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Surface waters as a potential source of Giardia and Cryptosporidium in Serbia
No full textGiardiasis and cryptosporidiosis are recognized by the WHO as important emerging diseases of the 21st century. Symptoms are similar and include diarrhoea and vomiting, which may be severe, even life-threatening, for the immunocompromised and children under five years of age. Between 2013 and 2017, the Institute for Public Health in Serbia recorded 10 waterborne epidemics that manifested as gastrointestinal disease. Routine testing for enteropathogenic bacteria and viruses did not identify the aetiological agents of these outbreaks. As water is not examined for the presence of protozoa in Serbia, we performed a pilot study to analyse samples from four major rivers and their tributaries using a newly implemented methodology for detection of Giardia and Cryptosporidium, based on the ISO 15553:2006 standard. Using immunofluorescence microscopy, Giardia was detected in 10 out of the 31 samples, Cryptosporidium in five, while two samples were positive for both. Presence of G. duodenalis gDNA was confirmed by amplification of the beta-giardin gene in eight samples, of which one and two, respectively, were identified by RFLP as potentially zoonotic assemblages A and B. The results suggest that surface water in Serbia may be a potential source of infection and call for more in-depth studies using sophisticated molecular tools
Ambulatory assessment of language use: Evidence on the temporal stability of Electronically Activated Recorder and stream of consciousness data
Get PDFThe ambulatory assessment offers a wide range of methods enabling researchers to investigate psychological, behavioral, emotional, and biological processes. These methods enable us to gather data on individual differences in language use for psychological research. Two studies were conducted with an aim to evaluate and compare the temporal stability of language measures extracted by LIWC software form data obtained by two frequently used methods for assessment of language use, i.e., Electronically Activated Recorder (EAR) and stream of consciousness (SOC) task. Additionally, we examined the amount of variance in language use (assessed by both methods) that can be attributed to intra-individual variability and stable individual differences. Study 1 was focused on investigating language use obtained from 74 respondents using the EAR for 3 consecutive days. Study 2 was conducted on 250 respondents participating in a SOC task where verbal production was collected at ten time points over a 2-month period. Results show that measures obtained using the SOC task have higher temporal stability and consistency, and to a certain extent enable better detection of individual differences. Taking into account certain situational variations improves the reliability of EAR measures
Effect of Hyperbaric Oxygen Therapy on Fatty Acid Composition and Insulin-like Growth Factor Binding Protein 1 in Adult Type 1 Diabetes Mellitus Patients: A Pilot Study
Get PDFObjective: Metabolic changes in type 1 diabetes mellitus (TIDM) impair vasodilation, and this leads to tissue hypoxia and microvascular pathology. Hyperbaric oxygen therapy (HBOT) can significantly improve the outcome of ischemic conditions in T1DM patients and reduce vascular complications. The aim of our study was to assess the effects of HBOT on plasma fatty acid (FA) composition, and expression of insulin-like growth factor binding protein 1 (IGFBP-1) in T1DM patients. Methods: Our study included 24 adult T1DM patients diagnosed with peripheral vascular complications. The patients were exposed to 10 sessions of 100% oxygen inhalation at 2.4 atmosphere absolute for 1 hour. Blood samples were collected at admission and after HBOT for measurement of metabolic parameters, FA composition and IGFBP-1. Measurement of plasma FA composition was determined by gas chromatography. Expression of IGFBP-1 in the serum was estimated by Western blot analysis. Results: HBOT decreased blood levels of total cholesterol (p lt 0.05), triglycerides (p lt 0.05) and low-density lipoprotein (p lt 0.05). HBOT increased plasma levels of individual FAs: palmitic acid (p lt 0.05), palmitoleic acid (p lt 0.05), docosapentaenoic acid (p lt 0.05) and docosahexaenoic acid (p lt 0.01), and decreased levels of stearic acid (p lt 0.05), alpha linolenic acid (p lt 0.05) and linoleic acid (p lt 0.01). Expression of IGFBP-1 (p lt 0.01) was increased, whereas the level of insulin (p lt 0.001) was decreased in the serum after HBOT. Conclusions: Our results indicate that HBOT exerts beneficial effects in TIDM patients by improving the lipid profile and altering FA composition
