Microfluidic high-throughput RT-qPCR measurements of the immune response of primary bovine mammary epithelial cells cultured from milk to mastitis pathogens.

Bovine mastitis, the inflammation of the udder, is a major problem for the dairy industry and for the welfare of the animals. To better understand this disease, and to implement two special techniques for studying mammary gland immunity in vitro, we measured the innate immune response of primary bovine mammary epithelial cells (pbMEC) from six Brown Swiss cows after stimulation with the heat-inactivated mastitis pathogens, Escherichia coli 1303 and Staphylococcus aureus 1027. The cells were extracted and cultivated from milk instead of udder tissue, which is usually done. The advantages of this technique are non-invasiveness and less contamination by fibroblasts. For the first time, pbMEC gene expression (GE) was measured with a microfluidic high-throughput real-time reverse transcription-quantitative PCR platform, the BioMark HD™ system from Fluidigm. In addition to the physiological analysis, the precision and suitability of this method was evaluated in a large data set. The mean coefficient of variance (± s.e.) between repeated chips was 4.3 ± 0.4% for highly expressed and 3.3 ± 0.4% for lowly expressed genes. Quantitative PCR (qPCR) replicate deviations were smaller than the cell culture replicate deviations, indicating that biological and cell culture differences could be distinguished from the background noise. Twenty-two genes (complement system, chemokines, inflammatory cytokines, antimicrobial peptides, acute phase response and toll-like receptor signalling) were differentially expressed (P < 0.05) with E. coli. The most upregulated gene was the acute phase protein serum amyloid A3 with 618-time fold. S. aureus slightly induced CCL5, IL10, TLR4 and S100A12 expression and failed to elicit a distinct overall innate immune response. We showed that, with this milk-derived pbMEC culture and the high-throughput qPCR technique, it is possible to obtain similar results in pbMEC expression as with conventional PCR and with satisfactory precision so that it can be applied in future GE studies in pbMEC.

Integration of the e-Learning into the medical university curricula.

OBJECTIVES: The aim of this contribution was to present the e-Learning introduction in the Slovak Medical University (SMU) with a focus on the implementation phase of the two blended courses - Healthcare Quality and Healthcare Professionals' Ethics. BACKGROUND: The introduction of the e-Learning was realized during the period 2008-2009 in the partnership of SMU and IBM Company, following strictly the project management approach. METHODS: The development of the e-module beta-versions was evaluated by the modules' authors using a structured interview. In a consequent pilot testing, the blended courses were evaluated by 23 students of the bachelor program in Rescue health care, and by 61 public health students at the master level program, respectively, using the standardized questionnaires. RESULTS: The tangible results included the documented SMU strategy for the e-Learning integration, six e-Learning modules and evaluation results. The authors' evaluation showed high scores for the experience in collaboration with IBM, as well as for the experience with the LMS environment. The students' evaluation showed a high acceptance of the e-Learning by both part-time and full-time students. The access to Internet was not recognized as a serious barrier. CONCLUSION: The first experience with the integration of the e-Learning into the curricula of the Slovak Medical University showed the advantage of the systematic approach. The experience with developing the strategy in an interdisciplinary/ intercultural team, the knowledge about specific characteristics of distance learning by the involved SMU staff, and the know-how and skills represented the important benefits. It was demonstrated that the blended learning is recommended as optimal for the education in medical environment (Tab. 4, Fig. 1, Ref. 22).

Stakeholder approach in strengthening the patients' rights in the Slovak Republic.

OBJECTIVES: The described project aimed to promote patients' rights in the Slovak Republic that are stipulated by law but in practice not observed fully. BACKGROUND: The project was based on a bi-lateral agreement between The Netherlands and the Slovak Republic in the framework of European Union pre-accession program and implemented in the period from January 2002 to June 2003. METHODS: Successful Dutch models of patients' rights promotion were used. They were applied under Slovak conditions and focused on the areas in the greatest need of attention, such as information campaigning and public awareness of patients' rights issues. The initiation of a cooperation of various stakeholders active in healthcare and national patients' right forums was among the most positive aspects of the project. RESULTS: The information campaign was constructed in order to educate citizens about their entitlements and rights within healthcare. Healthcare professionals were trained on respecting the rights of their clients. Pilots were started in several hospitals and in Healthy City projects, and patients' rights modules were made available at educational centers and various private and health organizations. Some of the cooperating healthcare professionals expressed their fear that the public might misuse the gained advocacy skills. This idea has originated from the current situation in Slovak healthcare, where inadequate financing, education, and management is a barrier in the physician's ability to serve the patient's best interest. CONCLUSIONS: The implemented project started wide public discussion on patients' rights and stimulated a cooperation of a large number of stakeholders in the promotion of patients' rights (Tab. 2, Ref. 13). Full Text (Free, PDF) www.bmj.sk.

