Science.gov

Sample records for primary mixed leucocyte

  1. The induction of suppressor cells in mixed leucocyte cultures and in mixed leucocyte-non-lymphoid cell cultures.

    PubMed Central

    Pawelec, G

    1980-01-01

    X-ray resistant porcine suppressor T cells expressing Ia-like antigens were obtained from mixed cultures of leucocytes and tissue cells (cultured kidney cells, liver cells, endothelial cells, fibroblasts or X-irradiated leucocytes), and were assayed by their ability to suppress lymphocyte proliferation in a second mixed culture. All tissues tested induced suppressor cells although quantitative differences existed between them. Suppressor cell induction was under genetic control by at least two loci, one of which was within the major histocompatibility (MHC) complex. Suppressor cell function was restricted by the MHC type of the responding cell but not the stimulating cell in the second culture. PMID:6445866

  2. Method for labelling leucocytes with indium In-111 oxine

    SciTech Connect

    Kaminsky, D.

    1992-03-03

    This patent describes an improved method for radio-labelling leucocytes with Indium In-111 oxine. It comprises separating the leucocytes from whole blood for obtaining separated leucocytes mixed with residual red blood cells; and then labelling the separated leucocytes with Indium In-111 oxine; wherein the improvement comprises the following further step: depleting residual red blood cells from the separated leucocytes by resuspending the leucocytes in an isotonic saline solution, then rocking the resuspended leucocytes for causing the leucocytes to preferentially settle out, and then removing residual red blood cells which remain suspended within the supernatant isotonic saline solution.

  3. Association of primary central nervous system vasculitis with the presence of specific human leucocyte antigen gene variant.

    PubMed

    Kraemer, Markus; Becker, Jana; Horn, Peter A; Schwitalla, Jan Claudius; Keyvani, Kathy; Metz, Imke; Wegner, Christiane; Brück, Wolfgang; Schlamann, Marc; Heinemann, Falko M; Berlit, Peter

    2017-09-01

    The etiology and genetic susceptibility of primary central nervous system vasculitis (PCNSV) are still unclear. We analyzed the DNA of 25 Caucasian patients with PCNSV for human leucocyte antigen genes HLA-A, HLA-B, HLA-DRB1, and HLA-DQB1, respectively. HLA-frequencies of the 25 patients with PCNSV were compared with HLA-frequencies of matched Caucasian controls. No statistically significant associations were found for HLA-B, HLA-DR1 and HLA-DQB1 variant. In the PCNSV group, only the HLA-A*69 variant was found more often than expected statistically. The results of this study indicate a potential association of HLA marker with PCNSV in Caucasian patients. Further studies are needed to elucidate the role of genes within the human major histocompatibility complex in the pathogenesis of this angiopathy. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Effects of dietary n-3 fatty acids on Toll-like receptor activation in primary leucocytes from Atlantic salmon (Salmo salar).

    PubMed

    Arnemo, Marianne; Kavaliauskis, Arturas; Andresen, Adriana Magalhaes Santos; Bou, Marta; Berge, Gerd Marit; Ruyter, Bente; Gjøen, Tor

    2017-03-09

    The shortage of the n-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on the international markets has led to increasing substitution of fish oil by plant oils in Atlantic salmon (Salmo salar) feed and thereby reducing the EPA and DHA content in salmon. However, the minimum required levels of these fatty acids in fish diets for securing fish health are unknown. Fish were fed with 0, 1 or 2% EPA or DHA alone or in combination of both over a period, growing from 50 to 400 g. Primary head kidney leucocytes were isolated and stimulated with Toll-like receptor (TLR) ligands to determine if EPA and DHA deficiency can affect expression of important immune genes and eicosanoid production. Several genes related to viral immune response did not vary between groups. However, there was a tendency that the high-level EPA and DHA groups expressed lower levels of IL-1β in non-stimulated leucocytes. These leucocytes were also more responsive to the TLR ligands, inducing higher expression levels of IL-1β and Mx1 after stimulation. The levels of prostaglandin E2 and leukotriene B4 in serum and media from stimulated leucocytes were lower in both low and high EPA and DHA groups. In conclusion, leucocytes from low EPA and DHA groups seemed to be less responsive towards immunostimulants, like TLR ligands, indicating that low levels or absence of dietary EPA and DHA may have immunosuppressive effects.

  5. Leucocyte migration inhibition test in coeliac disease - a reappraisal.

    PubMed Central

    Simpson, F G; Field, H P; Howdle, P D; Robertson, D A; Losowsky, M S

    1983-01-01

    Results of the direct leucocyte migration inhibition (LMI) test using gluten fraction III as antigen were unaffected by incorporation of puromycin into the culture medium at concentrations shown to prevent lymphokine mediated inhibition. Results of the LMI test performed with purified polymorphs were similar to and correlated with results of the standard LMI test using mixed leucocytes in both coeliacs and controls. The addition of purified T lymphocytes did not increase migration inhibition. Normal leucocytes incubated with serum from coeliac patients and washed showed marked migration inhibition when incubated with gluten fraction III. This sensitisation of normal leucocytes was prevented by preincubation with aggregated human IgG. These results suggest that leucocyte migration inhibition by gluten in coeliac disease is not due to lymphokine production by sensitised lymphocytes but is caused by cytophilic antibody. PMID:6832627

  6. Using urinary leucocyte esterase tests as an indicator of infection with gonorrhoea or chlamydia in asymptomatic males in a primary health care setting.

    PubMed

    Rahman, Md Saifur; Beever, Warwick; Skov, Steven; Boffa, John

    2014-02-01

    To evaluate a leucocyte esterase test as a predictor of gonorrhoea or chlamydia in asymptomatic Aboriginal males at the Central Australian Aboriginal Congress Male Clinic (Ingkintja), first-void urine samples and clinical information were collected from consecutive asymptomatic males presenting to the Ingkintja in Alice Springs between March 2008 and December 2009. Urine was tested immediately with a leucocyte esterase test dipstick and then by polymerase chain reaction for gonorrhoea and chlamydia. Among the 292 specimens from asymptomatic males, 15.4% were positive for gonorrhoea or chlamydia. In this group, compared with polymerase chain reaction result for gonorrhoea or chlamydia, leucocyte esterase test alone and in combination with age ≤35 years showed sensitivities of 66.7% and 60%, specificities of 90.7% and 94.7%, positive predictive values of 56.6% and 67.5%, negative predictive values of 93.7% and 92.8% and the area under receiver operating characteristics curve values of 0.79 and 0.85, respectively. Leucocyte esterase tests can reasonably be used as a basis for immediate empirical treatment for gonorrhoea or chlamydia in asymptomatic central Australian Aboriginal men under 35 years of age.

  7. Primary cicatricial alopecia: Other lymphocytic primary cicatricial alopecias and neutrophilic and mixed primary cicatricial alopecias.

    PubMed

    Bolduc, Chantal; Sperling, Leonard C; Shapiro, Jerry

    2016-12-01

    Primary cicatricial alopecias can be frustrating for both patients and physicians. Proper diagnosis guides more successful management of these challenging conditions. Part II will cover the remaining lymphocytic primary cicatricial alopecias, which include pseudopelade of Brocq, central centrifugal cicatricial alopecia, alopecia mucinosa, and keratosis follicularis spinulosa decalvans. It will also discuss the neutrophilic and mixed primary cicatricial alopecias, namely folliculitis decalvans, dissecting cellulitis, folliculitis keloidalis, folliculitis (acne) necrotica, and erosive pustular dermatosis.

  8. The effects of cyclic AMP on leucocyte inhibitory factor (LIF) production and on the inhibition of leucocyte migration.

    PubMed Central

    Lomnitzer, R; Rabson, A R; Koornhof, H J

    1976-01-01

    The effect of drugs known to increase intracellular levels of cyllic AMP were studied in the leucocyte migration ihibition system. It was found that cyclic AMP, dibutyryl cyclic AMP, theophyline, and prostaglandins E1 and E2 inhibited the production of leucocyte inhibiting factor by HA pulsed lymphocytes Inhibition only occured when the drugs were present during or after the PHA pulse. In addition it was found that these drugs enhanced the migration of polymorphonuclear leucocytes (PMN), in this system. Electrophoretic mobility of PMN cells was not altered by these drug indicating that the effect is not due to changes in membrane charge. However, granulocyte adhesion was reduced in the presence of these drug suggesting that adhesion is of primary importance in the migration of polymorphonuclear leucocytes out of capillary tubes. The findings show that cyclic AMP is important in modulating both cell-mediated and inflammatory responses. PMID:181189

  9. Effects of different antimicrobial treatments on serum acute phase responses and leucocyte counts in pigs after a primary and a secondary challenge infection with Actinobacillus pleuropneumoniae.

    PubMed

    Sjölund, M; Fossum, C; Martín de la Fuente, A J; Alava, M; Juul-Madsen, H R; Lampreave, F; Wallgren, P

    2011-07-16

    The susceptibility to an initial challenge and a re-challenge inoculation with Actinobacillus pleuropneumoniae was analysed in pigs that were treated with antimicrobials of different efficacies following the first exposure to A pleuropneumoniae. In brief, 30 nine-week-old specific pathogen-free pigs were allocated to five groups of six. After acclimatisation, four groups were inoculated with A pleuropneumoniae serotype 2. At the onset of clinical signs, three of the groups of pigs were treated with enrofloxacin, tetracycline or penicillin. A fourth group served as the inoculated control and the fifth group as a control group that had not been inoculated. On day 28, all five groups were re-challenged with the same strain of A pleuropneumoniae serotype 2 as had been used in the first inoculation. No treatments were carried out at this time. The acute phase responses and differential leucocyte counts were monitored in detail after both inoculations. Leucocytosis and acute phase responses in the forms of serum amyloid A, pig-major acute phase protein and haptoglobin were recorded in all of the inoculated groups after the onset of clinical signs following the first inoculation. A porcine mannan-binding lectin-A response was less evident in the pigs. Acute phase responses resembling those of the first inoculation were observed in the pigs that had not previously been inoculated and in the pigs treated with enrofloxacin. Acute phase responses were not recorded in the other three groups, where the pigs had seroconverted to A pleuropneumoniae serotype 2 following the first inoculation.

  10. Space maintenance in the primary and mixed dentitions.

    PubMed

    Simon, Thomas; Nwabueze, Ifechide; Oueis, Hassan; Stenger, James

    2012-01-01

    Loss of space in the primary dentition is considered one of the main causes of malocclusion in the permanent dentition. The purpose of this paper is to review ond summarize the indications and use of space maintainers in primary and mixed dentitions as preventive measures of future malocclusion. Two main types of space maintainers are used to maintain the space in primary ond mixed dentitions: fixed and removable appliances. Band and loop is the appliance of choice when a primary maxillary or mandibular first molar is prematurely lost. With the premature loss of a second primary molar, Nance or transpalatal (TPA) appliances can be used on the maxillary arch and the lower lingual holding arch (LLHA) for the mandibular arch.

  11. Designing A Mixed Methods Study In Primary Care

    PubMed Central

    Creswell, John W.; Fetters, Michael D.; Ivankova, Nataliya V.

    2004-01-01

    BACKGROUND Mixed methods or multimethod research holds potential for rigorous, methodologically sound investigations in primary care. The objective of this study was to use criteria from the literature to evaluate 5 mixed methods studies in primary care and to advance 3 models useful for designing such investigations. METHODS We first identified criteria from the social and behavioral sciences to analyze mixed methods studies in primary care research. We then used the criteria to evaluate 5 mixed methods investigations published in primary care research journals. RESULTS Of the 5 studies analyzed, 3 included a rationale for mixing based on the need to develop a quantitative instrument from qualitative data or to converge information to best understand the research topic. Quantitative data collection involved structured interviews, observational checklists, and chart audits that were analyzed using descriptive and inferential statistical procedures. Qualitative data consisted of semistructured interviews and field observations that were analyzed using coding to develop themes and categories. The studies showed diverse forms of priority: equal priority, qualitative priority, and quantitative priority. Data collection involved quantitative and qualitative data gathered both concurrently and sequentially. The integration of the quantitative and qualitative data in these studies occurred between data analysis from one phase and data collection from a subsequent phase, while analyzing the data, and when reporting the results. DISCUSSION We recommend instrument-building, triangulation, and data transformation models for mixed methods designs as useful frameworks to add rigor to investigations in primary care. We also discuss the limitations of our study and the need for future research. PMID:15053277

  12. Investigating Mixed-Ability Teaching in Taiwanese Primary Schools

    ERIC Educational Resources Information Center

    Huang, Ling-Ying

    2014-01-01

    In 2001, Taiwan reformed English language teaching in primary schools, and a mixed ability approach was taken as an organisational method for this. Many teachers claim that they encounter numerous difficulties in catering for different needs because of the large number of differences between students. However, the debate and comparisons between…

  13. Mixing and phase partitioning of primary and secondary organic aerosols

    NASA Astrophysics Data System (ADS)

    Asa-Awuku, A.; Miracolo, M. A.; Kroll, J. H.; Robinson, A. L.; Donahue, N. M.

    2009-08-01

    Predicting primary and secondary organic aerosol (POA and SOA) concentrations requires understanding the phase partitioning of semi-volatile organic species. A well-mixed single phase organic aerosol can absorb greater amounts of semi-volatile species but little experimental evidence exists on the phase distribution of particulate organics. We investigated the phase partitioning and mixing of semi-volatile POA and SOA in a smog chamber. Particle time of flight (PToF) data from an Aerodyne aerosol mass spectrometer (AMS) were used to quantify the extent of mixing. The SOA plus motor oil and diesel fuel combination produced a weakly mixed system, in which two particulate organic phases coexist. However, the POA in diesel exhaust readily mixed with SOA, forming a single phase after one hour. Although both POA types contain semi-volatile components, there is a fundamental difference in their partitioning behavior with SOA. The high resolution AMS data reveal minor differences in composition between the two types of POA. This work provides further evidence that there exists a set of unidentified components that influence particulate mixing that affect OA formation and suggests the extent of absorbent phase mixing (strong versus weak) can be observed and quantified with PToF data.

