High-Dimensional Analysis of Single-Cell Flow Cytometry Data Predicts Relapse in Childhood Acute Lymphoblastic Leukaemia

B-cell Acute Lymphoblastic Leukaemia is one of the most common cancers in childhood, with 20% of patients eventually relapsing. Flow cytometry is routinely used for diagnosis and follow-up, but it currently does not provide prognostic value at diagnosis. The volume and the high-dimensional character of this data makes it ideal for its exploitation by means of Artificial Intelligence methods. We collected flow cytometry data from 56 patients from two hospitals. We analysed differences in intensity of marker expression in order to predict relapse at the moment of diagnosis. We finally correlated this data with biomolecular information, constructing a classifier based on CD38 expression. Artificial intelligence methods may help in unveiling information that is hidden in high-dimensional oncological data. Flow cytometry studies of haematological malignancies provide quantitative data with the potential to be used for the construction of response biomarkers. Many computational methods from the bioinformatics toolbox can be applied to these data, but they have not been exploited in their full potential in leukaemias, specifically for the case of childhood B-cell Acute Lymphoblastic Leukaemia. In this paper, we analysed flow cytometry data that were obtained at diagnosis from 56 paediatric B-cell Acute Lymphoblastic Leukaemia patients from two local institutions. Our aim was to assess the prognostic potential of immunophenotypical marker expression intensity. We constructed classifiers that are based on the Fisher's Ratio to quantify differences between patients with relapsing and non-relapsing disease. We also correlated this with genetic information. The main result that arises from the data was the association between subexpression of marker CD38 and the probability of relapse

Advances in chemical and biological methods to identify microorganisms—from past to present

Fast detection and identification of microorganisms is a challenging and significant feature from industry to medicine. Standard approaches are known to be very time-consuming and labor-intensive (e.g., culture media and biochemical tests). Conversely, screening techniques demand a quick and low-cost grouping of bacterial/fungal isolates and current analysis call for broad reports of microorganisms, involving the application of molecular techniques (e.g., 16S ribosomal RNA gene sequencing based on polymerase chain reaction). The goal of this review is to present the past and the present methods of detection and identification of microorganisms, and to discuss their advantages and their limitations.C.F.R. would like to thank the Portuguese Foundation for Science and Technology (FCT–Portugal) for the C.F.R. for the project UID/EQU/00511/2019—Laboratory for Process Engineering, Environment, Biotechnology, and Energy—LEPABE funded by national funds through FCT/MCTES (PIDDAC) and N.M. for the Strategic project ref. UID/BIM/04293/2013 and “NORTE2020 - Programa Operacional Regional do Norte” (NORTE-01-0145-FEDER-000012)

Faculty Publications and Creative Works 1998

One of the ways in which we recognize our faculty at the University of New Mexico is through Faculty Publications & Creative Works. An annual publication, it highlights our faculty\u27s scholarly and creative activities and achievements and serves as a compendium of UNM faculty efforts during the 1998 calendar year. Faculty Publications & Creative Works strives to illustrate the depth and breadth of research activities performed throughout our University\u27s laboratories, studios and classrooms. We believe that the communication of individual research is a significant method of sharing concepts and thoughts and ultimately inspiring the birth of new ideas. In support of this, UNM faculty during 1998 produced over 2,457 works, including 1,990 scholarly papers and articles, 69 books, 98 book chapters, 119 reviews, 165 creative works and 16 patents. We are proud of the accomplishments of our faculty which are in part reflected in this book, which illustrates the diversity of intellectual pursuits in support of research and education at the University of New Mexico. Nasir Ahmed, Ph.D. Interim Associate Provost for Research and Dean of Graduate Studie

MULTI-DIMENSIONAL ANALYSIS APPROACHES FOR HETEROGENEOUS SINGLE-CELL DATA

Improvements in experimental techniques have led to an explosion of information in biology research. The increasing number of measurements comes with challenges in analyzing resulting data, as well as opportunities to obtain deeper insights of biological systems. Conventional average based methods are unfit to analyze high dimensional datasets since they fail to take full advantage of such rich information. More importantly, they are not able to capture the heterogeneity that is prevalent in biological systems. Sophisticated algorithms that are able to utilize all available measurements simultaneously are hence emerging rapidly. These algorithms excel at making full use of information within datasets and revealing detailed heterogeneity. However, there are several important disadvantages of existing algorithms. First, specific knowledge in statistics or machine learning is required to appropriately interpret and tune parameters in these algorithms for future use. This may result in misusage and misinterpretation. Second, using all measurements with equal weighting runs the risk of noise contamination. In addition, information overload has become more common in biology research, with a large volume of irrelevant measurements. Third, regardless of the quality of measurements, analysis methods that simultaneously use a large number of measurements need to avoid the “curse of dimensionality”, which warns that distance estimation and nearest neighbor estimation are not meaningful in high dimensional space. However, most current sophisticated algorithms involve distance estimation and/or nearest neighbor estimation. In this dissertation, my goal is to build analysis methods that are complex enough to capture heterogeneity and at the same time output results in a format that is easy to interpret and familiar to biologists and medical researchers. I tackle the dimension reduction problem by finding not the best subspace but dividing them into multiple subspaces and examine them one by one. I demonstrate my methods with three types of datasets: image-based high-throughput screening data, flow cytometry data, and mass cytometry data. From each dataset, I was able to discover new biological insights as well as re-validate well-established findings with my methods

