL Munich Leukemia Laboratory: Employment. Schnittger: MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Kern: MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Haferlach: MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Haferlach: MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Kohlmann: MLL Munich Leukemia Laboratory: Employment.

Abstract Abstract 1691 RUNX1 (runt-related transcription factor 1) mutations constitute a disease-defining molecular aberration in myelodysplastic syndromes (MDS) and acute myeloid leukemia (AML). Mechanistically, deregulations occur either through balanced translocations or molecular mutations. Importantly, patient-specific RUNX1 mutations have been proposed to represent clinically useful biomarkers to follow disease progression from MDS to s-AML, as well as to monitor minimal residual disease (MRD) during AML treatment. As such, unbiased methodologies are warranted to provide necessary diagnostic sensitivity and throughput. Here, we investigated 116 samples from 25 patients (18 AML and 7 MDS) using next-generation amplicon deep-sequencing. For a longitudinal analysis starting at diagnosis and following the course of treatment peripheral blood (n=20) or bone marrow specimens (n=96) were obtained between 11/2005 and 6/2010. PCR assays targeting the RUNX1 beta isoform were performed with 60 ng of genomic DNA, obtained from mononuclear cells. In median, 5 time points per patient were analyzed with a median time span of 14 months (range: 5 – 34 months). The median sampling interval was 3.2 months. For each patient, one or more molecular mutations were known from standard testing at diagnosis using a combination of denaturing high-performance liquid chromatography and direct Sanger sequencing. In 166 amplicons covering the full spectrum of RUNX1 mutations we applied the 454 small volume Titanium chemistry assay to perform ultra-deep sequencing of specific PCR products (454 Life Sciences, Branford, CT). In median, 3346 reads per amplicon were generated, thereby allowing a highly sensitive assessment of RUNX1 mutational burden in these patients. As such, at 5% diagnostic sensitivity, 167 reads would cover a certain molecular mutation. At a cut-off of 0.5% sensitivity in median 17 reads were remaining for evaluation. First, we evaluated the concordance of NGS and conventional methods for the samples being taken at initial diagnosis. In all 25 patients deep-sequencing analyses concordantly detected the mutations known from conventional methods, i.e. in total 9 missense mutations, 1 nonsense mutation, 2 in-frame alterations, and 13 frameshift alterations. At initial diagnosis, deep-sequencing detected in AML cases the mutations with a median burden of 44% sequencing reads, whereas in MDS cases in median 35% sequencing reads harbored the mutations, respectively. In 2/25 (8%) cases, deep-sequencing detected additional low-level mutations (0.9% and 3.2%) that were not observed by standard techniques. Secondly, we investigated whether the technique of ultra-deep sequencing would be superior to current routine testing methods during follow-up and in detecting MRD. In 7/25 (28%) patients, an increasing clone size was detectable earlier than by conventional methods. Clone sizes with mutations as low as 0.2% - 7.0% of reads were detectable by NGS up to 9 months earlier during course of disease than by conventional methods. In no case did NGS miss mutations known by conventional methods. Overall, in 12/25 (48%) patients, ultra-deep sequencing revealed additional subclones and enabled the quantitative assessment of their respective clone size. In 6/25 (24%) cases this ultra-deep sequencing approach allowed to then quantitatively monitor the changing composition of parallel subclones per patient during treatment and disease progression. In particular, in two MDS patients dominant clones were proven to disappear during course of the disease and existing low-level or novel clones were emerging at s-AML stage. Similarly, in two AML patients dominant clones were suppressed during chemotherapy. Previously existing low-level mutations, already observed at the stage of initial diagnosis, were then detected at relapse with much greater mutational burden. Finally, in 2/25 cases with mutations concomitantly occurring in the same amplicon deep-sequencing was able to delineate monoallelic or biallelic status of the mutation. In conclusion, RUNX1 mutations are useful biomarkers with clinical utility for the detection of MRD in patients with hematological malignancies. We here demonstrated that amplicon-based NGS is a suitable method to accurately detect and quantify the variety of RUNX1 aberrations with high sensitivity and enables an individualized monitoring of disease progression and treatment efficacy. Disclosures: Kohlmann: MLL Munich Leukemia Laboratory: Employment. Grossmann:MLL Munich Leukemia Laboratory: Employment. Schindela:MLL Munich Leukemia Laboratory: Employment. Kern:MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Haferlach:MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Haferlach:MLL Munich Leukemia Laboratory: Employment, Equity Ownership. Schnittger:MLL Munich Leukemia Laboratory: Employment, Equity Ownership.