  14. Primary malignant mixed tumor of bone: a case report

    PubMed Central

    Su, Zhansan; Li, Zhi; Liu, Baoan

    2015-01-01

    Background: An extremely rare primary mixed tumor occurring in left proximal femurs of a 47-year old female is reported. Case report: She had left hip pain for three months in April 2004. Radiological examinations revealed that a translucent expansive lesion in the left greater trochanter. She received the curettage of lesion and bone graft surgery. Curettage specimens were diagnosed as malignant mixed tumor, considered to be metastatic. Five months late the lesion recurred. She underwent obturator neurotomy plus total hip replacement of left hip. A long-term of more than ten years follow-up showed there were no evidence of disease recurrence or metastasis and no any signs of other tumor in her body. Discussion: The tumor contained myoepithelial component with positive immunostain of S-100 protein, p63, CK-pan, and vimentin, epithelial component confirmed by CK-pan, CK-LMW and cartilage, which indicated the tumor was a mixed tumor. Cellular atypia, relative high mitosis index, cartilage consistent with grade I chordrosarcoma, focal coagulative necrosis, and infiltration between trabeculae found in the tumor indicated that the tumor had a low grade malignant nature. During long-time follow-up there were no signs of any tumor found in the patient, which strongly suggested that the tumor be a primary one. PMID:26339414

  15. Use of leucocyte migration under agarose to study spontaneous and directed locomotion of leucocytes.

    PubMed Central

    Repo, H; Kostiala, A A; Kosunen, T U

    1978-01-01

    Three different cell attractants, together with the parallel use of the leucocyte migration agarose test (LMAT) and the leading front modification (LFM) of the Boyden chamber technique, were employed in studying whether the maximal migration of normal human polymorphonuclear leucocytes (PMNs) is higher toward an attractant (chemotaxis) than in the same attractant incorporated in the culture media (chemokinesis). Using LMAT, the maximal migration distance toward zymosan activated serum (ZAS) was found to be significantly longer than that under agarose mixed with ZAS, thus indicating a chemotactic effect exerted by ZAS. When bacterial culture filtrate (BCF) and casein were used as attractants, the corresponding difference was not significant, implying that the stimulatory effect of these substances on cell migration could be explained by increased random locomotion (chemokinesis) alone. In LFM, the migration rate was significantly higher along a casein gradient than without a gradient. Using ZAS, however, only chemokinesis could be demonstrated. BCF was found to attract PMNs into membrane filters only in the presence of human serum albumin. These observations give credence to the view that both LMAT and LFM are applicable to the in vitro assessment of chemotaxis and chemokinesis but the attractant of choice for this is different in each of the two methods. Images Figure 1 PMID:359465

  16. Mucopolysaccharides in Peripheral Leucocytes of Cancer Patients

    PubMed Central

    Riesco, Andres; Leyton, Cecilia

    1971-01-01

    The presence of mucopolysaccharides (MPS) in leucocytes of peripheral blood of 19 cancer patients, 13 patients with pulmonary tuberculosis and 14 normal controls, was studied histochemically. MPS was revealed in different proportions in polynuclears and mononuclears. According to the staining technics, the MPS appear to be mainly carboxylated and contain hyaluronic acid and chondroitinsulphate groups. The quantitative analysis revealed that MPS appeared only in around 3% of leucocytes of normal controls, while in the cancer patients 56% of polynuclear and 90% of mononuclears contained it. In the tuberculous patients, 90% of polynuclears and 86% of the mononuclears revealed MPS. The differences between the prevalence of leucocytes containing MPS in controls and in cancer or tuberculous patients are highly significant. The possibility that the difference in MPS content of leucocytes is related with low inmunological activity is postulated. PMID:4256006

  17. Improved leucocyte migration inhibition response of leucocytes from lepromatous leprosy patients with hapten modified M. leprae.

    PubMed Central

    Fotedar, A; Mustafa, A S; Narang, B S; Talwar, G P

    1982-01-01

    Two acetoacetylated derivatives of Mycobacterium leprae with variable hapten groups and a conjugate with tetanus toxoid were prepared. These were tested as antigens along with unmodified M. leprae in the leucocyte migration inhibition response of leucocytes from clinically, bacteriologically and histopathologically confirmed cases of lepromatous leprosy. LMI response was poor with M. leprae, but was significantly enhanced with acetoacetylated M. leprae. PMID:6751637

  18. PURE CULTURES OF LARGE MONONUCLEAR LEUCOCYTES

    PubMed Central

    Carrel, Alexis; Ebeling, Albert H.

    1922-01-01

    1. Pure strains of mononuclear leucocytes were isolated from the blood of adult chickens and keptin active condition for nearly 3 months. 2. The cultures were composed of large mononuclear leucocytes which migrated and proliferated in vitro at a slower rate than fibroblasts. The cells had no tendency to form a tissue, as do fibroblasts and epithelial cells. They were much less resistant than fibroblasts. 3. Differentiation of the large mononuclears into cells assuming the appearance of fibroblasts took place under certain conditions. 4. The activity of the large mononuclears was increased by embryonic tissue juice and inhibited by homologous serum. PMID:19868678

  19. [Leucocyte alkaline phosphatase in normal and pathological pregnancy (author's transl)].

    PubMed

    Stark, K H; Zaki, I; Sobolewski, K

    1981-01-01

    The activities of leucocyte alkaline phosphatase were determined in 511 patients with normal and pathological pregnancy. Mean values were compared and the enzyme followed up, and the conclusion was drawn that leucocyte alkaline phosphatase was no safe indicator of foetal condition. No direct relationship were found to exist between leucocyte alkaline phosphatase, total oestrogens, HSAP, HLAP, HPL, and oxytocinase.

  20. Stories From Three States: Changing to Mixed-Age Primaries.

    ERIC Educational Resources Information Center

    Severeide, Rebecca

    Changes in the structure of instruction in schools may affect a variety of participants in the educational system. These first-person narratives from teachers, staff, parents and students detail how each was affected by the change to mixed-age programming in kindergarten through third grade. Participants came from a number of schools in three…

  1. Fetal leucocyte count in rhesus disease.

    PubMed Central

    Davies, N P; Buggins, A G; Snijders, R J; Noble, P N; Layton, D M; Nicolaides, K H

    1992-01-01

    The effect of fetal anaemia on the total and differential leucocyte counts was studied by examining blood samples obtained by cordocentesis from 177 previously untransfused rhesus affected fetuses at 17-36 weeks' gestation. The mean fetal total leucocyte, lymphocyte, and monocyte counts were significantly lower than the corresponding values in normal controls and there were significant associations between the decrease in these cells and the degree of fetal anaemia. Possible mechanisms for leucopenia include (i) stimulation of erythroid progenitor production at the expense of production of myeloid progenitors, (ii) non-specific haemophagocytosis, or (iii) general suppression of haemopoiesis. Further understanding of the underlying mechanism and the implications of leucopenia as well as the previously reported thrombocytopenia and anaemia may provide a basis for improved antenatal and/or postnatal treatment. PMID:1586179

  2. Primary Biliary Mixed Adenoneuroendocrine Carcinoma (MANEC): A Short Review.

    PubMed

    Acosta, Andres M; Wiley, Elizabeth Louise

    2016-10-01

    Mixed adenoneuroendocrine carcinomas (MANECs) are composite neoplasms with areas of adenocarcinoma or squamous cell carcinoma intermingled with neuroendocrine carcinoma or neuroendocrine tumor, each composing at least 30% of the neoplasm. MANECs are very infrequent overall, and they are more commonly diagnosed in the appendix, colon, and stomach. Biliary MANECs are particularly rare, and their histogenesis is debated because neuroendocrine cells are seldom identified in the normal biliary tract. They can show one of the 3 different architectural patterns described in Lewin's original classification: collision tumors, combined lesions, or amphicrine neoplasms. The neuroendocrine component is usually of a high grade, with small or large cell cytomorphology, whereas the adenocarcinoma component is either an intestinal or biliary type. Clinical presentation is characterized by locally advanced disease at the time of initial diagnosis. Recent studies suggest that treatment should be guided by the most aggressive histologic component.

  3. Teams and seams: skill mix in primary care.

    PubMed

    Jenkins-Clarke, S; Carr-Hill, R; Dixon, P

    1998-11-01

    The study described in this paper is set against a background of rapid changes in primary (community) care delivery in the United Kingdom (UK) and consequently the methodology of the study has been shaped by three broad issues - workforce changes, increase in workload and changing roles and boundaries. Ten 'ordinary' general practices (general practitioners (GPs) and the 'attached' community nurses) participated in the study and a large amount of data were collected over a 2-week observation period. Three study objectives are described, relating to workload, delegation and attitudes to delegation. The characteristics of the workload of the three main groups of community nurses (practice nurses, district nurses and health visitors) are described and compared. Thirty-nine per cent of all the GP consultations (836) had a delegatable element and 17% were deemed to be delegatable in their entirety. General practitioners most frequently referred to delegation to practice nurses in the current team and nurse practitioners in an enhanced team. The study identified the activities most amenable to delegation to these two groups of nurses. Attitudes to delegation were sought through focus group discussions, with reservations being expressed by both doctors and community nurses. This study provides evidence that GPs are prepared to delegate a considerable proportion of their workload; this clearly has implications for the nursing profession.

  4. Regulation of leucocyte homeostasis in the circulation.

    PubMed

    Scheiermann, Christoph; Frenette, Paul S; Hidalgo, Andrés

    2015-08-01

    The functions of blood cells extend well beyond the immune functions of leucocytes or the respiratory and hemostatic functions of erythrocytes and platelets. Seen as a whole, the bloodstream is in charge of nurturing and protecting all organs by carrying a mixture of cell populations in transit from one organ to another. To optimize these functions, evolution has provided blood and the vascular system that carries it with various mechanisms that ensure the appropriate influx and egress of cells into and from the circulation where and when needed. How this homeostatic control of blood is achieved has been the object of study for over a century, and although the major mechanisms that govern it are now fairly well understood, several new concepts and mediators have recently emerged that emphasize the dynamism of this liquid tissue. Here we review old and new concepts that relate to the maintenance and regulation of leucocyte homeostasis in blood and briefly discuss the mechanisms for platelets and red blood cells. Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2015. For permissions please email: journals.permissions@oup.com.

  5. Regulation of leucocyte homeostasis in the circulation

    PubMed Central

    Scheiermann, Christoph; Frenette, Paul S.; Hidalgo, Andrés

    2015-01-01

    The functions of blood cells extend well beyond the immune functions of leucocytes or the respiratory and hemostatic functions of erythrocytes and platelets. Seen as a whole, the bloodstream is in charge of nurturing and protecting all organs by carrying a mixture of cell populations in transit from one organ to another. To optimize these functions, evolution has provided blood and the vascular system that carries it with various mechanisms that ensure the appropriate influx and egress of cells into and from the circulation where and when needed. How this homeostatic control of blood is achieved has been the object of study for over a century, and although the major mechanisms that govern it are now fairly well understood, several new concepts and mediators have recently emerged that emphasize the dynamism of this liquid tissue. Here we review old and new concepts that relate to the maintenance and regulation of leucocyte homeostasis in blood and briefly discuss the mechanisms for platelets and red blood cells. PMID:25750191

  6. A Mixed Methods Study of Technology Integration in Rural Primary and Secondary High Schools in Jamaica

    ERIC Educational Resources Information Center

    Malcolm-Bell, Arlene

    2010-01-01

    The purpose of this concurrent nested mixed methods study was to explore and describe the status of technology integration in primary and secondary high schools in a rural parish in Jamaica to determine whether and how technological innovations were being used in instruction and learning. Diffusion theory is recommended as a framework to guide…

  7. A Mixed Methods Study of Technology Integration in Rural Primary and Secondary High Schools in Jamaica

    ERIC Educational Resources Information Center

    Malcolm-Bell, Arlene

    2010-01-01

    The purpose of this concurrent nested mixed methods study was to explore and describe the status of technology integration in primary and secondary high schools in a rural parish in Jamaica to determine whether and how technological innovations were being used in instruction and learning. Diffusion theory is recommended as a framework to guide…

  8. Primary Health Care Experiences of Hispanics with Serious Mental Illness: A Mixed-Methods Study

    PubMed Central

    Cabassa, Leopoldo J.; Gomes, Arminda P.; Meyreles, Quisqueya; Capitelli, Lucia; Younge, Richard; Dragatsi, Dianna; Alvarez, Juana; Nicasio, Andel; Druss, Benjamin; Lewis-Fernández, Roberto

    2013-01-01

    This mixed-methods study examines the primary health care experiences of Hispanic patients with serious mental illness. Forty patients were recruited from an outpatient mental health clinic. Participants reported a combination of perceived discrimination and stigmatization when receiving medical care. They rated the quality of chronic illness care as poor and reported low levels of self-efficacy and patient activation. These indicators were positively associated with how patients viewed their relationships with primary care providers. A grounded model was developed to describe the structural, social, and interpersonal processes that shaped participants’ primary care experiences. PMID:24162079

  9. Primary health care experiences of hispanics with serious mental illness: a mixed-methods study.

    PubMed

    Cabassa, Leopoldo J; Gomes, Arminda P; Meyreles, Quisqueya; Capitelli, Lucia; Younge, Richard; Dragatsi, Dianna; Alvarez, Juana; Nicasio, Andel; Druss, Benjamin; Lewis-Fernández, Roberto

    2014-11-01

    This mixed-methods study examines the primary health care experiences of Hispanic patients with serious mental illness. Forty patients were recruited from an outpatient mental health clinic. Participants reported a combination of perceived discrimination and stigmatization when receiving medical care. They rated the quality of chronic illness care as poor and reported low levels of self-efficacy and patient activation. These indicators were positively associated with how patients viewed their relationships with primary care providers. A grounded model was developed to describe the structural, social, and interpersonal processes that shaped participants' primary care experiences.

  10. Miniaturized LED primary optics design used for short-distance color mixing.

    PubMed

    Lee, Tsung-Xian; Tsai, Meng-Che; Chang, Shuo-Chieh; Liu, Kuei-Chun

    2016-11-10

    Color-tunable LED light fixtures generally change colors by controlling LEDs of multiple colors. This type of light source requires additional secondary optics and light-mixing distances to deliver color-mixing functions and perform high color uniformity. However, the color-mixing elements increase the optics size, resulting in more difficulties in making tiny lighting fixtures. Therefore, in this study, we introduce a LED primary optics design method that retains standard LED package size while featuring a color-mixing chamber. This method combines a lens having a rotational symmetry with a freeform profile and a zigzag structure by using double total internal reflection to disperse light uniformly. In contrast to a typical hemispherical lens, our design effectively lowers the weighted average color difference from 0.03 to 0.0035, and maintains optical efficiency of at least 90% without using any optical diffuser.

  11. [Primary branch size of Pinus koraiensis plantation: a prediction based on linear mixed effect model].

    PubMed

    Dong, Ling-Bo; Liu, Zhao-Gang; Li, Feng-Ri; Jiang, Li-Chun

    2013-09-01

    By using the branch analysis data of 955 standard branches from 60 sampled trees in 12 sampling plots of Pinus koraiensis plantation in Mengjiagang Forest Farm in Heilongjiang Province of Northeast China, and based on the linear mixed-effect model theory and methods, the models for predicting branch variables, including primary branch diameter, length, and angle, were developed. Considering tree effect, the MIXED module of SAS software was used to fit the prediction models. The results indicated that the fitting precision of the models could be improved by choosing appropriate random-effect parameters and variance-covariance structure. Then, the correlation structures including complex symmetry structure (CS), first-order autoregressive structure [AR(1)], and first-order autoregressive and moving average structure [ARMA(1,1)] were added to the optimal branch size mixed-effect model. The AR(1) improved the fitting precision of branch diameter and length mixed-effect model significantly, but all the three structures didn't improve the precision of branch angle mixed-effect model. In order to describe the heteroscedasticity during building mixed-effect model, the CF1 and CF2 functions were added to the branch mixed-effect model. CF1 function improved the fitting effect of branch angle mixed model significantly, whereas CF2 function improved the fitting effect of branch diameter and length mixed model significantly. Model validation confirmed that the mixed-effect model could improve the precision of prediction, as compare to the traditional regression model for the branch size prediction of Pinus koraiensis plantation.

