Metabolic Glycoengineering in hMSC-TERT as a Model for Skeletal Precursors by Using Modified Azide/Alkyne Monosaccharides

Metabolic glycoengineering enables a directed modification of cell surfaces by introducing target molecules to surface proteins displaying new features. Biochemical pathways involving glycans differ in dependence on the cell type; therefore, this technique should be tailored for the best results. We characterized metabolic glycoengineering in telomerase-immortalized human mesenchymal stromal cells (hMSC-TERT) as a model for primary hMSC, to investigate its applicability in TERT-modified cell lines. The metabolic incorporation of N-azidoacetylmannosamine (Ac₄ManNAz) and N-alkyneacetylmannosamine (Ac₄ManNAl) into the glycocalyx as a first step in the glycoengineering process revealed no adverse effects on cell viability or gene expression, and the in vitro multipotency (osteogenic and adipogenic differentiation potential) was maintained under these adapted culture conditions. In the second step, glycoengineered cells were modified with fluorescent dyes using Cu-mediated click chemistry. In these analyses, the two mannose derivatives showed superior incorporation efficiencies compared to glucose and galactose isomers. In time-dependent experiments, the incorporation of Ac₄ManNAz was detectable for up to six days while Ac₄ManNAl-derived metabolites were absent after two days. Taken together, these findings demonstrate the successful metabolic glycoengineering of immortalized hMSC resulting in transient cell surface modifications, and thus present a useful model to address different scientific questions regarding glycosylation processes in skeletal precursors.     

Dynamic time warping in classification and selection of motion capture data

Multidimensional Systems and Signal Processing - The paper is a comprehensive study on classification of motion capture data on the basis of dynamic time warping (DTW) transform. It presents both...     

Magnetic Resonance Signal Amplification by Reversible Exchange of Selective PyFALGEA Oligopeptide Ligands Towards Epidermal Growth Factor Receptors

The biorelevant PyFALGEA oligopeptide ligand, which is selective towards the epidermal growth factor receptor (EGFR), has been successfully employed as a substrate in magnetic resonance signal amplification by reversible exchange (SABRE) experiments. It is demonstrated that PyFALGEA and the iridium catalyst IMes form a PyFALGEA:IMes molecular complex. The interaction between PyFALGEA:IMes and H₂ results in a ternary SABRE complex. Selective 1D EXSY experiments reveal that this complex is labile, which is an essential condition for successful hyperpolarization by SABRE. Polarization transfer from parahydrogen to PyFALGEA is observed leading to significant enhancement of the ¹H NMR signals of PyFALGEA. Different iridium catalysts and peptides are inspected to discuss the influence of their molecular structures on the efficiency of hyperpolarization. It is observed that PyFALGEA oligopeptide hyperpolarization is more efficient when an iridium catalyst with a sterically less demanding NHC ligand system such as IMesBn is employed. Experiments with shorter analogues of PyFALGEA, that is, PyLGEA and PyEA, show that the bulky phenylalanine from the PyFALGEA oligopeptide causes steric hindrance in the SABRE complex, which hampers hyperpolarization with IMes. Finally, a single-scan ¹H NMR SABRE experiment of PyFALGEA with IMesBn revealed a unique pattern of NMR lines in the hydride region, which can be treated as a fingerprint of this important oligopeptide.     

Estimation of heat transfer coefficient of forced-air cooling and its experimental validation in controlled processing of forgings

Estimation of the cooling efficiency of an accelerated air for the needs of cooling of die forgings is presented. Temperature dependence of heat transfer coefficient (HTC) was calculated for different cooling conditions varied by airflow velocity, covering the range from 18 to 48?m/s. Time–temperature measurements performed on a full-scale semi-industrial cooling line provided similarity to conditions typical of industrial conveyor, which gives the results utilitarian significance in design of controlled processing (of steel forged products). Acquired HTC values, ranging from 164.7 to 298?W/m²?·?K, were validated in numerical simulation of cooling complex-shape forgings and subject to experimental verification, indicating perfect agreement with physical measurements.     