Abstract Abstract 4972 Background: The growth of plasma cells in multiple myeloma (MM) is dependent on a complex interplay among various cytokines, adhesion molecules and other factors in the tumor microenvironment. Several cytokines, including Interleukin (IL)-6, IL-10 and IL-17 have been shown to promote myeloma cell growth in vitro. Furthermore, several investigators have shown the increase in levels of serum IL-6, IL-10, IL-17, and IL-18 in MM patients compared with healthy donors. Although many studies have shown that the dysregulation of these cytokines can be associated with MM development, there are a few reports showing the influence of polymorphisms in cytokine genes on the risk of MM. We examined the single nucleotide polymorphisms (SNPs) of these cytokines: IL-10 (rs1800870 − 1082 A/G, rs1800871 − 819 T/C, and rs1800872 − 592 A/C), IL-17A (rs2275913, −197G/A), IL-17F (rs763780, 7488 T/C), and IL-18(rs187238 −137G/C and rs1946518 −607 A/C) in MM patients, and analyzed the relationship between these SNPs and the susceptibility and clinical features. Patients and Methods: Ninety three patients [age range, 35–83 years; male/female 44/49; Durie and Salmon stage I (n=8), stage II (n=22), stage III (n=61), unknown (n=2); International staging system (ISS) 1 (n=21), 2 (n=21), 3 (n=29), unknown (n=22); IgG (n=55), IgA (n=15), IgD (n=2), non-secretory(n=3), Bence Jones(n=18)] with MM and 192 healthy race- and sex-matched healthy controls were examined. Genomic DNA was isolated from peripheral blood using the DNA extraction Kit. Genotyping of IL-10, IL-17A, and IL-17F polymorphisms were determined by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and the genotyping of the IL-18 polymorphism was determined by the allelic specific polymerase chain reaction technique. Genotype and allele frequencies were compared between the study groups using Χ2-test. The characteristics and laboratory features of MM patients with each polymorphisms were compared using Χ2-tests and student t-tests. The Kaplan-Meier method was used in the calculation of overall survival (OS). OS were compared with the log-rank test. Probability values <0. 05 were considered statistically significant. Results: Genotype and allele frequencies of cytokines in MM patients and the control: The frequencies of genotypes of cytokines in patients with MM were as follows: AA 92. 5% and AG 7. 5% for IL-10–1082; TT 43%, TC 48. 4% and CC 8. 6% for IL-10–819; AA 43%, AC 48. 4% and CC 8. 6% for IL-10–592; AA 19. 4%, AG 40. 9% and GG 39. 8% for IL-17A-197; TT 82. 8% and TC 17. 2% for IL-17F; GG 65. 6%, GC 26. 9% and CC 7. 5% for IL-18–137; AA 35. 5%, AC 47. 3% and CC 17. 2% for IL-18–607 loci. No significant differences were observed in the allele or genotype frequencies of IL-10 and IL-17F polymorphisms between MM patients and the control group. However, patients with MM had a significantly higher frequency of the IL-18–137 CC genotype compared to the control group (7. 5% vs. 2. 2%, P<0. 05). The number of IL-18–137 C alleles among the patients with MM was also higher than in the control group (21% vs. 13. 3%, p<0. 05). Furthermore, MM patients had a significantly lower frequency of the IL-17A A/G genotype compared to the control group (40. 9% vs. 58. 7%, P<0. 01). Patients' characteristics according to cytokine polymorphisms: IL-10 592 CC genotype (high producer type) was significantly associated with advanced ISS (P=0. 03) and higher β2 microglobulin level (CC vs non CC; 9. 81±4. 78 g/dL vs. 5. 27±3. 27g/dL, p<0. 05). IL-17A-197 AA genotype (high producer type) was also significantly associated with higher bone scale (66. 6% vs 44%, p=0. 05). IL-18–137 CC or GC genotype was significantly associated with advanced ISS (P<0. 05) and lower hemoglobin level (8. 8±2. 6 mg/dL vs. 9. 9±2. 4 mg/dL, p=0. 04). Although there was no significant difference in overall survival of IL17 A, IL-17F and IL-18 polymorphisms, patients with IL-10–592 CC or IL-18–607 AA genotype showedtendency to more unfavorable survival (p=0. 07). A multivariate analysis using cox proportional hazard model demonstrated that Bence Jones protein (p=0. 001), ISS stage III (p<0. 05), the use of new drugs (p=0. 001), IL-10–592CC genotype (P=0. 005) and IL-17 AA (P=0. 00001) were independent adverse prognostic factors. Conclusion: These results indicate that cytokine polymorphisms, including IL-10, IL-17 and IL-18, are associated with prevalence and clinical feature of MM in Japanese patients. Disclosures: No relevant conflicts of interest to declare.

