Anxiety, depression, somatization and psychological distress before and 2-6 years after a late termination of pregnancy due to fetal anomalies.

BACKGROUND: For many women, a late termination of pregnancy (TOP) can be an enormous psychological burden. Few studies have investigated the long-term psychological impact of late TOP. METHODS: N = 90 women answered a questionnaire containing questions about anxiety, depression and somatization (Brief-Symptom Inventory, BSI-18) shortly before (T1) and 2-6 years after (T4) their late termination of pregnancy. RESULTS: Prior to the late TOP, 57.8% of participants showed above-average levels of overall psychological distress (66.7% anxiety, 51.1% depression, 37.8% somatization). This number decreased significantly over time for all scales of the BSI-18. 2-6 years later, only 10.0% of women still reported above-average levels (17.8% anxiety, 11.1% depression, 10.0% somatization). CONCLUSIONS: Our results support those of previous research showing that late TOP has a substantial psychological impact on those experiencing it in the short-term. In the long-term, most women return to normal levels of psychological distress, although some still show elevated levels. Limitations of the study include monocentric data collection, drop-out between T1 and T4, and the relatively wide range of two to six years after TOP. Further research should be conducted in order to identify factors that impact the psychological processing of the experience.

Online Interaction Turns the Congeniality Bias Into an Uncongeniality Bias.

Online phenomena like echo chambers and polarization are believed to be driven by humans' penchant to selectively expose themselves to attitudinally congenial content. However, if like-minded content were the only predictor of online behavior, heated debate and flaming on the Internet would hardly occur. Research has overlooked how online behavior changes when people are given an opportunity to reply to dissenters. Three experiments (total N = 320; convenience student samples from Germany) and an internal meta-analysis show that in a discussion-forum setting where participants can reply to earlier comments larger cognitive conflict between participant attitude and comment attitude predicts higher likelihood to respond (uncongeniality bias). When the discussion climate was friendly (vs. oppositional) to the views of participants, the uncongeniality bias was more pronounced and was also associated with attitude polarization. These results suggest that belief polarization on social media may not only be driven by congeniality but also by conflict.

A randomized controlled trial of problem-solving training (PST) for hematopoietic stem cell transplant (HSCT) patients: Effects on anxiety, depression, distress, coping and pain.

The effect of problem-solving training (PST) on psychological distress, coping, pain, overall distress, and problem-solving behavior in hematopoietic stem cell transplant (HSCT) patients was investigated in a randomized controlled trial. The intervention was performed during aplasia and included five 1-hour individual sessions; it was started 2 days before HSCT. Forty-five patients were included in the intervention group (IG) and 46 in the control group (CG). The effects were measured at 11 (t2) and 21 days (t3) after HSCT. At t2, 31 patients remained in the IG and 36 patients in the CG. Patients > 18 years, with adequate cognitive performance, and with adequate command of the German language, were included. They were assessed with regard to anxiety and depression (Hospital Anxiety and Depression Scale), psychological distress (Symptom Checklist short version-9), coping (Brief Cope), problem-solving (Social Problem Solving Inventory-Revised), pain (Questions of Pain), and distress (National Comprehensive Cancer Network Distress Thermometer). The data were analyzed using analyses of covariance (general linear model) to examine the differences between the two conditions. Anxiety, psychological distress, pain, and general stress were reduced after PST in the IG compared with those in the CG. Active coping was reinforced in the IG. The IG patients were also better able to reduce negative problem orientation and improve problem implementation. The effect of the PST on these features was between Eta 2 =.13 and .45 and can thus be described as medium to strong. There were no changes in depression and social support. After careful interpretation of the results, it can be said that PST affects patients' mental health, problem-solving, and coping. Because these psychological changes occur shortly after the transplantation in the phase of aplasia, HSCT patients are likely to be given a more favorable starting position for the course of recovery.

Conception and Associated Evaluation of a Problem-Solving Training (PST) for Patients in the Hospital Context of Hematopoietic Stem Cell Transplantation (HSCT).