Personalisation of dexamethasone in childhood acute lymphoblastic leukaemia

PhD ThesisDexamethasone (dex) is a key treatment for childhood acute lymphoblastic leukaemia (ALL), but is associated with significant variability in terms of toxicity and efficacy. In this project, the following variables were assessed to better understand how dex personalisation may be achieved: pharmacokinetics, intracellular dex accumulation, glucocorticoid receptor (GR) posttranslational modifications and B-cell maturation state. For pharmacokinetic studies, samples were collected from 154 patients randomised to short (10mg/m2 x 14 days) or standard (6mg/m2 x 28 days) dex induction therapy, as part of the UKALL 2011 trial, and analysed using a validated LC/MS method. Wide pharmacokinetic variability was observed, with AUC0-12h and Cmax significantly higher on the short compared to standard arm. However there was substantial overlap between the two arms, with a number of patients on the standard arm exhibiting higher exposures than those on short therapy. The UKALL 2011 trial found no statistical difference in terms of steroid-related toxicity or MRD response between short and standard dosing. These data suggest that the considerable dex pharmacokinetic variation identified may be a more important factor than variation in dosing regimen. For cellular pharmacology experiments, cell lines, primagraft and primary patient samples were studied. Dex sensitivity was assessed using Alamar Blue assays and GI50 values ranged from 2-1000nM. Western blotting indicated wildtype GR in all samples. Dex accumulation was assessed by LC/MS and flow cytometric analysis of dex-FITC. While patient samples exhibited large variability, dex accumulation was not significantly different between sensitive and resistant cells. Differential dex sensitivity was not accounted for by differences in GR posttranslational modifications, assessed using capillary isoelectric focusing. However, assessment of B-cell maturation using mass cytometry revealed a relationship with dex resistance. Importantly, >50% of patient cell samples had dex GI50 values greater than plasma concentrations observed on either arm of the UKALL 2011 trial. A combined approach incorporating pharmacokinetic assessments and cellular response in ALL cells may allow a more comprehensive understanding of dex pharmacology to optimise its clinical utility.Cancer Research UK, for funding my project, and to Children with Cancer UK, the ECMC and NECCR for the funds they provided

Toxicology and molecular epidemiology of microbes detected in surface water in the Western Cape: The Impact of Informal Settlement

>Magister Scientiae - MScInformal settlements are often implicated in surface water pollution with faecal matter. In most instances faecal pollution in the associated surface waters persists despite improvements in sewage removal infrastructure. This study evaluates the importance of investigating the water quality of the Plankenbrug River before it reaches Khayamnandi settlement by comparing water quality in spring and in winter upstream (Pre-Khayamnandi) and downstream (Post- Khayamnandi) from the settlement. In this study, faecal indicator bacteria (Escherichia coli and total coliforms) were enumerated using Chromocult agar. E. coli was further characterized with analytical profiling index (API) and haemolysis assays. Both Pre- and Post-Khayamnandi were not significantly different from each other for both total coliforms and E. coli in winter. Pre-Khayamnandi had between 105 and 108 cfu/100 ml for total coliforms while Post-Khayamnandi had total coliform colony count between 106 and 107 cfu/100 ml. E. coli also exhibited a similar pattern with slightly higher counts at Post-Khayamnandi with colony counts from 104 to 107 and 105 to 107 cfu/100 ml. Spring microbial count demonstrated a significant difference to winter counts within each test site (p ≤ 0.01) and across the two sites (p ≤ 0.05). Both total coliforms and E. coli were 102 fold higher at Post-Khayamnandi than at Pre-Khayamnandi in spring. The API assay demonstrated significant difference (p ≤ 0.05) between the two test sites. Pre- Khayamnandi predominantly had two different profiles while Post-Khayamnandi had three. These profiles represented five distinct E. coli biotypes. Sorbitol and sucrose tests within the API assay demonstrated significant differences (p ≤ 0.05) between the two test sites. The prevalence of sorbitol fermenters at Pre-Khayamnandi was 100% while at Post-Khayamnandi it was 73%. Pre-Khayamnandi also demonstrated a significantly higher prevalence of sucrose fermenters than Post-Khayamnandi at 100% and 59% respectively. These differences indicated dissimilar sources of faecal contamination around these sites. Differences in the distributions of sorbitol and sucrose fermenting biotypes demonstrate different toxicity potentials across these two test sites. The haemolysis assay demonstrated that 9% of isolates were haemolytic with reference to both known α- and β-haemolyitic streptococci at Post-Khayamnandi. At Pre-Khayamnandi there was a higher percentage of α- and β-haemolyitic species, 29% and 28%, respectively. Post- Khayamnandi and Pre-Khayamnandi were significantly different from each other with reference to both α- and β-haemolysis (p ≤ 0.05). These haemolytic activities also demonstrate different toxicity potentials across the two sites. In conclusion Khayamnandi contributes to an already heavy faecal load in the Plankenbrug River. Thus remedial measures to maintain high surface water quality of Plankenbrug River should be directed upstream from the Khayamnandi settlement as well as within the settlement equally. This study recommends integration of microbial loads with programs such as the National Microbial Monitoring Program of South Africa to drive prioritization process in directing reclaiming of water quality, inter alia