[Basic assessment of needs for training in evidence-based medicine in Slovakia]. / Zakladny odhad potrieb vzdelavania v oblasti mediciny zalozenej na dokazoch na Slovensku.

BACKGROUND: The health care reform in Slovakia produces a desire for greater responsibility for and control of strategic decisions and to be better able to evaluate international knowledge and experience in the specific national social and professional contexts. Evidence based medicine (EBM) provides an increasingly organised and accessible database of international knowledge in health and health care, capable of informing decisions at the macro and micro levels. AIM: The aim of this pilot study was to assess education, training and other capacity building needs in EBM and evidence based health care. METHODS: This study was primarily qualitative and based on a triangular approach, which included: (1) The analysis of the situation in pre- and postgraduate education in Slovakia aiming to the estimation of needs in EBM and critical appraisal skills training; (2) The analysis of questionnaires distributed in a sample of 50 medical doctors and university educated public health workers undergoing a postgraduate training; (3) The discussion in focused groups. RESULTS: The findings and analysis uncovered a gap in knowledge and experience of EBM approaches, particularly of searching for evidence, utilising information technology, of undertaking critical appraisals of the validity and quality of external evidence and of knowledge of English. On the other hand the findings revealed a high access to information including the Internet access at the workplace, an increasing awareness of the need for up-date information, a demand for training and potential opportunities for action. CONCLUSIONS AND RECOMMENDATIONS: The effective implementation introduction of EBM approach would require changes in broader political, cultural and behavioural contexts, including changes in pre- and postgraduate systems of professional and managerial education, changes in professional and managerial attitudes and changes in emphasis in skills and capacity building and improvements in knowledge management systems at the national level.

Contribution to systematic education of quality management in Slovak health care.

THE AIM: Of the study was to contribute to quality improvement initiatives in Slovak health services through systematic approach to the education and training in quality management (QM). Consequently, the main objectives were to analyse the content of the education in QM abroad, to conduct an audit of perceived training needs in Slovakia, and to propose the design of QM training programme to be applied within CME scheme based on the study results. Triangular method in the design of the study was implemented. Review of relevant information, data from the questionnaire and semi-structured interview in the sample of 67 Slovak trainees from Health Management School and School of Public Health--were adopted in complementary fashion. BETWEEN FINDINGS: Highlighted in the survey are positive attitudes to training in quality management documented by the median score higher than 6 in all tested areas, on scale 0-10. No significant differences in profession groups as physicians, nurses, HC managers or among training institutions involved were displayed. However, potential obstacles were identified in deeper study using interviews. The absence of knowledge and skills in management in general and in quality management approaches especially are observed. Typically, the role of strategic planning is undermined. The large scale of quality management approaches is converted to problems of accreditation. Barriers to participative culture, innovation, devolution of accountability, resistance to change and to team based management are authentic findings as well. CONCLUSIONS: Drawn from the study were related to: fostering managers--"transformational leaders" for locally driven decision making in health care policy and practice; need of training activities for the continuing education in quality with respect to specific target groups interests and their level of knowledge in management; content of training oriented towards combination of rational utilization of information, critical analytical skills and planning for quality with human resource development-interpersonal skills, team building (soft skills), not just reduction of quality management tools to hard techniques (statistics, ISO norms); methods of education, where the usage of experiential learning methods, participative training inclusive action learning is highlighted; team training complemented with individual professional development support inclusive a coaching and mentoring scheme. AS IMPLICATIONS: Four types of CME training: Basic Module QM, Training for QM teams, Training Trainers Scheme and Guiding through Accreditation and Quality Award were proposed. (Tab. 9, Ref. 38.)

[Performance of Slovak hospitals as related to Porter's generic strategies]. / Výkon nemocníc na Slovensku vo vzt'ahu k uplatnovaným generickým stratégiám podla Portera.