It appears from empirical studies that the problem-solving ability of patients is associated with the experience of distress and the patients' mental state. The goals of this study were the (1) conception and (2) associated evaluation of the psychological short-time intervention "problem-solving training" (PST) for patients hospitalized for hematopoietic stem cell transplantation (HSCT). (1) The conception of the PST comprised a multi-stage development phase. An existing manual for outpatients diagnosed with cancer was adapted to the specific situation of a HSCT. This was followed by development of a manual, definition of the general framework, instruction of coaches, and implementation in a hospital setting. (2) The associated evaluation of PST was conducted from the patients' and the coaches' point of view. A total of 22 patients and five coaches evaluated the training. The training was evaluated by both patients and coaches as being well achievable with the exception of a limited time frame for the first module. The manual explanations were judged to be intelligible by all participants. Regarding on-topic alertness, patients were, on average, rated as "rather "to "very attentive." The patients evaluated the response to their needs as "good." They further assessed their overall condition due to the training as "good." This study provides preliminary evidence for the feasibility of PST by using the developed manual (Psychological Short-Term Intervention PST for Patients During HSCT). Based on this, it is conceivable to implement this intervention in similar situations to the advantage of a different patient clientele.

Identification of late assembly domains of the human endogenous retrovirus-K(HML-2).

BACKGROUND: Late assembly (L)-domains are protein interaction motifs, whose dysfunction causes characteristic budding defects in enveloped viruses. Three different amino acid motifs, namely PT/SAP, PPXY and YPX(n)L have been shown to play a major role in the release of exogenous retroviruses. Although the L-domains of exogenous retroviruses have been studied comprehensively, little is known about these motifs in endogenous human retroviruses. RESULTS: Using a molecular clone of the human endogenous retrovirus K113 that had been engineered to reverse the presumed non-synonymous postinsertional mutations in the major genes, we identified three functional L-domains of the virus, all located in the Gag p15 protein. A consensus PTAP tetrapeptide serves as the core of a main L-domain for the virus and its inactivation reduces virus release in HEK 293T cells by over 80%. Electron microscopy of cells expressing the PTAP mutant revealed predominantly late budding structures and budding chains at the plasma membrane. The fact that this motif determines subcellular colocalization with Tsg101, an ESCRT-I complex protein known to bind to the core tetrapeptide, supports its role as an L-domain. Moreover, two YPX(n)L motifs providing additional L-domain function were identified in the p15 protein. One is adjacent to the PTAP sequence and the other is in the p15 N-terminus. Mutations in either motif diminishes virus release and induces an L-domain phenotype while inactivation of all three L-domains results in a complete loss of particle release in HEK 293T cells. The flexibility of the virus in the use of L-domains for gaining access to the ESCRT machinery is demonstrated by overexpression of Tsg101 which rescues the release of the YPX(n)L mutants. Similarly, overexpression of Alix not only enhances release of the PTAP mutant by a factor of four but also the release of a triple mutant, indicating that additional cryptic YPX(n)L domains with a low affinity for Alix may be present. No L-domain activity is provided by the proline-rich peptides at the Gag C-terminus. CONCLUSIONS: Our data demonstrate that HERV-K(HML-2) release is predominantly mediated through a consensus PTAP motif and two auxiliary YPX(n)L motifs in the p15 protein of the Gag precursor.

Physical activity is inversely associated with multimorbidity in elderly men: results from the KORA-Age Augsburg Study.

OBJECTIVE: Physical activity is suggested to play a key role in the prevention of several chronic diseases. However, data on the association between physical activity and multimorbidity are lacking. METHODS: Using data from 1007 men and women aged 65-94 years who participated in the population-based KORA (Cooperative Health Research in the Region of Augsburg)-Age project conducted in Augsburg/Germany and two adjacent counties in 2008/09, 13 chronic conditions were identified, and physical activity scores were calculated based on the self-reported physical activity scale for the elderly (PASE). Multivariable sex-specific logistic regression was applied to determine the association of the continuous physical activity score with multimorbidity (≥ 2 out of 13 diseases). RESULTS: Physical activity (mean PASE score±SD) was higher in men (125.1 ± 59.2) than in women (112.2 ± 49.2). Among men, the odds ratio (OR) for multimorbidity was 0.73 (95% CI: 0.60-0.90) for a 1 standard deviation increase of the PASE score. No significant results could be observed for women (OR: 1.05; 95% CI: 0.83-1.33). CONCLUSION: We demonstrated an inverse association between physical activity and multimorbidity among men. Further prospective studies have to confirm the temporality of effects.

