Real-world outcomes of mepolizumab treatment in severe eosinophilic asthma patients - retrospective cohort study in Slovakia.

AIMS: Mepolizumab, a fully-humanized recombinant IgG1 kappa monoclonal antibody directed against IL-5, has shown improved asthma control and lung function in randomised controlled trials. The aim of this study was to evaluate real-world clinical experience in patients with severe eosinophilic asthma treated with mepolizumab in Slovakia. METHODS: A retrospective, non-interventional study based on medical records of all adult asthma patients initiating mepolizumab between November 1, 2017 and January 31, 2019, completing 12 months of treatment. At baseline, general and clinical profile data were recorded 12 months prior to treatment. Primary and secondary endpoints described the results of mepolizumab use at 2, 6, and 12 months after the initiation and compared to baseline. Statistical testing of individual change (in each patient) in selected parameters was performed. RESULTS: The cohort included 17 patients with particularly severe asthma at baseline, with frequent severe exacerbations (SE, median 5 [IQR 4-6]/patient/year), high blood eosinophil counts (median 0.6x109/L), frequent oral corticosteroid (OCS) dependence (82.35%), median dose 15 (IQR 7.5-20) mg/day, impaired lung function, and a spectrum of comorbidities. In a one-year follow-up, the data showed reductions in median SE (0 [IQR 0-1] patient/year, eosinophilia (median 0.175x109/L) and OCS maintenance dose (median 6.25 [IQR 2.5-20] mg/day), all statistically significant after 12 months on mepolizumab. Improved and stabilised lung functions throughout the cohort and a reduced incidence of nasal polyposis were observed. CONCLUSIONS: The results provide clinical evidence of mepolizumab efficacy in a real sample of patients with severe asthma when administered in routine care settings in Slovakia.

COVID-19 in the Czech Republic 2020 and 2021: comparative analysis of probable work-related transmission of the coronavirus SARS-CoV-2.

OBJECTIVES: The aim of the analysis was to determine the probable places of coronavirus transmission in association with the work and compare the situation between 2020 and 2021. METHODS: The work analysed data from the Information System of Infectious Diseases managed by the Institute of Health Information and Statistics of the Czech Republic in the period from March 2020 - December 2021. RESULTS: 2,483,219 COVID-19 cases were officially confirmed (732,202 during 2020 and 1,338,790 in 2021), from them 140,368 (6%) represented work-related disease, 520,830 cases (21%) work-related contact, and 1,822,021 (73%) out-of-work contact. There were identified 13 occupations with the highest incidence of COVID-19 in the observed period (458,341 cases), in descending order - clerk, machinist, teacher, craftsman, worker/agency worker, driver, sales worker/cashier, warehouse worker/expediter, nurse, manager, food worker, paramedic, and social worker. Comparing 2020 and 2021, there was a difference in the ranking of occupations by incidence of disease. In 2021, the risk of infection acquiring increased for the occupations clerk, machinist, craftsman, worker/agency worker, manager, and food worker, while it decreased for the health professions (nurse, other paramedic, physician) and for social worker; 5,514 cases of COVID-19 were recognized as an occupational disease in 2020 and 2021, from them 5,483 cases (99.4%) in the health and social care economic activity sector. CONCLUSION: The available data show probable exposures to an infectious agent (without proof of specific contact with the source of the infection), of which 27% cases of COVID-19 are related to work (cases of work-related disease and work-related contact represented together the closest relationship to work). Different relevant anti-epidemic measures in the workplace have considerable practical importance for epidemic control. The use of personal protection of the mouth and nose with respirators/muffs is essential to reduce the risk of airborne transmission.

Musculoskeletal disorders and working risk factors.

OBJECTIVES: The authors evaluate situation in the field of musculoskeletal disorders (MSDs) and working risk factors in the Czech Republic. METHODS: Analysis of selected data from the Czech National Registry of Occupational Diseases during 2008-2018, description of criteria for recognition of occupational MSDs and risk factors associated with MSDs. RESULTS: 6,152 cases of 6 selected recognized occupational musculoskeletal diseases represent 47% of all recognized occupational diseases. The main proportion formed diseases of peripheral nerves in the extremities in terms of compressive neuropathies due to long-term and stereotyped overload, diseases of tendons, tendon capsules or insertions of muscles or joints in the extremities due to long-term and stereotyped overload and diseases of peripheral nerves in the upper extremities in terms of ischaemic and compressive neuropathies due to work with vibrating tools and devices. The authors discussed decreasing trend of the exposure time in three main diagnoses and association of the incidence of MSDs with the risky work related to overload of the musculoskeletal system of the upper extremities. CONCLUSIONS: Important factors for the prevention of MSDs are directly related to the technical provision of production, the organization of work and the regime of work and rest, the length of working hours and the setting of production standards, and generally to the work related stress.