Comparison of growth and morphology of Borrelia burgdorferi sensu lato in BSK-H and BSK-II media stored for prolonged periods
No full textBarbour-Stoenner-Kelly II (BSK-II) and BSK-H media were used for cultivation and isolation of fastidiousBorreliaspecies - the causative agents of Lyme borreliosis. Culture media have a limited shelf life and require adequate storage. Our goal was to assess how the growth ofBorreliawould be affected by prolonged storage of media and inadequate storage conditions (BSK-H stored at +4 degrees C for 2.5 years and BSK-II stored at -20 degrees C for 11 years). Growth of differentBorrelia afzelii,Borrelia garinii,Borrelia lusitaniaeandBorrelia valaisianastrains was assessed during 2 weeks of incubation at 33 degrees C. Monitored parameters included cell count per mL, morphology and motility. The results of this study have shown weaker growth of borrelia strains in BSK-H at +4 degrees C (median final cell number of 1.5 x 10(6)/mL) than in BSK-II at -20 degrees C (median final cell number of 7.75 x 10(6)/mL) and in fresh BSK-H media (median final cell number of 8.95 x 10(6)/mL). Duration of storage of media had no impact onBorreliamorphology and motility. Our results indicate that temperature of -20 degrees C is optimal for long-term storage of medium, BSK-II stored for 11 years provided effective support to growth ofBorreliaand may be employed for cultivation
Specificity of 3D MSC Spheroids Microenvironment: Impact on MSC Behavior and Properties
No full textMesenchymal stem cells (MSC) have been considered the promising candidates for the regenerative and personalized medicine due to their self-renewal potential, multilineage differentiation and immunomodulatory capacity. Although these properties have encouraged profound MSC studies in recent years, the majority of research has been based on standard 2D culture utilization. The opportunity to resemble in vivo characteristics of cells native niche has been provided by implementation of 3D culturing models such as MSC spheroid formation assesed through cells self-assembling. In this review, we address the current literature on physical and biochemical features of 3D MSC spheroid microenvironment and their impact on MSC properties and behaviors. Starting with the reduction in the cells' dimensions and volume due to the changes in adhesion molecules expression and cytoskeletal proteins rearrangement resembling native conditions, through the microenvironment shifts in oxygen, nutrients and metabolites gradients and demands, we focus on distinctive and beneficial features of MSC in spheroids compared to cells cultured in 2D conditions. By summarizing the data for 3D MSC spheroids regarding cell survival, pluripotency, differentiation, immunomodulatory activities and potential to affect tumor cells growth we highlighted advantages and perspectives of MSC spheroids use in regenerative medicine. Further detailed analyses are needed to deepen our understanding of mechanisms responsible for modified MSC behavior in spheroids and to set future directions for MSC clinical application
Effect of extracellular hemoglobin of xenogeneic origin on functional characteristics of mesenchymal cells in vitro
Get PDFИстраживања обухваћена овом докторском дисертацијом имала су за циљ добијање података о утицају ванћелијског хемоглобина на функцијска својства мезенхимских ћелија in vitro. Свињски и говеђи хемоглобин коришћени су као модел ксеногених, лако доступних хемоглобина, који показују висок степен хомологије са хуманим хемоглобином. Као адекватни ћелијски модел системи одабрани су леукоцити периферне крви човека и мезенхимске матичне ћелије периферне крви човека (ПК-ММЋ), као ћелије са којима ванћелијски хемоглобин може доћи у контакт при инстраваскуларној хемолизи. Додатно, утицај хемоглобина испитан је и на четири ћелијске линије ATDC5, MC3T3-E1, 3T3-L1 и C2C12 које представљају униформније модел системе у односу на ПК-ММЋ за проучавање хондро-, остео-, адипо- и миогенезе. Добијени резултати показују да ванћелијски хемоглобин има значајан утицај на функцијска својства мезенхимских ћелија, који зависи од врсте организма из ког је хемоглобин изолован, примењене концентрације и типа ћелија на којима је ефекат испитиван. Ванћелијски хемоглобин модулише вијабилност, способност адхезије и продукције реактивних врста кисеоника испитиваних леукоцита, као и пролиферацију/диференцијацију опредељених матичних ћелија хематопоезе. Додатно, показано је да