e4176066In this paper we present a discussion about the implications of the theory of mental models in the teaching and learning process in Chemistry, with the purpose of discussing the use of stop motion animation as a data collection instrument to investigate the students' mental representations about chemical concepts, especially about their dynamic character. To achieve the objectives mentioned in the present work, we seek to relate the theory of mental models and their implications for the teaching and learning processes in chemistry, with the proposal of the triangulation of the three different representational levels of Chemistry (submicroscopic, symbolic and macroscopic), which are necessary to understand this Science. It is also noteworthy that this articulation was based on the main difficulties associated with the teaching and learning processes in Chemistry and was thought of the particularity involving the dynamics of the atomic-molecular level. The discussions carried out involving the different references used allowed us to discuss the potentialities associated with the use of stop motion as an instrument for investigating mental models on chemical concepts, highlighting the fact that it is a technique that is easy to apply, low cost and that allows the dynamic representation of chemical phenomena considering the submicroscopic level.ResumoNo presente trabalho apresentamos uma discussão acerca das implicações da teoria dos modelos mentais nos processos de ensino e aprendizagem em Química, com a finalidade de discorrer sobre o uso de animação em stop motion como um instrumento de coleta de dados para se investigar as representações mentais dos sujeitos sobre conceitos químicos, em especial sobre seu caráter dinâmico. Para atingir os objetivos mencionados no presente trabalho, buscamos relacionar a teoria dos modelos mentais e suas implicações para os processos de ensino e aprendizagem em Química, com a proposta da triangulação dos três diferentes níveis representacionais da Química (submicroscópico, simbólico e macroscópico), que são necessários para a compreensão dessa Ciência. Destacamos também, que tal articulação foi pautada nas principais dificuldades associadas aos processos de ensino e aprendizagem em Química e pensada na particularidade envolvendo a dinamicidade do nível atômico-molecular. 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Abstract Interstellar complex organic molecules (iCOMs) have been identified in different interstellar environments including star forming regions as well as cold dense molecular clouds. Laboratory studies show that iCOMs can be formed either in gas-phase or in the solid state, on icy grains, from ”non-energetic” (atom-addition/abstraction) or energetic (UV-photon, particle bombardments) processes. In this contribution, using a new experimental approach mixing matrix isolation technique, mass spectrometry, and infrared and EPR spectroscopies, we want to investigate the COM formation at 35 K from a complex mixture of ground state radicals trying to draw a general reaction scheme. We photolyse (121 nm) CH3OH diluted in Ar at low temperature (below 15 K) to generate ${H^.CO}$, ${HO^.CO}$, ${^.CH2OH}$, ${CH3O^.}$, ${^.OH}$, and ${^.CH3}$ radicals and ”free” H-atoms within the matrix. Radicals have been identified using infrared and EPR spectroscopies. With the disappearance of the Ar matrix (at 35 K), these unstable species are then free to react, forming new species in a solid film. Some recombination products have been detected using infrared spectroscopy and mass spectrometry in the solid film after Ar removal, namely methyl formate (CH3OCHO), glycolaldehyde (HOCH2CHO), ethylene glycol (HOCH2CH2OH), glyoxal (CHOCHO), ethanol (CH3CH2OH), formic acid (HCOOH), dimethyl ether (CH3OCH3), methoxymethanol (CH3OCH2OH) and CH4O2 isomers (methanediol and/or methyl hydroperoxide). The detected molecules are fully consistent with the radicals detected and strongly support the solid state scenario of iCOM formation in interstellar ices based on radical-radical recombination. We then discuss astrophysical implications of the radical pathways on the observed gas-phase iCOMs.