Relation between depressed mood, somatic comorbidities and health service utilisation in older adults: results from the KORA-Age study.

BACKGROUND: prior literature suggests that comorbidity with depression significantly worsens the health state of people with chronic diseases. OBJECTIVE: the present study examines whether depressed mood increased medical care use for patients with a comorbid physical disease. DESIGN, SETTING AND SUBJECTS: the study was a population-based study (KORA-Age), with 3,938 participants aged 64-94. METHODS: we investigated differences in health services use in participants with and without depressed mood (Geriatric Depression Scale). A further adjustment for disease was done and differences were examined with the Mann-Whitney U test. The incidence rate ratios (IRRs) for doctors' appointments or the number of days in hospital were explored with (zero-inflated) negative binomial regression models. RESULTS: there are increased self-neglecting behaviours and medical comorbidities in participants with depressed mood. Depressed mood increased participants' use of medical services (P < 0.0001). Among participants who visited the doctor during the last 3 months, those with depressed mood had more visits than those without depressed mood, irrespective of somatic comorbidities (P < 0.0001 and P < 0.05 for ill and healthy, respectively). Additionally, patients with coexisting depressed mood and physical disease visited the doctor's practice significantly more often. Having depressed mood significantly increases the likelihood for more doctor visits (IRR = 1.5, CI = 1.3-1.7) and longer hospital stays (IRR = 1.9, CI = 1.6-2.3). In participants with somatic comorbidities the risk is even greater (IRR = 1.6, CI = 1.3-2, for the number of doctors visits and IRR = 2, CI = 1.4-2.9, for the number of days in the hospital). CONCLUSIONS: results suggest that patients with depressed mood had increased use of health-care services overall, particularly those with somatic comorbidities.

Identification of the protease cleavage sites in a reconstituted Gag polyprotein of an HERV-K(HML-2) element.

BACKGROUND: The human genome harbors several largely preserved HERV-K(HML-2) elements. Although this retroviral family comes closest of all known HERVs to producing replication competent virions, mutations acquired during their chromosomal residence have rendered them incapable of expressing infectious particles. This also holds true for the HERV-K113 element that has conserved open reading frames (ORFs) for all its proteins in addition to a functional LTR promoter. Uncertainty concerning the localization and impact of post-insertional mutations has greatly hampered the functional characterization of these ancient retroviruses and their proteins. However, analogous to other betaretroviruses, it is known that HERV-K(HML-2) virions undergo a maturation process during or shortly after release from the host cell. During this process, the subdomains of the Gag polyproteins are released by proteolytic cleavage, although the nature of the mature HERV-K(HML-2) Gag proteins and the exact position of the cleavage sites have until now remained unknown. RESULTS: By aligning the amino acid sequences encoded by the gag-pro-pol ORFs of HERV-K113 with the corresponding segments from 10 other well-preserved human specific elements we identified non-synonymous post-insertional mutations that have occurred in this region of the provirus. Reversion of these mutations and a partial codon optimization facilitated the large-scale production of maturation-competent HERV-K113 virus-like particles (VLPs). The Gag subdomains of purified mature VLPs were separated by reversed-phase high-pressure liquid chromatography and initially characterized using specific antibodies. Cleavage sites were identified by mass spectrometry and N-terminal sequencing and confirmed by mutagenesis. Our results indicate that the gag gene product Pr74Gag of HERV-K(HML-2) is processed to yield p15-MA (matrix), SP1 (spacer peptide of 14 amino acids), p15, p27-CA (capsid), p10-NC (nucleocapsid) and two C-terminally encoded glutamine- and proline-rich peptides, QP1 and QP2, spanning 23 and 19 amino acids, respectively. CONCLUSIONS: Expression of reconstituted sequences of original HERV elements is an important tool for studying fundamental aspects of the biology of these ancient viruses. The analysis of HERV-K(HML-2) Gag processing and the nature of the mature Gag proteins presented here will facilitate further studies of the discrete functions of these proteins and of their potential impact on the human host.

