Dental professionals participating in a home visiting programme for first-time parents.

BACKGROUND: A home visiting programme was offered to first-time parents in the south of Sweden to reduce health inequalities among young children and support parents. The programme involved a collaboration between child healthcare nurses and midwives, social workers, and dental hygienists/dental nurses. It is unusual for dental professionals to participate in such programmes. OBJECTIVE: To describe the experiences of collaboration between child healthcare nurses and dental professionals and their views on the programme from a dental perspective. BASIC RESEARCH DESIGN: Qualitative semi-structured interview study with content analysis. PARTICIPANTS: Four child healthcare nurses, three dental hygienists, and two dental nurses. RESULTS: Analysis identified five themes: contribution of knowledge, reinforced oral health advice, family-based oral health advice, financial considerations, and future role of the dental care. CONCLUSIONS: Participants stressed the importance of dental professionals' knowledge, the need for child healthcare nurses and dental professional to conduct home visits together to deliver family-based and reinforced dental advice. They suggested a follow-up visit and the participation of the public dental service before a private dental care provider since most children will visit a public dentist later. The programme was perceived as worth the costs, but for the best utility, the resources should primarily be for non-native parents. Further research should focus on the effect of the home visiting programme on the children's oral health.

The undergraduate degree project--preparing dental students for professional work and postgraduate studies?

OBJECTIVES: The undergraduate degree project is a requisite part of higher education in Sweden, designed to prepare students for professional work and postgraduate studies. This article examines the extent to which the degree project in Swedish dental education helps students achieve these purposes. The focus was on the students' choice of topics and research methods as well as their ability to reflect on the implications of their results for dental practice. METHODS: Degree projects from three of the four Swedish dental schools were analysed using content analysis. RESULTS: The students' topics concerned clinical dentistry, biomedicine, educational issues and public oral health. Quantitative research methods were used more often than qualitative ones. Some of the degree projects were based on literature reviews. Students demonstrated shortcomings in their reflections on the implications of their results for dental practice. The level of reflection was particularly low in one of the schools; this may be because the students in this school were not expected to reflect on the results. CONCLUSIONS: The degree project gives the students an opportunity to develop their knowledge on a topic relevant to dentistry, to be trained in conducting research and to reflect on scientific knowledge in relation to dentistry. However, this study shows the need of assessment criteria that urge the students to reflect on the link between science and clinical work and motivate them to learn to reflect so they become critical thinkers. It is also suggested that dental students should learn more about qualitative research methods.

Undergraduate degree projects in the Swedish dental schools: a documentary analysis.

BACKGROUND: Undergraduate degree projects have currently been introduced into courses in the four Swedish dental schools. The rationale for research projects is that they enable students to develop research expertise skills and to show their ability to apply and develop knowledge relevant to professional practice. This paper reports a qualitative analysis of the curriculum documents and handbooks including the criteria used to assess the students' research reports. The aim was to investigate commonalities and differences in the design of degree projects between the four Swedish dental schools and to explore any inconsistencies within the documents. METHODS: The documentary analysis was based on the constant comparison method. RESULTS: Four overarching themes emerged from the analysis: (i) developing scientific expertise, (ii) developing professional expertise, (iii) following rules and (iv) fostering creativity. CONCLUSIONS: The documents from the four dental schools revealed similar views on the purposes of the projects and provided similar assessment criteria. The students were requested to formulate an odontological problem, apply a relevant scientific method, analyse texts and empirical data, express critical reflections and write a short thesis. The students were free to choose topics. There were differences between the dental schools on the emphasis placed on practical uses of the projects and theoretical background of the projects. Two of the schools insisted on rigid rules of completing and writing the project yet paradoxically emphasised creativity. There were wide variations in the required length of the project report. The report may prove useful to dental schools in other countries who are about to design undergraduate research projects.

Characterization of human clinical isolates of Dietzia species previously misidentified as Rhodococcus equi.

In this study, 16 human clinical isolates of Dietzia species previously misidentified as Rhodococcus equi were evaluated using phenotypic methods, including traditional and commercial (API Coryne) biochemical tests, antimicrobial susceptibility testing, and 16S rRNA gene and gyrB gene sequencing. Positive results for both the hydrolysis of adenine and Christie-Atkins-Munch-Petersen (CAMP) reaction allowed for differentiation between the Dietzia isolates and the type strain of Rhodococcus equi; however, traditional and commercial phenotypic profiles could not be used to reliably identify Dietzia species. The analysis of 16S rRNA gene and gyrB gene sequences could discriminate all Dietzia strains from the type strain of R. equi. Most Dietzia species had distinct 16S rRNA gene and gyrB gene sequences; however, the 16S rRNA gene sequences of the type strains of D. schimae and D. cercidiphylli were identical to D. maris and D. natronolimnaea, respectively. Based on comparative sequence analysis, five clinical isolates clustered with D. maris/D. schimae and nine with D. natronolimnaea/D. cercidiphylli. The two remaining isolates were found to be most closely related to the D. cinnamea/D. papillomatosis clade. Even though molecular analyses were not sufficiently discriminative to accurately identify all Dietzia species, the method was able to reliably identify isolates that were previously misidentified by phenotypic methods to the genus level.

