Mycobacterium tuberculosis Responds to Chloride and pH as Synergistic Cues to the Immune Status of its Host Cell

PubMed ID: 23592993This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

Probing host pathogen cross-talk by transcriptional profiling of both Mycobacterium tuberculosis and infected human dendritic cells and macrophages

This study provides the proof of principle that probing the host and the microbe transcriptomes simultaneously is a valuable means to accessing unique information on host pathogen interactions. Our results also underline the extraordinary plasticity of host cell and pathogen responses to infection, and provide a solid framework to further understand the complex mechanisms involved in immunity to M. tuberculosis and in mycobacterial adaptation to different intracellular environments

Prime movers : mechanochemistry of mitotic kinesins

Mitotic spindles are self-organizing protein machines that harness teams of multiple force generators to drive chromosome segregation. Kinesins are key members of these force-generating teams. Different kinesins walk directionally along dynamic microtubules, anchor, crosslink, align and sort microtubules into polarized bundles, and influence microtubule dynamics by interacting with microtubule tips. The mechanochemical mechanisms of these kinesins are specialized to enable each type to make a specific contribution to spindle self-organization and chromosome segregation

m6A potentiates Sxl alternative pre-mRNA splicing for robust Drosophila sex determination

N6-methyladenosine (m6A) is the most common internal modification of eukaryotic messenger RNA (mRNA) and is decoded by YTH domain proteins1, 2, 3, 4, 5, 6, 7. The mammalian mRNA m6A methylosome is a complex of nuclear proteins that includes METTL3 (methyltransferase-like 3), METTL14, WTAP (Wilms tumour 1-associated protein) and KIAA1429. Drosophila has corresponding homologues named Ime4 and KAR4 (Inducer of meiosis 4 and Karyogamy protein 4), and Female-lethal (2)d (Fl(2)d) and Virilizer (Vir)8, 9, 10, 11, 12. In Drosophila, fl(2)d and vir are required for sex-dependent regulation of alternative splicing of the sex determination factor Sex lethal (Sxl)13. However, the functions of m6A in introns in the regulation of alternative splicing remain uncertain3. Here we show that m6A is absent in the mRNA of Drosophila lacking Ime4. In contrast to mouse and plant knockout models5, 7, 14, Drosophila Ime4-null mutants remain viable, though flightless, and show a sex bias towards maleness. This is because m6A is required for female-specific alternative splicing of Sxl, which determines female physiognomy, but also translationally represses male-specific lethal 2 (msl-2) to prevent dosage compensation in females. We further show that the m6A reader protein YT521-B decodes m6A in the sex-specifically spliced intron of Sxl, as its absence phenocopies Ime4 mutants. Loss of m6A also affects alternative splicing of additional genes, predominantly in the 5′ untranslated region, and has global effects on the expression of metabolic genes. The requirement of m6A and its reader YT521-B for female-specific Sxl alternative splicing reveals that this hitherto enigmatic mRNA modification constitutes an ancient and specific mechanism to adjust levels of gene expression

Pathogenic mycobacteria achieve cellular persistence by inhibiting the Niemann-Pick Type C disease cellular pathway

Background. Tuberculosis remains a major global health concern. The ability to prevent phagosome-lysosome fusion is a key mechanism by which intracellular mycobacteria, including Mycobacterium tuberculosis, achieve long-term persistence within host cells. The mechanisms underpinning this key intracellular pro-survival strategy remain incompletely understood. Host macrophages infected with intracellular mycobacteria share phenotypic similarities with cells taken from patients suffering from Niemann-Pick Disease Type C (NPC), a rare lysosomal storage disease in which endocytic trafficking defects and lipid accumulation within the lysosome lead to cell dysfunction and cell death. We investigated whether these shared phenotypes reflected an underlying mechanistic connection between mycobacterial intracellular persistence and the host cell pathway dysfunctional in NPC.  Methods. The induction of NPC phenotypes in macrophages from wild-type mice or obtained from healthy human donors was assessed via infection with mycobacteria and subsequent measurement of lipid levels and intracellular calcium homeostasis. The effect of NPC therapeutics on intracellular mycobacterial load was also assessed.  Results. Macrophages infected with intracellular mycobacteria phenocopied NPC cells, exhibiting accumulation of multiple lipid types, reduced lysosomal Ca 2+ levels, and defects in intracellular trafficking. These NPC phenotypes could also be induced using only lipids/glycomycolates from the mycobacterial cell wall. These data suggest that intracellular mycobacteria inhibit the NPC pathway, likely via inhibition of the NPC1 protein, and subsequently induce altered acidic store Ca 2+ homeostasis. Reduced lysosomal calcium levels may provide a mechanistic explanation for the reduced levels of phagosome-lysosome fusion in mycobacterial infection. Treatments capable of correcting defects in NPC mutant cells via modulation of host cell calcium were of benefit in promoting clearance of mycobacteria from infected host cells.  Conclusion. These findings provide a novel mechanistic explanation for mycobacterial intracellular persistence, and suggest that targeting interactions between the mycobacteria and host cell pathways may provide a novel avenue for development of anti-TB therapies