  12. The effects of temporal variability of mixed layer depth on primary productivity around Bermuda

    NASA Technical Reports Server (NTRS)

    Bissett, W. Paul; Meyers, Mark B.; Walsh, John J.; Mueller-Karger, Frank E.

    1994-01-01

    Temporal variations in primary production and surface chlorophyll concentrations, as measured by ship and satellite around Bermuda, were simulated with a numerical model. In the upper 450 m of the water column, population dynamics of a size-fractionated phytoplankton community were forced by daily changes of wind, light, grazing stress, and nutrient availability. The temporal variations of production and chlorophyll were driven by changes in nutrient introduction to the euphotic zone due to both high- and low-frequency changes of the mixed layer depth within 32 deg-34 deg N, 62 deg-64 deg W between 1979 and 1984. Results from the model derived from high-frequency (case 1) changes in the mixed layer depth showed variations in primary production and peak chlorophyll concentrations when compared with results from the model derived from low-frequency (case 2) mixed layer depth changes. Incorporation of size-fractionated plankton state variables in the model led to greater seasonal resolution of measured primary production and vertical chlorophyll profiles. The findings of this study highlight the possible inadequacy of estimating primary production in the sea from data of low-frequency temporal resolution and oversimplified biological simulations.

  13. The effects of temporal variability of mixed layer depth on primary productivity around Bermuda

    NASA Technical Reports Server (NTRS)

    Bissett, W. Paul; Meyers, Mark B.; Walsh, John J.; Mueller-Karger, Frank E.

    1994-01-01

    Temporal variations in primary production and surface chlorophyll concentrations, as measured by ship and satellite around Bermuda, were simulated with a numerical model. In the upper 450 m of the water column, population dynamics of a size-fractionated phytoplankton community were forced by daily changes of wind, light, grazing stress, and nutrient availability. The temporal variations of production and chlorophyll were driven by changes in nutrient introduction to the euphotic zone due to both high- and low-frequency changes of the mixed layer depth within 32 deg-34 deg N, 62 deg-64 deg W between 1979 and 1984. Results from the model derived from high-frequency (case 1) changes in the mixed layer depth showed variations in primary production and peak chlorophyll concentrations when compared with results from the model derived from low-frequency (case 2) mixed layer depth changes. Incorporation of size-fractionated plankton state variables in the model led to greater seasonal resolution of measured primary production and vertical chlorophyll profiles. The findings of this study highlight the possible inadequacy of estimating primary production in the sea from data of low-frequency temporal resolution and oversimplified biological simulations.

  14. Turbulent Mixing of Primary and Secondary Flow Streams in a Rocket-Based Combined Cycle Engine

    NASA Technical Reports Server (NTRS)

    Cramer, J. M.; Greene, M. U.; Pal, S.; Santoro, R. J.; Turner, Jim (Technical Monitor)

    2002-01-01

    This viewgraph presentation gives an overview of the turbulent mixing of primary and secondary flow streams in a rocket-based combined cycle (RBCC) engine. A significant RBCC ejector mode database has been generated, detailing single and twin thruster configurations and global and local measurements. On-going analysis and correlation efforts include Marshall Space Flight Center computational fluid dynamics modeling and turbulent shear layer analysis. Potential follow-on activities include detailed measurements of air flow static pressure and velocity profiles, investigations into other thruster spacing configurations, performing a fundamental shear layer mixing study, and demonstrating single-shot Raman measurements.

  15. Complexes with mixed primary and secondary cellulose synthases are functional in Arabidopsis thaliana plants

    SciTech Connect

    Carroll, Andrew; Mansoori, N; Li, Shundai; Lei, Lei; Vernhettes, Samantha; Visser, Richard G. F.; Somerville, Chris R; Gu, Ying; Trindade, Luisa M.

    2012-10-01

    In higher plants, cellulose is synthesized by so-called rosette protein complexes with cellulose synthases (CESAs) as catalytic subunits of the complex. The CESAs are divided into two distinct families, three of which are thought to be specialized for the primary cell wall and three for the secondary cell wall. In this article, the potential of primary and secondary CESAs forming a functional rosette complex has been investigated. The membrane-based yeast two-hybrid and biomolecular fluorescence systems were used to assess the interactions between three primary (CESA1, CESA3, CESA6), and three secondary (CESA4, CESA7, CESA8) Arabidopsis (Arabidopsis thaliana) CESAs. The results showed that all primary CESAs can physically interact both in vitro and in planta with all secondary CESAs. Although CESAs are broadly capable of interacting in pairwise combinations, they are not all able to form functional complexes in planta. Analysis of transgenic lines showed that CESA7 can partially rescue defects in the primary cell wall biosynthesis in a weak cesa3 mutant. Green fluorescent protein-CESA protein fusions revealed that when CESA3 was replaced by CESA7 in the primary rosette, the velocity of the mixed complexes was slightly faster than the native primary complexes. CESA1 in turn can partly rescue defects in secondary cell wall biosynthesis in a cesa8ko mutant, resulting in an increase of cellulose content relative to cesa8ko. These results demonstrate that sufficient parallels exist between the primary and secondary complexes for cross-functionality and open the possibility that mixed complexes of primary and secondary CESAs may occur at particular times.

  16. Enumeration of semen leucocytes by fluorescence in situ hybridisation technique

    PubMed Central

    Conte, R A; Luke, S; Verma, R S

    1995-01-01

    Aim—To determine whether the fluorescent in situ hybridisation technique (FISH) using a total human DNA genomic probe can be used to enumerate semen leucocytes. Methods—Semen samples from five donors were subjected to a mild KC1 solution. These samples were then biotin labelled under FISH conditions using a total human DNA genomic probe and the leucocyte counts were determined. To check the accuracy of the technique a monoclonal antibody against the common leucocyte antigen CD45 [KC56(T-200)] served as a control. An isotypic control for [KC56(T-200)], the immunoglobulin [MsIgG1], served as a secondary control. Results—Semen leucocytes stained by the FISH technique were easily detected because of their distinct bright yellow colour, while the sperm cells were red. The leucocyte count ranged from 0·5 to 4·9 × 106 per ml of semen. KC56(T-200) and its isotypic control MsIgG1, which served as control for the FISH technique, accurately identified 94% and 97% of the semen leucocytes of a control donor, respectively. Conclusions—The FISH technique using a total human DNA probe can accurately and effectively enumerate the overall leucocyte population in semen. Images PMID:16696031

  17. Primary Student-Teachers' Conceptual Understanding of the Greenhouse Effect: A mixed method study

    NASA Astrophysics Data System (ADS)

    Ratinen, Ilkka Johannes

    2013-04-01

    The greenhouse effect is a reasonably complex scientific phenomenon which can be used as a model to examine students' conceptual understanding in science. Primary student-teachers' understanding of global environmental problems, such as climate change and ozone depletion, indicates that they have many misconceptions. The present mixed method study examines Finnish primary student-teachers' understanding of the greenhouse effect based on the results obtained via open-ended and closed-form questionnaires. The open-ended questionnaire considers primary student-teachers' spontaneous ideas about the greenhouse effect depicted by concept maps. The present study also uses statistical analysis to reveal respondents' conceptualization of the greenhouse effect. The concept maps and statistical analysis reveal that the primary student-teachers' factual knowledge and their conceptual understanding of the greenhouse effect are incomplete and even misleading. In the light of the results of the present study, proposals for modifying the instruction of climate change in science, especially in geography, are presented.

  18. The polyphosphoinositide content of the leucocyte, erythrocyte and macrophage

    PubMed Central

    Wieneke, Antonnette A.; Woodin, A. M.

    1967-01-01

    1. The polyphosphoinositide content of macrophages and the cell membranes of leucocytes and erythrocytes was determined by an extension of the `acid-hydrolysis' procedure of Dawson & Eichberg (1965). The estimation was controlled by adding a little highly radioactive polyphosphoinositide to the tissue extracts before fractionation. Several standard methods for determining polyphosphoinositides gave low recoveries when applied to leucocytes, and it is suggested that these cells contain materials that form complexes with the polyphosphoinositides and interfere with the assay. 2. The method for the preparation of leucocyte cell surface membranes has been modified. PMID:16742528

  19. Circulating leucocytes perpetuate stroke-induced aortic dysfunction.

    PubMed

    Asano, Shinichi; O'Connell, Grant C; Lemaster, Kent C; DeVallance, Evan R; Branyan, Kayla W; Simpkins, James W; Frisbee, Jefferson C; Barr, Taura L; Chantler, Paul D

    2017-07-24

    What is the central question of this study? Does a stroke event influence aortic endothelial function; and what is the role of peripheral circulating leucocytes in stroke on the vascular reactivity of the aorta? What is the main finding and its importance? In vitro co-culture experiments demonstrated that aortic endothelium-dependent relaxation was impaired when rat aortic rings were co-cultured with leucocytes stimulated with serum from stroke patients. Impaired vascular reactivity was not observed in aortic rings without leucocytes stimulated with serum from stroke patients or age-matched control patients with or without leucocytes. These data suggest that leucocyte-dependent altered aortic endothelium-dependent relaxation with stroke and the systemic consequences of stroke on vascular inflammation may occur in the aorta. Post-stroke inflammation has been linked to poor stroke outcomes. The vascular endothelium senses and responds to circulating factors, in particular inflammatory cytokines. Although stroke-associated local cerebrovascular dysfunction is well reported, the effects of a stroke on conduit artery function are not fully understood. We tested the hypothesis that serum from stroke patients triggers leucocyte-dependent aortic endothelial dysfunction that is associated with elevated concentrations of cytokines. Total leucocytes were isolated from healthy individuals, and the cells were incubated in serum from control subjects or stroke patients for 6 h. The quantity of cytokines in media was determined using an immunoassay. Vascular reactivity was determined by the rat aortic rings that were co-cultured with or without leucocytes and stimulated with serum samples from control subjects or stroke patients. Endothelium-dependent dilatation was significantly impaired in aortic rings co-cultured with leucocytes plus serum from stroke patients (50 ± 30 versus 85 ± 13%, P < 0.05) versus serum from control subjects. In contrast, no difference was

  20. In search of mixed anxiety-depressive disorder: a primary care study.

    PubMed

    Means-Christensen, Adrienne J; Sherbourne, Cathy D; Roy-Byrne, Peter P; Schulman, Martin C; Wu, Jennifer; Dugdale, David C; Lessler, Daniel; Stein, Murray B

    2006-01-01

    The diagnosis of mixed anxiety-depressive disorder, as proposed in DSM-IV, is intended to be useful in settings such as primary care, where low-level anxiety and depressive symptoms may cause clinically significant impairment but are undiagnosable using current criteria. Evidence of the prevalence of this diagnosis is, however, lacking, particularly since the publication of the proposed diagnostic criteria in DSM-IV. Our study examined symptoms of anxiety and depression in 65 primary care patients screened for anxiety and depression while visiting their doctor. Results indicated that of the 37 patients without a diagnosable anxiety or depressive disorder, none had symptoms of depression and anxiety accompanied by interference that the patient deemed significant and attributable to his or her symptoms. These data dispute the need for a mixed anxiety-depression category (beyond mood and anxiety syndromes currently in DSM-IV) in future editions of the DSM.

  1. Validation of ACG Case-mix for equitable resource allocation in Swedish primary health care

    PubMed Central

    Zielinski, Andrzej; Kronogård, Maria; Lenhoff, Håkan; Halling, Anders

    2009-01-01

    Background Adequate resource allocation is an important factor to ensure equity in health care. Previous reimbursement models have been based on age, gender and socioeconomic factors. An explanatory model based on individual need of primary health care (PHC) has not yet been used in Sweden to allocate resources. The aim of this study was to examine to what extent the ACG case-mix system could explain concurrent costs in Swedish PHC. Methods Diagnoses were obtained from electronic PHC records of inhabitants in Blekinge County (approx. 150,000) listed with public PHC (approx. 120,000) for three consecutive years, 2004-2006. The inhabitants were then classified into six different resource utilization bands (RUB) using the ACG case-mix system. The mean costs for primary health care were calculated for each RUB and year. Using linear regression models and log-cost as dependent variable the adjusted R2 was calculated in the unadjusted model (gender) and in consecutive models where age, listing with specific PHC and RUB were added. In an additional model the ACG groups were added. Results Gender, age and listing with specific PHC explained 14.48-14.88% of the variance in individual costs for PHC. By also adding information on level of co-morbidity, as measured by the ACG case-mix system, to specific PHC the adjusted R2 increased to 60.89-63.41%. Conclusion The ACG case-mix system explains patient costs in primary care to a high degree. Age and gender are important explanatory factors, but most of the variance in concurrent patient costs was explained by the ACG case-mix system. PMID:19765286

  2. In vitro interactions between Neoparamoeba spp. and salmonid leucocytes; The effect of parasite sonicate on anterior kidney leucocyte function

    USGS Publications Warehouse

    Gross, K.; Alcorn, S.; Murray, A.; Morrison, R.; Nowak, B.

    2006-01-01

    Sonicated Neoparamoeba spp. (Nspp) did not affect the in vitro respiratory burst response of leucocytes isolated from Atlantic salmon Salmo salar, rainbow trout Oncorhynchus mykiss and chinook salmon Oncorhynchus tshawytscha anterior kidneys (P > 0.05). Atlantic salmon and chinook salmon leucocytes pre-incubated with the parasites, however, responded to phorbol myristate acetate (PMA) stimulation with a greater response compared to cells incubated with PMA on its own (P < 0.05). Sonicated Nspp was not chemo-attractive for anterior kidney leucocytes isolated from all three fish species. ?? 2006 The Fisheries Society of the British Isles.

  3. Characteristics of the transport of ascorbic acid into leucocytes

    SciTech Connect

    Raghoebar, M.; Huisman, J.A.M.; van den Berg, W.B.; van Ginneken, C.A.M.

    1987-02-02

    The degree and the mode of association of (/sup 14/C)-ascorbic acid with leucocytes are examined. The degree of association of ascorbic acid with polymorphonuclear leucocytes (1-3 %) is dependent on cell type, extracellular concentration of ascorbic acid, incubation temperature, intactness of the cells and the extracellular pH. All experiments are performed according to strict protocols as these compounds are labile in aqueous solutions. Further it is noticed that in all experiments an outward gradient of leucocyte endogenic ascorbic acid exists. The results suggest that the association process comprises at least one saturable pathway. The activation of polymorphonuclear leucocytes by phorbol myristate acetate increases the accumulation of ascorbic acid threefold. 30 references, 7 figures, 3 tables.