Syphilis in composers and musicians--Mozart, Beethoven, Paganini, Schubert, Schumann, Smetana.

In the pre-antibiotics era, syphilis was an extremely common disease. The first well-recorded European outbreak of what is now known as syphilis occurred in 1494, when it appeared among French troops besieging Naples. Thereafter, the disease spread all over Europe and, in the 18th and 19th centuries, many artists became victims of syphilis, among them poets, painters, philosophers, and musicians and composers. This review presents biographies of several musicians and composers that probably suffered from syphilis.

Biotransformation and detoxification of selenite by microbial biogenesis of selenium-sulfur nanoparticles.

This study combines the interaction between the toxic oxyanions selenite and selenate and the plant growth promoting bacterium Azospirillum brasilense with a comprehensive characterization of the formed selenium particles. As selenium is an essential trace element, but also toxic in high concentrations, its state of occurrence in nature is of major concern. Growth of the bacterium was affected by selenite (1-5mM) only, observable as a prolonged growth lag-phase of 3days. Subsequently, selenite reduction occurred under aerobic conditions resulting in extracellularly formed insoluble Se0 particles. Complementary studies by microscopic and spectroscopic techniques revealed the particles to be homogeneous and stable Se8-nSn structured spheres with an average size of 400nm and highly negative surface charge of -18mV in the neutral pH range. As this is the first study showing Azospirillum brasilense being able to biotransform selenite to selenium particles containing a certain amount of sulfur, even if environmental waters supplemented with selenite were used, they may significantly contribute to the biogeochemical cycling of both elements in soil as well as to their soil-plant transfer. Therefore, microbial biotransformation of selenite under certain circumstances may be used for various bio-remediation and bio-technological applications.

Fatal pneumonia associated with human metapneumovirus (HMPV) in a patient with myeloid leukemia and adenocarcinoma in the lung.

We describe a clinical case of a 59 old caucasian male who was delivered to the hospital for severe pneumonia associated to human metapneumovirus. The patient suffered from a leukemia and an adenocarcinoma in the lung and died two weeks after submission due to fatal respiratory failure.

Phagocytosis and killing of Mycobacterium avium complex by human neutrophils.

Organisms belonging to the Mycobacterium avium complex (MAC) cause life-threatening bacteremia in immunocompromised patients. Monocytes and macrophages are thought to be responsible for ingestion and killing of MAC. However, it has been suggested that neutrophils may play a role in the early immune response to MAC infection. Here, neutrophils in autologous plasma were incubated (at 0 and 37 degrees C) with M. avium labeled with Auramine O, a potent fluorochrome. Neutrophil phagocytosis was measured by flow cytometry. Neutrophils incubated at 37 degrees C showed an increase in fluorescence over time with a maximum at 15 min, whereas neutrophils on ice showed no time-dependent increase in FL1. At 15 min Fl 1 at 37 degrees C was twice as high as FL1 at 0 degrees C. Examination under the fluorescent microscope showed multiple intracellular fluorescent mycobacteria. Results in nine independent experiments showed time-dependent decrease of colony-forming units in neutrophil-associated live M. avium. Significant killing was observed within 30 min and was complete by 120 min. Observation by electron microscopy clearly confirmed the presence of intraphagosomal MAC, both intact and with evidence of degradation. These data demonstrate that MAC is rapidly phagocytized and killed by human neutrophils. The newly established flow cytometry method should be useful in further studies of neutrophil function and of the role of G-CSF and other cytokines in MAC disease.

Spectroscopic evidence for selenium(IV) dimerization in aqueous solution.

The aqueous speciation of selenium(iv) was elucidated by a combined approach applying quantum chemical calculations, infrared (IR), Raman, and (77)Se NMR spectroscopy. The dimerization of hydrogen selenite (HSeO3(-)) was confirmed at concentrations above 10 mmol L(-1) by both IR and NMR spectroscopy. Quantum chemical calculations provided the assignment of vibrational bands observed to specific molecular modes of the (HSeO3)2(2-) ion. The results presented will provide a better understanding of the chemistry of aqueous Se(iv) which is of particular interest for processes occurring at mineral/water interfaces.