Porter's generic strategies characterize organizations in terms of their competitiveness, and are related to the performance of the organization. The aim of this study was to analyze the Porter's generic strategies and their effect on performance in the context of the Slovak hospital industry. Acute care hospitals with more than 30 beds were included into the study. National institutes providing specialized service were excluded from the study. Strategy and performance were evaluated on the basis of self-reported questionnaires, completed by chief administrators of hospitals (total 76 completed questionnaires were obtained, out of 81 distributed, i.e. 94% response rate). The cluster analysis was used for the identification of strategic orientation. Performance differences across strategic groups were tested using multivariate analysis of covariance (MANCOVA). The hierarchical cluster analysis uncovered a four-group taxonomy of hospitals: the group "Focused Cost Leadership" included 33% of hospitals, the group "Stuck-in-the middle" 49%, the group "Wait and See" 13% and the group "Cost leadership" 5%. Significant differences in performance were related to the Porter's pure, or hybrid strategies, respectively. In terms of industry evolution, the Slovak hospital industry could be characterized as fragmented, having a large number of small and medium size mainly state owned hospitals, with absence of market leaders, and with high exit barriers (mainly social and political) that hold back consolidation. (Tab. 1, Ref. 35.).

Performance implications of Porter's generic strategies in Slovak hospitals.

The aim of the study was to examine the use of Porter's generic strategies and their effect on performance in the context of the Slovak hospital industry. Using mail survey the study first identified the natural taxonomy of four strategic types of Slovak hospitals, based on their use of Porter's generic strategies in pure form and in combination. Next the study examined whether different strategic types were associated with different levels of organisational performance, while controlling for such variables as size and location, which have been argued to influence the hospital performance. The findings indicate that hospitals which follow a "stuck-in-the-middle" strategy, in general, have superior performance on all used performance measures, while hospitals that place only low emphasis on cost leadership, differentiation and focus, labelled "wait and see" in this study, perform the poorest. The study concludes that the research provided body of knowledge relevant for the Slovak hospital industry, that may be used by hospital managers in the strategy formulation process as well as by the researches in exploring the influence of different contingencies on hospitals' strategic orientation.

Basic estimate of needs for training in evidence-based medicine in Slovakia.

This study reports the findings of the first survey of the knowledge and needs for training in evidence based medicine (EBM) of health-care workers in Slovakia. This study was primarily qualitative and based on a triangular approach, which included: analysis of the situation in pre- and postgraduate education in Slovakia aimed at estimating needs in EBM and critical appraisal skills (CAS) training; analysis of questionnaires distributed in a sample of medical doctors and university educated public health workers undergoing postgraduate training; and focus group discussions. The findings revealed a real gap in knowledge in EBM and CAS in Slovakia and identified several areas as the focus for intervention. The results showed also some important behavioural and cultural aspects, including low individual responsibility for education; tendency to delegate responsibility to authorities (experts, top management, Ministry of Health); and persistence of the state paternalistic type of education. Concludes that managers planning to implement EBM in Slovakia should therefore consider a broader behavioural and cultural context for change, not just introduction of a training EBM module.

Predictors of mortality in bacteremic cancer patients: retrospective analysis of 64 deaths occurring among 262 bacteremic episodes.

A total of 262 bacteremic episodes were observed in cancer patients in a single cancer institution during the last 7 years, and the recorded outcome was death in 65. The 65 patients who died (24.8% overall mortality) were divided retrospectively into two subgroups: (a) those who died of underlying disease with bacteremia (45 cases, 16.9% crude mortality) and (b) those who died of bacteremia (20 patients, 7.7% attributable mortality). Comparison of several risk factors in subgroups of patients who achieved a cure (197 cases) and of those who died and whose deaths were attributable (20 cases) revealed six risk factors that were associated with attributable mortality: (1) chemotherapy-induced neutropenia (P < 0.03), (2) Acinetobacter/Stenotrophomonas spp. bacteremias (P < 0.001), (3) liver failure (P < 0.001), (4) inappropriate therapy (P < 0.0001), (5) organ complications (P < 0.003) and (6) multiresistant organisms (P < 0.001). Enterococci and Pseudomonas aeruginosa, surprisingly, were found more frequently in those who died of an underlying disease with bacteremia than among patients who were cured (17.6% vs 7.6%, P < 0.05 and 29.1% vs 13.8%, P < 0.02). Those who died of infection had higher numbers of positive blood cultures, with 2.05 per episode, than did those who died of underlying disease with bacteremia (1.82) or those who were cured (1.51). Other risk factors, such as underlying disease, type of chemotherapy, origin of bacteremia, age, and catheters did not predict either overall or attributable mortality within the study group.