  4. Screening mixed depression and bipolarity in the postpartum period at a primary health care center.

    PubMed

    Çelik, Sercan Bulut; Bucaktepe, Gamze Erten; Uludağ, Ayşegül; Bulut, İbrahim Umud; Erdem, Özgür; Altınbaş, Kürşat

    2016-11-01

    Mixed depression is a clinical condition accompanied by the symptoms of (hypo)mania and is considered to be a predictor for bipolar disorder. Compared to pure major depression, mixed depression is worse in progress. There are limited data on the prevalence of mixed depression since it is a relatively new entity. Therefore, the present study aimed to investigate the prevalence of mixed depression during the postpartum period which is risky for mood disorders. The study included 63 postpartum women. The participants were administered Beck Depression Scale, Edinburgh Postnatal Depression Scale (EPDS), Mood Disorders Questionnaire (MDQ), and Modified Hypomania Symptom Checklist-32 (mHCL-32). The MDQ scores of the women with expected depression according to the EPDS cut-off scores, were significantly higher than the women with lower EPDS scores (t=-4.968; p<0.001). The modified hypomania scores were significantly higher in the women with higher depression scores compared to the women under EPDS cut-off scores (t=-4.713; p<0.001). According to the EPDS and BDS results, 27 (42.9%) and 14 (22.2%) women needed additional clinical examination for depression, respectively. In addition, 3 (4.8%) women require additional clinical examination for bipolar disorder. The scores for the first item of MDQ were above the cut-off value in 11 (17.5%) women. According to the mHCL-32 results, 50 (79.4%) women had at least 1 symptom, 45 (71.4%) women had at least 3 symptoms, and 43 (68.3%) women had at least 5 symptoms of mixed depression. Postpartum mixed depression should be promptly diagnosed by using appropriate diagnostic tools, particularly by primary health care physicians. Patients with mixed depression should be closely monitored to avoid manic switch. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. Effectiveness of transdiagnostic Internet cognitive behavioural treatment for mixed anxiety and depression in primary care.

    PubMed

    Newby, Jill M; Mewton, Louise; Williams, Alishia D; Andrews, Gavin

    2014-08-01

    Internet-delivered cognitive behavioural treatment (iCBT) has been shown to be effective for the combined treatment of depression and anxiety in randomised controlled trials. The degree to which these findings generalise to patients in primary care awaits further investigation. Using an open-trial design, we investigated adherence to, and effectiveness of a 6-lesson therapist-assisted iCBT program for mixed anxiety and depression for patients (n = 707) who completed the program under the supervision of primary care clinicians (general practitioners, psychologists and other allied health professionals). Primary outcome measures were the PHQ-9 (depression), GAD-7 (generalised anxiety), K-10 (distress), WHODAS-II (disability), mini-SPIN (social anxiety) and panic disorder severity scale self-report version (PDSS). Adherence to the iCBT program was modest (47.3%), but within-subjects effect sizes ranged from medium (0.51 for PDSS) to large (1.20 for PHQ-9). The lack of control group, limited post-treatment data due to drop-out, and short follow-up period. iCBT is an effective treatment for mixed depression and anxiety when delivered in primary care settings. Methods to increase adherence are needed to optimise the benefits to patients. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. The primary and mixed dentition, post-eruptive enamel maturation and dental caries: a review.

    PubMed

    Lynch, Richard J M

    2013-12-01

    The mouth is in flux from the time the primary teeth begin to erupt, in the first year of life, through to the end of the 'mixed dentition' (i.e. the concurrent eruption of the permanent teeth and exfoliation of the primary teeth), at around 12 years of age. Primary teeth facilitate the development of the facial muscles and speech. They act as 'guides' for erupting permanent teeth. If lost prematurely, subsequent misalignment of permanent teeth can make them difficult to clean and possibly more caries-prone. During the mixed dentition phase, teeth are at relatively high risk of caries. Erupting teeth are difficult to clean and cleaning may be avoided because of tender gums and behavioural factors in children. Permanent enamel (and possibly primary enamel) undergoes post-eruptive maturation, accumulating fluoride, becoming harder, less porous and less caries-prone. Overall, primary teeth are more vulnerable to caries than permanent teeth. Widespread use of fluoride toothpaste has effected marked reductions in caries. Some evidence exists that fluoride delivered from toothpastes may be somewhat more effective in reducing caries in primary than in permanent teeth. However, caries remains a public health concern globally. New fluoride toothpaste formulations, optimised using in vivo fluoride delivery and efficacy studies, may improve the caries resistance of mineral deposited during post-eruptive maturation. Behaviour should not be ignored; new formulations will be more effective if used according to professionally endorsed recommendations based on sound science. Establishing good oral hygiene behaviour early in life can lead to lasting anti-caries benefits. © 2013 FDI World Dental Federation.

  7. Avian leucocyte counting using the hemocytometer

    USGS Publications Warehouse

    Dein, F.J.; Wilson, A.; Fischer, D.; Langenberg, P.

    1994-01-01

    Automated methods for counting leucocytes in avian blood are not available because of the presence of nucleated erythrocytes and thrombocytes. Therefore, total white blood cell counts are performed by hand using a hemocytometer. The Natt and Herrick and the Unopette methods are the most common stain and diluent preparations for this procedure. Replicate hemocytometer counts using these two methods were performed on blood from four birds of different species. Cells present in each square of the hemocytometer were counted. Counting cells in the corner, side, or center hemocytometer squares produced statistically equivalent results; counting four squares per chamber provided a result similar to that obtained by counting nine squares; and the Unopette method was more precise for hemocytometer counting than was the Natt and Herrick method. The Unopette method is easier to learn and perform but is an indirect process, utilizing the differential count from a stained smear. The Natt and Herrick method is a direct total count, but cell identification is more difficult.

  8. Connecting the Dots and Merging Meaning: Using Mixed Methods to Study Primary Care Delivery Transformation

    PubMed Central

    Scammon, Debra L; Tomoaia-Cotisel, Andrada; Day, Rachel L; Day, Julie; Kim, Jaewhan; Waitzman, Norman J; Farrell, Timothy W; Magill, Michael K

    2013-01-01

    Objective. To demonstrate the value of mixed methods in the study of practice transformation and illustrate procedures for connecting methods and for merging findings to enhance the meaning derived. Data Source/Study Setting. An integrated network of university-owned, primary care practices at the University of Utah (Community Clinics or CCs). CC has adopted Care by Design, its version of the Patient Centered Medical Home. Study Design. Convergent case study mixed methods design. Data Collection/Extraction Methods. Analysis of archival documents, internal operational reports, in-clinic observations, chart audits, surveys, semistructured interviews, focus groups, Centers for Medicare and Medicaid Services database, and the Utah All Payer Claims Database. Principal Findings. Each data source enriched our understanding of the change process and understanding of reasons that certain changes were more difficult than others both in general and for particular clinics. Mixed methods enabled generation and testing of hypotheses about change and led to a comprehensive understanding of practice change. Conclusions. Mixed methods are useful in studying practice transformation. Challenges exist but can be overcome with careful planning and persistence. PMID:24279836

  9. Connecting the dots and merging meaning: using mixed methods to study primary care delivery transformation.

    PubMed

    Scammon, Debra L; Tomoaia-Cotisel, Andrada; Day, Rachel L; Day, Julie; Kim, Jaewhan; Waitzman, Norman J; Farrell, Timothy W; Magill, Michael K

    2013-12-01

    To demonstrate the value of mixed methods in the study of practice transformation and illustrate procedures for connecting methods and for merging findings to enhance the meaning derived. An integrated network of university-owned, primary care practices at the University of Utah (Community Clinics or CCs). CC has adopted Care by Design, its version of the Patient Centered Medical Home. Convergent case study mixed methods design. Analysis of archival documents, internal operational reports, in-clinic observations, chart audits, surveys, semistructured interviews, focus groups, Centers for Medicare and Medicaid Services database, and the Utah All Payer Claims Database. Each data source enriched our understanding of the change process and understanding of reasons that certain changes were more difficult than others both in general and for particular clinics. Mixed methods enabled generation and testing of hypotheses about change and led to a comprehensive understanding of practice change. Mixed methods are useful in studying practice transformation. Challenges exist but can be overcome with careful planning and persistence. © Health Research and Educational Trust.

  10. Primary Care Resident Training for Obesity, Nutrition, and Physical Activity Counseling: A Mixed-Methods Study.

    PubMed

    Antognoli, Elizabeth L; Seeholzer, Eileen L; Gullett, Heidi; Jackson, Brigid; Smith, Samantha; Flocke, Susan A

    2017-09-01

    National guidelines have been established to support the role of primary care physicians in addressing obesity. Preparing primary care residents to recognize and treat overweight/obesity has been identified as an essential component of postgraduate medical training that is currently lacking. This study aims to identify how primary care residency programs are preparing physicians to counsel about obesity, nutrition, and physical activity (ONPA) and to examine program members' perspectives regarding the place of ONPA counseling in the curriculum, and its relevance in primary care training. Using mixed methods, we collected and analyzed data on 25 family medicine, internal medicine, and obstetrics/gynecology residency programs across Ohio. Programs averaged 2.8 hours of ONPA-related didactics per year. Ten programs (42%) taught techniques for health behavior counseling. Having any ONPA-related didactics was associated with greater counseling knowledge (p = .01) among residents but poorer attitudes (p < .001) and poorer perceived professional norms (p = .004) toward ONPA counseling. Findings from interview data highlighted similar perceived barriers to ONPA counseling across all three specialties but variation in perception of responsibility to provide ONPA counseling. While widespread expectations that primary care physicians counsel their overweight and obese patients prevail, few residency programs provide training to support such counseling.

  11. Antinuclear antibodies in scleroderma, mixed connective tissue disease and "primary" Raynaud's phenomenon.

    PubMed

    Cruz, M; Mejia, G; Lavalle, C; Cortes, J J; Reyes, P A

    1988-03-01

    The diversity of antibodies in patients with scleroderma, mixed connective tissue disease or "primary" Raynaud's phenomenon could be used as a laboratory aid in the clinical diagnosis. In serum samples of 75 patients we screened for antinuclear antibodies (HEp 2 cells), anti DNA, soluble nucleoprotein and extractable nuclear antigens (Sm, rRNP, U1-nRNP, SSA/Ro, SSB/La and Scl-70). Distinctive antinuclear antibodies pattern was identified in each group of patients. This immunologic profile is valuable for clinical diagnosis and the preferential association of certain autoantibodies with some diseases and not with others, suggest an antigen-driven stimulus for its production.

  12. Human leucocytes in asthenozoospermic patients: endothelial nitric oxide synthase expression.

    PubMed

    Buldreghini, E; Hamada, A; Macrì, M L; Amoroso, S; Boscaro, M; Lenzi, A; Agarwal, A; Balercia, G

    2014-12-01

    In a basic study at the Andrology Unit, Department of Clinical and Molecular Sciences, Polytechnic University of Marche, Ancona, Italy, we evaluated the pattern of mRNA endothelial nitric oxide synthase (eNOS) expression in human blood leucocytes isolated from normozoospermic fertile and asthenozoospermic infertile men to elucidate any pathogenic involvement in sperm cell motility. Forty infertile men with idiopathic asthenozoospermia and 45 normozoospermic fertile donors, age-matched, were included. Semen parameters were evaluated, and expression analysis of mRNA was performed in human leucocytes using reverse transcription polymerase chain reaction. Sperm volume, count, motility and morphology were determined, and eNOS expression and Western blotting analyses were performed. A positive correlation was observed between the concentrations of NO and the percentage of immotile spermatozoa. The mRNA of eNOS was more expressed in peripheral blood leucocytes isolated from asthenozoospermic infertile men versus those of fertile normozoospermic men (7.46 ± 0.38 versus 7.06 ± 0.56, P = 0.0355). A significant up-regulation of eNOS gene in peripheral blood leucocytes was 1.52-fold higher than that of fertile donors. It is concluded that eNOS expression and activity are enhanced in blood leucocytes in men with idiopathic asthenozoospermia.

  13. Characteristics of premaxillary supernumerary teeth in primary and mixed dentitions: a retrospective analysis of 212 cases.

    PubMed

    Shekhar, Madiraju G

    2012-08-01

    To investigate the characteristics and distribution of premaxillary supernumerary teeth affecting primary and mixed dentitions in Indian children. This retrospective analysis included 11 200 children, aged 3-12 years, who attended a pediatric dental clinic for dental care during the period 2007-2010. The children were divided into group I (aged 3-6 years) and group II (aged 7-12 years), and data regarding maxillary anterior supernumerary teeth, diagnosed both as isolated and/or as unexpected findings during routine clinical and radiological examinations, were gathered. Pearson's χ(2) -test, with a 0.05 level of significance, was used for the analysis. The overall prevalence of premaxillary supernumerary teeth in primary and mixed dentitions was 1.9%. Single supernumerary teeth (84.9%) and conical morphology (68.7%) were commonly seen in both groups. The most common sagittal position was palatal (92.3%) among both erupted and impacted maxillary anterior supernumerary teeth. The prevalence of premaxillary supernumerary teeth in Indian children found in this study was 1.9%, with an overall male to female ratio of 1.7:1. Single supernumerary teeth, conical morphology, and erupted supernumerary teeth were the most commonly seen. Supernumerary teeth associated with clinical complications were relatively low (48%), and axial rotation or displacement of maxillary incisors was the most common sequelae. © 2012 Blackwell Publishing Asia Pty Ltd.

  14. Human primary mixed brain cultures: preparation, differentiation, characterization and application to neuroscience research.

    PubMed

    Ray, Balmiki; Chopra, Nipun; Long, Justin M; Lahiri, Debomoy K

    2014-09-16

    Culturing primary cortical neurons is an essential neuroscience technique. However, most cultures are derived from rodent brains and standard protocols for human brain cultures are sparse. Herein, we describe preparation, maintenance and major characteristics of a primary human mixed brain culture, including neurons, obtained from legally aborted fetal brain tissue. This approach employs standard materials and techniques used in the preparation of rodent neuron cultures, with critical modifications. This culture has distinct differences from rodent cultures. Specifically, a significant numbers of cells in the human culture are derived from progenitor cells, and the yield and survival of the cells grossly depend on the presence of bFGF. In the presence of bFGF, this culture can be maintained for an extended period. Abundant productions of amyloid-β, tau and proteins make this a powerful model for Alzheimer's research. The culture also produces glia and different sub-types of neurons. We provide a well-characterized methodology for human mixed brain cultures useful to test therapeutic agents under various conditions, and to carry forward mechanistic and translational studies for several brain disorders.