Urothelial cells undergo epithelial-to-mesenchymal transition after exposure to muscle invasive bladder cancer exosomes.

Bladder cancer, the fourth most common noncutaneous malignancy in the United States, is characterized by high recurrence rate, with a subset of these cancers progressing to a deadly muscle invasive form of disease. Exosomes are small secreted vesicles that contain proteins, mRNA and miRNA, thus potentially modulating signaling pathways in recipient cells. Epithelial-to-mesenchymal transition (EMT) is a process by which epithelial cells lose their cell polarity and cell-cell adhesion and gain migratory and invasive properties to become mesenchymal stem cells. EMT has been implicated in the initiation of metastasis for cancer progression. We investigated the ability of bladder cancer-shed exosomes to induce EMT in urothelial cells. Exosomes were isolated by ultracentrifugation from T24 or UMUC3 invasive bladder cancer cell conditioned media or from patient urine or bladder barbotage samples. Exosomes were then added to the urothelial cells and EMT was assessed. Urothelial cells treated with bladder cancer exosomes showed an increased expression in several mesenchymal markers, including α-smooth muscle actin, S100A4 and snail, as compared with phosphate-buffered saline (PBS)-treated cells. Moreover, treatment of urothelial cells with bladder cancer exosomes resulted in decreased expression of epithelial markers E-cadherin and ß-catenin, as compared with the control, PBS-treated cells. Bladder cancer exosomes also increased the migration and invasion of urothelial cells, and this was blocked by heparin pretreatment. We further showed that exosomes isolated from patient urine and bladder barbotage samples were able to induce the expression of several mesenchymal markers in recipient urothelial cells. In conclusion, the research presented here represents both a new insight into the role of exosomes in transition of bladder cancer into invasive disease, as well as an introduction to a new platform for exosome research in urothelial cells.

Osteopontin mediates an MZF1-TGF-ß1-dependent transformation of mesenchymal stem cells into cancer-associated fibroblasts in breast cancer.

Interactions between tumor cells and cancer-associated fibroblasts (CAFs) in the tumor microenvironment significantly influence cancer growth and metastasis. Transforming growth factor-ß (TGF-ß) is known to be a critical mediator of the CAF phenotype, and osteopontin (OPN) expression in tumors is associated with more aggressive phenotypes and poor patient outcomes. The potential link between these two pathways has not been previously addressed. Utilizing in vitro studies using human mesenchymal stem cells (MSCs) and MDA-MB231 (OPN+) and MCF7 (OPN-) human breast cancer cell lines, we demonstrate that OPN induces integrin-dependent MSC expression of TGF-ß1 to mediate adoption of the CAF phenotype. This OPN-TGF-ß1 pathway requires the transcription factor, myeloid zinc finger 1 (MZF1). In vivo studies with xenotransplant models in NOD-scid mice showed that OPN expression increases cancer growth and metastasis by mediating MSC-to-CAF transformation in a process that is MZF1 and TGF-ß1 dependent. We conclude that tumor-derived OPN engenders MSC-to-CAF transformation in the microenvironment to promote tumor growth and metastasis via the OPN-MZF1-TGF-ß1 pathway.

Polymerase chain reaction for microsporidian DNA in gastrointestinal biopsy specimens of HIV-infected patients.

OBJECTIVE: To study the accuracy of polymerase chain reaction (PCR) for microsporidian DNA in gastrointestinal biopsy specimens of HIV-infected patients for the diagnosis of intestinal microsporidiosis. SETTING: Infectious disease in- and outpatient clinic of a university hospital in Cologne, Germany. PATIENTS: Forty-six HIV-infected patients with diarrhoea. METHODS: PCR and Southern blot hybridization were performed using DNA extracted from intestinal biopsy specimens with primers and probes from the small subunit rRNA gene of Enterocytozoon bieneusi and Septata intestinalis. Histological examination of intestinal biopsy specimens was performed using a fluorescence technique. Transmission electron microscopy of intestinal biopsy specimens was performed in 13 patients. RESULTS: Amplification and Southern blot hybridization with species-specific primers and probes gave positive results in 10 patients for E. bieneusi, and in 10 patients for S. intestinalis. Overall, five cases of double infection with E. bieneusi and S. intestinalis were seen when both primer pairs and probes were used. Histological examination showed microsporidian spores in all 15 cases, but light microscopy was unable to distinguish between species in almost all cases. CONCLUSIONS: PCR detection of microsporidian DNA in intestinal biopsy specimens can be used reliably for the diagnosis of intestinal microsporidiosis in HIV-infected patients and is also useful for species differentiation between microsporidia. Infections with S. intestinalis and double infections with two types of microsporidia appear to be more common than previously described.

