The Hippo-YAP/TAZ-TEAD axis promotes multipotency in bone marrow stromal cells.

The mechanisms by which bone marrow stromal cells (BMSCs) maintain multilineage potency in vitro remain elusive. To identify the transcriptional regulatory circuits that contribute to BMSC multipotency, we performed paired single-nucleus multiomics of the expansion of freshly isolated BMSCs and of BMSCs undergoing tri-lineage differentiation. By computationally reconstructing the regulatory programs associated with initial stages of differentiation and early expansion, we identified the TEAD family of transcription factors, which is inhibited by Hippo signaling, as highly active in the BMSC in vitro multipotent state. Pharmacological inhibition of TEAD enhanced BMSC osteogenic and adipogenic differentiation, whereas its activation maintained BMSCs in an undifferentiated state, supporting a model whereby isolation of BMSCs coincides with a TEAD-controlled transcriptional state linked to multipotency. Our study highlights the Hippo pathway as a pivotal regulator of BMSC multipotency, and our regulatory network inferences are a reservoir of testable hypotheses that link transcription factors and their regulons to specific aspects of BMSC behavior.

Influence of Accidental Impurities on the Spectroscopic and Luminescent Properties of ZnWO4 Crystal.

Special techniques for deep purification of ZnO and WO3 have been developed in this work. A ZnWO4 single crystal has been grown by the Czochralski method using purified ZnO and WO3 chemicals, along with the ZnWO4 crystal-etalon, which has been grown at the same conditions using commercially available 5N ZnO and WO3 chemicals. The actual accidental impurities compositions of both the initial chemicals and the grown crystals have been measured by inductively coupled plasma mass-spectrometry. A complex of comparative spectroscopic studies of the crystals has been performed, including optical absorption spectra, photo-, X-ray-, and cathodoluminescence spectra and decay kinetics, as well as the photoluminescence excitation spectra. The revealed differences in the measured properties of the crystals have been analyzed in terms of influence of the accidental impurities on these properties.

The Influence of Medium on Fluorescence Quenching of Colloidal Solutions of the Nd3+: LaF3 Nanoparticles Prepared with HTMW Treatment.

An original method was proposed to reduce the quenching of the NIR fluorescence of colloidal solutions of 0.1 at. % Nd3+: LaF3 nanoparticles (NPs) synthesized by aqueous co-precipitation method followed by hydrothermal microwave treatment. For this, an aqueous colloidal solution of NPs was precipitated by centrifugation and dissolved in the same volume of DMSO. The kinetics of static fluorescence quenching of Nd3+ donors of doped NPs dispersed in two solvents was analyzed to determine and to compare the concentrations of OH- quenching acceptors uniformly distributed throughout the volume of the NPs. The dependences of the relative fluorescence quantum yield φ of colloidal solutions on the concentration of OH- groups in the NPs were calculated and were also used to determine concentration of acceptors in the volume of NPs in different solvents. It was found that the concentration of OH- groups in NPs dispersed in DMSO is almost two times lower than in NPs dispersed in water. This gives an almost two-fold increase in the relative fluorescence quantum yield φ for the former. The sizes of synthesized NPs were monitored by common TEM and by applying a rapid procedure based on optical visualization of the trajectories of the Brownian motion of NPs in solution using a laser ultramicroscope. The use of two different methods made it possible to obtain more detailed information about the studied NPs.

Human knowledge models: Learning applied knowledge from the data.

Artificial intelligence and machine learning have demonstrated remarkable results in science and applied work. However, present AI models, developed to be run on computers but used in human-driven applications, create a visible disconnect between AI forms of processing and human ways of discovering and using knowledge. In this work, we introduce a new concept of "Human Knowledge Models" (HKMs), designed to reproduce human computational abilities. Departing from a vast body of cognitive research, we formalized the definition of HKMs into a new form of machine learning. Then, by training the models with human processing capabilities, we learned human-like knowledge, that humans can not only understand, but also compute, modify, and apply. We used several datasets from different applied fields to demonstrate the advantages of HKMs, including their high predictive power and resistance to noise and overfitting. Our results proved that HKMs can efficiently mine knowledge directly from the data and can compete with complex AI models in explaining the main data patterns. As a result, our study reveals the great potential of HKMs, particularly in the decision-making applications where "black box" models cannot be accepted. Moreover, this improves our understanding of how well human decision-making, modeled by HKMs, can approach the ideal solutions in real-life problems.