  15. Return on Investment in Electronic Health Records in Primary Care Practices: A Mixed-Methods Study

    PubMed Central

    Sanche, Steven

    2014-01-01

    Background The use of electronic health records (EHR) in clinical settings is considered pivotal to a patient-centered health care delivery system. However, uncertainty in cost recovery from EHR investments remains a significant concern in primary care practices. Objective Guided by the question of “When implemented in primary care practices, what will be the return on investment (ROI) from an EHR implementation?”, the objectives of this study are two-fold: (1) to assess ROI from EHR in primary care practices and (2) to identify principal factors affecting the realization of positive ROI from EHR. We used a break-even point, that is, the time required to achieve cost recovery from an EHR investment, as an ROI indicator of an EHR investment. Methods Given the complexity exhibited by most EHR implementation projects, this study adopted a retrospective mixed-method research approach, particularly a multiphase study design approach. For this study, data were collected from community-based primary care clinics using EHR systems. Results We collected data from 17 primary care clinics using EHR systems. Our data show that the sampled primary care clinics recovered their EHR investments within an average period of 10 months (95% CI 6.2-17.4 months), seeing more patients with an average increase of 27% in the active-patients-to-clinician-FTE (full time equivalent) ratio and an average increase of 10% in the active-patients-to-clinical-support-staff-FTE ratio after an EHR implementation. Our analysis suggests, with a 95% confidence level, that the increase in the number of active patients (P=.006), the increase in the active-patients-to-clinician-FTE ratio (P<.001), and the increase in the clinic net revenue (P<.001) are positively associated with the EHR implementation, likely contributing substantially to an average break-even point of 10 months. Conclusions We found that primary care clinics can realize a positive ROI with EHR. Our analysis of the variances in the

  16. Return on investment in electronic health records in primary care practices: a mixed-methods study.

    PubMed

    Jang, Yeona; Lortie, Michel A; Sanche, Steven

    2014-09-29

    The use of electronic health records (EHR) in clinical settings is considered pivotal to a patient-centered health care delivery system. However, uncertainty in cost recovery from EHR investments remains a significant concern in primary care practices. Guided by the question of "When implemented in primary care practices, what will be the return on investment (ROI) from an EHR implementation?", the objectives of this study are two-fold: (1) to assess ROI from EHR in primary care practices and (2) to identify principal factors affecting the realization of positive ROI from EHR. We used a break-even point, that is, the time required to achieve cost recovery from an EHR investment, as an ROI indicator of an EHR investment. Given the complexity exhibited by most EHR implementation projects, this study adopted a retrospective mixed-method research approach, particularly a multiphase study design approach. For this study, data were collected from community-based primary care clinics using EHR systems. We collected data from 17 primary care clinics using EHR systems. Our data show that the sampled primary care clinics recovered their EHR investments within an average period of 10 months (95% CI 6.2-17.4 months), seeing more patients with an average increase of 27% in the active-patients-to-clinician-FTE (full time equivalent) ratio and an average increase of 10% in the active-patients-to-clinical-support-staff-FTE ratio after an EHR implementation. Our analysis suggests, with a 95% confidence level, that the increase in the number of active patients (P=.006), the increase in the active-patients-to-clinician-FTE ratio (P<.001), and the increase in the clinic net revenue (P<.001) are positively associated with the EHR implementation, likely contributing substantially to an average break-even point of 10 months. We found that primary care clinics can realize a positive ROI with EHR. Our analysis of the variances in the time required to achieve cost recovery from EHR

  17. OBSERVATIONS ON THE PRODUCTION OF PYROGENIC SUBSTANCES BY RABBIT AND HUMAN LEUCOCYTES

    PubMed Central

    Fessler, J. H.; Cooper, K. E.; Cranston, W. I.; Vollum, R. L.

    1961-01-01

    1. The mechanism of release of a pyrogen from leucocytes has been studied in cells obtained from sterile rabbit peritoneal exudates and from rabbit blood. Attempts were made to induce human leucocytes—from blood—to release a pyrogen. 2. Rabbit leucocytes, kept below 4°C., were not pyrogenic and did not release any pyrogen when disintegrated. Incubating such cells, in various media, at 37°C. led to the formation of a pyrogen which was heat-labile. The maximum yield was attained after 1½ hours' incubation. 3. The formation of rabbit leucocytic pyrogen was prevented by freezing and thawing the leucocytes, by heating them to 56°C. for half an hour before incubation, and by ageing them in the cold. 4. Nitrofurazone (5-nitro-2-furaldehyde semicarbazone) prevents the formation of leucocytic pyrogen when given by mouth to the cell-donor animals, or when added to leucocytes in intro. 5. Leucocytes from rabbit blood formed leucocytic pyrogen, on incubation in saline, and this formation was also inhibited by nitrofurazone. 6. No leucocytic pyrogen was released from human leucocytes subjected to mechanical, osmotic, or thermal damage, and it was not formed when the cells were incubated in saline. 7. The source of rabbit leucocytic pyrogen, the action of nitrofurazone on leucocytes, and the supposed role of leucocytic pyrogen in fever are discussed. PMID:13699218

  18. ISOLATION, CHARACTERIZATION, AND DISTRIBUTION OF ACID MUCOPOLYSACCHARIDES IN RABBIT LEUCOCYTES

    PubMed Central

    Fedorko, Martha E.; Morse, Stephen I.

    1965-01-01

    Acid mucopolysaccharides have been extracted from whole rabbit polymorphonuclear leucocytes and from the cytoplasmic granules of these cells. The leucocyte acid mucopolysaccharides can be separated into two fractions by the solubility of their CPC complexes in solutions of differing salt concentration. One of these fractions appears to be identical with hyaluronic acid; the other appears to be an atypical chondroitin sulfate. On both a dry weight and total protein basis the polymorphonuclear leucocyte granule contains approximately 2.6 times as much acid mucopolysaccharide as does the whole cell. Hyaluronic acid is concentrated in the granules in particular; its function is unknown. These results do not indicate that all lysosomes contain abundant acid mucopolysaccharides, for no detectable carbohydrate of this class could be extracted from lysosome-rich alveolar macrophages. PMID:14253486

  19. Arteriovenous fistula stent infection diagnosed with radiolabelled leucocyte scintigraphy.

    PubMed

    Yoo, Jeong Rae; Heo, Sang Taek; Kim, Miyeon; Kim, Hyun Woo; Chang, Jee Won; Song, Heesung

    2015-07-01

    Infectious complications of haemodialysis in patients with arteriovenous fistula stent are rare. In addition, patients with low-grade infection are more difficult to diagnose. Here, we report the first case of low-grade infection of an arteriovenous fistula stent diagnosed using (99m)Tc-hexamethylpropylene amine oxime (HMPAO)-labelled leucocyte scintigraphy. A 62-year-old man with end-stage renal disease was referred for prolonged fever. We performed (99m)Tc-HMPAO-labelled leucocyte scintigraphy following a work-up according to fever of unknown origin. A focal uptake on the left forearm compatible with the arteriovenous fistula stent insertion site was shown, and the stent was removed. (99m)Tc-HMPAO-labelled leucocyte scintigraphy could be a suitable method for assessing vascular stent infection in low-grade fever.

  20. The sheep analogue of leucocyte common antigen (LCA).

    PubMed Central

    Maddox, J F; Mackay, C R; Brandon, M R

    1985-01-01

    The tissue distribution and immunochemical properties of antigens recognized by two monoclonal antibodies 1.11.32 and 1.28.124 define the sheep analogue of the leucocyte common antigen found in rat, man and mouse. Histological and immunofluorescent studies show that this antigen is found on all lymphocytes, as well as other leucocytes but is absent from non-leucocytic cells. Immunochemical data show that a series of proteins of high molecular weight (190,000-225,000) are recognized, and histological studies show the presence of this antigen on a subpopulation of fetal liver cells as early as Day 27-30 of gestation, and on all fetal thymocytes from Day 40 of gestation. Images Figure 2 Figure 3 Figure 4 PMID:3159652

  1. Role of Anatomic and Salivary Factors in Dental Calculus Formation in Primary and Mixed Dentition Stages.

    PubMed

    Afshar, Hossein; Ghandehari, Mehdi; Khorsand, Afshin; Ansari, Ghassem; Nahvi, Azam; Baniameri, Zahra

    2016-01-01

    Clinical experience shows that formation of calculus is a very rare phenomenon in primary teeth, but it increases as the permanent teeth erupt. The purpose of this study was to assess the relationship between dental calculus, dental anatomy, and salivary factors in primary and mixed dentition stages. A cross-sectional study was carried out to determine the buccolingual dimensions of the most concave and the most convex surfaces of the lingual aspect of mandibular central incisor crowns in a sample group of 120 three- to five-old children and 120 eight- to 10-year old children. Saliva samples were collected from 20 in each group. Data were analyzed using t tests. Significant differences were found between the ratio of the buccolingual dimensions of the most convex to the most concave areas of the lingual surfaces in primary and permanent incisors (P=0.028). Saliva analysis revealed significant differences in total protein (P=0.002), sodium (P=0.037), bicarbonate (P=0.003), and ammonia (P=0.025) between the two age groups. Anatomic and salivary factors may be important reasons for the differences in calculus formation.

  2. Lazy leucocyte syndrome--disorder of the granulocyte membrane?

    PubMed Central

    Pinkerton, P H; Robinson, J B; Senn, J S

    1978-01-01

    An adult with long-standing neutropenia had the functional granulocyte abnormalities typical of the lazy leucocyte syndrome. Scanning electron microscopy of the patient's neutrophils showed alteration in the surface configuration of the cell with coarsening of the normal fine ruffles and the appearance of knob-like projections. Similar functional and anatomical changes were induced in normal neutrophils by treatment with vinblastine. The lazy leucocyte syndrome may be a consequence of altered membrane microfilamentous protein structure or function, and undue rigidity of the affected neutrophils may explain the clinicopathological features of the disease. Images Fig. 3 Fig. 4 Fig. 1 Fig. 2 Fig. 5 Fig. 6 PMID:641207

  3. Skill mix, roles and remuneration in the primary care workforce: who are the healthcare professionals in the primary care teams across the world?

    PubMed

    Freund, Tobias; Everett, Christine; Griffiths, Peter; Hudon, Catherine; Naccarella, Lucio; Laurant, Miranda

    2015-03-01

    World-wide, shortages of primary care physicians and an increased demand for services have provided the impetus for delivering team-based primary care. The diversity of the primary care workforce is increasing to include a wider range of health professionals such as nurse practitioners, registered nurses and other clinical staff members. Although this development is observed internationally, skill mix in the primary care team and the speed of progress to deliver team-based care differs across countries. This work aims to provide an overview of education, tasks and remuneration of nurses and other primary care team members in six OECD countries. Based on a framework of team organization across the care continuum, six national experts compare skill-mix, education and training, tasks and remuneration of health professionals within primary care teams in the United States, Canada, Australia, England, Germany and the Netherlands. Nurses are the main non-physician health professional working along with doctors in most countries although types and roles in primary care vary considerably between countries. However, the number of allied health professionals and support workers, such as medical assistants, working in primary care is increasing. Shifting from 'task delegation' to 'team care' is a global trend but limited by traditional role concepts, legal frameworks and reimbursement schemes. In general, remuneration follows the complexity of medical tasks taken over by each profession. Clear definitions of each team-member's role may facilitate optimally shared responsibility for patient care within primary care teams. Skill mix changes in primary care may help to maintain access to primary care and quality of care delivery. Learning from experiences in other countries may inspire policy makers and researchers to work on efficient and effective teams care models worldwide.

  4. Anterior crossbite correction in primary and mixed dentition with removable inclined plane (Bruckl appliance).

    PubMed

    Jirgensone, Irena; Liepa, Andra; Abeltins, Andris

    2008-01-01

    Anterior crossbite correction in early mixed dentition is highly recommended as this kind of malocclusion do not diminish with age. Uncorrected anterior crossbite may lead to abnormal wear of the lower incisors, dental compensation of mandibular incisors leading to thinning of labial alveolar plate and/or gingival recession. There are several methods for solving this problem. In this article we would like to describe removable inclined plane. This is a removable simple functional appliance on the lower arch (jaw), which works as inclined plane. One of the advantage of the Bruckl appliance is that it can also be used as retention appliance after active treatment as well as it is possible to add acrylic teeth if necessary. Therefore it can be used as a removable partial denture in lower jaw in case where there is a premature loss of the primary teeth. The use of this appliance is illustrated with three cases.

  5. Mixed functional microarchitectures for orientation selectivity in the mouse primary visual cortex

    PubMed Central

    Kondo, Satoru; Yoshida, Takashi; Ohki, Kenichi

    2016-01-01

    A minicolumn is the smallest anatomical module in the cortical architecture, but it is still in debate whether it serves as functional units for cortical processing. In the rodent primary visual cortex (V1), neurons with different preferred orientations are mixed horizontally in a salt and pepper manner, but vertical functional organization was not examined. In this study, we found that neurons with similar orientation preference are weakly but significantly clustered vertically in a short length and horizontally in the scale of a minicolumn. Interestingly, the vertical clustering is found only in a part of minicolumns, and others are composed of neurons with a variety of orientation preferences. Thus, the mouse V1 is a mixture of vertical clusters of neurons with various degrees of orientation similarity, which may be the compromise between the brain size and keeping the vertical clusters of similarly tuned neurons at least in a subset of clusters. PMID:27767032

  6. Slow conduction in mixed cultured strands of primary ventricular cells and stem cell-derived cardiomyocytes

    PubMed Central

    Kucera, Jan P.; Prudat, Yann; Marcu, Irene C.; Azzarito, Michela; Ullrich, Nina D.

    2015-01-01

    Modern concepts for the treatment of myocardial diseases focus on novel cell therapeutic strategies involving stem cell-derived cardiomyocytes (SCMs). However, functional integration of SCMs requires similar electrophysiological properties as primary cardiomyocytes (PCMs) and the ability to establish intercellular connections with host myocytes in order to contribute to the electrical and mechanical activity of the heart. The aim of this project was to investigate the properties of cardiac conduction in a co-culture approach using SCMs and PCMs in cultured cell strands. Murine embryonic SCMs were pooled with fetal ventricular cells and seeded in predefined proportions on microelectrode arrays to form patterned strands of mixed cells. Conduction velocity (CV) was measured during steady state pacing. SCM excitability was estimated from action potentials measured in single cells using the patch clamp technique. Experiments were complemented with computer simulations of conduction using a detailed model of cellular architecture in mixed cell strands. CV was significantly lower in strands composed purely of SCMs (5.5 ± 1.5 cm/s, n = 11) as compared to PCMs (34.9 ± 2.9 cm/s, n = 21) at similar refractoriness (100% SCMs: 122 ± 25 ms, n = 9; 100% PCMs: 139 ± 67 ms, n = 14). In mixed strands combining both cell types, CV was higher than in pure SCMs strands, but always lower than in 100% PCM strands. Computer simulations demonstrated that both intercellular coupling and electrical excitability limit CV. These data provide evidence that in cultures of murine ventricular cardiomyocytes, SCMs cannot restore CV to control levels resulting in slow conduction, which may lead to reentry circuits and arrhythmias. PMID:26442264

  7. Quantifying subtropical North Pacific gyre mixed layer primary productivity from Seaglider observations of diel oxygen cycles

    NASA Astrophysics Data System (ADS)

    Nicholson, David P.; Wilson, Samuel T.; Doney, Scott C.; Karl, David M.

