Ramanovu spektroskopii lze v chemii použít nejen k určování chemického složení, ale také pro získávání dalších informací o struktuře materiálu. Ve spektrech semikrystalických polymerů lze nalézt vzájemně odlišné pásy charakteristické pro krystalickou nebo amorfní fázi, stanovit z nich krystalinitu a z ní odhadnout míru degradace polymeru. V předložené studii byly vyhodnoceny změny raménka pásu na vlnočtu 1733 cm−1 v Ramanových spektrech vlákna z poly(p-dioxanonu) podrobeného hydrolytické degradaci. Pro různě dlouhé doby degradace byly vypočteny obsahy ploch pod raménkem tohoto pásu a též byl proveden jeho modelový rozklad na předpokládané píky krystalické a amorfní fáze. Obsahy ploch pod raménkem i parametry modelových píků byly porovnány s hodnotami krystalinity získanými pomocí diferenční skenovací kalorimetrie, přičemž bylo dosaženo dobré shody. Tato práce ukazuje příklad využití Ramanovy spektroskopie při studiu hydrolytické degradace polymerů.
Raman spectroscopy can be used in chemistry not just to determine chemical composition, but also to obtain further information on the material structure. In the spectra of semi-crystalline polymers, distinct bands characteristic of the crystalline or the amorphous phase can be found, the degree of crystallinity determined from them, and the degree of polymer degradation estimated from the crystallinity. In the present study, changes in the 1733 cm−1 band shoulder in Raman spectra of poly(p-dioxanone) fibres subjected to hydrolytic degradation were evaluated. For different degradation periods, the areas under the shoulder of this band were calculated and a model deconvolution of this band into assumed crystalline and amorphous peaks was also performed. The areas under the shoulder, as well as the model peaks' parameters, were compared with the crystallinity values obtained by differential scanning calorimetry, achieving a good agreement. This work shows an example of using Raman spectroscopy when studying the hydrolytic degradation of polymers.
PURPOSE: Adhesive remnants removal is the last key step influencing orthodontic treatment outcomes. Four different clearance methods (CM) of orthodontic adhesive were evaluated to determine, which achieved the smoothest enamel surface in the shortest time. MATERIALS AND METHODS: 75 intact premolars extracted for orthodontic purposes were included, sixty had an orthodontic bracket bonded and subsequently removed, and fifteen served as the control group. Four CMs were used to clear the tooth surface of 15 premolars each: carbide bur (CB), carbide bur with titanium nitride surface treatment + fine carbide bur (CBCB), glass fiber-reinforced composite instrument (GFCB), zirconia bur + glass fiber-reinforced composite bur (ZBCB). The processing time was recorded. In ten premolars from each group, the enamel surface was evaluated by atomic force microscopy estimating mean roughness (Ra), roughness profile value (Rq), and roughness depth (Rt). Enamel Damage Index (EDI) was assessed with a scanning electron microscope on 5 remaining premolars. RESULTS: Significant differences were observed in all evaluated parameters - Ra (p<0.0001), Rq (p<0.0001), and Rt (p<0.0001). GFCB exhibited the smoothest surface in all parameters. The lowest EDI exhibited teeth treated by GFCB, however, the differences were not significant. Working with GFCB took the longest time (mean 116 s), and the shortest with CBCB (mean 49 s). CONCLUSION: Using CB is the fastest clearance method, but the enamel surface roughness was highest. Clearing with a set of instruments CBCB proved to be a fast method with satisfying remaining enamel roughness.
- Publikační typ
- časopisecké články MeSH
Although some clinical studies have reported increased mitochondrial respiration in patients with fatty liver and early non‐alcoholic steatohepatitis (NASH), there is a lack of in vitro models of non‐alcoholic fatty liver disease (NAFLD) with similar findings. Despite being the most commonly used immortalized cell line for in vitro models of NAFLD, HepG2 cells exposed to free fatty acids (FFAs) exhibit a decreased mitochondrial respiration. On the other hand, the use of HepaRG cells to study mitochondrial respiratory changes following exposure to FFAs has not yet been fully explored. Therefore, the present study aimed to assess cellular energy metabolism, particularly mitochondrial respiration, and lipotoxicity in FFA‐treated HepaRG and HepG2 cells. HepaRG and HepG2 cells were exposed to FFAs, followed by comparative analyses that examained cellular metabolism, mitochondrial respiratory enzyme activities, mitochondrial morphology, lipotoxicity, the mRNA expression of selected genes and triacylglycerol (TAG) accumulation. FFAs stimulated mitochondrial respiration and glycolysis in HepaRG cells, but not in HepG2 cells. Stimulated complex I, II‐driven respiration and β‐oxidation were linked to increased complex I and II activities in FFA‐treated HepaRG cells, but not in FFA‐treated HepG2 cells. Exposure to FFAs disrupted mitochondrial morphology in both HepaRG and HepG2 cells. Lipotoxicity was induced to a greater extent in FFA‐treated HepaRG cells than in FFA‐treated HepG2 cells. TAG accumulation was less prominent in HepaRG cells than in HepG2 cells. On the whole, the present study demonstrates that stimulated mitochondrial respiration is associated with lipotoxicity in FFA‐treated HepaRG cells, but not in FFA‐treated HepG2 cells. These findings suggest that HepaRG cells are more suitable for assessing mitochondrial respiratory adaptations in the developed in vitro model of early‐stage NASH.
