Porphyromonas gingivalis HmuY and Bacteroides vulgatus Bvu—A Novel Competitive Heme Acquisition Strategy

Human oral and gut microbiomes are crucial for maintenance of homeostasis in the human body. Porphyromonas gingivalis, the key etiologic agent of chronic periodontitis, can cause dysbiosis in the mouth and gut, which results in local and systemic infectious inflammatory diseases. Our previous work resulted in extensive biochemical and functional characterization of one of the major P. gingivalis heme acquisition systems (Hmu), with the leading role played by the HmuY hemophore-like protein. We continued our studies on the homologous heme acquisition protein (Bvu) expressed by Bacteroides vulgatus, the dominant species of the gut microbiome. Results from spectrophotometric experiments showed that Bvu binds heme preferentially under reducing conditions using Met145 and Met172 as heme iron-coordinating ligands. Bvu captures heme bound to human serum albumin and only under reducing conditions. Importantly, HmuY is able to sequester heme complexed to Bvu. This is the first study demonstrating that B. vulgatus expresses a heme-binding hemophore-like protein, thus increasing the number of members of a novel HmuY-like family. Data gained in this study confirm the importance of HmuY in the context of P. gingivalis survival in regard to its ability to cause dysbiosis also in the gut microbiome.     

UVA Radiation Enhances Lomefloxacin-Mediated Cytotoxic,Growth-Inhibitory and Pro-Apoptotic Effect in Human Melanoma Cells through Excessive Reactive Oxygen Species Generation

Melanoma, the most dangerous type of cutaneous neoplasia, contributes to about 75% of all skin cancer-related deaths. Thus, searching for new melanoma treatment options is an important field of study. The current study was designed to assess whether the condition of mild and low-dose UVA radiation augments the lomefloxacin-mediated cytotoxic, growth-inhibitory and pro-apoptotic effect of the drug in melanoma cancer cells through excessive oxidative stress generation. C32 amelanotic and COLO829 melanotic (BRAF-mutant) melanoma cell lines were used as an experimental model system. The combined exposure of cells to both lomefloxacin and UVA irradiation caused higher alterations of redox signalling pathways, as shown by intracellular reactive oxygen species overproduction and endogenous glutathione depletion when compared to non-irradiated but lomefloxacin-treated melanoma cells. The obtained results also showed that lomefloxacin decreased both C32 and COLO829 cells’ viability in a concentration-dependent manner. This effect significantly intensified when melanoma cells were exposed to UVA irradiation and the drug. For melanoma cells exposed to lomefloxacin or lomefloxacin co-treatment with UVA irradiation, the concentrations of the drug that decreased the cells’ viability by 50% (EC₅₀) were found to be 0.97, 0.17, 1.01, 0.18 mM, respectively. Moreover, we found that the redox imbalance, mitochondrial membrane potential breakdown, induction of DNA fragmentation, and changes in the melanoma cells’ cell cycle distribution (including G₂/M, S as well as Sub-G₁-phase blockade) were lomefloxacin in a dose-dependent manner and were significantly augmented by UVA radiation. This is the first experimental work that assesses the impact of excessive reactive oxygen species generation upon UVA radiation exposure on lomefloxacin-mediated cytotoxic, growth-inhibitory and pro-apoptotic effects towards human melanoma cells, indicating the possibility of the usage of this drug in the photochemotherapy of malignant melanoma as an innovative medical treatment option which could improve the effectiveness of therapy. The obtained results also revealed that the redox imbalance intensification mediated by the phototoxic potential of fluoroquinolones may be considered as a more efficient treatment model of malignant melanoma and may constitute the basis for the development of new compounds with a high ability to excessive oxidative stress generation upon UVA radiation in cancer cells.     

The Amino Acid Transporter Mct10/Tat1 Is Important to Maintain the TSH Receptor at Its Canonical Basolateral Localization and Assures Regular Turnover of Thyroid Follicle Cells in Male Mice