Aims: Pollution by wastewaters from various urban activities such as artisanal dyeing plants is a real problem for developing countries. The treatment of wastewater by the adsorption method is carried out by means of less expensive and available adsorbent media. Two techniques of the adsorption method are possible: adsorption in continuous mode (column adsorption) and adsorption in discontinuous mode (batch adsorption). The choice of the continuous adsorption technique is justified by its ability to process large volumes of solutions. In this study, dyes contained in wastewater from artisanal dyeing plants were removed by continuous adsorption in a fixed-bed column of deactivated lichen biomass (Parmotrema dilatatum). Study Design: Random design Place and Duration of Study: Laboratory of Thermodynamics and Environmental Physico-Chemistry (University Nangui Abrogoua, Ivory Coast) between May 2020 and October 2020. Methodology: Four (4) categories of wastewater were collected in artisanal cotton and leather dyeing plants through two municipalities of the city of Abidjan, economic capital of Ivory Coast. Two (2) wastewaters colored in blue from dyeing of cotton boubous and jeans and two (2) wastewaters colored in red from dyeing of leather jackets and bags. These wastewaters were treated through the fixed bed column of deactivated lichens. The column feed rate was set at 0, 07 L.min-1 and the adsorbent bed mass at 100 g. Results: The study showed that, regardless of the nature of the dyed object and regardless of the target dye, the amount of dye adsorbed was better with waters of higher initial concentration. Thus the best amount of adsorbed dye is 44.444 mg.g-1 and the best removal rate is 97.9%. These values are obtained with the red wastewater of bags (RWB) treatment which was the most concentrated wastewater. Conclusion: Good efficiency of deactivated lichen bed as adsorbent for the in situ removal of dyes from wastewater by continuous adsorption.

Introdução: O maior foco das pesquisas odontológicas nos últimos 60 anos tem sido a adesão e suas técnicas. Mais de 7000 artigos já foram publicados a este respeito. O desenvolvimento dos materiais odontológicos adesivos e as técnicas a eles relacionadas possuem uma história interessante, onde descobertas do passado ainda são usadas de alguma forma no presente. Objetivo: expor, através de uma revisão de literatura, um breve histórico sobre materiais e técnicas restauradoras, bem como o estágio atual da odontologia adesiva, com ênfase na tradução de evidências baseadas em pesquisas laboratoriais para a prática clínica. Materiais e Métodos: Foram selecionados livros de preferência do autor para a introdução de conceitos clássicos e artigos de revisão publicados nos últimos 10 anos, utilizando as cinco palavras-chave: “Dental Bonding” AND “Dental Cements” AND “Resin Cements” AND “Adhesives” AND “Ceramics”, sorteados pela melhor combinação na plataforma Pub/Med/MEDLINE. Resultados: Duzentos e um artigos, foram encontrados, sendo utilizados para análise qualitativa e quantitativa aqueles pertinentes ao direcionamento do autor, de acordo com o tema. Conclusão: Considerando as limitações do estudo, concluiu-se que a odontologia adesiva é uma área que segue em constante desenvolvimento, fundamental para a realização de restaurações minimamente invasivas e estéticas. Onde para que seja possível consequentemente longevidade clínica, os materiais utilizados e substrato dentário requerem conhecimento do profissional e fidelidade na execução de um correto pré-tratamento das superfícies, respeitando suas naturezas e composições.Descritores: Colagem Dentária; Cimentos Dentários; Cimentos de Resina; Adesivos; Cerâmica.ReferênciasVan Meerbeek B, De Munck J, Yoshida Y, Inoue S, Vargas M, Vijay P, et al. Buonocore memorial lecture. Adhesion to enamel and dentin: current status and future challenges. Oper Dent. 2003;28:215-35.Miyashita E, Fonseca AS. 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