Study of synthesis temperature effect onß-NaGdF4: Yb3+, Er3+ upconversion luminescence efficiency and decay time using maximum entropy method.

Upconversion materials have several advantages for many applications due to their great potential in converting infrared light to visible. For practical use, it is necessary to achieve high intensity of UC luminescence, so the studies of the optimal synthesis parameters for upconversion nanoparticles are still going on. In the present work, we analyzed the synthesis temperature effect on the efficiency and luminescence decay ofß-NaGd0.78Yb0.20Er0.02F4(15-25 nm) upconversion nanoparticles with hexagonal crystal structure synthesized by anhydrous solvothermal technique. The synthesis temperature was varied in the 290 °C-320 °C range. The synthesis temperature was shown to have a significant influence on the upconversion luminescence efficiency and decay time. The coherent scattering domain linearly depended on the synthesis temperature and was in the range 13.1-22.3 nm, while the efficiency of the upconversion luminescence increases exponentially from 0.02 to 0.10% under 1 W cm-2excitation. For a fundamental analysis of the reasons for the upconversion luminescence intensity dependence on the synthesis temperature, it was proposed to use the maximum entropy method for luminescence decay kinetics processing. This method does not require a preliminary setting of the number of exponents and, due to this, makes it possible to estimate additional components in the luminescence decay kinetics, which are attributed to different populations of rare-earth ions in different conditions. Two components in the green luminescence and one component in the red luminescence decay kinetics were revealed for nanoparticles prepared at 290 °C-300 °C. An intense short and a weak long component in green luminescence decay kinetics could be associated with two different populations of ions in the surface quenching layer and the crystal core volume. With an increase in the synthesis temperature, the second component disappears, and the decay time increases due to an increase in the number of ions in the crystal core volume and a more uniform distribution of dopants.

Novel substituted 5-methyl-4-acylaminoisoxazoles as antimitotic agents: Evaluation of selectivity to LNCaP cancer cells.

A series of novel antimitotic agents was designed using the replacement of heterocyclic cores in two tubulin-targeting lead molecules with the acylated 4-aminoisoxazole moiety. Target compounds were synthesized via heterocyclization of ß-aryl-substituted vinylketones by tert-butyl nitrite in the presence of water as a key step. 4-Methyl-N-[5-methyl-3-(3,4,5-trimethoxyphenyl)isoxazol-4-yl]benzamide (1aa) was found to stimulate partial depolymerization of microtubules of human lung carcinoma A549 cells at a high concentration of 100 µM and to totally inhibit cell growth (IC50 = 0.99 µM) and cell viability (IC50 = 0.271 µM) in the nanomolar to submicromolar concentration range. These data provide evidence of the multitarget profile of the cytotoxic action of compound 1aa. The SAR study demonstrated that the 3,4,5-trimethoxyphenyl residue is the key structural parameter determining the efficiency both towards tubulin and other molecular targets. The cytotoxicity of 3-methyl-N-[5-methyl-3-(3,4,5-trimethoxyphenyl)isoxazol-4-yl]benzamide (1ab) to the androgen-sensitive human prostate adenocarcinoma cancer cell line LNCaP (IC50 = 0.301 µM) was approximately one order of magnitude higher than that to the conditionally normal cells lines WI-26 VA4 (IC50 = 2.26 µM) and human umbilical vein endothelial cells (IC50 = 5.58 µM) and significantly higher than that to primary fibroblasts (IC50 > 75 µM).

Secreted frizzled related-protein 2 (Sfrp2) deficiency decreases adult skeletal stem cell function in mice.

