The Effects of Mild Intermittent Hypoxia Exposure on the Abdominal Subcutaneous Adipose Tissue Proteome in Overweight and Obese Men: A First-in-Human Randomized, Single-Blind, and Cross-Over Study

Adipose tissue (AT) oxygen tension (pO2) has been implicated in AT dysfunction and metabolic perturbations in both rodents and humans. Compelling evidence suggests that hypoxia exposure alters metabolism, at least partly through effects on AT. However, it remains to be elucidated whether mild intermittent hypoxia (MIH) exposure impacts the AT proteome. We performed a randomized, single-blind, and cross-over study to investigate the effects of seven consecutive days of MIH (FiO2 15%, 3x2h/d) compared to normoxia (FiO2 21%) exposure on the AT proteome in overweight/obese men. In vivo AT insulin sensitivity was determined by the gold standard hyperinsulinemic-euglycemic clamp, and abdominal subcutaneous AT biopsies were collected under normoxic fasting conditions following both exposure regimens (day 8). AT proteins were isolated and quantified using liquid chromatography-mass spectrometry. After correction for blood contamination, 1,022 AT protein IDs were identified, of which 123 were differentially expressed following MIH (p < 0.05). We demonstrate for the first time that MIH exposure, which markedly reduces in vivo AT oxygen tension, impacts the human AT proteome. Although we cannot exclude that a single differentially expressed protein might be a false positive finding, several functional pathways were altered by MIH exposure, also after adjustment for multiple testing. Specifically, differentially expressed proteins were involved in redox systems, cell-adhesion, actin cytoskeleton organization, extracellular matrix composition, and energy metabolism. The MIH-induced change in AT TMOD3 expression was strongly related to altered in vivo AT insulin sensitivity, thus linking MIH-induced effects on the AT proteome to metabolic changes in overweight/obese humans.

Sexual Dimorphism in Extracellular Matrix Composition and Viscoelasticity of the Healthy and Inflamed Mouse Brain

Magnetic resonance elastography (MRE) has revealed sexual dimorphism in brain stiffness in healthy individuals and multiple sclerosis (MS) patients. In the animal model of MS, experimental autoimmune encephalomyelitis (EAE), we showed previously that inflammation-induced brain softening was associated with alterations of the extracellular matrix (ECM). However, it remained unclear whether the brain ECM presents sex-specific properties that can be visualized by MRE. Therefore, we aimed here at quantifying sexual dimorphism in brain viscoelasticity in association with ECM changes in healthy and inflamed brains. Multifrequency MRE was applied to the midbrain of healthy and EAE mice of both sexes to quantitatively map regional stiffness. To define differences in brain ECM composition, gene expression of the key basement membrane components laminin (Lama4, Lama5), collagen (Col4a1, Col1a1) and fibronectin (Fn1) was investigated by RT-qPCR. We showed that the healthy male cortex expressed less Lama4, Lama5, Col4a1 but more Fn1 (all p < 0.05) than the healthy female cortex, which was associated with 9% softer properties (p = 0.044) in that region. At peak EAE, cortical softening was similar in both sexes compared to healthy tissue, with an 8% difference remaining between males and females during EAE (p < 0.001). Cortical Lama4, Lama5 and Col4a1 expression increased 2 to 3-fold in EAE in both sexes while Fn1 decreased only in males (all p < 0.05). No significant sex differences in stiffness were detected in other brain regions. In conclusion, sexual dimorphism in the ECM composition of cortical tissue in the mouse brain is reflected by in vivo stiffness measured with MRE and should be considered in future studies by sex-specific reference values.

Impact of Immune Cell Heterogeneity on HER2+ Breast Cancer Prognosis and Response to Therapy

Breast cancer is a heterogeneous disease with a high degree of diversity among and within tumors, and in relation to its different tumor microenvironment. Compared to other oncotypes, such as melanoma or lung cancer, breast cancer is considered a “cold” tumor, characterized by low T lymphocyte infiltration and low tumor mutational burden. However, more recent evidence argues against this idea and indicates that, at least for specific molecular breast cancer subtypes, the immune infiltrate may be clinically relevant and heterogeneous, with significant variations in its stromal cell/protein composition across patients and tumor stages. High numbers of tumor-infiltrating T cells are most frequent in HER2-positive and basal-like molecular subtypes and are generally associated with a good prognosis and response to therapies. However, effector immune infiltrates show protective immunity in some cancers but not in others. This could depend on one or more immunosuppressive mechanisms acting alone or in concert. Some of them might include, in addition to immune cells, other tumor microenvironment determinants such as the extracellular matrix composition and stiffness as well as stromal cells, like fibroblasts and adipocytes, that may prevent cytotoxic T cells from infiltrating the tumor microenvironment or may inactivate their antitumor functions. This review will summarize the state of the different immune tumor microenvironment determinants affecting HER2+ breast tumor progression, their response to treatment, and how they are modified by different therapeutic approaches. Potential targets within the immune tumor microenvironment will also be discussed.