The Involvement of Xanthone and (E)-Cinnamoyl Chromophores for the Design and Synthesis of Novel Sunscreening Agents

Excessive UV exposure contributes to several pathological conditions like skin burns, erythema, premature skin aging, photodermatoses, immunosuppression, and skin carcinogenesis. Effective protection from UV radiation may be achieved with the use of sunscreens containing UV filters. Currently used UV filters are characterized by some limitations including systemic absorption, endocrine disruption, skin allergy induction, and cytotoxicity. In the research centers all over the world new molecules are developed to improve the safety, photostability, solubility, and absorption profile of new derivatives. In our study, we designed and synthesized seventeen novel molecules by combining in the structures two chromophores: xanthone and (E)-cinnamoyl moiety. The ultraviolet spectroscopic properties of the tested compounds were confirmed in chloroform solutions. They acted as UVB or UVA/UVB absorbers. The most promising compound 9 (6-methoxy-9-oxo-9H-xanthen-2-yl)methyl (E)-3-(2,4-dimethoxyphenyl)acrylate) absorbed UV radiation in the range 290–369 nm. Its photoprotective activity and functional photostability were further evaluated after wet milling and incorporation in the cream base. This tested formulation with compound 9 possessed very beneficial UV protection parameters (SPF_{in vitro} of 19.69 ± 0.46 and UVA PF of 12.64 ± 0.32) which were similar as broad-spectrum UV filter tris-biphenyl triazine. Additionally, compound 9 was characterized by high values of critical wavelength (381 nm) and UVA/UVB ratio (0.830) thus it was a good candidate for broad-spectrum UV filter and it might protect skin against UVA-induced photoaging. Compound 9 were also shown to be photostable, non-cytotoxic at concentrations up to 50 µM when tested on five cell lines, and non-mutagenic in Ames test. It also possessed no estrogenic activity, according to the results of MCF-7 breast cancer model. Additionally, its favorable lipophilicity (miLogP = 5.62) does not predispose it to penetrate across the skin after topical application.     

Elaboration of small‐diameter vascular prostheses—Selection of appropriate sterilisation method

The aim of study is the elaboration of semi‐biodegradable, multilayered tubular structures as substitutes for the reconstruction of small diameter vascular prostheses (<6 mm). The inert external layer of the prostheses will be fabricated via the melt electrospinning of poly (l ‐lactide‐co‐glycolide) (PLGA). The middle layer will be constructed from polypropylene (PP); the first prototype will be produced via melt electrospinning and the second using the melt blowing technique. The general aim of this stage of the research is the selection of a sterilisation technique that is appropriate for semi‐biodegradable, multilayered tubular structures. For this purpose, single tubular structures created via the melt electrospinning of PLGA or PP and melt blown tubular structures of PP were elaborated. The influence of steam, ethylene‐oxide (EO), and radiation sterilisation techniques on the elaborated microstructure of tubular structures was analyzed during this study. The effect of each sterilisation technique was evaluated using differential scanning calorimetry (DSC), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy/energy‐dispersive X‐ray spectroscopy analysis (SEM/EDS). The changes in average molecular weight (M_w) and crystallinity index (CI) of the PLGA tubular structures after EO and steam sterilisation were evaluated. The EO and steam sterilisation resulted in the complete destruction of PLGA tubular structures. Only the radiation sterilisation (accelerated electrons) did not influence on PLGA tubular structures morphology as well as thermal and chemical properties. FTIR and SEM/EDS analysis indicated that no changes in the chemical properties of PP tubular structures after each sterilisation occurred. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 40812.     