In a previous transcriptomic study of human bone marrow stromal cells (BMSCs, also known as bone marrow-derived "mesenchymal stem cells"), SFRP2 was highly over-represented in a subset of multipotent BMSCs (skeletal stem cells, SSCs), which recreate a bone/marrow organ in an in vivo ectopic bone formation assay. SFRPs modulate WNT signaling, which is essential to maintain skeletal homeostasis, but the specific role of SFRP2 in BMSCs/SSCs is unclear. Here, we evaluated Sfrp2 deficiency on BMSC/SSC function in models of skeletal organogenesis and regeneration. The skeleton of Sfrp2-deficient (KO) mice is overtly normal; but their BMSCs/SSCs exhibit reduced colony-forming efficiency, reflecting low SSC self-renewal/abundancy. Sfrp2 KO BMSCs/SSCs formed less trabecular bone than those from WT littermates in the ectopic bone formation assay. Moreover, regeneration of a cortical drilled hole defect was dramatically impaired in Sfrp2 KO mice. Sfrp2-deficient BMSCs/SSCs exhibited poor in vitro osteogenic differentiation as measured by Runx2 and Osterix expression and calcium accumulation. Interestingly, activation of the Wnt co-receptor, Lrp6, and expression of Wnt target genes, Axin2, C-myc and Cyclin D1, were reduced in Sfrp2-deficient BMSCs/SSCs. Addition of recombinant Sfrp2 restored most of these activities, suggesting that Sfrp2 acts as a Wnt agonist. We demonstrate that Sfrp2 plays a role in self-renewal of SSCs and in the recruitment and differentiation of adult SSCs during bone healing. SFRP2 is also a useful marker of BMSC/SSC multipotency, and a factor to potentially improve the quality of ex vivo expanded BMSC/SSC products.

A monoclinic semiorganic molecular crystal GUHP for terahertz photonics and optoelectronics.

In this paper we describe the properties of the crystal of guanylurea hydrogen phosphate (NH[Formula: see text])[Formula: see text]CNHCO(NH[Formula: see text])H[Formula: see text]PO[Formula: see text] (GUHP) and propose its application in terahertz photonics and optoelectronics. GUHP crystal has a wide window of transparency and a high optical threshold in the visible and NIR spectral regions and narrow absorption bands in the terahertz frequency range. The spectral characteristics of absorption and refraction in the THz range were found to be strongly dependent on crystal temperature and orientation. Computer simulations made it possible to link the nature of the resonant response of the medium at THz frequencies with the molecular structure of the crystal, in particular, with intermolecular hydrogen bonds and the layered structure of the lattice. The possibility of application of the crystal under study for the conversion of femtosecond laser radiation from visible an NIR to terahertz range was demonstrated. It was shown that dispersion properties of the crystal allow the generation of narrow band terahertz radiation, whose spectral properties are determined by conditions close to phase matching. The properties of the generated terahertz radiation under various temperatures suggest the possibility of phonon mechanism of enhancement for nonlinear susceptibility of the second order.

Can Radiographic Tumor Volume of Oral Squamous Cell Carcinoma Help Predict Clinical and Pathological Tumor Features?

PURPOSE: Radiographic tumor volume (RTV) of oral squamous cell carcinoma (SCC) is seldom measured in practice. Aims of the study are to estimate RTV of SCC and to investigate its relationship with clinical and pathological stage, tumor margin status, recurrence, and need for chemo/radiation. METHODS: The design is a retrospective cohort study. The predictor variable is SCC RTV. The primary outcome variables are clinical and pathological tumor size. The secondary outcomes are margin status and postoperative chemo/radiation. Tumor dimensions were measured on preoperative maxillofacial or neck computer tomography images with contrast. Information on patient and tumor characteristics was obtained. Pearson correlation, t test, ANOVA and log rank test were used for statistical analysis. The significance level was set at .05. RESULTS: Thirty-six subjects aged 36 to 86 were included in the study. Positive association was found between clinical T stage and RTV (P = .0003) and between pathologic T stage and RTV (P = .002). Mean value of RTV was significantly higher in the group with positive margins (P = .0004). RTV was significantly higher in cancers requiring adjuvant chemo/radiation (P = .033). Mean RTV for patients with recurrence was 1.86 cm3 as compared to 1.29 cm3 for patients with no recurrence. Higher tumor volumes were more likely to be associated with recurrence. CONCLUSIONS: RTV is a variable that is readily available to head and neck surgeons. RTV is associated with clinical and pathological tumor sizes, margin status, need for adjuvant chemo/radiation and tumor recurrence.