    2015-05-01

    Using autonomous underwater gliders, we quantified diurnal periodicity in dissolved oxygen, chlorophyll, and temperature in the subtropical North Pacific near the Hawaii Ocean Time-series (HOT) Station ALOHA during summer 2012. Oxygen optodes provided sufficient stability and precision to quantify diel cycles of average amplitude of 0.6 µmol kg-1. A theoretical diel curve was fit to daily observations to infer an average mixed layer gross primary productivity (GPP) of 1.8 mmol O2 m-3 d-1. Cumulative net community production (NCP) over 110 days was 500 mmol O2 m-2 for the mixed layer, which averaged 57 m in depth. Both GPP and NCP estimates indicated a significant period of below-average productivity at Station ALOHA in 2012, an observation confirmed by 14C productivity incubations and O2/Ar ratios. Given our success in an oligotrophic gyre where biological signals are small, our diel GPP approach holds promise for remote characterization of productivity across the spectrum of marine environments.

  8. Building COPD care on shaky ground: a mixed methods study from Swedish primary care professional perspective.

    PubMed

    Lundell, Sara; Tistad, Malin; Rehn, Börje; Wiklund, Maria; Holmner, Åsa; Wadell, Karin

    2017-07-10

    Chronic obstructive pulmonary disease (COPD) is a public health problem. Interprofessional collaboration and health promotion interventions such as exercise training, education, and behaviour change are cost effective, have a good effect on health status, and are recommended in COPD treatment guidelines. There is a gap between the guidelines and the healthcare available to people with COPD. The aim of this study was to increase the understanding of what shapes the provision of primary care services to people with COPD and what healthcare is offered to them from the perspective of healthcare professionals and managers. The study was conducted in primary care in a Swedish county council during January to June 2015. A qualitatively driven mixed methods design was applied. Qualitative and quantitative findings were merged into a joint analysis. Interviews for the qualitative component were performed with healthcare professionals (n = 14) from two primary care centres and analysed with qualitative content analysis. Two questionnaires were used for the quantitative component; one was answered by senior managers or COPD nurses at primary care centres (n = 26) in the county council and the other was answered by healthcare professionals (n = 18) at two primary care centres. The questionnaire data were analysed with descriptive statistics. The analysis gave rise to the overarching theme building COPD care on shaky ground. This represents professionals driven to build a supportive COPD care on 'shaky' organisational ground in a fragmented and non-compliant healthcare organisation. The shaky ground is further represented by uninformed patients with a complex disease, which is surrounded with shame. The professionals are autonomous and pragmatic, used to taking responsibility for their work, and with limited involvement of the management. They wish to provide high quality COPD care with interprofessional collaboration, but they lack competence and are hindered by

  9. Comparison of Effects of Running and Playing Exercises on Differential Leucocyte Count in Young Elite Athletes

    ERIC Educational Resources Information Center

    Cenikli, Abdullah

    2016-01-01

    The aims of the present research are to test the effects of running and playing exercises on leucocyte and differential leucocyte accounts, and to test the possible differences between running and playing exercises in terms of leucocyte accounts. They were thirty two male young soccer players. Participants arrived at the laboratory after a 12-hour…

  10. Effects of repetitive low-pressure explosive blast on primary neurons and mixed cultures.

    PubMed

    Zander, Nicole E; Piehler, Thuvan; Banton, Rohan; Benjamin, Richard

    2016-09-01

    Repetitive mild traumatic brain injury represents a considerable health concern, particularly for athletes and military personnel. For blast-induced brain injury, threshold shock-impulse levels required to induce such injuries and cumulative effects with single and/or multiple exposures are not well characterized. Currently, there is no established in vitro experimental model with blast pressure waves generated by live explosives. This study presents results of primary neurons and mixed cultures subjected to our unique in vitro indoor experimental platform that uses real military explosive charges to probe the effects of primary explosive blast at the cellular level. The effects of the blast on membrane permeability, generation of reactive oxygen species (ROS), uptake of sodium ions, intracellular calcium, and release of glutamate were probed 2 and 24 hr postblast. Significant changes in membrane permeability and sodium uptake among the sham, single-blast-injured, and triple-blast-injured samples were observed. A significant increase in ROS and glutamate release was observed for the triple-blast-injured samples compared with the sham. Changes in intracellular calcium were not significant. These results suggest that blast exposure disrupts the integrity of the plasma membrane, leading to the upset of ion homeostasis, formation of ROS, and glutamate release. Published 2016. †This article is a U.S. Government work and is in the public domain in the USA.

  11. Influence of provider mix and regulation on primary care services supplied to US patients.

    PubMed

    Richards, Michael R; Polsky, Daniel

    2016-04-01

    Access to medical care and how it differs for various patients remain key policy issues. While existing work has examined clinic structure's influence on productivity, less research has explored the link between provider mix and access for different patient types - which also correspond to different service prices. We exploit experimental data from a large field study spanning 10 US states where trained audit callers were randomly assigned an insurance status and then contacted primary care physician practices seeking new patient appointments. We find clinics with more non-physician clinicians are associated with better access for Medicaid patients and lower prices for office visits; however, these relationships are only found in states granting full practice autonomy to these providers. Substituting more non-physician labor in primary care settings may facilitate greater appointment availability for Medicaid patients, but this likely rests on a favorable policy environment. Relaxing regulations for non-physicians may be an important initiative as US health reforms continue and also relevant to other countries coping with greater demands for medical care and related financial strain.

  12. Neoplastic cells obtained from Hodgkin's disease are potent stimulators of human primary mixed lymphocyte cultures.

    PubMed

    Fisher, R I; Bostick-Bruton, F; Sauder, D N; Scala, G; Diehl, V

    1983-06-01

    Neoplastic cells obtained from the pleural effusion of a patient with Hodgkin's disease have been maintained in culture since 1978. These tumor cells have been shown to have the cytologic features, cytochemical staining, and cell surface markers of Reed-Sternberg cells. In this study we demonstrate that the cell line termed L428 is a potent stimulator of the primary human mixed lymphocyte reaction. Significant proliferation occurred when mononuclear leukocytes obtained from normal donors were stimulated with radiated L428 cells at responder:stimulator ratios varying from 200:1 to 20:1. Proliferative responses occurred between days 3 and 6 of the cultures with maximal proliferation on day 5. Under optimal culture conditions, mean net proliferative response of 14 normal donors was 51,000 +/- 10,600 dpm. The mixed lymphocyte response was totally blocked by concentrations of monoclonal anti-Ia antibody that had no effect on concanavalin A-induced proliferation. However, the mixed lymphocyte response was not blocked by an anti-K562 cell monoclonal antibody of the same immunoglobulin subclass that binds to the L428 cells. Antigen processing by responder monocytes or Ia-positive cells was not required for the MLC. When responder T cells from two normals were depleted of Ia-bearing cells and monocytes, the mixed lymphocyte reaction between the two normals was eliminated, yet the stimulation of each normal by the L428 cells was not reduced. The cells that proliferated in response to stimulation by the L428 cells were T cells, primarily of the helper subset. No IL 1 activity could be detected in concentrated supernatants of L428 cultures after stimulation of L428 cells by mitogens, phorbol esters, or muramyl dipeptide, or in the MLC. All of these cultures contain fetal calf serum. However, the L428 cells are capable of producing IL 1, because IL 1 was detected when the L428 cells were stimulated with LPS in the absence of fetal calf serum. These neoplastic cells, obtained

  13. Client/patient perceptions of achieving equity in primary health care: a mixed methods study.

    PubMed

    Akhavan, Sharareh; Tillgren, Per

    2015-08-12

    To provide health care on equal terms has become a challenge for the health system. As the front line in health services, primary care has a key role to play in developing equitable health care, responsive to the needs of different population groups. Reducing inequalities in care has been a central and recurring theme in Swedish health reforms. The aim of this study is to describe and assess client/patient experiences and perceptions of care in four primary health care units (PHCUs) involved in Sweden's national Care on Equal Terms project. Mixed Method Research (MMR) was chosen to describe and assess client/patient experiences and perceptions of health care with regard to equity. There was a focus group discussion, and individual interviews with 21 clients/patients and three representatives of patient associations. Data from the Swedish National Patient Survey (NPS), conducted in 2011 and followed up in 2013, were also used. The interview data were divided into two main categories and three subcategories. The first category "Perception of equitable health care" had two subcategories, namely "Health care providers' perceptions" and "Fairness and participation". The second category "To achieve more equitable health care" had four subcategories: "Encounter", "Access", "Interpreters and bilingual/diverse health care providers" and "Time pressure and continuity". Results from the NPS showed that two of the PHCUs improved in some aspects of patient perceived quality of care (PPQC) while two were not so successful. Clients/patients perceived health care providers' perceptions of their ethnic origin and mental health status as important for equitable health care. Discriminatory perceptions may lead to those in need of care refraining from seeking it. More equitable care means longer consultations, better accessibility in terms of longer opening hours, and ways of communicating other than just via voice mail. It also involves continuity in care and access to an interpreter

  14. Impact of a quality improvement program on primary healthcare in Canada: a mixed-method evaluation.

    PubMed

    Harris, Stewart B; Green, Michael E; Brown, Judith Belle; Roberts, Sharon; Russell, Grant; Fournie, Meghan; Webster-Bogaert, Susan; Paquette-Warren, Jann; Kotecha, Jyoti; Han, Han; Thind, Amardeep; Stewart, Moira; Reichert, Sonja; Tompkins, Jordan W; Birtwhistle, Richard

    2015-04-01

    Rigorous comprehensive evaluations of primary healthcare (PHC) quality improvement (QI) initiatives are lacking. This article describes the evaluation of the Quality Improvement and Innovation Partnership Learning Collaborative (QIIP-LC), an Ontario-wide PHC QI program targeting type 2 diabetes management, colorectal cancer (CRC) screening, access to care, and team functioning. This article highlights the primary outcome results of an external retrospective, multi-measure, mixed-method evaluation of the QIIP-LC, including: (1) matched-control pre-post chart audit of diabetes management (A1c/foot exams) and rate of CRC screening; (2) post-only advanced access survey (third-next available appointment); and (3) post-only semi-structured interviews (team functioning). Chart audit data was collected from 34 consenting physicians per group (of which 88% provided access data). Between-group differences were not statistically significant (A1c [p=0.10]; foot exams [p=0.45]; CRC screening [p=0.77]; advanced access [p=0.22]). Qualitative interview (n=42) themes highlighted the success of the program in helping build interdisciplinary team functioning and capacity. The rigorous design and methodology of the QIIP-LC evaluation utilizing a control group is one of the most significant efforts thus far to demonstrate the impact of a QI program in PHC, with improvements over time in both QIIP and control groups offering a likely explanation for the lack of statistically significant primary outcomes. Team functioning was a key success, with team-based chronic care highlighted as pivotal for improved health outcomes. Policy makers should strive to endorse QI programs with proven success through rigorous evaluation to ensure evidence-based healthcare policy and funding. Copyright © 2014 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.

  15. Effect of mixing process on microleakage of glass ionomer cements used in atraumatic restorative treatment on primary molars.

    PubMed

    Ferreira, Fernanda de Morais; do Vale, Miriam Pimenta Parreira; Jansen, Wellington Corrêa; Paiva, Saul Martins; Pordeus, Isabela Almeida

    2007-01-01

    Aiming to assess the effect of mixing process on microleakage, 40 primary molars were filled with encapsulated glass ionomer cements (GICs) (Vidrion, RCaps and Fuji, IXGPFAST) or with GICs stored in bottles (Vidrion, R and Fuji, IX). Dye penetration was assessed using scores. Encapsulation and mechanical mixing have reduced significantly marginal microleakage levels in class II restorations performed with conventional GICs if compared to the values obtained by their bottled correspondents (p=0.000).

  16. Involvement of rainbow trout leucocytes in the pathogenesis of infectious hematopoietic necrosis

    USGS Publications Warehouse

    Chilmonczyk, S.; Winton, J.R.

    1994-01-01

    Rainbow trout Oncorhynchus myluss leucocytes were tested for their ability to support replication of infectious hematopoietic necrosis virus (IHNV). Viral replication occurred in vitro uslng leucocytes cultured from peripheral blood, kidney, and thymus where viral titers peaked at 2 to 4 d post-inoculation. Leucocytes collected from trout following waterborne challenge with IHNV were cocultured on EPC cell monolayers. These assays detected IHNV in leucocytes infected in vivo as early as 6 h post-exposure before the challenge virus had undergone replication. These data showed that leucocyte populations could serve as target cells in the initial phase of IHNV infection.

  17. UK NEQAS for leucocyte immunophenotyping: the first 10 years

    PubMed Central

    Reilly, J; Barnett, D

    2001-01-01

    In the past decade, cellular immunophenotyping has become a new discipline in diagnostic haematology and immunology, and is invaluable in the rapid diagnosis of leukaemia and monitoring disease progression in human immunodeficiency virus infected individuals. The introduction of bench top flow cytometers has meant that immunophenotyping is now also used for the quantitation of CD34+ peripheral blood stem cells (PBSCs) to ensure the correct timing and adequacy of haematopoietic progenitor cell harvests. Furthermore, flow cytometry has become an important tool for the counting of leucocytes in blood components after leucocyte depletion. Because this new discipline is now such a major diagnostic and prognostic tool in the clinical arena, its use must be subject to both internal and external quality control. Such a requirement was first recognised as early as 1986 when an Inter-Regional Quality Assessment Scheme (IRQAS) was initiated for laboratories that undertook the immunocytochemical diagnosis of leukaemia using the alkaline phosphates anti-alkaline phosphatase technique. This programme began with around 25 UK laboratories. In 1990, after the introduction of two more programmes (one for leukaemia diagnosis using UV microscopy and latterly flow cytometry, and one for the enumeration of CD4+ T cells) the IRQAS achieved UK National External Quality Assessment Scheme (UK NEQAS) status and changed its title to UK NEQAS for Leucocyte Immunophenotyping. In the past decade the once small IRQAS programme has evolved into the largest international scheme of its kind, providing EQA to over 650 laboratories world wide for leukaemia immunophenotyping, lymphocyte subset analysis, PBSCs, and more recently low level leucocyte counting. Over the years, this EQA programme has highlighted important problems, such as the inappropriate use of fluorochromes and antibody titre, and the identification of effective gating strategies, all of which have contributed directly to the high

  18. A scoring system for appraising mixed methods research, and concomitantly appraising qualitative, quantitative and mixed methods primary studies in Mixed Studies Reviews.

