Search results for: Lior Pachter

Authors: Catherine Felce, Lior Pachter

Abstract:

Current methods for building phylogenetic trees from gene expression data consider mean expression levels. With single-cell technologies, we can leverage more information about cell dynamics by considering the entire distribution of gene expression across cells. Using biophysical modeling, we propose a method for constructing phylogenetic trees from scRNA-seq data, building on Felsenstein's method of continuous characters. This method can highlight genes whose level of expression may be unchanged between species, but whose rates of transcription/decay may have evolved over time.

Keywords: phylogenetics, single-cell, biophysical modeling, transcription

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Authors: Fan Gao, Lior Pachter

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The primary tool currently used to pre-process 10X Chromium single-cell ATAC-seq data is Cell Ranger, which can take very long to run on standard datasets. To facilitate rapid pre-processing that enables reproducible workflows, we present a suite of tools called scATAK for pre-processing single-cell ATAC-seq data that is 15 to 18 times faster than Cell Ranger on mouse and human samples. Our tool can also calculate chromatin interaction potential matrices, and generate open chromatin signal and interaction traces for cell groups. We use scATAK tool to explore the chromatin regulatory landscape of a healthy adult human brain and unveil cell-type specific features, and show that it provides a convenient and computational efficient approach for pre-processing single-cell ATAC-seq data.

Keywords: single-cell, ATAC-seq, bioinformatics, open chromatin landscape, chromatin interactome

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Authors: Lior Davidovitch

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Traditional training based on static models and case studies is the backbone of most teaching and training programs of engineering education. However, project management learning is characterized by dynamics models that requires new and enhanced learning method. The results of empirical experiments evaluating the effectiveness and efficiency of using cognitive simulator as a new training technique are reported. The empirical findings are focused on the impact of keeping and reviewing learning history in a dynamic and interactive simulation environment of engineering education. The cognitive simulator for engineering project management learning had two learning history keeping modes: manual (student-controlled), automatic (simulator-controlled) and a version with no history keeping. A group of industrial engineering students performed four simulation-runs divided into three identical simple scenarios and one complicated scenario. The performances of participants running the simulation with the manual history mode were significantly better than users running the simulation with the automatic history mode. Moreover, the effects of using the undo enhanced further the learning process. The findings indicate an enhancement of engineering students’ learning and decision making when they use the record functionality of the history during their engineering training process. Furthermore, the cognitive simulator as educational innovation improves students learning and training. The practical implications of using simulators in the field of engineering education are discussed.

Keywords: cognitive simulator, decision making, engineering learning, project management

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Authors: Kan Yu, Khui Hung Lee, Eben Afrifa-Yamoah, Jing Guo, Katrina Harrison, Jack Goldblatt, Nicholas Pachter, Jitian Xiao, Guicheng Brad Zhang

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Background and Significance of the Study: Congenital Heart Defects (CHDs) are the most common malformation at birth and one of the leading causes of infant death. Although the exact etiology remains a significant challenge, epigenetic modifications, such as DNA methylation, are thought to contribute to the pathogenesis of congenital heart defects. At present, no existing DNA methylation biomarkers are used for early detection of CHDs. The existing CHD diagnostic techniques are time-consuming and costly and can only be used to diagnose CHDs after an infant was born. The present study employed a machine learning technique to analyse genome-wide methylation data in children with and without CHDs with the aim to find methylation biomarkers for CHDs. Methods: The Illumina Human Methylation EPIC BeadChip was used to screen the genome‐wide DNA methylation profiles of 24 infants diagnosed with congenital heart defects and 24 healthy infants without congenital heart defects. Primary pre-processing was conducted by using RnBeads and limma packages. The methylation levels of top 600 genes with the lowest p-value were selected and further investigated by using a random forest approach. ROC curves were used to analyse the sensitivity and specificity of each biomarker in both training and test sample sets. The functionalities of selected genes with high sensitivity and specificity were then assessed in molecular processes. Major Findings of the Study: Three genes (MIR663, FGF3, and FAM64A) were identified from both training and validating data by random forests with an average sensitivity and specificity of 85% and 95%. GO analyses for the top 600 genes showed that these putative differentially methylated genes were primarily associated with regulation of lipid metabolic process, protein-containing complex localization, and Notch signalling pathway. The present findings highlight that aberrant DNA methylation may play a significant role in the pathogenesis of congenital heart defects.