Deaf awareness workshop for medical students - an evaluation.

Background: Due to a lack of communication strategies and knowledge about the Deaf community, health care professionals are often not prepared to provide deaf or hard of hearing patients with accessible and adapted healthcare. Methods: In the present study, a workshop was designed to determine the effect of deaf awareness training on medical students concerning their gain of knowledge regarding deafness and their competence in providing adapted communication and healthcare for deaf and hard of hearing patients. 95 medical students were evaluated in an online survey prior to as well as following the workshop. The workshop was held online in three consecutive sessions. Results: Students reported a substantially more confident approach to working with hearing impaired patients and indicated that an online learning environment is a suitable and helpful alternative to face-to-face teaching. Participants improved significantly in all evaluated items concerning their knowledge and competence (p<0.001). Furthermore, measurements revealed interaction effects between students' current period of study and the point in time of self-evaluation before and after the workshop. Preclinical students not only catch up but even surpass their clinical peers concerning their learning outcome. Conclusion: Reviewing the results obtained by this study, we are optimistic with respect to all participants' highly positive experiences and learning outcomes. Deaf awareness training should be included in the curriculum of all medical faculties.

COMSKIL: a communication skills training program for medical students.

Objectives: Training communication skills has come to be recognized as a vital aspect of medical school education. A medical communication course based on the COMSKIL Communication Skills Training (CST) Program was developed, integrated into the core curriculum, and evaluated at the Leipzig University Medical School. Methods: Between October 2016 and July 2017, 312 medical students (mean age 21.80 years; 62% male) participated in the medical communication course. Each course unit was evaluated via questionnaires specifically designed to address the theoretical and practical content of the curriculum. The items correspond to the material covered in each course unit. Students responded using a 5-point-Likert scale (1="not at all helpful", 5="extremely helpful") to rate the degree to which the course helped them learn about the subject matter and train the skills covered in the curriculum. Results: The average score for the first part of the course (theoretical foundations) was M=3.69 (SD=0.35). The second part received a similar rating (M=3.84; SD=0.73). The role play exercises with actor-patients received a score of M=4.27 (SD=0.62). In an overall evaluation at the end of the course, students rated the administration of the course (setting, etc), knowledge gained, and skills trained with a score of M=4.11 (SD=0.66). The role play exercises received an overall score of M=4.36 (SD=0.61). Conclusion: A new curriculum for teaching medical students patient-physician communication skills based on the COMSKIL CST program was established at the University of Leipzig. The goal of this course is to teach students about the kinds of communication scenarios they will encounter in their future working lives as care providers and equip them with the fundamental communication techniques and skills they need to successfully handle those situations. A formal evaluation of the program resulted in satisfactory findings, indicating that it is well suited for use in medical universities.

The Longitudinal Communication Curriculum at Leipzig University, Medical Faculty - implementation and first experiences.

Purpose: Communication skills are an essential instrument for building a sustainable patient-doctor-relationship for future doctors. They are learnable and teachable. The learning should be facilitated with the help of a longitudinal curriculum, which is planned at Leipzig University. Project: At the Medical Faculty of Leipzig University, the Longitudinal Communication Curriculum is established since 2016/17. Up to now, the curriculum consists of four parts in which students repeatedly practise their communication skills in curricular and extracurricular courses. Several formats help to teach an integrated learning of communication and physical examination skills. Assessment of communication skills is also performed. Curricular implementation is accompanied by concomitant evaluation. Results: Three parts of the curriculum already have taken place. Students report an increase in communication skills. Students rate the units as instructive and helpful. The assessment of communication skills occurs in two clinical practical examinations (OSCEs). Together with summative assessment a formative feedback was implemented. Students judge this practice as highly positive. Discussion: The curriculum is part of undergraduate medical education in Leipzig. It would be beneficial to add another simulated patient encounter, as well as interprofessional units. Student questionnaires will be evaluated and results will help to develop the curriculum. Conclusion: Consolidation of the curriculum accompanied by evaluation and adaption of content can help to assure the quality of the curriculum. Additional professions and study units shall be integrated in the Longitudinal Communication Curriculum in the future.