Fluorescent bacterial biosensor E. coli/pTdcR-TurboYFP sensitive to terahertz radiation.

A fluorescent biosensor E. coli/pTdcR-TurboYFP sensitive to terahertz (THz) radiation was developed via transformation of Escherichia coli (E. coli) cells with plasmid, in which the promotor of the tdcR gene controls the expression of yellow fluorescent protein TurboYFP. The biosensor was exposed to THz radiation in various vessels and nutrient media. The threshold and dynamics of fluorescence were found to depend on irradiation conditions. Heat shock or chemical stress yielded the absence of fluorescence induction. The biosensor is applicable to studying influence of THz radiation on the activity of tdcR promotor that is involved in the transport and metabolism of threonine and serine in E. coli.

Bone Marrow Stromal Cell Assays: In Vitro and In Vivo.

Populations of bone marrow stromal cells (BMSCs, also known as bone marrow-derived "mesenchymal stem cells") contain a subset of cells that are able to recapitulate the formation of a bone/marrow organ (skeletal stem cells, SSCs). It is now apparent that cells with similar but not identical properties can be isolated from other skeletal compartments (growth plate, periosteum). The biological properties of BMSCs, and these related stem/progenitor cells, are assessed by a variety of assays, both in vitro and in vivo. Application of these assays in an appropriate fashion provide a great deal of information on the role of BMSCs, and the subset of SSCs, in health and in disease.

The Post-Translational Modifications, Localization, and Mode of Attachment of Non-Covalently Bound Glucanosyltransglycosylases of Yeast Cell Wall as a Key to Understanding their Functioning.

Glucan linked to proteins is a natural mega-glycoconjugate (mGC) playing the central role as a structural component of a yeast cell wall (CW). Regulation of functioning of non-covalently bound glucanosyltransglycosylases (ncGTGs) that have to remodel mGC to provide CW extension is poorly understood. We demonstrate that the main ncGTGs Bgl2 and Scw4 have phosphorylated and glutathionylated residues and are represented in CW as different pools of molecules having various firmness of attachment. Identified pools contain Bgl2 molecules with unmodified peptides, but differ from each other in the presence and combination of modified ones, as well as in the presence or absence of other CW proteins. Correlation of Bgl2 distribution among pools and its N-glycosylation was not found. Glutathione affects Bgl2 conformation, probably resulting in the mode of its attachment and enzymatic activity. Bgl2 from the pool of unmodified and monophosphorylated molecules demonstrates the ability to fibrillate after isolation from CW. Revealing of Bgl2 microcompartments and their mosaic arrangement summarized with the results obtained give the evidence that the functioning of ncGTGs in CW can be controlled by reversible post-translational modifications and facilitated due to their compact localization. The hypothetical scheme of distribution of Bgl2 inside CW is represented.

Study on the effects of terahertz radiation on gene networks of Escherichia coli by means of fluorescent biosensors.

Three novel fluorescent biosensors sensitive to terahertz (THz) radiation were developed via transformation of Escherichia coli (E. coli) cells with plasmids, in which a promotor of genes matA, safA, or chbB controls the expression of a fluorescent protein. The biosensors were exposed to THz radiation from two sources: a high-intensity pulsed short-wave free electron laser and a low-intensity continuous long-wave IMPATT-diode-based device. The threshold and dynamics of fluorescence were found to depend on radiation parameters and exposure time. Heat shock or chemical stress yielded the absence of fluorescence induction. The biosensors are evaluated to be suitable for studying influence of THz radiation on the activity of gene networks related with considered gene promoters.

Temperature Sensing in the Short-Wave Infrared Spectral Region Using Core-Shell NaGdF4:Yb3+, Ho3+, Er3+@NaYF4 Nanothermometers.