ванћелијски хемоглобин у концентрацији од 0,1 μM остварује ефекат на вијабилност, миграторни и имуномодулаторни капацитет мезенхимских ћелија и инхибира њихову диференцијацију ка хондрогеној, остеогеној, адипогеној и миогеној линији, модулишући експресију специфичних генских маркера. Ови резултати доприносе бољем сагледавању утицаја ванћелијског хемоглобина у условима интраваскуларне хемолизе на физиологију мезенхимских ћелија, као и његове потенцијалне примене као дијететског суплемента или адитива медијума за култивацију ћелија.The main goal of this doctoral dissertation was the investigation of the effect of extracellular hemoglobin on the functional properties of mesenchymal cells in vitro. Porcine and bovine hemoglobin were used as models of xenogeneic, readily available hemoglobins, showing a high degree of homology with human hemoglobin. Human peripheral blood leukocytes and human peripheral blood mesenchymal stem cells (PB-MSCs) were selected as adequate cell model systems, since extracellular hemoglobin may come into contact with these cells during instravascular hemolysis. In addition, the effects of hemoglobin were also tested on four cell lines ATDC5, MC3T3-E1, 3T3-L1, and C2C12 as more uniform model systems compared to PB-MSCs to study chondro-, osteo-, adipo-, and myogenesis.The obtained results show that extracellular hemoglobin has major effect on the functional properties of mesenchymal cells,which depends on the species of organism from which hemoglobin was isolated, hemoglobin concentration used and the type of cells tested. Extracellular hemoglobin modulates viability, adhesion and synthesis ability of reactive oxygen species of leukocytes examined, as well as proliferation/differentiation of committed hematopoietic stem cells. Furthermore, at a concentration of 0.1 μM extracellular hemoglobin had the effect on viability, migratory and immunomodulatory capacity of mesenchymal cells and inhibited their differentiation toward chondro-, osteo-, adipo-and myogenic lineages, modulating the expression of specific gene markers.These results contribute to a better understanding of the effects of extracellular hemoglobin during intravascular hemolysis on the physiology of mesenchymal cells, as well as its potential applications as a dietary supplement or cell culture media additive
Prospective analysis of gait characteristics in chronic inflammatory demyelinating polyradiculoneuropathy
No full textChronic inflammatory demyelinating polyradiculoneuropathy (CIDP) is a disorder that may lead to functional impairment, including gait abnormalities. Our aim was to analyze gait characteristics in patients with CIDP compared to healthy controls (HC). Moreover, we sought to determine changes of gait parameters after six-month follow-up period. Twenty-four patients with CIDP and 24 HCs performed basic walking task, dual-motor task, dual-mental task, and combined task using the same GAITRite system. Lower limb MRC-SS and lower limb INCAT disability score were assessed. Fourteen patients were retested after six months. Majority of gait parameters showed significant differences in all experimental conditions when compared between CIDP and HCs. The most consistent findings in CIDP were shorter stride length (SL), prolonged cycle time (CT) and double support time (DS), as well as increased variation of SL and of swing time (ST) (p lt 0.05). During follow-up, INCAT improved in nine (64.3%) of 14 patients and MRC-SS improved in eight (57.1%) patients. Six-month changes of CT and its variation during combined task significantly differentiated patients with improved vs. non-improved INCAT (p lt 0.05). In conclusion, patients with CIDP had slower gait with prolonged DS and with shorter SL compared to HCs. Increased variation of SL and of ST in CIDP may suggest a potential risk for instability and falls. Shorter CT duration and less CT variation during time correlated well with improvement in disability
Transstadial Transmission and Replication Kinetics of West Nile Virus Lineage 1 in Laboratory Reared Ixodes ricinus Ticks
