Mutational Landscape of Bone Marrow CD19 and CD138 Cells in Waldenström Macroglobulinemia (WM) and IgM Monoclonal Gammopathy of Undetermined Significance (IgM MGUS).

Background: Despite recurrent and activating mutations, including MYD88, CXCR4, ARID1A, KMT2D, and CD79B were identified, the genetic basis for Waldenström's Macroglobulinemia (WM) and the risk of progression of IgM MGUS to WM remain to be fully elucidated.

Methods: We investigated the mutation status of WM (n = 8), sWM (n = 7), and IgM MGUS (n = 5) patients, by performing high-throughput targeted AmpliSeq NGS on 117 target genes. Specifically, we analyzed the CD19+ cells from 15 WM/sWM patients and five IgM MGUS patients.

Clothing and individual equipment for the female soldier: developing a framework to improve the evidence base which informs future design and evaluation.

The development of inclusive equipment and clothing is a priority across national defence departments that are part of The Technical Cooperation Programme. As such, a collaborative effort has been established to inform the development of clothing and equipment for women. This invited review provides an overview of an ongoing collaborative project presented at the sixth International Congress on Soldiers Physical Performance.

Rho-dependent transcriptional switches regulate the bacterial response to cold shock.

Binding of the bacterial Rho helicase to nascent transcripts triggers Rho-dependent transcription termination (RDTT) in response to cellular signals that modulate mRNA structure and accessibility of Rho utilization (Rut) sites. Despite the impact of temperature on RNA structure, RDTT was never linked to the bacterial response to temperature shifts. We show that Rho is a central player in the cold-shock response (CSR), challenging the current view that CSR is primarily a posttranscriptional program.

Clothing and Equipment Fit Among Male and Female Canadian Armed Forces Members.

Introduction: The fit of military clothing and equipment is essential for the health and safety of military operators. Given the aim of increasing the proportion of women and the known biological and morphological differences between male and female soldiers, an understanding of fit across different items of kit is needed. The aim of this study was to quantify subjective fit ratings of 8 items of military clothing and equipment, including combat shirt, combat pants, rucksack, small pack, tactical vest, fragmentation vest, helmet, and ballistic eyewear as a function of relative stature and occupational group among male and female Canadian Armed Forces members.

Transcription-driven DNA supercoiling counteracts H-NS-mediated gene silencing in bacterial chromatin.

In all living cells, genomic DNA is compacted through interactions with dedicated proteins and/or the formation of plectonemic coils. In bacteria, DNA compaction is achieved dynamically, coordinated with dense and constantly changing transcriptional activity. H-NS, a major bacterial nucleoid structuring protein, is of special interest due to its interplay with RNA polymerase.

Dynamics of macrophage polarization support persistence in a whole living organism.

Numerous intracellular bacterial pathogens interfere with macrophage function, including macrophage polarization, to establish a niche and persist. However, the spatiotemporal dynamics of macrophage polarization during infection within host remain to be investigated. Here, we implement a model of persistent Typhimurium infection in zebrafish, which allows visualization of polarized macrophages and bacteria in real time at high resolution.

Immunogenicity of PE18, PE31, and PPE26 proteins from in humans and mice.

Introduction: The large family of PE and PPE proteins accounts for as much as 10% of the genome of . In this study, we explored the immunogenicity of three proteins from this family, PE18, PE31, and PPE26, in humans and mice.

Methods: The investigation involved analyzing the immunoreactivity of the selected proteins using sera from TB patients, IGRA-positive household contacts, and IGRA-negative BCG vaccinated healthy donors from the TB endemic country Mozambique.

A Nine-Gene Expression Signature Distinguished a Patient with Chronic Lymphocytic Leukemia Who Underwent Prolonged Periodic Fasting.

: This study aimed to investigate the causes of continuous deep fluctuations in the absolute lymphocyte count (ALC) in an untreated patient with Chronic Lymphocytic Leukemia (CLL), who has had a favorable prognosis since the time of diagnosis. Up until now, the patient has voluntarily chosen to adopt a predominantly vegetarian and fruitarian diet, along with prolonged periods of total fasting (ranging from 4 to 39 days) each year. : For this purpose, we decided to analyze the whole transcriptome profiling of peripheral blood (PB) CD19+ cells from the patient (#1) at different time-points vs.

Cordycepin (3'dA) Induces Cell Death of AC133 Leukemia Cells via Re-Expression of and Down-Modulation of .

Wnt/β-catenin signaling is critically required for the development and maintenance of leukemia stem cells (LSCs) in acute myeloid leukemia (AML) by constitutive activation of myeloid regeneration-related pathways. Cell-intrinsic activation of canonical Wnt signaling propagates in the nucleus by β-catenin translocation, where it induces expression of target oncogenes such as , and . As the Wnt/β-catenin pathway is now well established to be a key oncogenic signaling pathway promoting leukemic myelopoiesis, targeting it would be an effective strategy to impair LSC functionality.

The effect of two sessions of combined jump and sprint training per week on fitness parameters in soccer players. A randomized controlled trial.

This study aimed to investigate the effect of a combined jump and sprint training program, two sessions a week for 6 weeks, on sprinting, change of directions (COD) and jumping performance in semi-professional soccer players. Twenty soccer players were enrolled in this randomized controlled trial (age 20 ± 2 years, body mass 74.3 ± 5.

Comparison of In-service Reduced vs. Full Torso Coverage Armor for Females.

Introduction: Body armor and torso-borne equipment are critical to the survivability and operational effectiveness of a soldier. Historically, in-service designs have been predominantly designed for males or unisex, which may be disadvantageous for females who are shaped differently and, on average, smaller in stature and mass than their male counterparts. This study assesses the biomechanical and performance impact of two Canadian in-service armors and fighting load conditions on females.