- MeSH
- buněčné linie MeSH
- buňky Hep G2 MeSH
- dýchání MeSH
- kyseliny mastné neesterifikované MeSH
- lidé MeSH
- mitochondrie MeSH
- nealkoholová steatóza jater * MeSH
- triglyceridy MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Although Holder pasteurization is the recommended method for processing breast milk, it does affect some of its nutritional and biological properties and is ineffective at inactivating spores. The aim of this study was to find and validate an alternative methodology for processing breast milk to increase its availability for newborn babies and reduce the financial loss associated with discarding milk that has become microbiologically positive. We prepared two series of breast milk samples inoculated with the Bacillus cereus (B. cereus) strain to verify the effectiveness of two high-pressure treatments: (1) 350 MPa/5 min/38 °C in four cycles and (2) cumulative pressure of 350 MPa/20 min/38 °C. We found that the use of pressure in cycles was statistically more effective than cumulative pressure. It reduced the number of spores by three to four orders of magnitude. We verified that the method was reproducible. The routine use of this method could lead to an increased availability of milk for newborn babies, and at the same time, reduce the amount of wasted milk. In addition, high-pressure treatment preserves the nutritional quality of milk.
- Publikační typ
- časopisecké články MeSH
The lack of physiological parity between 2D cell culture and in vivo culture has led to the development of more organotypic models, such as organoids. Organoid models have been developed for a number of tissues, including the liver. Current organoid protocols are characterized by a reliance on extracellular matrices (ECMs), patterning in 2D culture, costly growth factors and a lack of cellular diversity, structure, and organization. Current hepatic organoid models are generally simplistic and composed of hepatocytes or cholangiocytes, rendering them less physiologically relevant compared to native tissue. We have developed an approach that does not require 2D patterning, is ECM independent, and employs small molecules to mimic embryonic liver development that produces large quantities of liver-like organoids. Using single-cell RNA sequencing and immunofluorescence, we demonstrate a liver-like cellular repertoire, a higher order cellular complexity, presenting with vascular luminal structures, and a population of resident macrophages: Kupffer cells. The organoids exhibit key liver functions, including drug metabolism, serum protein production, urea synthesis and coagulation factor production, with preserved post-translational modifications such as N-glycosylation and functionality. The organoids can be transplanted and maintained long term in mice producing human albumin. The organoids exhibit a complex cellular repertoire reflective of the organ and have de novo vascularization and liver-like function. These characteristics are a prerequisite for many applications from cellular therapy, tissue engineering, drug toxicity assessment, and disease modeling to basic developmental biology.
- MeSH
- hepatocyty MeSH
- játra * MeSH
- kultivované buňky MeSH
- lidé MeSH
- myši MeSH
- organoidy * MeSH
- tkáňové inženýrství MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- elektronová mikroskopie MeSH
- lidé MeSH
- ortodontické zámky škodlivé účinky MeSH
- premolár MeSH
- zubní nástroje MeSH
- zubní sklovina * ultrastruktura MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- klinická studie MeSH
BACKGROUND & AIMS: Lymphedema cholestasis syndrome 1 or Aagenaes syndrome is a condition characterized by neonatal cholestasis, lymphedema, and giant cell hepatitis. The genetic background of this autosomal recessive disease was unknown up to now. METHODS: A total of 26 patients with Aagenaes syndrome and 17 parents were investigated with whole-genome sequencing and/or Sanger sequencing. PCR and western blot analyses were used to assess levels of mRNA and protein, respectively. CRISPR/Cas9 was used to generate the variant in HEK293T cells. Light microscopy, transmission electron microscopy and immunohistochemistry for biliary transport proteins were performed in liver biopsies. RESULTS: One specific variant (c.-98G>T) in the 5'-untranslated region of Unc-45 myosin chaperone A (UNC45A) was identified in all tested patients with Aagenaes syndrome. Nineteen were homozygous for the c.-98G>T variant and seven were compound heterozygous for the variant in the 5'-untranslated region and an exonic loss-of-function variant in UNC45A. Patients with Aagenaes syndrome exhibited lower expression of UNC45A mRNA and protein than controls, and this was reproduced in a CRISPR/Cas9-created cell model. Liver biopsies from the neonatal period demonstrated cholestasis, paucity of bile ducts and pronounced formation of multinucleated giant cells. Immunohistochemistry revealed mislocalization of the hepatobiliary transport proteins BSEP (bile salt export pump) and MRP2 (multidrug resistance-associated protein 2). CONCLUSIONS: c.-98G>T in the 5'-untranslated region of UNC45A is the causative genetic variant in Aagenaes syndrome. IMPACT AND IMPLICATIONS: The genetic background of Aagenaes syndrome, a disease presenting with cholestasis and lymphedema in childhood, was unknown until now. A variant in the 5'-untranslated region of the Unc-45 myosin chaperone A (UNC45A) was identified in all tested patients with Aagenaes syndrome, providing evidence of the genetic background of the disease. Identification of the genetic background provides a tool for diagnosis of patients with Aagenaes syndrome before lymphedema is evident.