A Pro Bono Physical Therapy Clinic’s Pandemic Pivot to Telehealth and Its Impact on Student Readiness for a First Full-Time Clinic Experience

Purpose: The COVID-19 pandemic of 2020 led to a multitude of adjustments in physical therapist education. This article will describe the delivery model pivot that a student-run pro bono clinic made to sustain client care and student experience. The change in delivery model also led to a change in care model. The purpose of this study is to explore the impact that the change in delivery and care model within the student-run pro bono clinic had on student readiness for a first formal clinical education experience. Methods: This qualitative investigation utilized participant journals and a focus group to capture participants’ reflections and experiences in the first four weeks of their full-time clinical experience. Content analysis guided the research team in the data analysis. Triangulation, an audit trail, reflexivity, and member checking further enhanced confirmability of findings. Results: Seven participants kept journals and participated in the focus group. Six categories of impact emerged, three because of the change in delivery to telehealth and three due to the change in care model which led to increased continuity of care. The three categories related to telehealth included 1) impact on clinical skills, 2) facilitating communication, and 3) window into their home. The three categories specific to increased continuity of care included 1) clinical reasoning skills, 2) documentation, and 3) client rapport. Conclusions: Telehealth and the increased continuity of care presented advantages and disadvantages to student readiness. Post pandemic, student leaders should consider ways in which they might retain the positive outcomes of the switch in delivery and care model while resuming care in-person

Inland Waterway Operational Model & Simulation Along the Ohio River

The inland waterway system of the U.S. is a vital network for transporting key goods and commodities from the point of production to manufacturers and consumers. Shipping materials via the inland waterways is arguably the most economical and environmentally friendly option (compared to hauling freight by trains or railways). Despite the advantages the inland waterways enjoys over competing modes, key infrastructure – such as locks and dams, which help to control water levels on a number of rivers and make navigation possible – is declining. Limited funds have been allocated to make the necessary repairs to lock and dam facilities. Over the past 10 years Inland Waterways Trust Fund resources (which historically funded maintenance and improvement projects) has steadily declined. Locks and dams are of particular importance, because they assist in the maintenance of navigable depths on many of the major inland waterways (Ohio River, Upper Mississippi River, Tennessee River). To better understand the operation of the inland waterway system, this report examines a portion of the Ohio River (extending from Markland Locks and Dam to Lock 53). The specific focus is to determine what delays barge tows as they attempt to lock through these critical facilities. The Ohio River is a particularly important study area. In many ways it is representative of the conditions present throughout the inland waterways system. The average age of the lock and dam facilities exceed 50 years along our study segment. Most of these facilities are operating beyond their intended design life. As locks age, they increasingly demand more scheduled and unscheduled maintenance activities. Maintenance activities often require temporarily shuttering a lock chamber and diverting traffic through another onsite chamber (often of smaller capacity). All of the facilities included in the research area have two lock chambers ‐ thus, if one goes down for maintenance all vessels are diverted through the second chamber. In many cases this situation can produce extensive delays, which precludes cargo from reaching the destination in a timely manner. Recently, the aggregate number of hours that shippers and carriers lose due to delays has escalated. Although the U.S. Army Corps of Engineers – the agency responsible for the management and oversight of locks and dams – has worked to keep traffic flowing on the river, tightening budgets hamper efforts. For shippers and carriers to make informed decisions about when and where to deploy freight on the river, they require knowledge that illuminates factors that are most significant in affecting transit times. In particular this applies to certain conditions that are likely to create delays at lock and dam facilities. The purpose of this report is to 1) develop a comprehensive profile of the Ohio River that provides an overview of how it is integral to U.S. economic security 2) identify salient river characteristics or externally‐driven variables that influence the amount of water flowing through the main channel which consequently impacts vessels’ capacity to navigate 3) use this information (along with a 10‐year data set encompassing over 600,000 observations) to develop an Inland Waterways Operational Model (IWOM). The IWOM objective is to provide the U.S. Army Corps of Engineers, shippers, carriers, and other interested parties with access to8 a robust method that aids in the prediction of where and when conditions will arise on the river that have the potential to significantly impact lockage times and queue times (i.e. how long a vessel has to wait after it arrives at a facility to lock through). After qualitatively reviewing different features of the river system that affect vessel traffic, this report outlines two approaches to modeling inland waterway system behavior – a discrete event simulation (DES) model which uses proprietary software, and the IWOM. Although the DES produced robust findings that aligned with the historical data (because it relies upon proprietary software), it does not offer an ideal platform to distribute knowledge to stakeholders. Indeed, this is the major drawback of the DES given a critical objective of this project is to generate usable information for key stakeholders who are involved with inland waterway operations. Conversely, the IWOM is a preferable option given it relies on statistical analysis – in this sense, it is more of an open‐source solution. The IWOM uses linear regression to determine key variables affecting variation in lockage time. The final model accounts for over two‐thirds of the observed variation in lockage times from 2002‐2012, which is our study period. Practically, this means that the difference between predicted values and observed delay times is significantly less than how the delays vary around the composite average seen in the river system (R2 = 0.69). The IWOM confirms that variations in river conditions significantly affect vessel travel times. For example, river discharge ‐ the direction a vessel moves up or down a river ‐ meaningfully influences lockage times. The freight amount a vessel carries, which is represented by the amount of draft and newness of a vessel, influences lockage times. Larger vessels with more draft tend to wait longer and take longer to complete their lockage. The IWOM is less successful at predicting delay times. Because there is greater instability in this data only a modest amount of variation is explained by the model (R2 = 0.23). This, in turn, partly reflects in spillover from one vessel to the next that is difficult for the simulation to impose and account for therefore requiring additional logic. Once completed, the IWOM was used to parameterize a simulation model. This provided a graphical representation of vessels moving along the river. Users have the capability of adjusting the effects of different variables to anticipate how the system may react, and what changes in vessel traffic patterns emerge. This information will be of great use for stakeholders wanting to gain a better understanding of what conditions lockage times will increase or decrease, why delays emerge, and consequently how these impact traffic flows on the river. In programming a simulation model, users are able to visualize and intuit what causes vessel travel times to vary. Although the regression model accomplishes this, for many users this would prove unwieldy and difficult to grasp beyond a conceptual, abstract level. Matching up regression results with a visual counterpart lets users gain immediate and intimate knowledge of river and vessel behavior – this in turn can positively affect shipper and carrier modal choices. The report concludes with some recommendations for IWOM implementation and thoughts on future research needs. Also discussed are the implications results from the present study have for improving our ability to safely, securely, and swiftly move freight on the inland waterways network