Generating Libraries of Random or Gene Fusions in the Bacterial Chromosome: Screening for Differentially Expressed Fusions.

Transposable elements engineered to generate random gene fusions in the bacterial chromosome are valuable tools in the study of gene expression. In this protocol, we describe the use of a new series of transposons designed to obtain random fusions to either the operon or the gene for superfolder green fluorescent protein (sfGFP). Transposition is achieved through the activity of the hyperactive variant of Tn5 transposase (Tnp) whose gene is positioned in with respect to the transposable module and under the control of the anyhydrotetracycline (AHTc)-inducible P promoter.

Use of Transposable Reporters in the Analysis of Bacterial Regulatory Networks.

Transposable elements are genetic entities that have the capacity to promote their own translocation from one site to another within a genome. Initially discovered in by Barbara McClintock at the Cold Spring Harbor Laboratory, transposable elements have been found to populate the genomes of all forms of life. In bacteria, the discovery of transposons significantly enhanced genetic analyses; they have been widely used to make insertion mutants and have inspired elegant strategies for strain construction and in vivo genome engineering.

Mapping Transposon Insertion Sites by Inverse Polymerase Chain Reaction and Sanger Sequencing.

Inverse polymerase chain reaction (PCR) is a method designed to amplify a segment of DNA for which only a portion of the sequence is known. The method consists of circularizing the DNA fragment by self-ligation and performing PCR with primers annealing inside the known sequence but pointing away from each other (hence the technique is also called "inside-out PCR"). Here we describe how inverse PCR can be used to identify the site of transposon insertion in the bacterial chromosome.

Construction of Single-Copy Fluorescent Protein Fusions by One-Step Recombineering.

We describe a simple recombineering-based procedure for generating single-copy gene fusions to superfolder GFP (sfGFP) and monomeric Cherry (mCherry). The open reading frame (orf) for either protein is inserted at the targeted chromosomal location by λ Red recombination using an adjacent drug-resistance cassette ( or ) for selection. The drug-resistance gene is flanked by flippase (Flp) recognition target (FRT) sites in direct orientation, which allows removal of the cassette by Flp-mediated site-specific recombination once the construct is obtained, if desired.

Converting an FRT-Tagged Gene into a Fluorescent Protein Gene Fusion by Flp-Mediated Site-Specific Recombination.

This protocol uses conditional plasmids carrying the open reading frame (orf) of either superfolder green fluorescent protein (sfGFP) or monomeric Cherry (mCherry) fused to a flippase (Flp) recognition target (FRT) site. In cells expressing the Flp enzyme, site-specific recombination between the plasmid-borne FRT and an FRT "scar" in a target gene in the bacterial chromosome results in chromosomal integration of the plasmid with the concomitant in-frame fusion of the target gene to the fluorescent protein orf. This event can be positively selected using an antibiotic-resistance marker ( or ) present on the plasmid.

DNA Recombineering Applications.

The ability to manipulate the bacterial genome is an obligatory premise for the study of gene function and regulation in bacterial cells. The λ red recombineering technique allows modification of chromosomal sequences with base-pair precision without the need of intermediate molecular cloning steps. Initially conceived to construct insertion mutants, the technique lends itself to a wide variety of applications including the creation of point mutants, seamless deletions, reporter, and epitope tag fusions and chromosomal rearrangements.

Recombineering 101: Making an in-Frame Deletion Mutant.

DNA recombineering uses phage λ Red recombination functions to promote integration of DNA fragments generated by polymerase chain reaction (PCR) into the bacterial chromosome. The PCR primers are designed to have the last 18-22 nt anneal on either side of the donor DNA and to carry 40- to 50-nt 5' extensions homologous to the sequences flanking the chosen insertion site. The simplest application of the method results in knockout mutants of nonessential genes.

Scarless DNA Recombineering.

The method described here allows editing of the bacterial genome without leaving any secondary changes (scars) behind. This method uses a tripartite selectable and counterselectable cassette comprising an antibiotic-resistance gene ( or ) and the repressor gene linked to a P promoter- toxin gene fusion. In the absence of induction, the gene product represses the P promoter, preventing expression.

[Late preterm newborn: follow-up recommendations].

The WHO states that prematurity rates have increased mainly due to late preterm births. Since these babies are usually born with appropriate weight for their gestational age, their risk for morbidities such as neurodevelopmental delays, behavioral problems and organ systems immaturity are overlooked. Further, these clinical findings have an impact on short and long term outcomes (i.

Preparing Bacterial Genomic DNA.

We describe two alternative procedures for purifying bacterial chromosomal DNA. The first procedure incorporates the use of a commercial kit based on silica membrane technology. This approach relies on the selective binding of DNA to a silica-based column in the presence of chaotropic salts (guanidine salts).

Working with Phage P22.

Transduction experiments in and are usually performed with virulent phage variants. A widely used P1 mutant, called P1 , carries one or more uncharacterized mutations that prevent formation of lysogens. In the case of P22, by far the most frequently used variant is named P22 HT105/1 This phage has a high transducing (HT) frequency due to a mutant nuclease with lower specificity for the sequence.

Preparing Plasmid DNA from Bacteria.

The most common method for isolating plasmid DNA is derived from an alkaline lysis procedure. The procedure exploits the differential partitioning of plasmid and chromosomal DNA when denatured by alkali and subsequently renatured by neutralization of the medium. The circular covalently closed nature of plasmid DNA allows the denatured DNA strands to quickly find each other and reanneal during the renaturation step.