Deciphering the physiological response of Escherichia coli under high ATP demand

One long‐standing question in microbiology is how microbes buffer perturbations in energy metabolism. In this study, we systematically analyzed the impact of different levels of ATP demand in Escherichia coli under various conditions (aerobic and anaerobic, with and without cell growth). One key finding is that, under all conditions tested, the glucose uptake increases with rising ATP demand, but only to a critical level beyond which it drops markedly, even below wild‐type levels. Focusing on anaerobic growth and using metabolomics and proteomics data in combination with a kinetic model, we show that this biphasic behavior is induced by the dual dependency of the phosphofructokinase on ATP (substrate) and ADP (allosteric activator). This mechanism buffers increased ATP demands by a higher glycolytic flux but, as shown herein, it collapses under very low ATP concentrations. Model analysis also revealed two major rate‐controlling steps in the glycolysis under high ATP demand, which could be confirmed experimentally. Our results provide new insights on fundamental mechanisms of bacterial energy metabolism and guide the rational engineering of highly productive cell factories

Adenomyoepithelial tumours and myoepithelial carcinomas of the breast – a spectrum of monophasic and biphasic tumours dominated by immature myoepithelial cells

BACKGROUND: Adenomyoepithelial tumours and myoepithelial carcinomas of the breast are primarily defined by the presence of neoplastic cells with a myoepithelial immunophenotype. Current classification schemes are based on purely descriptive features and an assessment of individual prognosis is still problematic. METHODS: A series of 27 adenomyoepithelial tumours of the breast was analysed immunohistochemically with antibodies directed against various cytokeratins, p63, smooth muscle alpha-actin (SMA) and vimentin. Additionally, double immunofluorescence and comparative genomic hybridisation (CGH) was performed. RESULTS: Immunohistochemically, all the tumours showed a constant expression of high molecular weight cytokeratins (Ck) Ck5 and Ck14, p63, SMA and vimentin. With exception of one case diagnosed as myoepithelial carcinoma, all tested tumours expressed low molecular weight cytokeratin Ck18 in variable proportions of cells. Even in monophasic tumours lacking obvious glandular differentiation in conventional staining, a number of neoplastic cells still expressed those cytokeratins. Double immunofluorescence revealed tumour cells exclusively staining for Ck5/Ck14 in the presence of other cell populations that co-expressed high molecular weight Ck5/Ck14 as well as either low molecular weight Ck8/18 or SMA. Based on morphology, we assigned the series to three categories, benign, borderline and malignant. This classification was supported by a stepwise increase in cytogenetic alterations on CGH. CONCLUSION: Adenomyoepithelial tumours comprise a spectrum of neoplasms consisting of an admixture of glandular and myoepithelial differentiation patterns. As a key component SMA-positive cells co-expressing cytokeratins could be identified. Although categorisation of adenomyoepithelial tumours in benign, borderline and malignant was supported by results of CGH, any assessment of prognosis requires to be firmly based on morphological grounds. At present it is not yet clear, if and to what extent proposed Ck5-positive progenitor cells contribute to the immunohistochemical and morphological heterogeneity of these neoplasms of the breast

Deciphering a subgroup of breast carcinomas with putative progression of grade during carcinogenesis revealed by comparative genomic hybridisation (CGH) and immunohistochemistry

Distinct parallel cytogenetic pathways in breast carcinogenesis could be identified in recent years. Nevertheless, it remained unclear as to which tumours may have progressed in grade or which patterns of cytogenetic alteration may define the switch from an in situ towards an invasive lesion. In order to gain more detailed insights into cytogenetic mechanisms of the pathogenesis of breast cancer, the chromosomal imbalances of 206 invasive breast cancer cases were characterised by means of comparative genomic hybridisation (CGH). CGH data were subjected to hierarchical cluster analysis and the results were further compared with immunohistochemical findings on tissue arrays from the same breast cancer cases. The combined analysis of immunohistochemical and cytogenetic data provided evidence that carcinomas with gains of 7p, and to a lesser extent losses of 9q and gains of 5p, are a distinct subgroup within the spectrum of ductal invasive grade 3 breast carcinomas. These aberrations were associated with a high degree of cytogenetic instability (16.6 alterations per case on average), 16q-losses in over 70% of these cases, strong oestrogen receptor expression and absence of strong expression of p53, c-erbB2 and Ck 5. These characteristics provide strong support for the hypothesis that these tumours may develop through stages of well- and perhaps intermediately differentiated breast cancers. Our results therefore underline the existence of several parallel and also stepwise progression pathways towards breast cancer