    PubMed

    Pluye, Pierre; Gagnon, Marie-Pierre; Griffiths, Frances; Johnson-Lafleur, Janique

    2009-04-01

    A new form of literature review has emerged, Mixed Studies Review (MSR). These reviews include qualitative, quantitative and mixed methods studies. In the present paper, we examine MSRs in health sciences, and provide guidance on processes that should be included and reported. However, there are no valid and usable criteria for concomitantly appraising the methodological quality of the qualitative, quantitative and mixed methods studies. To propose criteria for concomitantly appraising the methodological quality of qualitative, quantitative and mixed methods studies or study components. A three-step critical review was conducted. 2322 references were identified in MEDLINE, and their titles and abstracts were screened; 149 potentially relevant references were selected and the full-text papers were examined; 59 MSRs were retained and scrutinized using a deductive-inductive qualitative thematic data analysis. This revealed three types of MSR: convenience, reproducible, and systematic. Guided by a proposal, we conducted a qualitative thematic data analysis of the quality appraisal procedures used in the 17 systematic MSRs (SMSRs). Of 17 SMSRs, 12 showed clear quality appraisal procedures with explicit criteria but no SMSR used valid checklists to concomitantly appraise qualitative, quantitative and mixed methods studies. In two SMSRs, criteria were developed following a specific procedure. Checklists usually contained more criteria than needed. In four SMSRs, a reliability assessment was described or mentioned. While criteria for quality appraisal were usually based on descriptors that require specific methodological expertise (e.g., appropriateness), no SMSR described the fit between reviewers' expertise and appraised studies. Quality appraisal usually resulted in studies being ranked by methodological quality. A scoring system is proposed for concomitantly appraising the methodological quality of qualitative, quantitative and mixed methods studies for SMSRs. This

  19. Evaluation of Leucocyte Functions Six Years after Tumour Autograft in Human Mammary Cancer

    PubMed Central

    Anderson, J. Maxwell; Kelly, F.; Wood, Suzanne E.; Rodger, K. D.; Freshney, R. Ian

    1973-01-01

    Mammary cancer directed and nonspecific immunoassays were made in 3 groups of female patients. One group had primary mammary cancer treated by mastectomy and postoperative radiotherapy plus an autograft of irradiated tumour (AIT) 40-66 months previously. A second age-matched group had mammary cancer comparable to the first group in clinical presentation and treatment except that no AIT was given. The third group consisted of non-cancer-bearing age-matched females. The migration of leucocytes from autografted patients was significantly inhibited in the presence of allogeneic mammary cancer cells from a standardized panel, compared with leucocytes from either non-autograft patients or non-cancer bearers. Selected data from a lymphocyte cytotoxicity test revealed a significantly greater kill of allogeneic mammary cancer target cells by autograft lymphocytes than by those of other groups. These indications of increased cancer directed cell mediated immunity in respect of sensitivity and toxicity in association with AIT require further elucidation under strictly controlled conditions. PMID:4807858

  20. Mixed anxiety and depressive disorder outcomes: prospective cohort study in primary care.

    PubMed

    Walters, Kate; Buszewicz, Marta; Weich, Scott; King, Michael

    2011-06-01

    Mixed anxiety and depressive disorder (MADD) is common yet ill-defined, with little known about outcomes. To determine MADD outcomes over 1 year. We recruited 250 adults attending seven London general practices with mild-moderate distress. Three groups were defined using a diagnostic interview: MADD, other ICD-10 psychiatric diagnosis, no psychiatric diagnosis. We assessed symptoms of distress (General Health Questionnaire-28), quality of life (12-item Short Form Health Survey), general practitioner (GP) diagnosis and consultation rate at baseline, 3 months and 1 year. Two-thirds of participants with MADD had no significant psychological distress at 3 months (61%) or 1 year (69%). However, compared with those with no diagnosis, individuals had twice the risk of significant distress (incidence rate ratio 2.39, 95% CI 1.29-4.42) at 3 months but not 1 year, and persistently lower quality of life (mental health functioning). There was no significant difference in GP consultation rate/diagnosis. The majority with MADD improved, but individuals had an increased risk of significant distress at 3 months and a lower quality of life. As we cannot currently predict those with a poorer prognosis these patients should be actively monitored in primary care.

  1. Nurse practitioner screening for childhood adversity among adult primary care patients: A mixed-method study.

    PubMed

    Kalmakis, Karen A; Chandler, Genevieve E; Roberts, Susan Jo; Leung, Katherine

    2017-01-01

    Researchers have demonstrated an association between experiencing childhood abuse and multiple chronic health conditions in adulthood, yet this evidence has not been routinely translated to practice. The purpose of this research study was to examine nurse practitioner (NP) practices, skills, attitudes, and perceived barriers associated with screening adult patients for childhood abuse to determine the extent to which evidence of the association between childhood abuse and negative health outcomes has been translated to NP practice. A mixed-method approach with web-based questionnaires and online focus groups was used to examine NP screening for histories of childhood abuse. A total of 188 complete NP surveys were analyzed along with data from focus groups with 12 NPs. One third of the NPs regularly screened for childhood abuse and believed screening was their responsibility. Six barriers, including insufficient time and lack of confidence when inquiring about abuse, were significantly associated with NP screening practices. The focus group participants discussed how and when one should ask about childhood abuse, and the need for education about screening. Time constraints and NPs' lack of confidence in their ability to screen for histories of childhood abuse must be addressed to encourage routine screening in primary care practice. ©2016 American Association of Nurse Practitioners.

  2. Evaluation of facial morphology and sagittal relationship between dental arches in primary and mixed dentition.

    PubMed

    Traldi, Aline; Valdrighi, Heloísa Cristina; de Souza, Luciane Zanin; Vedovello, Silvia Amélia Scudeler

    2015-01-01

    To assess facial morphology (Pattern) and sagittal relationship between dental arches (Class), and establish a potential association between them and the variables sex, age and ethnicity, among schoolchildren aged between 4 and 9 years old (mean age of 6.7 years) in primary and mixed dentitions. The sample comprised 875 children (457 males and 418 females) attending schools in Descalvado, São Paulo, Brazil. An attempt was made with a view to establish a potential association between children's morphological features with sex, age and ethnicity. Descriptive analysis revealed a predominance of facial Pattern I (69.9 %) and Class I (67.4 %). Statistical tests (p < 0.001) showed that Class I was more frequent among Pattern I children, whereas Class II prevailed among Pattern II, and Class III was frequent among Pattern I and III children. Ethnicity was the only variable associated with facial pattern. Results suggest that facial pattern and sagittal relationship between dental arches tend to be correlated. Ethnicity was associated with facial pattern, with Pattern I being the most recurrent among Caucasians and facial Pattern II being recurrent among Afro-descendant subjects.

  3. Evaluation of facial morphology and sagittal relationship between dental arches in primary and mixed dentition

    PubMed Central

    Traldi, Aline; Valdrighi, Heloísa Cristina; de Souza, Luciane Zanin; Vedovello, Silvia Amélia Scudeler

    2015-01-01

    OBJECTIVE: To assess facial morphology (Pattern) and sagittal relationship between dental arches (Class), and establish a potential association between them and the variables sex, age and ethnicity, among schoolchildren aged between 4 and 9 years old (mean age of 6.7 years) in primary and mixed dentitions. METHODS: The sample comprised 875 children (457 males and 418 females) attending schools in Descalvado, São Paulo, Brazil. An attempt was made with a view to establish a potential association between children's morphological features with sex, age and ethnicity. RESULTS: Descriptive analysis revealed a predominance of facial Pattern I (69.9 %) and Class I (67.4 %). Statistical tests (p < 0.001) showed that Class I was more frequent among Pattern I children, whereas Class II prevailed among Pattern II, and Class III was frequent among Pattern I and III children. Ethnicity was the only variable associated with facial pattern. CONCLUSIONS: Results suggest that facial pattern and sagittal relationship between dental arches tend to be correlated. Ethnicity was associated with facial pattern, with Pattern I being the most recurrent among Caucasians and facial Pattern II being recurrent among Afro-descendant subjects. PMID:26352847

  4. Histoplasma capsulatum inhibits apoptosis and Mac-1 expression in leucocytes.

    PubMed

    Medeiros, A I; Bonato, V L D; Malheiro, A; Dias, A R V; Silva, C L; Faccioli, L H

    2002-10-01

    Histoplasma capsulatum is a fungus found intracellularly in neutrophils and peripheral blood mononuclear cells (PBMCs), suggesting that it is capable of evading damage and survives inside these cells. In this study, we report that neutrophils from H. capsulatum-infected mice, and human neutrophils and mononuclear cells exposed to H. capsulatum presented less apoptosis than those from noninfected animals or cells exposed to medium only. Moreover, cells harvested from infected animals are resistant to apoptosis induced by dexamethasone - a proapoptotic stimulant. We also show that neutrophils harvested from infected mice and PBMCs from humans exposed to the fungus had a greatly decreased Mac-1 expression. We conclude that H. capsulatum induces an antiapoptotic state on leucocytes, which correlates with decreased cell-surface Mac-1 expression. These facts may represent an escape mechanism for the fungus by delaying cell death and allowing the fungus to survive inside leucocytes.

  5. Impact of financial incentives on alcohol intervention delivery in primary care: a mixed-methods study.

    PubMed

    O'Donnell, Amy; Haighton, Catherine; Chappel, David; Shevills, Colin; Kaner, Eileen

    2016-11-25

    Local and national financial incentives were introduced in England between 2008 and 2015 to encourage screening and brief alcohol intervention delivery in primary care. We used routine Read Code data and interviews with General Practitioners (GPs) to assess their impact. A sequential explanatory mixed-methods study was conducted in 16 general practices representing 106,700 patients and 99 GPs across two areas in Northern England. Data were extracted on screening and brief alcohol intervention delivery for 2010-11 and rates were calculated by practice incentive status. Semi-structured interviews with 14 GPs explored which factors influence intervention delivery and recording in routine consultations. Screening and brief alcohol intervention rates were higher in financially incentivised compared to non-incentivised practices. However absolute rates were low across all practices. Rates of short screening test administration ranged from 0.05% (95% CI: 0.03-0.08) in non-incentivised practices to 3.92% (95% CI: 3.70-4.14) in nationally incentivised practices. For the full AUDIT, rates were also highest in nationally incentivised practices (3.68%, 95% CI: 3.47-3.90) and lowest in non-incentivised practices (0.17%, 95% CI: 0.13-0.22). Delivery of alcohol interventions was highest in practices signed up to the national incentive scheme (9.23%, 95% CI: 8.91-9.57) and lowest in non-incentivised practices (4.73%, 95% CI: 4.50-4.96). GP Interviews highlighted a range of influences on alcohol intervention delivery and subsequent recording including: the hierarchy of different financial incentive schemes; mixed belief in the efficacy of alcohol interventions; the difficulty of codifying complex conditions; and GPs' beliefs about patient-centred practice. Financial incentives have had some success in encouraging screening and brief alcohol interventions in England, but levels of recorded activity remain low. To improve performance, future policies must prioritise alcohol

  6. MHC class I antigens and tumour-infiltrating leucocytes in laryngeal cancer: long-term follow-up.

    PubMed Central

    Esteban, F.; Redondo, M.; Delgado, M.; Garrido, F.; Ruiz-Cabello, F.

    1996-01-01

    Alteration in MHC class I expression may be used by cancer cells to avoid immune destruction. Much experimental evidence supports this idea, although survival studies are very scarce. To investigate whether the presence or absence of HLA-A, -B and -C antigens in laryngeal carcinoma influences survival, a series of 60 primary laryngeal tumours treated surgically and normal tissues were evaluated in frozen sections for the expression of MHC class I antigens and tumour-infiltrating leucocytes (CD3, CD4, CD8, CD11b, CD1, CD20 and CD16), using monoclonal antibodies and the APAAP, technique. Long-term follow-up from the patients is available, ranging from 6 to 10 years. Thirteen tumours presented total HLA-ABC loss, five selective losses of HLA-A antigens and one absence of HLA-B antigens. Total losses were statistically associated with several clinical and pathological parameters, but there were no differences regarding tumour-infiltrating leucocytes. After conducting a prospective study, only T and N staging and scoring according to Glanz's malignancy classification were found to be independently related to patients' outcome. From our data, we conclude that neither complete loss of HLA class I antigens nor tumour-infiltrating leucocytes appear to influence survival in squamous cell carcinoma of the larynx. PMID:8956796

  7. The main catechin of green tea, (-)-epigallocatechin-3-gallate (EGCG), reduces bleomycin-induced DNA damage in human leucocytes.

    PubMed

    Glei, M; Pool-Zobel, B L

    2006-04-01

    Interest in the beneficial effects of green tea has led to investigations on activities by the main catechin (-)-epigallocatechin-3-gallate (EGCG). This antioxidative compound could contribute to cancer chemoprevention by acting antigenotoxic. To further explore this hypothesis we investigated antigenotoxic potentials of low EGCG concentrations in human peripheral leucocytes. Leucocytes isolated from whole blood were (1) stimulated with phytohaemagglutinin, (2) damaged with genotoxic bleomycin, and (3) post-incubated to allow DNA repair. After each phase DNA integrity was measured with the comet assay. EGCG (2, 20, 100 microM) was added either during phases 1, 2 or 3 or during the whole process (1-3), to delineate mechanisms of antigenotoxicity reflecting induction of detoxification (phase 1), scavenging of radicals (phase 2), stimulation of repair (phase 3), respectively. Bleomycin induced breaks and endonuclease III specific damage, but EGCG did not affect damage or repair of these lesions when added during phases 1, 2 or 3. However, the application of EGCG during phases 1 and 2 significantly reduced both bleomycin-induced breaks and endonuclease III sensitive sites. EGCG added during all phases impaired persistence of damage. Our studies show that the continuous presence of EGCG can reduce radical-induced DNA damage in primary leucocytes, possibly due to a combination of different mechanisms. Together the findings support the hypotheses that EGCG acts protective in human cells.