Immature mast cells exhibit rolling and adhesion to endothelial cells and subsequent diapedesis triggered by E- and P-selectin, VCAM-1 and PECAM-1.

Mast cell numbers are markedly increased at sites of chronic inflammation. However, the underlying mechanisms of mast cell accumulation including mast cell progenitor trafficking remain to be identified in detail. Thus, the aim of this study was to identify the adhesion molecules involved in rolling, firm adhesion and transendothelial diapedesis of murine bone marrow-derived cultured mast cells (BMCMC) as a model for immature mast cells. We could show that BMCMCs exhibit in vivo rolling on skin vessel walls and strong adhesion to skin endothelial cells (ECs) in vitro under static and flow conditions. Interestingly, interaction of BMCMC with the EC adhesion molecules E- and P-selectin, vascular cell adhesion molecule-1 (VCAM-1) and platelet endothelial cell adhesion molecule-1 (PECAM-1) is required to mediate rolling and firm adhesion to ECs. The adhesion of BMCMCs to skin ECs is further enhanced by TNF, IL-4, IL-15 and vascular endothelial cell growth factor. Furthermore, BMCMCs exhibit directed and dose-dependent transmigration across an endothelial barrier, mediated by a PECAM-1-dependent mechanism. Our results demonstrate that BMCMCs can undergo a tightly regulated extravasation cascade consisting of rolling on and adhesion to endothelium and followed by directed diapedesis and reveal selectins, VCAM-1 and PECAM-1 as required adhesion molecules. These processes may contribute to mast cell accumulation in chronic inflammatory skin diseases and reveal opportunities to modulate peripheral tissue numbers of mast cells.

Clinical application of genomic high-throughput data: Infrastructural, ethical, legal and psychosocial aspects.

Genomic high-throughput technologies (GHTT) such as next-generation sequencing represent a fast and cost-effective tool toward a more comprehensive understanding of the molecular background of complex diseases. However, technological advances contrast with insufficient application in clinical practice. Thus, patients, physicians, and other professionals are faced with tough challenges that forestall the efficient and effective implementation. With the increasing application of genetic testing, it is of paramount importance that physicians and other professionals in healthcare recognize the restrictions and potential of GHTT, in order to understand and interpret the complex data in the context of health and disease. At the same time, the growing volume and complexity of data is forever increasing the need for sustainable infrastructure and state-of-the-art tools for efficient data management, including their analysis and integration. The large pool of sensitive information remains difficult to interpret and fundamental questions spanning from billing to legal, social, and ethical issues have still not been resolved. Here we summarize and discuss these obstacles in an interdisciplinary context and suggest ways to overcome them. Continuous discussion with clinicians, data managers, biostatisticians, systems medicine experts, ethicists, legal scholars, and patients illuminates the strengths, weakness, and current practices in the pipeline from biomaterial to sequencing and data management. This discussion also highlights the new, cross-disciplinary working collaborations to realize the wide-ranging challenges in clinical genomics including the exceptional demands placed on the staff preparing and presenting the data, as well as the question as to how to report the data and results to patients.

International Peer-teaching: the LernKlinik Leipzig "Erasmus-Week" for Incoming Erasmus Students.

Introduction: This project report describes the development, pilot phase, evaluation and implementation of a preparatory course week for incoming Erasmus medical students at the LernKlinik Leipzig, the Skills and Simulation Centre of the Medical Faculty, University of Leipzig. Project description: The aim of this project is to prepare Erasmus students for their year abroad using peer-assisted teaching as the method of choice. Major intended outcomes were support of language and clinical practical skill competency development, as well as enhancement of integration among international and German-speaking peer tutors. The methodological framework of Ross and Cameron [1] was used in planning the Erasmus-Week. For planning the 2012 pilot project, a survey among Erasmus students of the academic year 2011/12 was performed. All succeeding cohorts were asked to participate in pre- and post-surveys which were analyzed quantitatively and qualitatively. Results: Between 2012 and 2017, n=173 European medical students spent their Erasmus year in Leipzig. Of these, n=148 (86%) participated in the Erasmus-Week. The country of origin of most Erasmus students was Rumania (20,3%). Among the most positively received aspects of the Erasmus-Week, the active use of German for medical purposes and the use of peer-teaching as the method of choice to learn and repeat basic medical examination skills were mentioned. Students emphasized their wish for being able to participate in further course offers. Conclusions: Offering a preparatory course week for incoming Erasmus students focusing on language and clinical practical skills training using peer-teaching as the method of choice may facilitate the integration of Erasmus students into their foreign medical curriculum. Further studies are underway to elucidate if these experiences influence later professional careers and choice of employment.

