OBJECTIVE: Radiographic assessment of sacroiliac joints (SIJs) according to the modified New York (mNY) criteria is key in the classification of axial spondyloarthritis but has moderate interreader agreement. We aimed to investigate the improvements of the reliability in scoring SIJ radiographs after applying an online real-time iterative calibration (RETIC) module, in addition to a slideshow and video alone. METHODS: Nineteen readers, randomized to 2 groups (A or B), completed 3 calibration steps: (1) review of manuscripts, (2) review of slideshow and video with group A completing RETIC, and (3) re-review of slideshow and video with group B completing RETIC. The RETIC module gave instant feedback on readers' gradings and continued until predefined reliability ([Formula: see text]) targets for mNY positivity/negativity were met. Each step was followed by scoring different batches of 25 radiographs (exercises I to III). Agreement ([Formula: see text]) with an expert radiologist was assessed for mNY positivity/negativity and individual lesions. Improvements by training strategies were tested by linear mixed models. RESULTS: In exercises I, II, and III, mNY [Formula: see text] were 0.61, 0.76, and 0.84, respectively, in group A; and 0.70, 0.68, and 0.86, respectively, in group B (ie, increasing, mainly after RETIC completion). Improvements were observed for grading both mNY positivity/negativity and individual pathologies, both in experienced and, particularly, inexperienced readers. Completion of the RETIC module in addition to the slideshow and video caused a significant [Formula: see text] increase of 0.17 (95% CI 0.07-0.27; P = 0.002) for mNY-positive and mNY-negative grading, whereas completion of the slideshow and video alone did not ([Formula: see text] = 0.00, 95% CI -0.10 to 0.10; P = 0.99). CONCLUSION: Agreement on scoring radiographs according to the mNY criteria significantly improved when adding an online RETIC module, but not by slideshow and video alone.
- MeSH
- Axial Spondyloarthritis * diagnostic imaging MeSH
- Humans MeSH
- Observer Variation * MeSH
- Radiography * methods MeSH
- Reproducibility of Results MeSH
- Sacroiliac Joint * diagnostic imaging MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Randomized Controlled Trial MeSH
IMUNOR is an oral biotherapeutic drug that had been developed, registered, and approved in 1997 in the Czech Republic and Slovakia. IMUNOR is a dialyzable leukocyte extract (DLE) prepared from swine leukocytes. It is characterized as a mixture of small peptides with molecular weights smaller than 12 kDa and a specific portion of nucleotides. The medical uses of IMUNOR include therapeutic applications within its registered range of indications, primarily for the treatment of immunodeficiencies, allergies, and certain acute or relapsing bacterial infections in adults and children. Despite the long-term clinical application of DLE, with strong evidence of positive therapeutic effects and no serious side effects, a detailed physicochemical specification of this mixture was lacking. We developed several methods for more in-depth physicochemical characterization of IMUNOR, including a spectrophotometric method for quantification of the total protein concentration and total DNA concentration in a mixture, several chromatographic methods for identification of individual components present in significant concentrations in IMUNOR, such as HPLC methods and the Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis method, and characterization of amino acid composition of this mixture. For the investigation of the variability among different batches of IMUNOR, five to nine representative batches from a standard manufacturing process on an industrial scale were utilized. Using the analytical methods, we verified and confirmed the batch-to-batch reproducibility of the biological product IMUNOR.
- Publication type
- Journal Article MeSH
Spruce wood and Typha (wetland plant) derived biochars pyrolyzed at 350 °C and 600 °C were tested for their sorption affinity for organic pollutants (diclofenac, methylparaben, benzotriazole and sodium 1-decanesulfonate) and nutrients (nitrate, ammonium, phosphate and boron) commonly found in greywater. Batch and column studies combined with molecular dynamics modelling determined the sorption capacity, kinetics, and described the underlying mechanisms. The spruce biochar (600 °C) exhibited the highest sorption capacity mainly for the tested organics. The dynamic test performed for spruce biochar (600 °C) showed that the magnitude of desorption was low, and the desorbed amount ranged between 3 and 11 %. Molecular dynamics modelling (a computational tool for elucidating molecular-level interactions) indicated that the increased sorption of nitrate and boron on spruce biochar (600 °C) could be attributed to hydrophobic interactions. The molecular dynamics shows that predominant adsorption of organic pollutants was governed by π-π stacking, with a minor role of hydrogen-bonding on the biochar surface. In summary, higher pyrolysis temperature biochar yielded greater adsorption capacity greywater borne contaminants and the reaction temperature (10-34 °C) and presence of anionic surfactant had a limited effect on the adsorption of organic pollutants, suggesting efficacious application of biochar in general for greywater treatment in nature-based systems.