MaNGIA: 10 000 mock galaxies for stellar population analysis

Context. Modern astronomical observations give unprecedented access to the physical properties of nearby galaxies, including spatially resolved stellar populations. However, observations can only give a present-day view of the Universe, whereas cosmological simulations give access to the past record of the processes that galaxies have experienced in their evolution. To connect the events that happened in the past with galactic properties as seen today, simulations must be taken to a common ground before being compared to observations. Therefore, a dedicated effort is needed to forward-model simulations into the observational plane. Aims. We emulate data from the Mapping Nearby Galaxies at Apache Point Observatory (MaNGA) survey, which is the largest integral field spectroscopic galaxy survey to date with its 10000 nearby galaxies of all types. For this, we use the latest hydro-cosmological simulations IllustrisTNG to generate MaNGIA (Mapping Nearby Galaxies with IllustrisTNG Astrophysics), a mock MaNGA sample of similar size that emulates observations of galaxies for stellar population analysis. Methods. We chose TNG galaxies to match the MaNGA sample selection in terms of mass, size, and redshift in order to limit the impact of selection effects. We produced MaNGA-like datacubes from all simulated galaxies, and processed them with the stellar population analysis code pyPipe3D. This allowed us to extract spatially resolved maps of star formation history, age, metallicity, mass, and kinematics, following the same procedures used as part of the official MaNGA data release. Results. This first paper presents the approach used to generate the mock sample and provides an initial exploration of its properties. We show that the stellar populations and kinematics of the simulated MaNGIA galaxies are overall in good agreement with observations. Specific discrepancies, especially in the age and metallicity gradients in low- to intermediate-mass regimes and in the kinematics of massive galaxies, require further investigation, but are likely to uncover new physical understanding. We compare our results to other attempts to mock similar observations, all of smaller datasets. Conclusions. Our final dataset is released with this publication, consisting of ≲10000 post-processed datacubes analysed with pyPipe3D, along with the codes developed to create it. Future work will employ modern machine learning and other analysis techniques to connect observations of nearby galaxies to their cosmological evolutionary past

Achilles tendon mechanosensitivity is preserved in old age: In vivo evidence from a 1.5 years long resistance training intervention

Introduction Ageing deteriorates musculoskeletal system structure and function and limits its adaptability to mechanical loading. Medium-term (12-14 weeks) exercise interventions in older adults have been shown to increase tendon stiffness by increasing the tendon’s Young’s modulus [1], rather than the tendon’s cross-sectional area (CSA). However, little is known about the time-adaptive response relationship of the tendon in long-term (years) interventions involving alteration in mechanical loading. Therefore, we investigated whether the older human Achilles tendon (AT) demonstrates mechanosensitivity and alterations in material and/or size in response to long-term mechanical loading. Methods Thirty-four older female adults (age: 65±7 y) were recruited to a medium-term (14 weeks; n=21) strength training intervention with high AT strain cyclic loading (five sets of four repetitions of isometric plantarflexion contractions 3 times a week with 90% of MVC as in [2]) or a control group (n=13), with a sub-group of the intervention group (n=12) continuing exercise for 1.5 years. AT stiffness and Young’s modulus were quantified in vivo using ultrasonography and dynamometry. Tendon CSA was measured along the whole free AT by means of magnetic resonance imaging. Results Following 14 weeks of resistance training, the intervention group showed a significant (p<.05) increase in ankle plantarflexor muscle strength (141.5±36.2 vs 116.3±30.8 Nm at baseline), along with a 23% increase in AT stiffness (598.2±141.2 Nmm-1 vs 488.4±136.9 Nmm-1 at baseline), 20% increase in Young’s modulus (1.63±0.46 GPa vs 1.37±0.39 GPa at baseline) and a homogenous hypertrophy by about 6% along the entire free AT. However, continuing the exercise training for 1.5 years did not cause any further changes in muscle strength and tendon properties. The control group did not show any differences in muscle and tendon functional and structural properties between time points. Discussion The AT seems to have the capability to increase its stiffness in response to 14 weeks of mechanical loading exercise by altering both its material and size, and may thereby tolerate higher mechanical loading by reducing both the strain and stress it experiences during tensile loading. Continuing strength training appears to maintain, but not cause any further adaptive changes in tendons, which implies that the time-adaptive response relationship to mechanical loading is non-linear in ageing tendons. References 1. Reeves et al. (2003). J Physiol, 548, 971–981. 2. Arampatzis et al. (2007). J Exp Biol, 210, 2743–2753