Activity of Moxifloxacin Against Biofilms Formed by Clinical Isolates of Staphylococcus aureus Differing by Their Resistant or Persister Character to Fluoroquinolones

Staphylococcus aureus biofilms are poorly responsive to antibiotics. Underlying reasons include a matrix effect preventing drug access to embedded bacteria, or the presence of dormant bacteria with reduced growth rate. Using 18 clinical isolates previously characterized for their moxifloxacin-resistant and moxifloxacin-persister character in stationary-phase culture, we studied their biofilm production and matrix composition and the anti-biofilm activity of moxifloxacin. Biofilms were grown in microtiter plates and their abundance quantified by crystal violet staining and colony counting; their content in polysaccharides, extracellular DNA and proteins was measured. Moxifloxacin activity was assessed after 24 h of incubation with a broad range of concentrations to establish full concentration-response curves. All clinical isolates produced more biofilm biomass than the reference strain ATCC 25923, the difference being more important for those with high relative persister fractions to moxifloxacin, most of which being also resistant. High biofilm producers expressed icaA to higher levels, enriching the matrix in polysaccharides. Moxifloxacin was less potent against biofilms from clinical isolates than from ATCC 25923, especially against moxifloxacin-resistant isolates with high persister fractions, which was ascribed to a lower concentration of moxifloxacin in these biofilms. Time-kill curves in biofilms revealed the presence of a moxifloxacin-tolerant subpopulation, with low multiplication capacity, whatever the persister character of the isolate. Thus, moxifloxacin activity depends on its local concentration in biofilm, which is reduced in most isolates with high-relative persister fractions due to matrix effects, and insufficient to kill resistant isolates due to their high MIC.

Determination of changes in the mechanical properties of polymer composites with the addition of agglomerates of WC nanopowder

Recently, there has been a growing interest in the development of new composite materials with improved characteristics. The article presents the results of tests of composite specimens based on aramid fabrics modified with WC nanopowder agglomerates obtained from carbide manufacturing waste. The following mechanical characteristics were investigated: transverse bending resistance, fracture resistance and energy absorption during contact with a physical body at high speed. According to the results, the transverse bending resistance increased by 35% at a WC concentration of 5%. When 3% WC powder was added to the matrix composition, the total crack length after impact was almost halved. The largest increase in energy absorption of the samples was about 30% at 1% additive concentration. The significant increase in the investigated parameters can be explained by the complex morphology of the embedded particles. In further investigations it is planned to study in detail the mechanism of distribution of nanodispersed WC powder additive in the volume of the modified material.

Testing of Conductive Carbon Fiber Reinforced Polymer Composites Using Current Impulses Simulating Lightning Effects

Carbon fiber reinforced polymer (CFRP) composites are lightweight and an increasingly used material with good mechanical properties. In the aviation industry, they are also required to have specific electrical properties that guarantee resistance to the direct and indirect lightning effects. The paper is focused on the description of a test stand and development of a method used to determine the electrical characteristics of conductive CFRP laminate samples with the use of high current impulses of lightning nature. Samples of three laminates (square format with side 30 × 30 cm) with a different composition were tested on the constructed stand, confirming the possibility of characterizing this type of laminate sample in terms of electrical conductivity and resistance to the effects of lightning current. It was possible to observe the impulse current flow (with a peak value up to 15 kA and a rise time above 6 µs) from the high voltage electrode placed in the center of the sample in all directions towards the edge. The optical fiber measuring system was used to record the voltage and current time waveforms. The energy stored in the impulse current generator was sufficient to simulate the mechanical damage, such as burnout and delamination, that accompanies the direct lightning strike to structural elements made of CFRP. The influence of the matrix composition used for laminate fabrication on the test results describing the electrical properties of the tested CFRP samples was noted. The experimental setup allows the testing of specimens with a maximum width and length of 50 × 50 cm and any thickness with a peak current of up to 50 kA.