Inhalační podání suchých práškových částic je využívanou aplikační cestou pro dosažení lokálního i systémového působení léčiv. U plicních onemocnění je žádoucí depozice léčiv v místě účinku. Pro efektivní léčbu jsou tak zásadní parametry inhalovaných částic, a to především jejich velikost, tvar nebo aerosolizační parametry. Vhodných parametrů je možné dosáhnout volbou metody přípravy nebo pomocných látek (nosičů, porogenů nebo aerosolizačních činidel). Cílem experimentu byla příprava jedenácti šarží práškových směsí sprejovým sušením, které se lišily použitým nosičem, množstvím leucinu či porogenu. Cílem bylo optimalizovat složení pro navázání léčiva s ohledem na požadavky pro plicní podání. Připravené částice byly zhodnoceny z hlediska morfologie, tokových vlastností, pórovitosti, geometrického i aerodynamického průměru. Bylo zjištěno, že se zvyšující se koncentrací leucinu se sypná hustota částic snižovala a zároveň rostla hodnota FPF. Stejně tak docházelo i ke snižování MMAD. Jako vhodná se jevila šarže obsahující 15 % leucinu. Při stanovení optimální koncentrace porogenu (hydrogenuhličitan amonný) u manitolových částic dosahovala nejlepších výsledků šarže s jeho 1% zastoupením, vzhledem k vyhovující velikosti částic oproti ostatním šaržím (MMAD 5,92 ± 1,32 μm), vhodné pórovitosti a obecně přijatelné morfologii částic. Za účelem formulace částic s navázaným léčivem by tedy bylo vhodné snížit aerodynamický průměr částic např. úpravou procesních parametrů sprejového sušení.
Inhalation administration of dry powder particles is a common application route to achieve local and systemic drug effects. For pulmonary diseases, the deposition of drugs at the site of action is desirable. Thus, the parameters of the inhaled particles, especially their size, shape, or aerosolization, are essential for effective treatment. Suitable parameters can be achieved by choice of preparation method or excipients (carriers, porogens, or aerosolizing agents). This experiment aimed to prepare 11 batches of powder mixtures by spray drying, which differed in the carrier used and the amount of leucine or porogen. The aim was to optimize the formulation for drug binding concerning the requirements for pulmonary administration. The prepared particles were evaluated in terms of morphology, flow properties, porosity, and geometric and aerodynamic diameter. It was found that with increasing concentration of leucine, the bulk density of the particles decreased while the FPF value increased. Similarly, there was a decrease in MMAD. The batch containing 15% leucine was the most suitable. In determining the optimum porogen concentration for mannitol particles, the batch with its 1% gave the best results due to its adequate particle size compared to the other batches (MMAD 5.92 ± 1.32 μm), suitable porosity, and particle morphology. Thus, to formulate drug-loaded particles, it would be advisable to reduce the aerodynamic diameter of the particles, e.g., by spray drying process parameters.
Salmonella spp. is a common zoonotic pathogen, causing gastrointestinal infections in people. Pigs and pig meat are a major source of infection. Although farm biosecurity is believed to be important for controlling Salmonella transmission, robust evidence is lacking on which measures are most effective. This study enrolled 250 pig farms across nine European countries. From each farm, 20 pooled faecal samples (or similar information) were collected and analysed for Salmonella presence. Based on the proportion of positive results, farms were categorised as at higher or lower Salmonella risk, and associations with variables from a comprehensive questionnaire investigated. Multivariable analysis indicated that farms were less likely to be in the higher-risk category if they had '<400 sows'; used rodent baits close to pig enclosures; isolated stay-behind (sick) pigs; did not answer that the hygiene lock/ anteroom was easy to clean; did not have a full perimeter fence; did apply downtime of at least 3 days between farrowing batches; and had fully slatted flooring in all fattener buildings. A principal components analysis assessed the sources of variation between farms, and correlation between variables. The study results suggest simple control measures that could be prioritised on European pig farms to control Salmonella.
- MeSH
- Biosecurity MeSH
- Animal Husbandry methods MeSH
- Farms MeSH
- Swine Diseases * epidemiology prevention & control MeSH
- Swine MeSH
- Salmonella MeSH
- Salmonella Infections, Animal * epidemiology prevention & control MeSH
- Animals MeSH
- Check Tag
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Europe MeSH
The use of knowledge from technetium radiochemistry (even from nuclear medicine applications) allows us to select an sorbent for 99mTc radionuclide sorption, which is hydroxyapatite. Using radioisotope indication, the 99mTcO4- sorption process on synthetic hydroxyapatite was studied by the batch method in the presence of SnCl2 and FeSO4 reducing agents. The complexing organic ligands' effect on the 99mTcO4- sorption under reducing conditions was investigated. In the presence of Sn2+ ions without the addition of organic ligand, the sorption percentage reached above 90% independently of the environment. In the presence of Fe2+ ions without the addition of organic ligand, the sorption of 99mTcO4- was significantly lower and was at approximately 6%, depending on the concentration of Fe2+ ions in solution. The effect of complexing organic ligands on the 99mTcO4- sorption on hydroxyapatite from the aqueous solution, acetate buffer and phosphate buffer decreases in the following order for Sn2+: oxalic acid > ethylenediaminetetraacetic acid > ascorbic acid. In the presence of Fe2+ ions without organic ligands, the sorption reached up to 15% depending on the composition of the solution. The addition of oxalic acid and ascorbic acid increased the sorption up to 80%. The ethylenediaminetetraacetic acid had no significant effect on the sorption of technetium on hydroxyapatite.
