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[Autumn Pitz]

Basallike breast cancer (BLBC) or triple-negative breast cancer (TNBC) is an aggressive and highly metastatic subtype of human breast cancer. The present study aimed to elucidate the potential tumor-suppressive function of MATR3, an abundant nuclear protein, in BLBC/TNBC, whose cancer-relevance has not been characterized.

We analyzed in vitro tumorigenecity by cell proliferation and soft agar colony formation assays, apoptotic cell death by flow cytometry and Poly (ADP-ribose) polymerase (PARP) cleavage, epithelial-mesenchymal transition (EMT) by checking specific EMT markers with real-time quantitative PCR and in vitro migration and invasion by Boyden Chamber assays. To elucidate the underlying mechanism by which MATR3 functions as a tumor suppressor, we performed Tandem affinity purification followed by mass spectrometry (TAP-MS) and pathway analysis. We also scrutinized MATR3 expression levels in the different subtypes of human breast cancer and the correlation between MATR3 expression and patient survival by bioinformatic analyses of publicly available transcriptome datasets.

MATR3 suppressed in vitro tumorigenecity, promoted apoptotic cell death and inhibited EMT, migration, and invasion in BLBC/TNBC cells. Various proteins regulating apoptosis were identified as MATR3-binding proteins, and YAP/TAZ pathway was suppressed by MATR3. MATR3 expression was inversely correlated with the aggressive and metastatic nature of breast cancer. Moreover, high expression levels of MATR3 were associated with a good prognosis of breast cancer patients.

Our data demonstrate that MATR3 functions as a putative tumor suppressor in BLBC/TNBC cells. Also, MATR3 potentially plays a role as a biomarker in predicting chemotherapy-sensitivity and patient survival in breast cancer patients.

INTRODUCTION: As crucial as clinical laboratories are to preventing, identifying and managing resistance problems, laboratory scientists are among the most overlooked stakeholders. This review outlines the contributions that diagnostic laboratory systems should make toward all five of the World Health Organization's 2015 strategic objectives for antimicrobial resistance containment.

LABORATORY SYSTEMS IN RESISTANCE CONTAINMENT: Antimicrobial susceptibility testing and surveillance are central to antibacterial resistance management and control and need to be implemented more commonly and closer to sick patients. However, the scope of tests that promote judicious antimicrobial use extend beyond susceptibility testing. Laboratory tests for pathogens or their associated biomarkers confirm or rule out specific causes of signs and symptoms associated with infection. Laboratory systems also provide critical support to infection control programmes. All of these functions promote rational antimicrobial use and contain the spread of resistance. Routine laboratory data supports the development of vaccines and other technologies that could ease the pressure placed by antimicrobials. Laboratories are also a rich source of information for health professionals, policymakers and the general public about the urgency of the resistance problem and progress in containing it.

CONCLUSION: Laboratory systems are integral to antimicrobial resistance containment and contributions from African laboratories to addressing resistance need to be enhanced.

Antimicrobial resistance is a grand global challenge that is reducing the success and increasing the cost of treating infections. As with many complex problems, there are many stakeholders in resistance - patients, prescribers and dispensers are often cited.1 However, much less visible but high on the list are those stakeholders that develop and deploy diagnostic tests. Diagnostic testing is key to resistance containment and may in fact be the most important tool for increasing appropriate access to antimicrobials while simultaneously containing resistance.2,3 When appropriate tests are judiciously used within the context of an effective laboratory system, they lower selective pressure for resistance by promoting targeted and rational use of antimicrobials. They also lower drug and healthcare costs and can identify treatment failure caused by resistance, thereby unveiling the nature and scope of resistance and preventing its spread.4,5

Laboratory systems and laboratory professionals are key to identifying, mapping, quantifying and communicating about resistance. Patients with resistant infections can only be effectively and efficiently managed with laboratory support. The definitive evidence that resistance is a problem, as well as the fine description of the problem, comes from laboratories.6 Most critically, attempts to treat infections or presumed infections without laboratory input drive resistance by increasing unwarranted antimicrobial use, which is a needless selective pressure for resistant strains. Infectious patients, treated without laboratory support, are also more likely to remain infectious for longer and therefore spread their diseases. Laboratory system strengthening for resistance containment is needed everywhere on the globe, but particularly in African countries, where infection is the leading cause of disease and death.

It is now universally acknowledged, within and outside the scientific community, that deliberate and forceful steps across different sectors must be taken to contain resistance. The recent World Health Organization (WHO) global action plan on antimicrobial resistance7 outlines five strategic objectives for resistance containment, as follows:

Most readers of the plan will appreciate that clinical laboratories and laboratory professionals will lead the second objective of surveillance and research. However, even though their role toward goals are less prominent, as summarised in Table 1, laboratory systems are essential for meeting every one of these objectives, as discussed below.