A BRIDGE partnership model for health management education in the Slovak Republic.

An innovative Health Management Education Partnership (HMEP) was initiated to develop management education initiatives through the exchange of information and ideas. Health education efforts, projects and activities exist between the University of Scranton and three strategic partners in the Slovak Republic: Trnava University, the Health Management School and the University of Matej Bel. The BRIDGE model (Building Relationships in Developing and Growing Economies) utilizes several innovative educational initiatives and strategic projects including a professional journal, faculty development, professional development, curriculum development, certification and accreditation, faculty-students exchange and development of educational materials and modules. The BRIDGE organizational structure is reviewed as well as specific workplan objectives to operationalize the HMEP encouraging mutual cooperation, collaboration and sustainability of efforts. The model stresses implementation, monitoring, and evaluation of all initiatives through a strong community effort, focus on research, deployment of educational resources, curriculum modification, development of interpartnership activities, conferences, workshops, fieldwork experiences and study tours. Applied management practices enhance market-oriented solutions to health care delivery problems emphasizing a focus on privatization and entrepreneurship through education.

Qualitative evaluation of a health management education partnership.

Results of ongoing cooperation of four educational institutions in the field of Health Management Education in Slovakia are reviewed. The specific benefit of multilateral collaboration for the situation in central Europe is discussed in the context of a Health Management Education Partnership (HMEP). The project is based on a grant from the United States Agency for International Development (USAID) with assistance from the American International Health Alliance (AIHA). The U.S. partner is the University of Scranton, Scranton, Pennsylvania. Three organizations are involved in the partnership from the Slovak Republic: Trnava University, University of Matej Bel in Banska Bystrica and the Health Management School (HMS) in Bratislava, each having specific priority and focus in health management education. The HMEP program is designed to reach about 30 teachers of health administration as well as 200 undergraduates, senior and middle managers from health service organizations every year. The collaborative endeavor stimulates the communication and creation of personal and human relations, not only on an individual and institutional level, but on a community as well as national and international level. The project supports the development of knowledge and skills, professional identity and educational processes in health management. The emphasis is on increasing the quality of education and training in Slovakia through multilateral collaboration, curricula development, professional development, symposia, support of consultancies and accreditation. Specific areas of content are designed to address health policy, law, ethics, insurance and quality improvement.

Accreditation processes in the Slovak Republic.

This article provides a conceptual framework for understanding changes occurring in the accreditation process for institutions of higher learning in the Slovak Republic. Health sector transformation is occurring rapidly throughout the Central and Eastern European region as former socialist states are experimenting with market-oriented approaches. Educators and universities are playing an important role in the development of knowledge, skills and abilities of current and future health practitioners. Accreditation is viewed as an important process in the continuing development of quality health service management curricula and preparing competent practitioners for the health care field. Historical considerations are examined as well as future directions for accreditation in the Slovak Republic.

Analysis of 553 episodes of monomicrobial bacteraemia in cancer patients: any association between risk factors and outcome to particular pathogen?

Relationships between aetiology, various risk factors (such as neutropenia, catheter insertion, endoscopy, therapy with corticosteroids, therapeutic use of antimicrobials, antibiotic prophylaxis, source of infection), symptomatology and outcome were studied in 553 monomicrobial bacteraemic episodes in cancer patients observed within 7 years at the National Cancer Institute of the Slovak Republic. The ratio of gram-positive to gram-negative bacteraemia was 1:1 (43.5% vs 43.8%), and yeasts caused 7.2% of monomicrobial episodes. The highest mortality was associated with Pseudomonas aeruginosa (19.2%), non-albicans Candida yeasts (25%) and Bacteroides fragilis (22.6%). Independent risk factors for particular pathogens were investigated by a computerized logistic regression model. The only independent risk factor for staphylococcal and enterococcal bacteraemia was vascular catheter insertion (OR = 1.95 and 2.05, CI = 95%, P = 0.035 and 0.044, respectively). However, there were no independent specific risk significant factors for viridans streptococcal bacteraemia and bacteraemia due to Enterobacteriaceae or Ps. aeruginosa. Neutropenia was found to be an independent predictor for development of Acinetobacter spp. bacteraemia (OR = 3.84, CI = 95%, P = 0.044). Prior therapy with third-generation cephalosporines was a predictive, independent risk factor for the development of fungaemia (OR = 1.99, CI = 95%, P = 0.028) but not of enterococcal bacteraemia. We also did not observe any association between prior therapy with imipenem and Stenotrophomonas maltophilia bacteraemias. Multivariate analysis confirmed that fungaemia may be independently associated with higher mortality than bacteraemia caused by Enterobacteriaceae and staphylococci. However, the mortality of fungaemia was statistically no different from that of Ps. aeruginosa, Stenotrophomonas spp. and viridans streptococci bacteraemias.