Formulace mikročástic složených ze směsi nosičů představuje inovativní přístup pro podání léčiv do plic ve formě suchého prášku. Použité nosiče mohou významně ovlivnit výsledné vlastnosti mikročástic, jako je velikost, tvar, povrch, hygroskopicita či agregace, a tím zlepšit aerosolizaci léčiv po jejich inhalaci. Zmíněné vlastnosti jsou klíčové pro efektivní pulmonální terapii. Kombinací nosičů povahy sacharidů a gelujících látek je výhodné pro řízené uvolňování léčiva. Cílem experimentální práce bylo sprejovým sušením připravit a následně zhodnotit několik šarží mikročástic složených z nosičů na bázi cukrů (manitol, maltodextrin, dextran) a gelujících sacharidů (chitosan, chondroitin-sulfát) a vybrat vhodnou kombinaci pro navazující experimentální práce zaměřené na inkorporaci léčiva do mikročásticové matrice. Nejvhodnější parametry vykazovaly šarže, jejichž aerodynamický průměr se blížil 5 μm, a to částice připravené z kombinace manitolu a dextranu, chitosanu a chondroitinu nebo maltodextrinu a chondoitinu. U těchto šarží byla také naměřena nejvyšší hodnota frakce jemných částic (> 43 %). Z pohledu zpracovatelnosti je vhodná šarže se zastoupením maltodextrinu a chondroitinu vzhledem k nižší viskozitě vstupní disperze a pravidelnějšímu tvaru finálních mikročástic.
The formulation of microparticles composed of a mixture of carriers represents an innovative approach for lung drug delivery of dry powder. The carriers used can significantly influence the properties of the microparticles, such as size, shape, surface area, hygroscopicity, or aggregation, thus improving the aerosolization of the drugs after inhalation. The properties mentioned above are crucial for effective pulmonary therapy. The combination of carriers of a carbohydrate nature and gelling agents is advantageous for controlled drug release. The experimental work aimed to prepare by spray drying and subsequently evaluate ten batches of microparticles composed of sugar-based carriers (mannitol, maltodextrin, dextran) and gelling polymers (chitosan, chondroitin sulfate) and to select a suitable combination for follow-up experimental work aimed at drug incorporation into the microparticle matrix. The most suitable parameters were exhibited by batches whose aerodynamic diameter was close to 5 μm, particles prepared from a combination of mannitol and dextran, chitosan and chondroitin, or maltodextrin and chondroitin. These batches also showed the highest fine particle fraction value (> 43%). From a processability point of view, the batch with maltodextrin and chondroitin is preferable due to the lower viscosity of the dispersion and the more regular shape of the final microparticles.
- MeSH
- Administration, Inhalation MeSH
- Pharmaceutical Research MeSH
- Humans MeSH
- Microplastics MeSH
- Drug Carriers * MeSH
- Spray Drying MeSH
- Check Tag
- Humans MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
Hibernating amphibians are suitable for the research on the adaptation of gut microbiota to long-term fasting and cold stresses. However, the previous studies mainly focus on the large or whole gut microbiota but not the small gut microbiota. To test the structural discrepancy between the small and large gut microbiota during hibernation, we performed two independent batches of 16S rRNA gene amplicon sequencing to profile the small and large gut microbiota of hibernating Asiatic toad (Bufo gargarizans) from two wild populations. Both batches of data revealed that Proteobacteria, Bacteroidetes, and Firmicutes were the three most dominant phyla in the small and large gut microbiota. Three core OTUs with 100% occurrence in all gut microbiotas were annotated as Pseudomonas. A significant structural discrepancy was detected between the small and large gut microbiota. For instance, Proteobacteria assembled in the small intestine with a higher proportion than it did in the large intestine, but Bacteroidetes and Firmicutes assembled in the large intestine with a higher proportion than they did in the small intestine. The large gut microbiota exhibited higher diversity than the small gut microbiota. Nevertheless, a severe batch effect existed in the structural analysis of the gut microbiotas. The large gut microbiota showed a better resistance to the batch effect than the small gut microbiota did. This study provides preliminary evidence that microbes assemble in the small and large intestines of amphibians with discrepant patterns during hibernation.