Dissemination of information about antimicrobial resistance is key to effecting the behavioural change necessary to contain resistance.8 Communications from laboratorians are easily evidence-based. Diagnostic laboratory professionals in general, and microbiologists in particular, are in an excellent position to disseminate information on antimicrobial resistance to health professionals and the general public. Resistance is a health problem but is also a microbiological phenomenon brought about by the genetic change and selective pressure from antibiotics. Resistant organisms circulate among humans but also in other domesticated and wild animal species, as well as in the environment. Microbiologists can and should highlight the mechanisms by which resistance evolves and spreads and how these connect to risk factors for resistant infections in both humans and animals. Some microbiologists have played a pivotal role in public engagement projects and educational initiatives to drive home the message that antimicrobials must be conserved.8,9 For example, the UK Microbiology Society focused one issue of its comic, 'Marvellous Microbes', on antimicrobial resistance.10 Microbiologists and other scientists need to do more to communicate the urgency of the problem as well as the pivotal contributions that laboratories make to contain it. This is particularly true in Africa, where much of the general public and a significant proportion of health professionals have very little awareness of what diagnostic laboratories actually do, or of the biological basis for antimicrobial resistance.

Antimicrobial susceptibility testing by minimum inhibitory concentration (MIC) measurement or disc diffusion is the gold standard for identifying resistance.11,12 For bacteria, once a laboratory is capable of isolating and identifying pathogens from clinical specimens, very few additional resources are needed to obtain a susceptibility profile. MICs are measured in a broth dilution assay or on solid media by agar dilution.13 To obtain MICs, standardised test and control cultures are inoculated into or onto media containing different dilutions of antimicrobial agent. For diagnostic testing, doubling dilutions are the most commonly-used format, although any geometric or arithmetic progression can be used to broaden the test range or increase precision. The MIC is the minimum concentration preventing growth under test conditions. Media composition, incubation temperature, inoculum size and quality, antibiotic format and incubation time can all affect the MIC and must be standardised.

Disc tests are the most commonly-used methods of bacterial susceptibility testing worldwide. They are simple and reproducible tests that determine whether a bacterial isolate is sensitive or resistant to a given drug or drug class. Standard discs containing specific concentrations of each antibacterial are placed on a calibrated lawn of bacteria. After incubation, a clear zone of no growth appears if the concentration of antibiotic around the disc is greater than the MIC. As the concentration of antibiotic in the medium decreases with distance of diffusion through the agar from the disc, the level of resistance is inversely proportional to the diameter of the clear zone of inhibition. Diffusion kinetics of the antibacterial in the test medium are also determinants of zone size, therefore test standardisation is key. Earlier challenges with reproducibility have been overcome by standardising methodologies and interpretations and by using commercially-available media, discs and inoculum standardisation tools.14,15 Thus today, disc diffusion tests are straightforward protocols that are easy to quality assure. If diffusion from a calibrated strip containing graded concentration of antibiotic, rather than a circular disc, an MIC can be obtained by a diffusion protocol. MIC strips are a somewhat costly but cost-effective and simple way to obtain an MIC, particularly in laboratories that are set up for disc testing.

The resistance crisis makes it impossible to continue to argue that antibiotic susceptibility testing is superfluous, a point that was made in many venues, even by experts, as recently as a decade ago.16,17 However, a common misconception in some parts of Africa is that antibacterial susceptibility testing is a reference laboratory-level technique. Susceptibility testing is best supported from central facilities and requires external quality assurance, but susceptibility testing of aerobic bacteria is designed to be performed at routine microbiology laboratories with minimal bacteriology facilities.18 The closer the site for culture and susceptibility testing is to the patient and the provider, the greater the chance that it will impact care for that specific patient and affect empiric prescribing at the relevant facility. Disc diffusion tests in particular are robust, reproducible and capacity can easily be built.19 Disc diffusion tests can even be rigorously performed in field laboratories equipped with a portable autoclave and a small incubator. Compiling and disseminating facility-level data, in the process of providing prescribers with a valuable tool for resistance control, can also help to garner the support of clinicians for laboratory services. Simple software, such as WHONET,20,21 can be used at the facility level to aggregate and analyse susceptibility data and have the added advantage of allowing these data to be forwarded to and integrated with national, regional and global surveillance data. WHONET will run on the most basic of computers, independent of platform, and capacity to enter, retrieve and analyse data can be built with ease. The software is available free, can be used on- or off-line, and the WHO collaborating centre for antimicrobial resistance provides free technical support.

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