Small diameter tubular structure design using solvent‐free textile techniques

The aim of research was to elaborate the non‐biodegradable (made of polypropylene (PP)) and resorbable (made of polylactide (PLA)) tubular fibrous structures for the reconstruction of the vascular vessels. For the mentioned structures design, nonconventional manufacturing techniques such as melt blown, melt electrospinning, and melt electroblowing were used. Three techniques were chosen as methods allowing on the fibrous structures manufacture containing fibers in nano‐ or submicro‐size diameter. Other advantages of free‐solvent technique use is the reduction in the clinical adverse events associated with solvent resided in the fibrous structure during the fabrication. The tubular fibrous structures of PP and PLA using above‐mentioned techniques were designed. In first stage, the analysis of the processing parameters influence on the nonbiodegradable and biodegradable tubular structures fiber diameter was performed. Subsequently, the validation step was the analysis of the influence of processing parameters on PP and PLA structural properties for each manufacturing techniques was investigated. The research results confirmed the ability of the tubular structures manufacture with various fiber diameter depending on the applied technique and processing parameters. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 40147.     

A survey on the incidence of Prototheca mastitis in dairy herds in Lublin province,Poland

Prototheca mastitis has recently become an emerging disease; although its incidence is increasing steadily, its epidemiology remains largely understudied. The aim of this work was to investigate the prevalence of Prototheca spp. in dairy cows and their environment in Lublin province, covering most of southeastern Poland. Between December 2015 and July 2016, a total of 172 milking cows from 10 dairy farms were inspected for mastitis using clinical examination and the California Mastitis Test (CMT). Quarter milk samples (QMS, n = 179) and body site swabs (n = 151) from CMT-positive cows were collected for microbiological culture. In addition, we evaluated QMS and body site swabs from 23 healthy cows, along with 91 environmental samples. Of 100 CMT-positive cows, 71 had at least one QMS positive for microbial growth. In 8 (11.3%) of these cows, originating from 7 dairy farms, Prototheca spp. were cultured. The average somatic cell count of the Prototheca-containing milk was 4.02 × 10⁶ cells/mL compared with 0.13 × 10⁶ cells/mL of the Prototheca-free milk (collected from control animals). No significant differences were observed between mastitis and control cows with respect to counts of total white blood cells, lymphocytes, neutrophils, and eosinophils. Half of the cows with Prototheca spp. in their milk did not yield the algae from other anatomical sites. Eight cows were negative for the presence of Prototheca spp. in their milk but positive for the algae in swabs from anatomical sites. Among the environmental sources that were positive for Prototheca growth were watering troughs, manure, feed, and mud. All (45) Prototheca isolates recovered in this study were subjected to species- and genotype-level molecular identification. All QMS and most of the animal swabs (90%) yielded Prototheca zopfii genotype (gen.) 2. Of the animal samples, P. zopfii gen. 1 and Prototheca blaschkeae were isolated only from feces and rectum. Environmental samples grew either P. zopfii gen. 2 (67%) or P. zopfii gen. 1 (33%). This study demonstrates that P. zopfii gen. 2 is the third most common pathogen of mastitis in cattle in southeast Poland, with an overall incidence of 4.6%. Finding Prototheca spp., including P. zopfii gen. 1 and 2 and P. blaschkeae, in stool and rectal swabs from healthy animals may suggest their role as nonpathogenic microflora of bovine gut.     

Real-time 3D microtubule gliding simulation accelerated by GPU computing

A microtubule gliding assay is a biological experiment observing the dynamics of microtubules driven by motor proteins fixed on a glass surface. When appropriate microtubule interactions are set up on gliding assay experiments, microtubules often organize and create higher-level dynamics such as ring and bundle structures. In order to reproduce such higher-level dynamics on computers, we have been focusing on making a real-time 3D microtubule simulation. This real-time 3D microtubule simulation enables us to gain more knowledge on microtubule dynamics and their swarm movements by means of adjusting simulation parameters in a real-time fashion. One of the technical challenges when creating a real-time 3D simulation is balancing the 3D rendering and the computing performance. Graphics processor unit (GPU) programming plays an essential role in balancing the millions of tasks, and makes this real-time 3D simulation possible. By the use of general-purpose computing on graphics processing units (GPGPU) programming we are able to run the simulation in a massively parallel fashion, even when dealing with more complex interactions between microtubules such as overriding and snuggling. Due to performance being an important factor, a performance model has also been constructed from the analysis of the microtubule simulation and it is consistent with the performance measurements on different GPGPU architectures with regards to the number of cores and clock cycles.