- MeSH
- Hibernation * MeSH
- RNA, Ribosomal, 16S genetics MeSH
- Bufonidae genetics microbiology MeSH
- Gastrointestinal Microbiome * MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
Animal models are essential in understanding of the mechanisms of sepsis moreover the development and the assessment of emerging therapies. In clinically relevant porcine model, however, a significant variability in the host response has been observed among animals. Thus, there is a strong demand to better understand the potential sources of this heterogeneity. In this study, we compared faecal microbiome composition of 12 animals. Three samples were collected at different time points from each animal. Bacteriome was subjected to 16S rDNA profiling. A significant difference in bacterial composition was associated with the season (p < 0.001) but not with the sex of the pig (p = 0.28), the timing of sample collection (p = 0.59), or interactions thereof (all p > 0.3). The season batch explained 55% of the total variance in the bacteriome diversity. The season term was highly significant from the high-resolution level of the bacterial amplicon sequencing variants up to the level of phylum. The diversity of the microbiome composition could significantly influence experimental model of sepsis, and studies are warranted to demonstrate the effects of gut microbiome diversity on the host-response. If confirmed, control of the gut microbiome should become a standard part of the pre-clinical sepsis experiments.
Reakce štěpu proti hostiteli (graft versus host disease – GvHD) patří mezi nejzávažnější komplikace alogenní transplantace hematopoetických kmenových buněk. Přestože se díky optimalizaci transplantačních postupů mortalita v důsledku GvHD neustále snižuje, nadále existuje početná skupina pacientů, u kterých se rozvine život ohrožující forma GvHD, případně kterým GvHD zásadně snižuje kvalitu života po transplantaci. Jedním z inovativních přístupů, které by mohly zlepšit prognózu pacientů, je adoptivní podání některých buněčných populací, které mají imunomodulační potenciál. Na našem pracovišti jsme v rámci klinické studie podávali léčivé přípravky z mezenchymálních kmenových buněk (MSC) v léčbě steroidně-refrakterní GvHD (SR-GvHD). Nyní ověřujeme možnost využití invariantních NKT lymfocytů (iNKT) v této indikaci. Tyto iNKT lymfocyty jsou schopné tlumit patologické imunitní reakce a rozvoj příznaků GvHD a současně podporovat protinádorovou imunitu – tzv. reakci štěpu proti leukemii (graft versus leukemia – GvL). V rámci preklinického testování jsme porovnali několik šarží kultivovaných a expandovaných iNKT a MSC od různých dárců z hlediska jejich imunofenotypu a imunomodulačního potenciálu. Průtokovou cytometrií jsme kvantifikovali expresi aktivačního znaku CD25 na nespecificky stimulovaných mononukleárních buňkách po kokultivaci s MSC nebo s iNKT. Studie prokázala, že je možné připravit čistou populaci iNKT buněk s dostatečnou imunomodulační kapacitou srovnatelnou s MSC. Zdá, že iNKT buňky představují slibnou léčebnou strategii v rámci terapie potransplantačních komplikací, nicméně vzhledem k výrazné heterogenitě jejich subpopulací bude nutné provést hlubší stanovení funkce iNKT a optimalizovat protokol pro jejich přípravu.
Graft versus host disease (GvHD) is one of the most serious complications of allogeneic hematopoietic stem cell transplantation. Although GvHD mortality is steadily declining due to the optimalization of transplant procedures, there remains a large group of patients who develop life-threatening forms of GvHD, or whose quality of life after transplantation is reduced due to GvHD. The adoptive administration of immunomodulatory cell populations is an innovative approach that could improve the prognosis of these patients. We had previously used mesenchymal stem cells (MSC) in steroid-refractory GvHD (SR-GvHD) in a clinical trial and we are currently verifying the possibility of using invariant NKT lymphocytes (iNKT) in SR-GvHD treatment. iNKT are a cell population that is able to suppress pathological immune responses and the development of GvHD symptoms. At the same time, iNKT are able to promote anti-tumour immunity – the graft versus leukaemia effect (GvL). In a preclinical study, we compared several batches of cultured and expanded iNKT and MSCs from different donors in terms of their immunophenotype and immunomodulatory potential. We quantified the expression of the activation marker CD25 on non-specifically stimulated mononuclear cells after co-cultivation with MSC or iNKT. The study showed that it is possible to prepare a pure population of iNKT with sufficient immunomodulatory capacity comparable to MSCs. While iNKT cells appear to be a promising treatment strategy for post-transplant complications, the significant heterogeneity of their subpopulations will require a deeper assessment of iNKT function and optimisation of their production protocols.
