BackgroundCultures of enterocytes and colonocytes represent valuable tools to study growth and differentiation of epithelial cells. In vitro models may be used to evaluate passage or toxicity of drugs, interactions of enteropathogenes bacteria strains with intestinal epithelium and other physiologic or pathologic phenomenon involving the digestive tract.ResultsCultures of bovine colonocytes and jejunocytes were obtained from organoid-enriched preparations, using a combination of enzymatic and mechanical disruption of the intestine epithelium, followed by an isopicnic centrifugation discarding most single cells.Confluent cell monolayers arising from plated organoids exhibited epithelium typical features, such as the pavement-like structure, the presence of apical microvilli and tight junctions. Accordingly, cells expressed several markers of enterocyte brush border (i.e. maltase, alkaline phosphatase and fatty acid binding protein) as well as an epithelial cytoskeleton component (cytokeratin 18). However, enterocyte primocultures were also positive for the vimentin immunostaining (mesenchyme marker). Vimentin expression studies showed that this gene is constitutively expressed in bovine enterocytes. Comparison of the vimentin expression profile with the pattern of brush border enzymes activities, suggested that the decrease of cell differentiation level observed during the enterocyte isolation procedure and early passages of the primoculture could result from a post-transcriptional de-repression of vimentin synthesis. The low differentiation level of bovine enterocytes in vitro could partly be counteracted adding butyrate (1–2 mM) or using a glucose-deprived culture medium.ConclusionThe present study describes several complementary approaches to characterize bovine primary cultures of intestinal cells. Cultured cells kept their morphologic and functional characteristics during several generations.
a b s t r a c tIn the present study we developed an enzymatic approach (through the use of collagenase and dispase) to isolate bovine intestinal epithelial cells. Using this method, freshly isolated jejunocytes could be distinguished from simultaneously isolated colonocytes, as the jejunocytes specifically exhibited the small intestinal peptidase gene transcript, as well as an active alkaline phosphatase. The transformation of both types of cell suspension was performed by retroviral infection, using reproduction-defective viruses bearing the gene coding for the large T antigen of the leukaemia simian virus (SV40). The success of the transfection was demonstrated by (1) a significant increase in cell passage numbers (52-53 vs. 7 passages for nontransfected cells), (2) the detection of both the large T transcript and the large T antigen in transformed cells. Possible contamination and progressive substitution of bovine primocultures by non-bovine lineages available in the laboratory was excluded, as the transformed cells presented a bovine typical karyotype. Most transfected cells kept an epithelial morphology after transformation. They also maintained the expression of FABP and enterocyte specific enzymes (brush-border associated maltase and IAP). However, levels of specific activity of these enzymes were low, suggesting that cell differentiation is not completely achieved under the applied culture conditions.
Background: Long-term sickness absence is a growing concern in Belgium and other European countries. Since 2017, Belgian physicians of the sickness funding organisations are required to assess the re-integration possibilities within the first two months of sickness absence. Given the shortage of physicians in the assessment of work disability and the growing number of people in sickness absence, there is a need for a triage tool, allowing to assign return-to work support to patients having a high-risk profile not to resume work.Methods/design: The current study comprises a comprehensive validation process of a screening tool that supports Belgian physicians in guiding people back to work. The study consists of a theoretical construct validation (face validity and content validity), and an empirical construct validation (concurrence validity, factorial validity, predictive validity, hypothesis testing validity and known- group validity).Expected impact of the study for Public Health: The screening instrument assessing the risk for long-term sickness absence is a tool developed to support physicians who work for sickness funds and for occupational health and safety organisations. Both professionals play an important role in the return to work process and the prevention of long-term sickness absence. The screening tool aims at making a distinction between people who will resume their work independently and people who will need support to do so. Generation of this prediction model will help physicians to focus effort and resources in the high-risk group. Results may also help understand the relationship between the biopsychosocial model and long-term sick-leave.Significance for public healthIn this research, we tested a generic instrument to screen for long-term sickness absence, regardless the cause of the sickness absence or the political context. Both biomedical factors and psychosocial factors (such as the patients’ own prediction) are questioned in the prediction model, which is thus adapted to the modern view on sick leave. A screening method to detect high risk of long-term sickness absence among the large group of sick employees might help to use resources (e.g. money, services) in a more efficient way. Physicians will be able to focus on patients with a high risk on long-term sick leave, and the return to work process of employees at high risk can start much earlier. The instrument will, next to the physicians’ prognosis, offer support in prioritizing patients’ files. Hence, a lot of patients will resume their work spontaneously. Patients who need support will experience shorter follow-up periods, and better quality of care. In addition, the relationship between predicting factors of the biopsychosocial model and long-term sickness absence will be examined.
BackgroundThis study assessed the psychosocial determinants as explanatory variables for the length of the work disability period. The aim was to estimate the predictive value of a selected set of psychosocial determinants from the Quickscan questionnaire for the length of the sick leave period. A comparison was also made with the most common biomedical determinant: diagnosis.MethodsIn a cohort study of 4 981 insured Belgian patients, the length of the sick leave was calculated using Kaplan–Meier. Predictive psychosocial determinants were selected using backward conditional selection in Cox regression and using concordance index values (C-index) we compared the predictive value of the biomedical to the psychosocial model in a sample subset.ResultsFourteen psychosocial determinants were significantly (p<0.10) related to the length of the sick leave: health perception of the patient, physical workload, social support management, social support colleagues, work–health interference, psychological distress, fear of colleagues’ expectations, stressful life-events, autonomy, learning and development opportunities, job satisfaction, workload, work expectations and expectation to return to work. The C-index of this biopsychosocial model including gender, age and labour status was 0.80 (CI: 0.78; 0.81) (n=4 981). In the subset of 2 868 respondents with diagnostic information, the C-index for the same model was .73 (CI: 0.71; 0.76) compared with 0.63 (CI: 0.61; 0.65) for the biomedical model.ConclusionsA set of 14 psychosocial determinants showed good predictive capacity (C-index: 0.80). Also, in a subset of the sample, the selected determinants performed better compared with diagnostic information to predict long-term sick leave (>6 months).
Objective:Increasing long-term sickness absence in many countries asks for specific measures regarding return-to work.Methods:The risk of long-term sickness absence was assessed using a questionnaire containing work-related, function-related, stressful life-events-related, and person-related factors. Additionally, workers’ occupational health physician estimated the worker's chances for work resumption. Reliability, construct, and criterion validity of the questionnaire were measured.Results:Two hundred seventy-six patients and 35 physicians participated in the study. The reliability was satisfying (α > 0.70) for all scales, except for perfectionism (α = 0.62). The results of the CFAs showed that the hypothesized factor models fitted the data well. Criterion validity tests showed that eight predictors significantly related to the estimation of the occupational physicians (ρ < 0.05).Conclusions:The scales of the questionnaire are reliable and valid, and may be implemented to assess sick-listed workers at risk who might benefit from a rehabilitation program.
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