Get PDFWest Nile virus (WNV) is a mosquito-borne agent that has also been isolated from several tick species. Vector competence of Ixodes ricinus, one of the most common tick species in Europe, has been poorly investigated for WNV to date. As such, to evaluate the vector competence, laboratory reared Ixodes ricinus nymphs were in vitro fed with WNV lineage 1 infectious blood, allowed to molt, and the resulting females artificially fed to study the virus transmission. Furthermore, we studied the kinetics of WNV replication in ticks after infecting nymphs using an automatic injector. Active replication of WNV was detected in injected nymphs from day 7 post-infection until 28 dpi. In the nymphs infected by artificial feeding, the transstadial transmission of WNV was confirmed molecularly in 46.7% of males, while virus transmission during in vitro feeding of I. ricinus females originating from infected nymphs was not registered. The long persistence of WNV in I. ricinus ticks did not correlate with the transmission of the virus and it is unlikely that I. ricinus represents a competent vector. However, there is a potential reservoir role that this tick species can play, with hosts potentially acquiring the viral agent after ingesting the infected ticks
Extracellular xenogeneic hemoglobin suppresses the capacity for C2C12 myoblast myogenic differentiation
Get PDFFunctional characteristics of satellite cells (SCs) that act as myogenesis initiators and have emerged as a promising target for cell therapy, are dependent on their microenvironment. The aim of this study was to investigate the effect of cell-free hemoglobin, as a part of the microenvironment of SCs, on their functional characteristics. The C2C12 cell line served as the experimental model of SCs; hemoglobin isolated from porcine (PHb) and bovine (BHb) slaughterhouse blood served as the experimental model for extracellular hemoglobin. The proliferation rate of C2C12 cells was assessed by the MTT test, migration capacity by the scratch assay, and myogenic differentiation capacity by histochemical staining and RT-PCR analysis of the expression of genes specific for myogenic lineage. The effect of hemoglobin on the proliferation and migration of C2C12 cells was dependent on its concentration and the animal species it was isolated from, but the effect of BHb was more prominent. Both PHb and BHb decreased the expression levels of myogenin and muscle specific creatine kinase at a 10 mu M concentration. While PHb had no effect on the morphometric parameters of C2C12 myotubes, BHb modified the area and length of C2C12 myotubes cultivated in DMEM/2% horse serum and DMEM/10% fetal calf serum. While PHb and BHb had no effect on heme oxygenase 1 (Hmox1) expression, they stimulated the expression of hypoxia-inducible factor 1-alpha (Hif1 alpha) at a concentration of 10 mu M. The mainly inhibitory effect of cell-free hemoglobin on myogenic differentiation suggests that it could be a relevant factor in the outcome of cell therapy of muscle injury
Community Pharmacists' Attitudes and Professional Practice in Relation to the Patient Safety Incidents
Get PDFBackground: Medicines dispensing is an error-prone activity, therefore potentially jeopardizing patient safety. This study aimed to assess the community pharmacists' attitudes towards the causes of dispensing errors and preventive measures, as well as their practice in incidents reporting. Materials and Methods: A cross-sectional survey was performed by distributing an adopted and validated questionnaire to a nationwide sample of community pharmacists in Serbia. The questionnaire included sections related to the participants' socio-demographic characteristics, their attitudes towards factors causing dispensing errors and corrective actions, as well as their practice in reporting. Statistical analyses were conducted using SPSS Statistics software ver. 21.0. The associations between categorical variables were analyzed using Chi-square test. Results: The study included 1,004 participants, mainly female (94.9%), with the mean age 40.9 +/- 9.9 years and mean work experience 14.3 +/- 10.0 years. More than a third of the participants (35.4%) indicated an increasing risk of dispensing errors. The main causes included illegible prescriber's handwriting (44.3%) and interruptions during dispensing (39.2%), while the major corrective actions were providing pharmacists with education in clinical pharmacy (71%) and reducing the interruptions during dispensing (63.9%). The majority of respondents (85.2%) stated that they routinely reported dispensing incidents. However, even 16.5% of them admitted to having fear sometimes or always. Additionally, only 58.1% of participants would use voluntary dispensing error reporting system. Conclusion: Serbian community pharmacists are aware of the existing risk in medicines dispensing and the corrective actions identified should be put into practice so as to manage them prospectively. Although the results indicate good practice in incidents reporting, conducting tailored educations and building of safety culture is necessary to improve patient safety