Keywords: biomarker, congenital heart defects, DNA methylation, random forest

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Authors: Lior Rake, Shmulik Pinkert

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Methane hydrate has been found in significant quantities in soils offshore within continental margins and in permafrost within arctic regions where low temperature and high pressure are present. The mechanical parameters for geotechnical engineering are commonly evaluated in geomechanical laboratories adapted to simulate the environmental conditions of methane hydrate-bearing sediments (MHBS). Due to the complexity and high cost of natural MHBS sampling, most laboratory investigations are conducted on artificially formed samples. MHBS artificial samples can be formed using different hydrate formation methods in the laboratory, where methane gas and water are supplied into the soil pore space under the methane hydrate phase conditions. The most commonly used formation method is the excess gas method which is considered a relatively simple, time-saving, and repeatable testing method. However, there are several differences in the procedures and techniques used to produce the hydrate using the excess gas method. As a result of the difference between the test facilities and the experimental approaches that were carried out in previous studies, different measurement criteria and analyses were proposed for MHBS geomechanics. The lack of uniformity among the various experimental investigations may adversely impact the reliability of integrating different data sets for unified mechanical model development. In this work, we address some fundamental aspects relevant to reliable MHBS geomechanical investigations, such as hydrate homogeneity in the sample, the hydrate formation duration criterion, the hydrate-saturation evaluation method, and the effect of temperature measurement accuracy. Finally, a set of recommendations for repeatable and reliable MHBS formation will be suggested for future standardization of MHBS geomechanical investigation.

Keywords: experimental study, laboratory investigation, excess gas, hydrate formation, standardization, methane hydrate-bearing sediment

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Authors: Ketaki D. Belsare, Nicholas F. Polizzi, Lior Shtayer, William F. DeGrado

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Nature utilizes metalloproteins to perform chemical transformations with activities and selectivities that have long been the inspiration for design principles in synthetic and biological systems. The chemical reactivities of metalloproteins are directly linked to local environment effects produced by the protein matrix around the metal cofactor. A complete understanding of how the protein matrix provides these interactions would allow for the design of functional metalloproteins. The de novo computational design of proteins have been successfully used in design of active sites that bind metals like di-iron, zinc, copper containing cofactors; however, precisely designing active sites that can bind small molecule ligands (e.g., substrates) along with metal cofactors is still a challenge in the field. The de novo computational design of a functional metalloprotein that contains a purposefully designed substrate binding site would allow for precise control of chemical function and reactivity. Our research strategy seeks to elucidate the design features necessary to bind the cofactor protoporphyrin IX (hemin) in close proximity to a substrate binding pocket in a four helix bundle. First- and second-shell interactions are computationally designed to control orientation, electronic structure, and reaction pathway of the cofactor and substrate. The design began with a parameterized helical backbone that positioned a single histidine residue (as an axial ligand) to receive a second-shell H-bond from a Threonine on the neighboring helix. The metallo-cofactor, hemin was then manually placed in the binding site. A structural feature, pi-bulge was introduced to give substrate access to the protoporphyrin IX. These de novo metalloproteins are currently being tested for their activity towards hydroxylation and epoxidation. The de novo designed protein shows hydroxylation of aniline to 4-aminophenol. This study will help provide structural information of utmost importance in understanding de novo computational design variables impacting the functional activities of a protein.