The short-wave infrared region (SWIR) is promising for deep-tissue visualization and temperature sensing due to higher penetration depth and reduced scattering of radiation. However, the strong quenching of luminescence in biological media and low thermal sensitivity of nanothermometers in this region are major drawbacks that limit their practical application. Nanoparticles doped with rare-earth ions are widely used as thermal sensors operating in the SWIR region through the luminescence intensity ratio (LIR) approach. In this study, the effect of the shell on the sensitivity of temperature determination using NaGdF4 nanoparticles doped with rare-earth ions (REI) Yb3+, Ho3+, and Er3+ coated with an inert NaYF4 shell was investigated. We found that coating the nanoparticles with a shell significantly increases the intensity of luminescence in the SWIR range, prevents water from quenching luminescence, and decreases the temperature of laser-induced heating. Thermometry in the SWIR spectral region was demonstrated using synthesized nanoparticles in dry powder and in water. The core-shell nanoparticles obtained had intense luminescence and made it possible to determine temperatures in the range of 20-40 °C. The relative thermal sensitivity of core-shell NPs was 0.68% °C-1 in water and 4.2% °C-1 in dry powder.

Optimization of upconversion luminescence excitation mode for deeper in vivo bioimaging without contrast loss or overheating.

Upconversion nanoparticles have attracted considerable attention as luminescent markers for bioimaging and sensing due to their capability to convert near-infrared (NIR) excitation into visible or NIR luminescence. However, the wavelength of about 970 nm is commonly used for the upconversion luminescence excitation, where the strong absorption of water is observed, which can lead to laser-induced overheating effects. One of the strategies for avoiding such laser-induced heating involves shifting the excitation into shorter wavelengths region. However, the influence of wavelength change on luminescent images quality has not been investigated yet. In this work, we compare wavelengths of 920, 940 and 970 nm for upconversion luminescence excitation in the thickness of biological tissues in terms of detected signal intensity and obtained image quality (contrast and signal-to-background ratio). Studies on biological tissue phantoms with various scattering and absorbing properties were performed to analyze the influence of optical parameters on the depth and contrast of the images obtained under 920-970 nm excitation. It was shown that at the same power the excitation wavelength shift reduces the detected signal intensity and the resulting image contrast. Visualization of biological tissue samples using shorter excitation wavelengths 920 and 940 nm also reduces signal-to-background ratio (S/B) of the obtained images. The S/B of the obtained images amounted to 2, 6 and 8 for 920, 940 and 970 nm, respectively. It was demonstrated that pulse-periodic excitation mode is preferable for obtaining high quality luminescent images of biological tissues deep layers and minimize overheating. Short pulse durations (duty cycle 20%) don't result in heating even for 1 W cm-2 pumping power density and allow obtaining high luminescence intensity, which provides good images quality.

Labyrinth Metasurface for Biosensing Applications: Numerical Study on the New Paradigm of Metageometries.

The use of metasurfaces operating in the terahertz regime as biosensor devices has attracted increased interest in recent years due to their enhanced sensitivity and more accurate detection capability. Typical designs are based on the replica of relatively simple unit cells, usually called metaatoms. In a previous paper, we proposed a new paradigm for ultrasensitive thin-film sensors based on complex unit cells, called generically metageometries or labyrinth metasurfaces. Here, we extend this concept towards biosensing, evaluating the performance of the labyrinth as a fungi detector. The sensing capabilities are numerically evaluated and a comparison with previous works in this field is performed, showing that metageometries improve the performance compared to metaatoms both in sensitivity and figure of merit, by a factor of more than four. In particular, we find that it is able to detect five fungi elements scattered on the unit cell, equivalent to a concentration of only 0.004/µm2.

OCT-elastography-based optical biopsy for breast cancer delineation and express assessment of morphological/molecular subtypes.