Viridans streptococcal bacteraemia due to penicillin-resistant and penicillin-sensitive streptococci: analysis of risk factors and outcome in 60 patients from a single cancer centre before and after penicillin is used for prophylaxis.

60 patients with 60 viridans streptococcal bacteraemic episodes (42 due to penicillin-sensitive and 18 due to penicillin-resistant viridans streptococci) were analysed in a population of 12,185 admissions and 1,380 bacteraemic episodes during a 7-year period in a National Cancer Institute. The incidence of viridans streptococci among bacteraemias decreased from 11.5% in 1989 to 2.5% in 1995 after penicillin was introduced for prophylaxis of febrile neutropenia in acute leukaemia in 1993. However, the proportion of penicillin-resistant viridans streptococcal bacteraemias increased from 0 in 1989 and 1990 before any prophylaxis was given, to 12.9-16.7% after quinolones were used for prophylaxis in 1991 and 1992, and to 44.4-81.8% in 1993-1995 after penicillin was added to the quinolones. Mortality rate was higher in the subgroup of penicillin-resistant viridans streptococcal bacteraemias (p < 0.05). Statistically significant risk factors in patients with penicillin-resistant (compared with penicillin-sensitive) viridans streptococcal bacteraemia were: acute leukaemia (p < 0.03), high doses of cytarabine (p < 0.05), mucocutaneous lesions (p < 0.004), breakthrough bacteraemia during prophylaxis with ofloxacine plus penicillin (p < 0.001). Multiple logistic regression analysis showed that only acute leukaemia (OR 2.05, CI 0.85-1.85, p < 0.00452) and penicillin-resistance (OR 0.71, CI 0.103-4.887, p < 0.0209) were significant independent predictors of inferior outcome. Breakthrough bacteraemia during empiric therapy with vancomycine occurred in 5 of 116 patients treated with vancomycine, and during therapy with ampicillin plus gentamicin in 6 patients of 18 treated.

Bacteremia and fungemia occurring during antimicrobial prophylaxis with ofloxacin in cancer patients: risk factors, etiology and outcome.

The authors analyzed 27 breakthrough bacteremias occurring during ofloxacin prophylaxis in afebrile neutropenia over 7 years in 9989 admissions and 979 bacteremic and fungemic episodes in a National Cancer Center in Bratislava, Slovak Republic. The most frequently isolated organisms in breakthrough bacteremias were gram-positive (71.3%), mainly coagulase-negative staphylococci (41.3%), enterococci (9.2%) and Corynebacteria (9.2%), followed by gram-negative rods-Pseudomonas aeruginosa (13.2%) and Stenotrophomonas maltophilia (9.2%). The outcome of breakthrough bacteremias during ofloxacin prophylaxis was not associated with the underlying disease, neutropenia, catheter insertion or resistance, but only with multiple risk factors. A higher failure rate was observed in those patients having a catheter infected with a resistant organism and during neutropenia. No patients with Hickman catheter were included in the study. Patients with mixed breakthrough bacteremia due to gram-negative and gram-positive organisms had higher failure rates than those with monomicrobial bacteremia. Catheter extraction and rapid institution of intravenous antibiotics in combination should be administered in breakthrough bacteremia.

Frank electrocardiograms.

Frank electrocardiograms were recorded in 426 schoolchildren (225 girls, 201 boys) aged 10 to 14 years, planar vectorcardiograms projected onto the horizontal and left sagittal planes were constructed. Spatial magnitude of instantaneous QRS vector end-points in 10 ms intervals of QRS duration, and of maximum QRS vector, as well as X, Y, Z coordinates of instantaneous QRS vector end-points in 10 ms intervals of QRS duration, and of the maximum QRS vector were analyzed in relation to weight, height and BMI. The initial part of VCG loop was not influenced by age and anthropometric parameters, respectively. The spatial QRS vector magnitude at the 30 ms of QRS decreased significantly with age and height. From the 40 ms of QRS duration the VCG parameters were significantly related to anthropometric parameters. Vectorcardiographic parameters were more influenced by the anthropometric parameters then by age.