Cathepsin K-mediated thyroglobulin proteolysis contributes to thyroid hormone (TH) liberation, while TH transporters like Mct8 and Mct10 ensure TH release from thyroid follicles into the blood circulation. Thus, thyroid stimulating hormone (TSH) released upon TH demand binds to TSH receptors of thyrocytes, where it triggers Gα_q-mediated short-term effects like cathepsin-mediated thyroglobulin utilization, and Gα_s-mediated long-term signaling responses like thyroglobulin biosynthesis and thyrocyte proliferation. As reported recently, mice lacking Mct8 and Mct10 on a cathepsin K-deficient background exhibit excessive thyroglobulin proteolysis hinting towards altered TSH receptor signaling. Indeed, a combination of canonical basolateral and non-canonical vesicular TSH receptor localization was observed in Ctsk^−/−/Mct8^−/y/Mct10^−/− mice, which implies prolonged Gα_s-mediated signaling since endo-lysosomal down-regulation of the TSH receptor was not detected. Inspection of single knockout genotypes revealed that the TSH receptor localizes basolaterally in Ctsk^−/− and Mct8^−/y mice, whereas its localization is restricted to vesicles in Mct10^−/− thyrocytes. The additional lack of cathepsin K reverses this effect, because Ctsk^−/−/Mct10^−/− mice display TSH receptors basolaterally, thereby indicating that cathepsin K and Mct10 contribute to TSH receptor homeostasis by maintaining its canonical localization in thyrocytes. Moreover, Mct10^−/− mice displayed reduced numbers of dead thyrocytes, while their thyroid gland morphology was comparable to wild-type controls. In contrast, Mct8^−/y, Mct8^−/y/Mct10^−/−, and Ctsk^−/−/Mct8^−/y/Mct10^−/− mice showed enlarged thyroid follicles and increased cell death, indicating that Mct8 deficiency results in altered thyroid morphology. We conclude that vesicular TSH receptor localization does not result in different thyroid tissue architecture; however, Mct10 deficiency possibly modulates TSH receptor signaling for regulating thyrocyte survival.     

Characterization of Microstructure and Mechanical Properties of Mg–Y–Zn Alloys with Respect to Different Content of LPSO Phase

The Mg–Y–Zn alloys with different contents of alloying elements are extruded at an extrusion ratio of 4:1 at 350 °C. The microstructure of the alloys is of an inhomogeneous character showing fine grains produced due to dynamic recrystallization and coarse original grains elongated along the extrusion direction (ED). Moreover, Y and Zn form a long‐period stacking‐ordered (LPSO) phase whose volume fraction increases with their increasing content in the alloy. All investigated alloys exhibit distinct fiber textures with basal planes oriented parallel to ED. It is seen that increasing content of alloying elements leads to a weaker texture. Compression tests with concurrent acoustic emission (AE) measurements are performed along ED at room temperature and a constant strain rate in order to reveal active deformation mechanisms in the alloys and to relate them to their mechanical properties. The AE response is also discussed with respect to the volume fraction of the LPSO phase.     

Ketoprofen Combined with UVA Irradiation Exerts Higher Selectivity in the Mode of Action against Melanotic Melanoma Cells than against Normal Human Melanocytes

Malignant melanoma is responsible for the majority of skin cancer-related deaths. The methods of cancer treatment include surgical removal, chemotherapy, immunotherapy, and targeted therapy. However, neither of these methods gives satisfactory results. Therefore, the development of new anticancer therapeutic strategies is very important and may extend the life span of people suffering from melanoma. The aim of this study was to examine the effect of ketoprofen (KTP) and UVA radiation (UVAR) therapy on cell proliferation, apoptosis, and cell cycle distribution in both melanotic melanoma cells (COLO829) and human melanocytes (HEMn-DP) in relation to its supportive effect in the treatment of melanoma. The therapy combining the use of pre-incubation with KTP and UVAR causes a significant increase in the anti-proliferative properties of ketoprofen towards melanoma cells and the co-exposure of melanotic melanoma cells induced apoptosis shown as the mitochondrial membrane breakdown, cell-cycle deregulation, and DNA fragmentation. Moreover, co-treatment led to GSH depletion showing its pro-apoptotic effect dependent on ROS overproduction. The treatment did not show a significant effect on normal cells—melanocytes—which indicates its high selectivity. The results suggest a possible benefit from the use of the ketoprofen and ultraviolet A irradiation as a new concept of melanotic melanoma therapy.     