Keywords: metalloproteins, protein design, de novo protein, biocatalysis

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Authors: Lior B. Navon

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The objective of this study is to investigate how technology, such as social media, can influence the biopower hierarchy between the medical knowledge about the body and women’s bodily knowledge through the case study of the hashtag 'notpee'. In January 2015, the hashtag #notpee, relating to a feminine physiological phenomenon called female ejaculation (FE) or squirting (SQ) started circulating on twitter. This hashtag, born as a reaction to a medical study claiming that SQ is essentially involuntary emission of urine during sexual activity, sparked an unusual public discourse about FE, a phenomenon that is usually not discussed or referred to in socio-legitimate public spheres. This unusual backlash got the attention of women’s magazines and blogs, as well as more mainstream large and respected outlets such as The Guardian and CNN. Both the tweets on twitter, as well as the media coverage of them, were mainly aimed at rejecting the research’s findings. While not offering an alternative and choosing to define the phenomenon by negation, women argued that the fluid extracted was not pee based on their personal experiences. Based on a critical discourse analysis of 742 tweets with the hashtag 'notpee' between January 2015 and January 2016, and of 15 articles covering the backlash, this study suggests that the #notpee backlash challenged the power balance between the medical knowledge about the feminine body and the feminine bodily knowledge through two different, yet related, forms of resistance to biopower. The first resistance is to the authority over knowledge production — who has the power to produce 'true' statements when it comes to the body? Is it the women who experience the phenomenon, or is it the medical institution? The second resistance to biopower has to do with what we regard as facts or veracity. A critical discourse analysis reveals that while both the scientific field, as well as the women arguing against its findings, use empirical information, they, nevertheless, rely on two dichotomic databases- while the scientific research relies on samples from the 'dead like body', these woman are relying on their lived subjective senses as a source for fact making. Nevertheless, while #notpee is asking to change the power relations between the feminine subjective bodily knowledge and the seemingly objective masculine medical knowledge about the body, it by no means dismisses it. These women are essentially asking the medical institution to take into consideration the subjective body as well as the objective one while acknowledging and accepting the power of the latter over knowledge production.

Keywords: biopower, female ejaculation, new media, bodily knowledge

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Authors: Tamar Lin, Nufar Buchshtab, Yifat Elharar, Julian Nicenboim, Rotem Edgar, Iddo Weiner, Lior Zelcbuch, Ariel Cohen, Sharon Kredo-Russo, Inbar Gahali-Sass, Naomi Zak, Sailaja Puttagunta, Merav Bassan

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Background: Atopic dermatitis (AD) is a chronic, relapsing inflammatory skin disorder that is characterized by dry skin and flares of eczematous lesions and intense pruritus. Multiple lines of evidence suggest that AD is associated with increased colonization by Staphylococcus aureus, which contributes to disease pathogenesis through the release of virulence factors that affect both keratinocytes and immune cells, leading to disruption of the skin barrier and immune cell dysfunction. The aim of the current study is to develop a bacteriophage-based product that specifically targets S. aureus. Methods: For the discovery of phage, environmental samples were screened on 118 S. aureus strains isolated from skin samples, followed by multiple enrichment steps. Natural phages were isolated, subjected to Next-generation Sequencing (NGS), and analyzed using proprietary bioinformatics tools for undesirable genes (toxins, antibiotic resistance genes, lysogeny potential), taxonomic classification, and purity. Phage host range was determined by an efficiency of plating (EOP) value above 0.1 and the ability of the cocktail to completely lyse liquid bacterial culture under different growth conditions (e.g., temperature, bacterial stage). Results: Sequencing analysis demonstrated that the 118 S. aureus clinical strains were distributed across the phylogenetic tree of all available Refseq S. aureus (~10,750 strains). Screening environmental samples on the S. aureus isolates resulted in the isolation of 50 lytic phages from different genera, including Silviavirus, Kayvirus, Podoviridae, and a novel unidentified phage. NGS sequencing confirmed the absence of toxic elements in the phages’ genomes. The host range of the individual phages, as measured by the efficiency of plating (EOP), ranged between 41% (48/118) to 79% (93/118). Host range studies in liquid culture revealed that a subset of the phages can infect a broad range of S. aureus strains in different metabolic states, including stationary state. Combining the single-phage EOP results of selected phages resulted in a broad host range cocktail which infected 92% (109/118) of the strains. When tested in vitro in a liquid infection assay, clearance was achieved in 87% (103/118) of the strains, with no evidence of phage resistance throughout the study (24 hours). A S. aureus host was identified that can be used for the production of all the phages in the cocktail at high titers suitable for large-scale manufacturing. This host was validated for the absence of contaminating prophages using advanced NGS methods combined with multiple production cycles. The phages are produced under optimized scale-up conditions and are being used for the development of a topical formulation (BX005) that may be administered to subjects with atopic dermatitis. Conclusions: A cocktail of natural phages targeting S. aureus was effective in reducing bacterial burden across multiple assays. Phage products may offer safe and effective steroid-sparing options for atopic dermatitis.

Keywords: atopic dermatitis, bacteriophage cocktail, host range, Staphylococcus aureus

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