Effect of biopassive and bioactive surface-coatings on the hemocompatibility of membrane oxygenators.

Postoperative complications associated with cardiopulmonary bypass (CPB) surgery and extracorporeal circulation (ECC) procedures are still a major clinical issue. Improving the hemocompatibility of blood contacting devices used for ECC procedures may ameliorate various postpump syndromes. In a simulated CPB model using human blood, we investigated the hemocompatibility, fibrinogen adsorption, and platelet receptor (GPIIb-IIIa) binding capacity of surface-modified membrane oxygenators (Jostra Quadrox). Three groups were compared: (i) biopassive protein coatings (SafeLine), (ii) bioactive heparin coatings (BioLine), and (iii) noncoated controls. During the 2 h recirculation period, plasma concentrations of activation markers for platelets (beta-thromboglobulin), inflammation (elastase), complement (C5a), and coagulation (prothrombin fragment 1+2, thrombin-antithrombin III) were lower in the groups with biopassive and bioactive coatings compared to the noncoated group (p < 0.01). These parameters did not significantly differ between the two surface-coated groups, except for complement activation: C5a levels were higher in the biopassive group compared to the bioactive group (p < 0.01). Moreover, surface-coated oxygenators showed less fibrinogen adsorption, GPIIb-IIIa binding, and platelet/leukocyte adhesion (p < 0.01). We assume that fewer fibrinogen and platelet receptor molecules bound to the surface-coated oxygenator surfaces results in fewer platelet adhesion and activation, which will significantly contribute to the improved hemocompatibility of the biopassive and bioactive oxygenators. Our results suggest that the application of bioactive oxygenators (BioLine) during CPB surgery may reduce postoperative complications for the patient more effectively than biopassive oxygenators (SafeLine).

Thrombospondin 1 as possible key factor in the hemocompatibility of endocoronary prostheses.

Intracoronary stenting has markedly improved the patency of native coronary arteries after percutaneous transluminal coronary angioplasty (PTCA). Advances in stent technology and design, including drug releasing stents, have contributed to reduce the long-term restenosis rate. However, stenosis caused by neointimal hyperplasia, vascular remodeling and thrombosis is still a major problem after endocoronary stent procedures. This study focuses on differential gene expression of circulating peripheral blood cells after 90 min exposure to stents to search for initially activated cellular pathways, which may foster restenosis. Fresh human whole blood (1 IU heparin/ml), taken from non-medicated healthy volunteers, was incubated under flow conditions in an in vitro closed-loop stent-testing model (modified Chandler-Loop). Differential gene expression compared to resting conditions and to the experimental controls was investigated by a DNA-microarray technique encoding for over 17,000 genes simultaneously. As expected, a large variety of genes showed differential gene expression. Interestingly, Thrombospondin 1 (TSP-1), which plays a key role in initial immune defense, was found to be the most markedly up-regulated gene. We propose TSP-1 expression as an early indicator for the activation of immune responses following intracoronary stenting. After clarifying the participation of TSP-1 in vivo, future studies will therefore focus on TSP-1 as a potential prognostic factor, which may also help to develop and control new surface materials with an improved biocompatibility.

Neighborhood and healthy aging in a German city: distances to green space and senior service centers and their associations with physical constitution, disability, and health-related quality of life.