Virological treatment failure of protease inhibitor therapy in an unselected cohort of HIV-infected patients.

OBJECTIVE: To determine the rate of virological treatment failure with protease inhibitor therapy in unselected patients and to assess underlying risk factors. DESIGN AND SETTING: Retrospective study in two German tertiary care treatment centres. PATIENTS: A total of 198 HIV-infected patients treated with protease inhibitors in 1996. MAIN OUTCOME MEASURES: Levels of HIV RNA 1-6 months after start of treatment; definition of treatment failure of < 1 log10 reduction in plasma HIV RNA within 6 months after starting protease inhibitor therapy; multivariate analysis of risk factors for treatment failures. RESULTS: A total of 226 treatment episodes with protease inhibitors were evaluable (saquinavir, 83; ritonavir, 47; indinavir, 96). The rate of virological treatment failure was 44% (saquinavir, 64%; ritonavir, 38%; indinavir, 30%). In a multivariate analysis, the following independent risk factors for virological failure were found: CD4 cell count, pretreatment with antiretroviral drugs (number), and protease inhibitor (compound). The relative risk reduction for each CD4 cell count increase was 0.997 (P = 0.012), 2.64 for pretreatment with one or two drugs versus no drug (P = 0.05), 2.97 for pretreatment with more than two drugs versus no drug (P = 0.05), and 4.62 for treatment with saquinavir versus indinavir (P = 0.001). CONCLUSION: An unexpectedly high rate of virological treatment failure of protease inhibitor therapy was found in an unselected cohort of HIV-infected patients. Response to antiretroviral combination therapy in normal clinical practice may considerably differ from results of randomized clinical trials. Further studies are warranted to find optimal treatment strategies for both initial and salvage therapy.

Salvage therapy with regimens containing ritonavir and saquinavir in extensively pretreated HIV-infected patients.

OBJECTIVE: To evaluate the efficacy and toxicity of salvage regimens containing ritonavir and saquinavir in patients failing highly active antiretroviral therapy (HAART), and to correlate outcome with plasma concentrations of protease inhibitors. DESIGN: Prospective, non-randomized interventional study. SUBJECTS AND METHODS: Thirty extensively pretreated HIV-infected patients with virological failure under HAART were treated with ritonavir (400 mg twice daily) and saquinavir (600 mg twice daily) and at least one reverse transcriptase inhibitor. HIV-RNA, CD4 cell counts and plasma concentrations of protease inhibitors were determined, and patients were monitored for toxicity at monthly intervals. RESULTS: Six patients showed complete virological success (HIV-RNA <200 copies/ml at week 12) which was sustained for a median follow-up of 6.3 months. Partial virological response (decrease of HIV-RNA of >1 log10 at week 12) was achieved by a further three patients. Patients with a virological response had significantly higher CD4 cell increases than patients without virological response (mean increase at week 12: 66x10(6) cells/l versus 6x10(6) cells/l; P = 0.01). No clinical events were observed during 6 months of follow-up. Neither the use of a non-nucleoside reverse transcriptase inhibitor (NNRTI) nor the number of newly introduced drugs influenced the virological response. Plasma concentrations of protease inhibitors did not statistically differ between patients with and without success. Toxicity included gastrointestinal disturbances, lipid abnormalities and liver dysfunction. CONCLUSIONS: In extensively pretreated patients, salvage regimens containing ritonavir and saquinavir had only limited and short-term anti-HIV activity and were associated with substantial toxicity. Plasma concentrations of saquinavir were not predictive for virological response.

Microsporidiosis: human diseases and diagnosis.

Microsporidia are considered opportunistic pathogens in humans because they are most likely to cause diseases if the immune status of a host is such that the infection cannot be controlled. A wide spectrum of diseases has been reported among persons infected with microsporidia and different diagnostic techniques have been developed during the last decade.

T-lymphocyte subsets in the duodenal lamina propria of patients infected with the human immunodeficiency virus type 1 and influence of high-dose immunoglobulin therapy.