Testing Hypoxia in Pig Meniscal Culture: Biological Role of the Vascular-Related Factors in the Differentiation and Viability of Neonatal Meniscus

Menisci play an essential role in shock absorption, joint stability, load resistance and its transmission thanks to their conformation. Adult menisci can be divided in three zones based on the vascularization: an avascular inner zone with no blood supply, a fully vascularized outer zone, and an intermediate zone. This organization, in addition to the incomplete knowledge about meniscal biology, composition, and gene expression, makes meniscal regeneration still one of the major challenges both in orthopedics and in tissue engineering. To overcome this issue, we aimed to investigate the role of hypoxia in the differentiation of the three anatomical areas of newborn piglet menisci (anterior horn (A), central body (C), and posterior horn (P)) and its effects on vascular factors. After sample collection, menisci were divided in A, C, P, and they were cultured in vitro under hypoxic (1% O2) and normoxic (21% O2) conditions at four different experimental time points (T0 = day of explant; T7 = day 7; T10 = day 10; T14 = day 14); samples were then evaluated through immune, histological, and molecular analyses, cell morpho-functional characteristics; with particular focus on matrix composition and expression of vascular factors. It was observed that hypoxia retained the initial phenotype of cells and induced extracellular matrix production resembling a mature tissue. Hypoxia also modulated the expression of angiogenic factors, especially in the early phase of the study. Thus, we observed that hypoxia contributes to the fibro-chondrogenic differentiation with the involvement of angiogenic factors, especially in the posterior horn, which corresponds to the predominant weight-bearing portion.

Unidirectional fibre reinforced geopolymer matrix composites

<p>Geopolymers have been suggested in the literature as matrix materials for fibre reinforced composites due to a unique combination of low-temperature synthesis and high temperature stability. This study investigated several key aspects of fibre reinforced geopolymer matrix composites in order to improve the basic knowledge of these materials. It was demonstrated that geopolymer matrix composites show great potential as fire-resistant materials for near room temperature applications. In particular, basalt fibre composites were of great interest due to their comparatively low cost and good mechanical performance. Microstructural investigations indicated that basalt fibres can potentially be used in geopolymer matrices up to 600°C. However, the success of the application of geopolymer matrix composites at higher temperatures is seen as critical and depends on further development of suitable matrices. Several compositions within a sodium-metahalloysite model matrix system were evaluated in order to identify a suitable formulation for composite fabrication. An average compressive strength of ~ 79 MPa and flexural strength and modulus of ~ 10 MPa and 8.5 GPa, respectively, were achieved for the best batch of the main matrix composition. By optimising the matrix composition, the mechanical properties could be significantly improved, achieving an extremely high maximum compressive strength value of 145 MPa. Issues with reproducibility and the influence of various aspects of the fabrication process are discussed. The room temperature flexural properties of unidirectional fibre reinforced composite bars with basalt, carbon and alumina fibres were investigated. Besides the fibre type, the effects of several other parameters including fibre sizing, matrix strength, span-to-depth ratio and specimen dimensions on the flexural properties and the failure behaviour of the composites were studied. Significant improvements to the mechanical properties were achieved with all fibre types. However, the mechanical behaviour was highly influenced by the elastic modulus of the fibre. Furthermore, it was shown that the composite properties were affected by the overall sample dimensions, the testing span and the mixing time of the geopolymer binder. The alumina fibre composites achieved the highest flexural stress with a maximum value of 470 MPa and a fibre content of ~ 30 vol.-%. Basalt and carbon fibre composites showed maximum flexural strength values around 200 MPa. Although all composite types displayed considerable post-fracture strength, only the basalt composites failed in tensile mode. The applicability of the weak matrix composites (WMC) concept to describe the mechanical behaviour of geopolymer matrix composites was discussed. The fibre-matrix interactions were analysed between room temperature and 1000°C by means of electron microscopy, EDS and x-ray diffraction. All fibres were found to be chemically stable under the highly alkaline conditions of the geopolymer synthesis and showed no significant reaction with the geopolymer matrix at room temperature. The results indicate that basalt fibre composites may be used up to 600°C without significant degradation of the fibre. The heating of the carbon fibre composites to 600°C had drastic effect on the strength and integrity of the composite, in particular, when using sized carbon fibres. The alumina fibres showed good wetting and bonding behaviour but otherwise little reaction with the matrix even after heating to 1000°C.</p>