Pressure Pain Sensitivity: Marker for Stress Affecting General Health.

OBJECTIVE: The study examined the links between pressure pain sensitivity (PPS) measured during preventive medical examination of men, their health status and occupation. METHOD: A one year (2015-2016) convenience sampling technique was used to gain primary data from actual medical examination, health records (personal and occupational history) and pressure pain sensitivity (PPS) measured by Ull Meter equipment during preventive medical examination of 371 men of different occupations (average age 43.6±10.4 years, range 19-66 years). Measured PPS values of 60 or more indicate high PPS, contrary PPS values to 40 indicate low PPS. RESULTS: 345 men (93%) were considered healthy (without diagnosis of a disease), 26 men (7%) had positive personal history/symptoms of disease. The average of measured PPS values for the whole group was 36.6±9.5 (first measurement) and 36.7±8.5 (second, repeated measurement), suggesting a high reproducibility of the measurements (r=0.80). Decreased PPS values were measured in men without reported diagnosis of a disease compared to men with diagnosed diseases. Elevated PPS values were measured in men with symptoms of neurocirculatory asthenia (NCA) compared to asymptomatic men and also to men with different diagnosis. The group of men with other than NCA symptoms did not differ significantly in PPS values compared to group of asymptomatic men. Road drivers (177 men) did not differ significantly compared to other occupations (194 men). CONCLUSION: The increased neuropsychological load/stress is connected with increased pain sensitivity to pressure. The PPS method is objective, reliable, simple, and noninvasive evaluation of the impact of stress and may be helpful in assessing medical fitness to work.

Anatomy and classification of the posterior tibial fragment in ankle fractures.

INTRODUCTION: The aim of this study was to analyze the pathoanatomy of the posterior fragment on the basis of a comprehensive CT examination, including 3D reconstructions, in a large patient cohort. MATERIALS AND METHODS: One hundred and forty one consecutive individuals with an ankle fracture or fracture-dislocation of types Weber B or Weber C and evidence of a posterior tibial fragment in standard radiographs were included in the study. The mean patient age was 49 years (range 19-83 years). The exclusion criteria were patients below 18 years of age, inability to provide written consent, fractures of the tibial pilon, posttraumatic arthritis and pre-existing deformities. In all patients, post-injury radiographs were obtained in anteroposterior, mortise and lateral views. All patients underwent CT scanning in transverse, sagittal and frontal planes. 3D CT reconstruction was performed in 91 patients. RESULTS: We were able to classify 137 cases into one of the following four types with constant pathoanatomic features: type 1: extraincisural fragment with an intact fibular notch, type 2: posterolateral fragment extending into the fibular notch, type 3: posteromedial two-part fragment involving the medial malleolus, type 4: large posterolateral triangular fragment. In the 4 cases it was not possible to classify the type of the posterior tibial fragment. These were collectively termed type 5 (irregular, osteoporotic fragments). CONCLUSION: It is impossible to assess the shape and size of the posterior malleolar fragment, involvement of the fibular notch, or the medial malleolus, on the basis of plain radiographs. The system that we propose for classification of fractures of the posterior malleolus is based on CT examination and takes into account the size, shape and location of the fragment, stability of the tibio-talar joint and the integrity of the fibular notch. It may be a useful indication for surgery and defining the most useful approach to these injuries.

The use of new surface-modified poly(2-hydroxyethyl methacrylate) hydrogels in tissue engineering: treatment of the surface with fibronectin subunits versus Ac-CGGASIKVAVS-OH, cysteine, and 2-mercaptoethanol modification.

Superporous poly(2-hydroxyethyl methacrylate) is successfully used as a scaffold material for tissue engineering; however, it lacks functional groups that support cell adhesion. The objective of this study was to investigate the cell-adhesive properties of biomimetic ligands, such as laminin-derived Ac-CGGASIKVAVS-OH (SIKVAV) peptide and fibronectin subunits (Fn), as well as small molecules exemplified by 2-mercaptoethanol (ME) and cysteine (Cys), immobilized on a copolymer of 2-hydroxyethyl methacrylate (HEMA) with 2-aminoethyl methacrylate (AEMA) by a maleimide-thiol coupling reaction. The maleimide group was introduced to the P(HEMA-AEMA) hydrogels by the reaction of their amino groups with N-γ-maleimidobutyryl-oxysuccinimide ester (GMBS). Mesenchymal stem cells (MSCs) were used to investigate the cell adhesive properties of the modified hydrogels. A significantly larger area of cell growth as well as a higher cell density were found on Fn- and SIKVAV-modified hydrogels when compared to the ME- and Cys-modified supports or neat P(HEMA-AEMA). Moreover, Fn-modification strongly stimulated cell proliferation. The ability of MSCs to differentiate into adipocytes and osteoblasts was maintained on both Fn- and SIKVAV-modifications, but it was reduced on ME-modified hydrogels and neat P(HEMA-AEMA). The results show that the immobilization of SIKVAV and Fn-subunits onto superporous P(HEMA-AEMA) hydrogels via a GMBS coupling reaction improves cell adhesive properties. The high proliferative activity observed on Fn-modified hydrogels suggests that the immobilized Fn-subunits maintain their bioactivity and thus represent a promising tool for application in tissue engineering.