T-lymphocyte subsets in duodenal biopsies of human immunodeficiency virus type 1 (HIV-1)-infected patients were studied by immunocytochemical staining to determine the alterations of CD4- and CD8-cell subsets in comparison with HIV-1 antibody-negative controls and to examine the association with stage, gastrointestinal symptoms, and peripheral lymphocyte subsets and the influence of high-dose intravenous immunoglobulins. A significant decrease in duodenal CD4 cells (p less than 0.001) and CD4/CD8 ratio (p less than 0.001) follows HIV-1 infection when compared to HIV-1-negative controls, more accentuated both in patients of stage WR6 and suffering from diarrhea than in those of stages WR1-5 or without diarrhea. In addition, a significant increase in CD8 cells (p less than 0.01) could be found in HIV-1-infected patients, again with lower levels in patients of stages WR6 than WR1-5. A strong correlation was found between the intestinal and peripheral CD4/CD8 ratio (R = 0.80), but the correlation was weak if HIV-1-negative persons were excluded from analysis (R = 0.29). Treatment with high-dose intravenous immunoglobulins improved diarrhea in four of five patients; two patients gained weight. Diffuse lymphocytic infiltration of the lamina propria, villous atrophy, CD4- and CD8-cell percentage, and CD4/CD8 ratio were not influenced.

Tissue diagnosis of intestinal microsporidiosis using a fluorescent stain with Uvitex 2B.

AIMS: To detect intestinal microsporidiosis in paraffin wax embedded biopsy specimens using a fluorescence technique incorporating optical brighteners. METHODS: Eight HIV infected patients with confirmed intestinal microsporidiosis (six with Enterocytozoon bieneusi, one with Encephalitozoon intestinalis and one with Encephalitozoon cuniculi infection) and 10 without infection were studied. Tissue sections of paraffin wax embedded duodenal biopsy specimens were stained with 1% Uvitex 2B, coded and analysed independently by two investigators. RESULTS: In all eight cases with confirmed intestinal microsporidian infection, spores could be detected easily in tissue sections using the fluorescence technique. Spores or other elements consistent with microsporidiosis were not found in the 10 patients without infection. CONCLUSION: Staining of tissue sections from paraffin wax embedded intestinal biopsy specimens with stains incorporating Uvitex 2B is a rapid and easy technique for the diagnosis of intestinal microsporidiosis.

Cryptosporidia and microsporidia--waterborne diseases in the immunocompromised host.

Cryptosporidia and microsporidia are emerging parasitic pathogens in immunocompetent and immunocompromised individuals. Cryptosporidium infects several wild and domestic animals that excrete oocysts into the environment and contaminated water represents the major source of infection for humans. Waterborne transmission of Cryptosporidium is a major risk for humans and appropriate measures have to be taken to protect immunocompetent and immunocompromised individuals to become infected. For microsporidia, the sources and ways of transmission are not well documented. Although several animal hosts have been identified recently, the relevant reservoirs of human microsporidia are still unknown. Also, the routes of spreading are unknown. Is microsporidiosis a zoonotic disease that will be transmitted through close contact with infected animals or is contaminated surface water responsible for transmission and represents a relevant reservoir? This review is designed to give information on these two emerging intestinal parasites in a format that will be useful to clinical microbiologists, physicians interested in infectious diseases, and public health personnel.

EPR probes with well-defined, long distances between two or three unpaired electrons

The synthesis of rod- and star-shaped compounds carrying two or three spin labels as end groups is described. The unpaired electrons are 2.8-5.1 nm apart from each other. The shape-persistent scaffolds were obtained through Pd-Cu-catalyzed alkynyl-aryl coupling and Pd-Cu-catalyzed alkyne dimerization in the presence of oxygen using p-phenyleneethynylene as the basic shape-persistent building block. The spin label 1-oxyl-2,2,5,5-tetramethylpyrroline-3-carboxylic acid (4) was attached through esterification of the terminal phenolic OH groups of the scaffold.

Intestinal microsporidiosis with Septata intestinalis in a patient with AIDS--response to albendazole.

Microsporidiosis is a common finding in HIV-infected patients who have diarrhoea. The species most commonly causing gastrointestinal disease is Enterocytozoon bieneusi. Recently Septata intestinalis has been described as a cause of diarrhoea and disseminated infection in patients with AIDS. A 44-year-old homosexual man with severe immunodeficiency (CD4 cell count 40/microliters) had a history of watery diarrhoea for 2 weeks. Microsporidian spores measuring 1.2 to 1.5 x 2.5 to 3.0 microns were found in stool samples. Electron microscopy of duodenal biopsies confirmed the diagnosis of intestinal microsporidiosis and showed parasitophorous vacuoles with the typical ultrastructure of S. intestinalis. The patient was treated with albendazole (400 mg twice daily) and became asymptomatic within 4 days. No spores could be detected in stool samples after a treatment period of 14 days. About 25 infections with S. intestinalis have been reported to date, and the case presented here is the first in a German patient.