The composition of the residential environment may have an independent influence on health, especially in older adults. In this cross-sectional study, we examined the associations between proximity to two features of the residential environment (green space and senior service centers) and three aspects of healthy aging (self-rated physical constitution, disability, and health-related quality of life). We included 1711 inhabitants from the city of Augsburg, Germany, aged 65 years or older, who participated in the KORA-Age study conducted in 2008/2009. We calculated the Euclidian distances between each participant's residential address and the nearest green space or senior service center, using a geographic information system. Multilevel logistic regression models were fitted to analyze the associations, controlling for demographic and socioeconomic factors. Contrary to expectations, we did not find clear associations between the distances to the nearest green space or senior service center and any of the examined aspects of healthy aging. The importance of living close to green space may largely depend on the study location. The city of Augsburg is relatively small (about 267,000 inhabitants) and has a high proportion of greenness. Thus, proximity to green space may not be as important as in a densely populated metropolitan area. Moreover, an objectively defined measure of access such as Euclidian distance may not reflect the actual use. Future studies should try to assess the importance of resources of the residential environment not only objectively, but also from the resident's perspective.

A PCR system for detection of species and genotypes of the Echinococcus granulosus-complex, with reference to the epidemiological situation in eastern Africa.

We describe the development of a specific and sensitive PCR/semi-nested PCR system for the rapid diagnosis of Echinococcus granulosus genotype G1, E. granulosus genotype G6/7, and Echinococcus ortleppi (G5). Diagnosis of G1 and the group G5/6/7 is performed by a simple PCR, while discrimination between E. ortleppi (G5) and G6/7 involves a subsequent semi-nested PCR step. The target sequence for amplification is part of the mitochondrial 12S rRNA gene. Specificity of the PCRs was 100% when evaluated with isolates of 16 species of cestodes, including Echinococcus multilocularis, Echinococcus equinus, E. ortleppi and three strains of E. granulosus (G1, G6 and G7). Sensitivity threshold was 0.25pg of DNA. This new approach was compared with published protocols of restriction fragment length polymorphism-PCR and sequencing of mitochondrial cytochrome c oxidase subunit 1 and NADH dehydrogenase 1 genes using Echinococcus isolates of human, sheep, goat, camel, cattle and pig origin from Kenya and Sudan. Additionally, two internal DNA probes were developed, one hybridising only with G1, the other with G5, G6 and G7 amplification products. Preliminary epidemiological results obtained with this PCR approach include the detection of a camel strain (G6) infection for the first time in a human patient from eastern Africa, and the first reports of E. ortleppi (G5) in livestock from Kenya and the Sudan.

Hemocompatibility of PMEA coated oxygenators used for extracorporeal circulation procedures.

An inflammatory response to cardiopulmonary bypass (CPB) caused by bioincompatibility of extracorporeal circuits is one of the major clinical issues in cardiac surgery. Recently a new coating material, poly-2-methoxyethylacrylate (PMEA), was developed to improve the biocompatibility of blood contacting surfaces. In a simulated cardiopulmonary bypass model, using fresh human whole blood, 15 membrane oxygenators (Capiox SX18, Terumo Corp., Tokyo, Japan) were compared. Five of them had the PMEA coating, five had a heparin-coated surface, and five had no surface treatment. Blood samples were taken at several time-points during a 90 minute circulation period. Changes in coagulation, complement, and blood cell alteration factors were measured by ELISA methods, plasma bradykinin levels were measured by radioimmunoassay, and expression of genes encoding cytokines TNF-alpha, interleukin-1beta, interleukin-6, and interleukin-8 was determined by semiquantitative real time RT-PCR. Platelet adhesion was significantly reduced in both the PMEA and the heparin coated circuits. Release of platelet activation marker beta-thromboglobulin was significantly higher in the uncoated control group (p < 0.01). After 5 minutes of blood circulation bradykinin levels significantly increased in all three groups (p < 0.01); however, the group with the PMEA coated oxygenators showed the lowest values. Expression of genes encoding proinflammatory cytokines in monocytes was increased in all groups, with the lowest being in the PMEA coated group. PMEA coated CPB surfaces in an in vitro experimental model showed an improved thrombogenicity, reduced bradykinin release, less platelet activation and less proinflammatory cytokines gene expression in comparison with a noncoated group. The authors assume that PMEA coating may ameliorate some of intra- and postperfusion syndromes, particularly hypotension, unspecific inflammation, hyperfibrinolysis, and blood loss.