Understanding the Photothermal and Photocatalytic Mechanism of Polydopamine Coated Gold Nanorods

Localized surface plasmon resonance (LSPRs) shown by gold nanorods (AuNRs) has several applications in photocatalysis, sensing, and biomedicine. The combination of AuNRs with Polydopamine (PDA) shells results in a strong photo-thermal effect, making them appealing nanomaterials for biomedical applications. However, the precise roles and relative contributions of plasmonic effects in gold, and light-to-heat conversion in PDA are still debated. Herein, a hybrid nanoplatform made by an AuNR core surrounded by a polydopamine (PDA) shell is synthesized, and its photocatalytic behavior is studied. Synthesis is based on a seed-mediated growth followed by the further self-polymerization of dopamine hydrochloride (DA) on the surface of the AuNRs, and the effect of the thickness of the PDA shell on the plasmon response of the composite is the main examined parameter. Photocatalytic performance is tested toward Rhodamine 6G (Rh6G), with the nanocomposites achieving better performance than bare AuNRs and bare PDA nanoparticles. The degradation of 54% of Rh6G initial concentration is achieved within 60 min of irradiation with a catalyst concentration of 7.4 µg mL⁻¹. Photodegradation kinetics, time-resolved spectroscopy, and finite-element-method simulations of plasmons show that AuNRs plasmons, coupled with the low thermal conductivity of PDA, provide slow thermalization, while enhancing the charge carrier transfer.     

A Low-Temperature Batch Process for the Deposition of High-Quality Conformal Alumina Thin Films for Electronic Applications

High-quality, alumina thin films are extensively used as dielectrics, passivation layers, and barrier layers in electronics and many other applications. However, to achieve optimum stoichiometry and thus performance, the layers are often grown at elevated temperatures (>200 °C) using techniques such as atomic layer deposition (ALD). This is problematic for substrates or structures with low thermal budgets. Herein, alumina thin films are grown on 200 mm silicon substrates employing a versatile deposition method known as MVD at low deposition temperatures (35–150 °C). The chemical composition of the resulting films is investigated postdeposition using X-ray photoelectron spectroscopy (XPS) and variable angle spectroscopic ellipsometry, with fully stoichiometric Al₂O₃ achieved at deposition temperatures as low as 100 °C. Dielectric measurements confirm outstanding dielectric properties compared to typical thermal ALD layers deposited at much higher temperatures. This low-temperature deposition performance by considering the MVD reactor design and the “pump-type” regime of precursor delivery versus the “flow-type” regime of ALD is rationalized and understood. The results clearly demonstrate that alumina thin films grown with MVD are highly versatile for electronic applications and are of particular relevance and interest for the high-volume processing of dielectric, passivation, and barrier layers at low temperatures.     

Electrospun Ca3Co4−xO9+δ nanofibers and nanoribbons: Microstructure and thermoelectric properties

Oxide-based ceramics offer promising thermoelectric (TE) materials for recycling high-temperature waste heat, generated extensively from industrial sources. To further improve the functional performance of TE materials, their power factor should be increased. This can be achieved by nanostructuring and texturing the oxide-based ceramics creating multiple interphases and nanopores, which simultaneously increase the electrical conductivity and the Seebeck coefficient. The aim of this work is to achieve this goal by compacting electrospun nanofibers of calcium cobaltite Ca₃Co_4−xO_9+δ, known to be a promising p-type TE material with good functional properties and thermal stability up to 1200 K in air. For this purpose, polycrystalline Ca₃Co_4−xO_9+δ nanofibers and nanoribbons were fabricated by sol–gel electrospinning and calcination at intermediate temperatures to obtain small primary particle sizes. Bulk ceramics were formed by sintering pressed compacts of calcined nanofibers during TE measurements. The bulk nanofiber sample pre-calcined at 973 K exhibited an improved Seebeck coefficient of 176.5 S cm⁻¹ and a power factor of 2.47 μW cm⁻¹ K⁻² similar to an electrospun nanofiber-derived ceramic compacted by spark plasma sintering.     

Electron transport in nanocluster films with random voids

Nanocluster films are modeled as a network of tunnel junctions in which random voids have been introduced. The effects of network size and void distribution on electron transport are studied using Monte Carlo simulations of the Coulomb blockade mediated transport. The random void distributions of the model networks are systematically varied by randomly deleting junctions along each row and between the different rows. The nonlinearity and voltage threshold of the I-V curves are calculated for different void topologies and network sizes. Both the threshold voltage and nonlinearity are sensitive to lateral fluctuations of the conduction paths due to the random voids. The nonlinearity is found to be maximized for network aspect ratios of unity or larger and for particular network topologies. Both the nonlinearity and voltage threshold scale with network size. The behaviors seen in simulation are found to correspond well to Au nanocluster I-V measurements.