Application of compressional optical coherence elastography (OCE) for delineation of tumor and peri-tumoral tissue with simultaneous assessment of morphological/molecular subtypes of breast cancer is reported. The approach is based on the ability of OCE to quantitatively visualize stiffness of studied samples and then to perform a kind of OCE-based biopsy by analyzing elastographic B-scans that have sizes ~several millimeters similarly to bioptates used for "gold-standard" histological examinations. The method relies on identification of several main tissue constituents differing in their stiffness in the OCE scans. Initially the specific stiffness ranges for the analyzed tissue components (adipose tissue, fibrous and hyalinized tumor stroma, lymphocytic infiltrate and agglomerates of tumor cells) are determined via comparison of OCE and morphological/molecular data. Then assessment of non-tumor/tumor regions and tumor subtypes is made based on percentage of pixels with different characteristic stiffness ("stiffness spectrum") in the OCE image, also taking into account spatial localization of different-stiffness regions. Examples of high contrast among benign (or non-invasive) and several subtypes of invasive breast tumors in terms of their stiffness spectra are given.

Reduced vs. standard dose native E. coli-asparaginase therapy in childhood acute lymphoblastic leukemia: long-term results of the randomized trial Moscow-Berlin 2002.

PURPOSE: Favorable outcomes were achieved for children with acute lymphoblastic leukemia (ALL) with the first Russian multicenter trial Moscow-Berlin (ALL-MB) 91. One major component of this regimen included a total of 18 doses of weekly intramuscular (IM) native Escherichia coli-derived asparaginase (E. coli-ASP) at 10000 U/m2 during three consolidation courses. ASP was initially available from Latvia, but had to be purchased from abroad at substantial costs after the collapse of Soviet Union. Therefore, the subsequent trial ALL-MB 2002 aimed at limiting costs to a reasonable extent and also at reducing toxicity by lowering the dose for standard risk (SR-) patients to 5000 U/m2 without jeopardizing efficacy. METHODS: Between April 2002 and November 2006, 774 SR patients were registered in 34 centers across Russia and Belarus, 688 of whom were randomized. In arm ASP-5000 (n = 334), patients received 5000 U/m2 and in arm ASP-10000 (n = 354) 10 000 U/m2 IM. RESULTS: Probabilities of disease-free survival, overall survival and cumulative incidence of relapse at 10 years were comparable: 79 ± 2%, 86 ± 2% and 17.4 ± 2.1% (ASP-5000) vs. 75 ± 2% and 82 ± 2%, and 17.9 ± 2.0% (ASP-10000), while death in complete remission was significantly lower in arm ASP-5000 (2.7% vs. 6.5%; p = 0.029). CONCLUSION: Our findings suggest that weekly 5000 U/m2E. coli-ASP IM during consolidation therapy are equally effective, more cost-efficient and less toxic than 10000 U/m2 for SR patients with childhood ALL.

In Vivo Formation of Stable Hyaline Cartilage by Naïve Human Bone Marrow Stromal Cells with Modified Fibrin Microbeads.

Osteoarthritic and other types of articular cartilage defects never heal on their own. Medicinal and surgical approaches are often ineffective, and the supply of autologous chondrocytes for tissue engineering is very limited. Bone marrow stromal cells (BMSCs, also known as bone marrow-derived mesenchymal stem cells) have been suggested as an adequate cell source for cartilage reconstruction. However, the majority of studies employing BMSCs for cartilage tissue engineering have used BMSCs predifferentiated into cartilage prior to implantation. This strategy has failed to achieve formation of stable, hyaline-like cartilage, resistant to hypertrophy in vivo. We hypothesized that in vitro predifferentiation of BMSCs is not necessary when cells are combined with an adequate scaffold that supports the formation of stable cartilage in vivo. In this study, naïve (undifferentiated) human BMSCs were attached to dehydrothermally crosslinked stable fibrin microbeads (FMBs) without and with other scaffolds and implanted subcutaneously into immunocompromised mice. Optimal formation of abundant, hypertrophy-resistant, ectopic hyaline-like cartilage was achieved when BMSCs were attached to FMBs covalently coated with hyaluronic acid. The cartilage that was formed was of human origin and was stable for at least 28 weeks in vivo. Stem Cells Translational Medicine 2019;8:586-592.