The combination of mesenchymal stem cells and a bone scaffold in the treatment of vertebral body defects.

PURPOSE: Vertebral body defects represent one of the most common orthopedic challenges. In order to advance the transfer of stem cell therapies into orthopedic clinical practice, we performed this study to evaluate the safety and efficacy of a composite bioartificial graft based on a hydroxyapatite bone scaffold (CEM-OSTETIC(®)) combined with human mesenchymal stem cells (MSCs) in a rat model of vertebral body defects. METHODS: Under general isoflurane anesthesia, a defect in the body of the L2 vertebra was prepared and left to heal spontaneously (group 1), implanted with scaffold material alone (group 2), or implanted with a scaffold together with 0.5 million MSCs (group 3) or 5 million MSCs (group 4). The rats were killed 8 weeks after surgery. Histological and histomorphometrical evaluation of the implant as well as micro-CT imaging of the vertebrae were performed. RESULTS: We observed a significant effect on the formation of new bone tissue in the defect in group 4 when compared to the other groups and a reduced inflammatory reaction in both groups receiving a scaffold and MSCs. We did not detect any substantial pathological changes or tumor formation after graft implantation. CONCLUSIONS: MSCs in combination with a hydroxyapatite scaffold improved the repair of a model bone defect and might represent a safe and effective alternative in the treatment of vertebral bone defects.

Highly efficient magnetic targeting of mesenchymal stem cells in spinal cord injury.

The transplantation of mesenchymal stem cells (MSC) is currently under study as a therapeutic approach for spinal cord injury, and the number of transplanted cells that reach the lesioned tissue is one of the critical parameters. In this study, intrathecally transplanted cells labeled with superparamagnetic iron oxide nanoparticles were guided by a magnetic field and successfully targeted near the lesion site in the rat spinal cord. Magnetic resonance imaging and histological analysis revealed significant differences in cell numbers and cell distribution near the lesion site under the magnet in comparison to control groups. The cell distribution correlated well with the calculated distribution of magnetic forces exerted on the transplanted cells in the subarachnoid space and lesion site. The kinetics of the cells' accumulation near the lesion site is described within the framework of a mathematical model that reveals those parameters critical for cell targeting and suggests ways to enhance the efficiency of magnetic cell delivery. In particular, we show that the targeting efficiency can be increased by using magnets that produce spatially modulated stray fields. Such magnetic systems with tunable geometric parameters may provide the additional level of control needed to enhance the efficiency of stem cell delivery in spinal cord injury.

The use of dopamine-hyaluronate associate-coated maghemite nanoparticles to label cells.

Sodium hyaluronate (HA) was associated with dopamine (DPA) and introduced as a coating for maghemite (γ-Fe(2)O(3)) nanoparticles obtained by the coprecipitation of iron(II) and iron(III) chlorides and oxidation with sodium hypochlorite. The effects of the DPA anchorage of HA on the γ-Fe(2)O(3) surface on the physicochemical properties of the resulting colloids were investigated. Nanoparticles coated at three different DPA-HA/γ-Fe(2)O(3) and DPA/HA ratios were chosen for experiments with rat bone marrow mesenchymal stem cells and human chondrocytes. The nanoparticles were internalized into rat bone marrow mesenchymal stem cells via endocytosis as confirmed by Prussian Blue staining. The efficiency of mesenchymal stem cell labeling was analyzed. From among the investigated samples, efficient cell labeling was achieved by using DPA-HA-γ-Fe(2)O(3) nanoparticles with DPA-HA/γ-Fe(2)O(3) = 0.45 (weight/ weight) and DPA/HA = 0.038 (weight/weight) ratios. The particles were used as a contrast agent in magnetic resonance imaging for the labeling and visualization of cells.

Flt3 ligand synergizes with granulocyte-colony-stimulating factor in bone marrow mobilization to improve functional outcome after spinal cord injury in the rat.