- MeSH
- 5' nepřekládaná oblast genetika MeSH
- cholestáza * genetika MeSH
- HEK293 buňky MeSH
- intracelulární signální peptidy a proteiny * genetika MeSH
- lidé MeSH
- lymfedém * diagnóza genetika metabolismus MeSH
- myosiny genetika metabolismus MeSH
- novorozenec MeSH
- transportní proteiny genetika MeSH
- Check Tag
- lidé MeSH
- novorozenec MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Due to a broad spectrum of endodontic rotary instruments on the market and no standardised protocol for comparing their mechanical properties, it can be challenging for clinician to choose proper instruments. In vitro studies using resin blocks with artificial canals can offer many valuable information because of their uniformity compared to studies performed on extracted teeth. To improve precision and reproducibility of artificial canals, 3D printing was used in this study to manufacture endodontic test block samples. 20 commercially available endodontic blocks Endo-Training-Bloc-J by Dentsply Sirona were tested. The mean values of the measured parameters were used for a 3D CAD model of their replicas. 20 copies of the endodontic training blocks were printed from acrylic resin (VeroClear-RGD810, Stratasys, Eden Prairie, USA) using the 3D printer Objet30 Pro (Stratasys, Eden Prairie, USA). The key dimensions of the commercial blocks and the 3D printed blocks were measured under and compared using t - test and Levene's test for equality of variances. The profiles of the 3D printed artificial canals showed significantly lower dimensional variability when compared with the commercial blocks. 3D polyjet printing proved to be a precise and reproducible method for production of blocks for testing endodontic rotary instruments.
- MeSH
- 3D tisk MeSH
- endometrióza * MeSH
- extrakce zubů MeSH
- lidé MeSH
- reprodukovatelnost výsledků MeSH
- výzkumný projekt * MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Publikační typ
- abstrakt z konference MeSH
OBJECTIVES: A stent is a mesh tube inserted into a natural passage in the body to prevent disease induction. Self-expandable esophageal nitinol stents such as SX-ELLA Stent Esophageal HV (HV Stent Plus) can be indicated for palliation of malignant esophageal strictures, for the treatment of benign esophageal strictures that are refractory to standard therapy and for the treatment of esophago-respiratory fistulas. A silicone-stent coating is used for tumor in-growth prevention and esophago-respiratory fistula occlusion. The thickness of the stent and the overall integrity of the silicone coating of all wires indicate the overall mechanical properties of the esophageal stent and the resistance to external adverse events such as corrosion and mechanical and chemical resistance. METHODS: The polymer multicomponent epoxy resin - a mixture of Epon and Durcupan - was used as a method for robust sample stabilization. A cutting system using a thin water beam with a powder (Blue Line) was chosen as the best variant to obtain 6 samples for both-sided measurement (10 measuring sides). The optical microscopic reflective light method was used to examine wire crossing points in the sections. Fifty values were measured on either sample side for the internal, external and mesh thickness of the silicone stent layer. The wire crossing points were selected so that the silicone layer structure could be clearly seen, and the wires approached each other most closely. Only approximately 4 to 8 crossing points in each section could be measured when applying this approach. The resolution of the microscope and calibration (based on the camera used) was 0.677 μm/pixel. RESULTS: Additional data could be obtained on 8 planes. Two boundary samples were destroyed by the cutting process. Whole coating of the stent was around all mesh wires, especially in areas with higher mechanical stress (wire crossing). The minimum detectable and admissible value determined for all 3 measuring areas (internal, external, mesh) on the wire crossings was 6.77 μm, i.e., 10 pixels, based on the microscope resolution and manufacturer's methodology. The results were characterized by p < 0.001 for all 3 parameters. We tested opposite samples in each section to verify the section quality and data consistency. For the 4 areas, the data were significantly different, but the thickness differences were only on the order of units percent, so the measurements were not appreciably affected. We assume that the material cutting loss, making up 1-2 mm, contributed to the differences in the sections. CONCLUSION: We examined the overall integrity of the silicone coating of the esophageal stent. The method of HV stent anchoring in a polymeric bath followed by cutting with a waterjet and sample measurement under an optical microscope proved to be very simple and reliable. Sufficient thicknesses of the silicone layer on the wire cross sections were verified. The coated silicone layer thickness appeared to be significantly different along the stent from the proximal part to the distant part, presumably due to the manufacturing technology.