The effect of the oil resin on the properties of solution of the petroleum wax treated in an ultrasonic field

It was found that the complex treatment of ultrasonic followed by the addition of 0.3% by weight. petroleum resins, a more efficient method of inhibiting sedimentation processes than just ultrasonic or addition of 0,3% by weight. petroleum resins. According to the obtained data, fragments of aliphatic petroleum resins are adsorbed on the high molecular hydrocarbons of normal structure and prevent their aggregation thus the inhibition of sedimentation occurs

Erratum to: 36th International Symposium on Intensive Care and Emergency Medicine

[This corrects the article DOI: 10.1186/s13054-016-1208-6.]

Global analysis of X-chromosome dosage compensation

BACKGROUND: Drosophila melanogaster females have two X chromosomes and two autosome sets (XX;AA), while males have a single X chromosome and two autosome sets (X;AA). Drosophila male somatic cells compensate for a single copy of the X chromosome by deploying male-specific-lethal (MSL) complexes that increase transcription from the X chromosome. Male germ cells lack MSL complexes, indicating that either germline X-chromosome dosage compensation is MSL-independent, or that germ cells do not carry out dosage compensation. RESULTS: To investigate whether dosage compensation occurs in germ cells, we directly assayed X-chromosome transcripts using DNA microarrays and show equivalent expression in XX;AA and X;AA germline tissues. In X;AA germ cells, expression from the single X chromosome is about twice that of a single autosome. This mechanism ensures balanced X-chromosome expression between the sexes and, more importantly, it ensures balanced expression between the single X chromosome and the autosome set. Oddly, the inactivation of an X chromosome in mammalian females reduces the effective X-chromosome dose and means that females face the same X-chromosome transcript deficiency as males. Contrary to most current dosage-compensation models, we also show increased X-chromosome expression in X;AA and XX;AA somatic cells of Caenorhabditis elegans and mice. CONCLUSION: Drosophila germ cells compensate for X-chromosome dose. This occurs by equilibrating X-chromosome and autosome expression in X;AA cells. Increased expression of the X chromosome in X;AA individuals appears to be phylogenetically conserved