BACKGROUND AIMS: The effect of granulocyte-colony-stimulating factor (G-CSF) and/or the cytokine fms-like thyrosin kinase 3 (Flt3) ligand on functional outcome and tissue regeneration was studied in a rat model of spinal cord injury (SCI). METHODS: Rats with a balloon-induced compression lesion were injected with G-CSF and/or Flt3 ligand to mobilize bone marrow cells. Behavioral tests (Basso-Beattie-Bresnahan and plantar test), blood counts, morphometric evaluation of the white and gray matter, and histology were performed 5 weeks after SCI. RESULTS: The mobilization of bone marrow cells by G-CSF, Flt3 ligand and their combination improved the motor and sensory performance of rats with SCI, reduced glial scarring, increased axonal sprouting and spared white and gray matter in the lesion. The best results were obtained with a combination of G-CSF and Flt3. G-CSF alone or in combination with Flt3 ligand significantly increased the number of white blood cells, but not red blood cells or hemoglobin content, during and after the time-course of bone marrow stimulation. The combination of factors led to infiltration of the lesion by CD11b(+) cells. CONCLUSIONS: The observed improvement in behavioral and morphologic parameters and tissue regeneration in animals with SCI treated with a combination of both factors could be associated with a prolonged time-course of mobilization of bone marrow cells. The intravenous administration of G-CSF and/or Flt3 ligand represents a safe and effective treatment modality for SCI.

The use of superporous Ac-CGGASIKVAVS-OH-modified PHEMA scaffolds to promote cell adhesion and the differentiation of human fetal neural precursors.

Modifications of poly(2-hydroxyethyl methacrylate) (PHEMA) with laminin-derived Ac-CGGASIKVAVS-OH peptide sequences have been developed to construct scaffolds that promote cell adhesion and neural differentiation. Radical copolymerization of 2-hydroxyethyl methacrylate with 2-aminoethyl methacrylate (AEMA) and ethylene dimethacrylate in the presence of ammonium oxalate crystals resulted in the formation of superporous P(HEMA-AEMA) hydrogels. They were reacted with gamma-thiobutyrolactone to yield 2-(4-sulfanylbutanamido)ethyl methacrylate (P(HEMA-AEMA)-SH) unit. The Ac-CGGASIKVAVS-OH peptide was immobilized to the sulfhydryl groups of P(HEMA-AEMA)-SH by 2,2'-dithiodipyridine linking reagent via 2-[4-(2-pyridyldisulfanyl)butanamido]ethyl methacrylate (P(HEMA-AEMA)-TPy). The adhesion and morphology of rat mesenchymal stem cells were investigated on the Ac-CGGASIKVAVS-OH-modified P(HEMA-AEMA) as well as on PHEMA, P(HEMA-AEMA)-SH and P(HEMA-AEMA)-TPy hydrogels. Superporous Ac-CGGASIKVAVS-OH-modified PHEMA scaffolds significantly increased the number of attached cells and their growth area on the hydrogel surface in the absence and in the presence of serum in the culture medium. Additionally, the Ac-CGGASIKVAVS-OH peptide supported the attachment, proliferation, differentiation and process spreading of human fetal neural stem cells during the first two weeks of expansion and contributed to the formation of a high percentage of more mature neural cells after four weeks of expansion. The Ac-CGGASIKVAVS-OH modification of superporous P(HEMA-AEMA) hydrogels improves cell adhesive properties and promotes neural stem cell differentiation.

Properties and growth of human bone marrow mesenchymal stromal cells cultivated in different media.

BACKGROUND AIMS: Human mesenchymal stromal cells (hMSC) are a promising tool for future clinical application, but their use requires rapid cell expansion in media suitable for clinical use. Therefore, we tested the influence of several culture media on colony formation, population doubling (PD) time, cell cycle and surface marker expression. METHODS: hMSC isolated from human bone marrow (BM) obtained from healthy donors were seeded and expanded in different culture media: alpha-minimum essential medium (MEM) supplemented with 2.5%, 5%, 10% or 20% fetal bovine serum (FBS), 5% or 10% human cord blood serum (hCBS), 5% or 10% human blood serum from AB adult donors (hABS), or mesenchymal stem cell growth medium (MSCGM). The number, diameter and total area of the colonies formed and PD time were determined, and the cell cycle and 16 surface markers were analyzed. RESULTS: Colony-forming efficiency was best in alpha-MEM/hCBS and alpha-MEM/hABS, good in MSCGM and worst in alpha-MEM/FBS. The shortest PD time was achieved in media enriched with human sera or MSCGM, while the time was increased in alpha-MEM/FBS. The largest proliferating fraction was seen in MSCGM followed by media enriched with human sera; the fraction was smallest in alpha-MEM/FBS. Staining for CD34, CD45, CD235a and CD271 was negative, while that for CD29, CD44, CD73, CD90, CD105 and human leukocyte antigen (HLA)-A, -B, -C was positive in all media tested. Media with human serum did not adversely affect the differentiation potential of hMSC, and differentiation into osteoblasts was enhanced. CONCLUSIONS: The choice of serum influences hMSC expansion and cell properties; alpha-MEM supplemented with hABS seems to be a promising candidate for clinical use.