Cerebral activations related to ballistic, stepwise interrupted and gradually modulated movements in parkinson patients

Patients with Parkinson's disease (PD) experience impaired initiation and inhibition of movements such as difficulty to start/stop walking. At single-joint level this is accompanied by reduced inhibition of antagonist muscle activity. While normal basal ganglia (BG) contributions to motor control include selecting appropriate muscles by inhibiting others, it is unclear how PD-related changes in BG function cause impaired movement initiation and inhibition at single-joint level. To further elucidate these changes we studied 4 right-hand movement tasks with fMRI, by dissociating activations related to abrupt movement initiation, inhibition and gradual movement modulation. Initiation and inhibition were inferred from ballistic and stepwise interrupted movement, respectively, while smooth wrist circumduction enabled the assessment of gradually modulated movement. Task-related activations were compared between PD patients (N = 12) and healthy subjects (N = 18). In healthy subjects, movement initiation was characterized by antero-ventral striatum, substantia nigra (SN) and premotor activations while inhibition was dominated by subthalamic nucleus (STN) and pallidal activations, in line with the known role of these areas in simple movement. Gradual movement mainly involved antero-dorsal putamen and pallidum. Compared to healthy subjects, patients showed reduced striatal/SN and increased pallidal activation for initiation, whereas for inhibition STN activation was reduced and striatal-thalamo-cortical activation increased. For gradual movement patients showed reduced pallidal and increased thalamo-cortical activation. We conclude that PD-related changes during movement initiation fit the (rather static) model of alterations in direct and indirect BG pathways. Reduced STN activation and regional cortical increased activation in PD during inhibition and gradual movement modulation are better explained by a dynamic model that also takes into account enhanced responsiveness to external stimuli in this disease and the effects of hyper-fluctuating cortical inputs to the striatum and STN in particular

Engineered antibodies: new possibilities for brain PET?

International audienceAlmost 50 million people worldwide are affected by Alzheimer's disease (AD), the most common neurodegenerative disorder. Development of disease-modifying therapies would benefit from reliable, non-invasive positron emission tomography (PET) biomarkers for early diagnosis, monitoring of disease progression, and assessment of therapeutic effects. Traditionally, PET ligands have been based on small molecules that, with the right properties, can penetrate the blood-brain barrier (BBB) and visualize targets in the brain. Recently a new class of PET ligands based on antibodies have emerged, mainly in applications related to cancer. While antibodies have advantages such as high specificity and affinity, their passage across the BBB is limited. Thus, to be used as brain PET ligands, antibodies need to be modified for active transport into the brain. Here, we review the development of radioligands based on antibodies for visualization of intrabrain targets. We focus on antibodies modified into a bispecific format, with the capacity to undergo transferrin receptor 1 (TfR1)-mediated transcytosis to enter the brain and access pathological proteins, e.g. amyloid-beta. A number of such antibody ligands have been developed, displaying differences in brain uptake, pharmacokinetics, and ability to bind and visualize the target in the brain of transgenic mice. Potential pathological changes related to neurodegeneration, e.g. misfolded proteins and neuroinflammation, are suggested as future targets for this novel type of radioligand. Challenges are also discussed, such as the temporal match of radionuclide half-life with the ligand's pharmacokinetic profile and translation to human use. In conclusion, brain PET imaging using bispecific antibodies, modified for receptor-mediated transcytosis across the BBB, is a promising method for specifically visualizing molecules in the brain that are difficult to target with traditional small molecule ligands