  8. Leucocyte adhesion deficiency presenting as a chronic ileocolitis.

    PubMed Central

    D'Agata, I D; Paradis, K; Chad, Z; Bonny, Y; Seidman, E

    1996-01-01

    CD11/CD18 leucocyte glycoprotein deficiency is a rare, congenital adhesion molecule disorder which, in its severe form, is usually fatal. Leucocytes in affected subjects have abnormal migration and adherence, rendering patients susceptible to life threatening infections. The CD11/CD18 integrins, and other adhesion molecules, are considered essential to the normal inflammatory response. It has been postulated that adhesion molecules may be responsible for mediating in part, the inflammatory changes observed in inflammatory bowel diseases and related disorders. This report describes the first case of CD11/CD18 deficiency characterised by a chronic ileocolitis. Bone marrow transplantation completely resolved the gastrointestinal symptoms, supporting a role for neutrophil dysfunction in the pathogenesis of the gut lesions. This case suggests that specific blockade of CD11/CD18 integrins alone may not halt the chronic inflammatory response observed in immune mediated bowel disorders, and that abnormalities of leucocyte function must be included in the differential diagnosis of paediatric Crohn's disease. Images p606-a PMID:8944573

  9. Role of interleukin 8 on leucocyte-endothelial cell adhesion in intestinal inflammation.

    PubMed Central

    Arndt, H; Bolanowski, M A; Granger, D N

    1996-01-01

    BACKGROUND: An important action of interleukin 8 (IL8) is stimulation of granulocytes. The object of this study was to assess the contribution of IL8 to the leucocyte-endothelial cell interactions associated with intestinal inflammation in the rat. METHODS: Two indomethacin injections (48 and 24 hours prior to the experiments) induced a longlasting ileitis in rats. The number of adherent and emigrated leucocytes, leucocyte rolling velocity, and shear rate were monitored in normal and inflamed mesenteric postcapillary venules. Some animals received a monoclonal antibody (MAb) against IL8 or CD11b/CD18 at 24 and 12 hours prior to the experiment. RESULTS: Indomethacin elicited a seven-fold increase in leucocyte adherence and a 5.4-fold increase in leucocyte emigration, while leucocyte rolling velocity was reduced by nearly 80%. The indomethacin induced increases in leucocyte adherence and emigration were significantly reduced (by 57% and 67%, respectively) while leucocyte rolling velocity was increased (to 63% of control) by the IL8-specific MAb. The level of inhibition seen with the IL8 MAb was similar to that associated with administration of a MAb directed against the leucocyte adhesion molecule CD11b/CD18. CONCLUSIONS: IL8 contributes to the leucocyte-endothelial cell interactions elicited in mesenteric venules by indomethacin. PMID:8984032

  10. Carbon footprint of patient journeys through primary care: a mixed methods approach

    PubMed Central

    Andrews, Elizabeth; Pearson, David; Kelly, Charlotte; Stroud, Laura; Rivas Perez, Martin

    2013-01-01

    Background The NHS has a target of cutting its carbon dioxide (CO2) emissions by 80% below 1990 levels by 2050. Travel comprises 17% of the NHS carbon footprint. This carbon footprint represents the total CO2 emissions caused directly or indirectly by the NHS. Patient journeys have previously been planned largely without regard to the environmental impact. The potential contribution of ‘avoidable’ journeys in primary care is significant. Aim To investigate the carbon footprint of patients travelling to and from a general practice surgery, the issues involved, and potential solutions for reducing patient travel. Design and setting A mixed methods study in a medium-sized practice in Yorkshire. Method During March 2012, 306 patients completed a travel survey. GIS maps of patients’ travel (modes and distances) were produced. Two focus groups (12 clinical and 13 non-clinical staff) were recorded, transcribed, and analysed using a thematic framework approach. Results The majority (61%) of patient journeys to and from the surgery were made by car or taxi; main reasons cited were ‘convenience’, ‘time saving’, and ‘no alternative’ for accessing the surgery. Using distances calculated via ArcGIS, the annual estimated CO2 equivalent carbon emissions for the practice totalled approximately 63 tonnes. Predominant themes from interviews related to issues with systems for booking appointments and repeat prescriptions; alternative travel modes; delivering health care; and solutions to reducing travel. Conclusion The modes and distances of patient travel can be accurately determined and allow appropriate carbon emission calculations for GP practices. Although challenging, there is scope for identifying potential solutions (for example, modifying administration systems and promoting walking) to reduce ‘avoidable’ journeys and cut carbon emissions while maintaining access to health care. PMID:23998839

  11. Harms from discharge to primary care: mixed methods analysis of incident reports

    PubMed Central

    Williams, Huw; Edwards, Adrian; Hibbert, Peter; Rees, Philippa; Prosser Evans, Huw; Panesar, Sukhmeet; Carter, Ben; Parry, Gareth; Makeham, Meredith; Jones, Aled; Avery, Anthony; Sheikh, Aziz; Donaldson, Liam; Carson-Stevens, Andrew

    2015-01-01

    Background Discharge from hospital presents significant risks to patient safety, with up to one in five patients experiencing adverse events within 3 weeks of leaving hospital. Aim To describe the frequency and types of patient safety incidents associated with discharge from secondary to primary care, and commonly described contributory factors to identify recommendations for practice. Design and setting A mixed methods analysis of 598 patient safety incident reports in England and Wales related to ‘Discharge’ from the National Reporting and Learning System. Method Detailed data coding (with 20% double-coding), data summaries generated using descriptive statistical analysis, and thematic analysis of special-case sample of reports. Incident type, contributory factors, type, and level of harm were described, informing recommendations for future practice. Results A total of 598 eligible reports were analysed. The four main themes were: errors in discharge communication (n = 151; 54% causing harm); errors in referrals to community care (n = 136; 73% causing harm); errors in medication (n = 97; 87% causing harm); and lack of provision of care adjuncts such as dressings (n = 62; 94% causing harm). Common contributory factors were staff factors (not following referral protocols); and organisational factors (lack of clear guidelines or inefficient processes). Improvement opportunities include developing and testing electronic discharge methods with agreed minimum information requirements and unified referrals systems to community care providers; and promoting a safety culture with ‘safe discharge’ checklists, discharge coordinators, and family involvement. Conclusion Significant harm was evident due to deficits in the discharge process. Interventions in this area need to be evaluated and learning shared widely. PMID:26622036

  12. Harms from discharge to primary care: mixed methods analysis of incident reports.

    PubMed

    Williams, Huw; Edwards, Adrian; Hibbert, Peter; Rees, Philippa; Prosser Evans, Huw; Panesar, Sukhmeet; Carter, Ben; Parry, Gareth; Makeham, Meredith; Jones, Aled; Avery, Anthony; Sheikh, Aziz; Donaldson, Liam; Carson-Stevens, Andrew

    2015-12-01

    Discharge from hospital presents significant risks to patient safety, with up to one in five patients experiencing adverse events within 3 weeks of leaving hospital. To describe the frequency and types of patient safety incidents associated with discharge from secondary to primary care, and commonly described contributory factors to identify recommendations for practice. A mixed methods analysis of 598 patient safety incident reports in England and Wales related to 'Discharge' from the National Reporting and Learning System. Detailed data coding (with 20% double-coding), data summaries generated using descriptive statistical analysis, and thematic analysis of special-case sample of reports. Incident type, contributory factors, type, and level of harm were described, informing recommendations for future practice. A total of 598 eligible reports were analysed. The four main themes were: errors in discharge communication (n = 151; 54% causing harm); errors in referrals to community care (n = 136; 73% causing harm); errors in medication (n = 97; 87% causing harm); and lack of provision of care adjuncts such as dressings (n = 62; 94% causing harm). Common contributory factors were staff factors (not following referral protocols); and organisational factors (lack of clear guidelines or inefficient processes). Improvement opportunities include developing and testing electronic discharge methods with agreed minimum information requirements and unified referrals systems to community care providers; and promoting a safety culture with 'safe discharge' checklists, discharge coordinators, and family involvement. Significant harm was evident due to deficits in the discharge process. Interventions in this area need to be evaluated and learning shared widely. © British Journal of General Practice 2015.

  13. Carbon footprint of patient journeys through primary care: a mixed methods approach.

    PubMed

    Andrews, Elizabeth; Pearson, David; Kelly, Charlotte; Stroud, Laura; Rivas Perez, Martin

    2013-09-01

    The NHS has a target of cutting its carbon dioxide (CO2) emissions by 80% below 1990 levels by 2050. Travel comprises 17% of the NHS carbon footprint. This carbon footprint represents the total CO2 emissions caused directly or indirectly by the NHS. Patient journeys have previously been planned largely without regard to the environmental impact. The potential contribution of 'avoidable' journeys in primary care is significant. To investigate the carbon footprint of patients travelling to and from a general practice surgery, the issues involved, and potential solutions for reducing patient travel. A mixed methods study in a medium-sized practice in Yorkshire. During March 2012, 306 patients completed a travel survey. GIS maps of patients' travel (modes and distances) were produced. Two focus groups (12 clinical and 13 non-clinical staff) were recorded, transcribed, and analysed using a thematic framework approach. The majority (61%) of patient journeys to and from the surgery were made by car or taxi; main reasons cited were 'convenience', 'time saving', and 'no alternative' for accessing the surgery. Using distances calculated via ArcGIS, the annual estimated CO2 equivalent carbon emissions for the practice totalled approximately 63 tonnes. Predominant themes from interviews related to issues with systems for booking appointments and repeat prescriptions; alternative travel modes; delivering health care; and solutions to reducing travel. The modes and distances of patient travel can be accurately determined and allow appropriate carbon emission calculations for GP practices. Although challenging, there is scope for identifying potential solutions (for example, modifying administration systems and promoting walking) to reduce 'avoidable' journeys and cut carbon emissions while maintaining access to health care.

  14. THE DEFORMABILITY AND THE WETTING PROPERTIES OF LEUCOCYTES AND ERYTHROCYTES.

    PubMed

    Mudd, S; Mudd, E B

    1931-07-20

    The resistance to deformation of polymorphonuclear neutrophile leucocytes under the conditions of our observations has been shown to be on the average considerably less than the resistance to deformation of large mononuclear leucocytes. It is recognized of course that the viscosity of leucocytes, as of other cells, may be markedly influenced by osmotic conditions (17), by the reaction of the suspending medium (18, 19), by temperature, or by injury (20, 21). Although the conditions of our observations were quite different from those of the body, they were nevertheless closely similar to those of simultaneous phagocytosis experiments in which the cells functioned exceedingly well (3). Moreover E. R. and E. L. Clark (22) have noted that polymorphonuclear leucocytes in the tails of living tadpoles were more fluid than the macrophages. And Goss (23) in microdissecting human polymorphonuclear neutrophiles reports that they are more fluid than the clasmatocytes and monocytes studied by Chambers and Borquist (24). Other types of leucocytes have in our experience seemed to fall between the large mononuclear and the polymorphonuclear leucocytes in their average resistance to the interfacial tensions. The leucocyte of each type studied is surrounded by an exceedingly delicate membrane. This membrane appears under the dark-field microscope as a pale, silvery line not distinguishable by inspection alone from a simple phase boundary between two immiscible liquids. That this is a membrane, however, and not a mere interface between immiscible phases, seems certain. In the first place the cell cytoplasm and the suspending medium are not immiscible. When the cell organization is broken down by the interfacial tension the greater part of the cell contents is immediately dissolved or dispersed. Goss (23) has noted that when the membrane is torn with a microdissection needle disintegration at once spreads over the membrane and the cytoplasm undergoes profound change. Moreover it is

  15. Establishment of primary mixed cell cultures from spontaneous canine mammary tumors: Characterization of classic and new cancer-associated molecules

    PubMed Central

    Gentile, Luciana B.; Nagamine, Marcia K.; Biondi, Luiz R.; Sanches, Daniel S.; Toyota, Fábio; Giovani, Tatiane M.; de Jesus, Isis P.; da Fonseca, Ivone I. M.; Queiroz-Hazarbassanov, Nicolle; Diaz, Bruno L.; Salles Gomes, Cristina de O. Massoco

    2017-01-01

    There are many factors which make canine cancer like cancer in humans. The occurrence of spontaneous mammary tumors in pet dogs, tumor genetics, molecular targets and exposure to the same environmental risk factors are among these factors. Therefore, the study of canine cancer can provide useful information to the oncology field. This study aimed to establish and characterize a panel of primary mixed cell cultures obtained from spontaneous canine mammary tumors. Eight established cell cultures obtained from one normal mammary gland, one complex adenoma, one mixed adenoma, two complex carcinomas and two mixed carcinomas were analyzed. The gene expression levels of classic molecular cancer players such as fibroblast growth factor receptor (FGFR) 2, breast cancer (BRCA) 1, BRCA2 and estrogen receptor (ESR) 1 were evaluated. For the first time, three orphan nuclear receptors, estrogen-related receptors (ERRs) α, β and γ were studied in canine mammary cancer. The highest expression level of ERRα was observed in complex carcinoma-derived cell culture, while the highest levels of ERRβ and γ were observed in cells derived from a mixed carcinoma. Meanwhile, complex carcinomas presented the highest levels of expression of ESR1, BRCA1 and FGFR2 among all samples. BRCA2 was found exclusively in complex adenoma. The transcription factor GATA3 had its highest levels in mixed carcinoma samples and its lowest levels in complex adenoma. Proliferation assays were also performed to evaluate the mixed cell cultures response to ER ligands, genistein and DES, both in normoxia and hypoxic conditions. Our results demonstrate that morphological and functional studies of primary mixed cell cultures derived from spontaneous canine mammary tumors are possible and provide valuable tool for the study of various stages of mammary cancer development. PMID:28945747

  16. Establishment of primary mixed cell cultures from spontaneous canine mammary tumors: Characterization of classic and new cancer-associated molecules.

    PubMed

    Gentile, Luciana B; Nagamine, Marcia K; Biondi, Luiz R; Sanches, Daniel S; Toyota, Fábio; Giovani, Tatiane M; de Jesus, Isis P; da Fonseca, Ivone I M; Queiroz-Hazarbassanov, Nicolle; Diaz, Bruno L; Salles Gomes, Cristina de O Massoco; Dagli, Maria Lucia Z

    2017-01-01

    There are many factors which make canine cancer like cancer in humans. The occurrence of spontaneous mammary tumors in pet dogs, tumor genetics, molecular targets and exposure to the same environmental risk factors are among these factors. Therefore, the study of canine cancer can provide useful information to the oncology field. This study aimed to establish and characterize a panel of primary mixed cell cultures obtained from spontaneous canine mammary tumors. Eight established cell cultures obtained from one normal mammary gland, one complex adenoma, one mixed adenoma, two complex carcinomas and two mixed carcinomas were analyzed. The gene expression levels of classic molecular cancer players such as fibroblast growth factor receptor (FGFR) 2, breast cancer (BRCA) 1, BRCA2 and estrogen receptor (ESR) 1 were evaluated. For the first time, three orphan nuclear receptors, estrogen-related receptors (ERRs) α, β and γ were studied in canine mammary cancer. The highest expression level of ERRα was observed in complex carcinoma-derived cell culture, while the highest levels of ERRβ and γ were observed in cells derived from a mixed carcinoma. Meanwhile, complex carcinomas presented the highest levels of expression of ESR1, BRCA1 and FGFR2 among all samples. BRCA2 was found exclusively in complex adenoma. The transcription factor GATA3 had its highest levels in mixed carcinoma samples and its lowest levels in complex adenoma. Proliferation assays were also performed to evaluate the mixed cell cultures response to ER ligands, genistein and DES, both in normoxia and hypoxic conditions. Our results demonstrate that morphological and functional studies of primary mixed cell cultures derived from spontaneous canine mammary tumors are possible and provide valuable tool for the study of various stages of mammary cancer development.

  17. Social Orders and Interactions among Children in Age-Mixed Classes in Primary Schools--New Perspectives from a Synthesis of Ethnographic Data

    ERIC Educational Resources Information Center

    Huf, Christina; Raggl, Andrea

    2015-01-01

    The article synthesises data from two ethnographic projects, which both explore interactions of children in age-mixed groups in primary schools. It illuminates critical perspectives on social orders and children's interactions in age-mixed classes by showing how pupils in age-mixed groups become involved in power relations and how the teacher's…

  18. Preliminary results of lab-scale investigations of products of incomplete combustion during incineration of primary and mixed digested sludge.

    PubMed

    Braguglia, C M; Bagnuolo, G; Gianico, A; Mininni, G; Pastore, C; Mascolo, G

    2016-03-01

    Separation between primary and secondary sludge treatment could be a valuable solution for sludge management. According to this approach, secondary sludge can be conveniently used in agriculture while primary sludge could be easily dried and incinerated. It follows that some concern may arise from incinerating primary sludge with respect to the current practice to incinerate mixed digested sludge. Incineration of primary and mixed digested municipal sludge was investigated with a lab-scale equipment in terms of emissions of products of incomplete combustion (PICs) during incineration failure modes. PICs can be grouped in three sub-categories, namely aliphatic hydrocarbons (alkanes and alkenes), compounds with a single aromatic ring, and polycyclic aromatic hydrocarbons (PAHs). After-burning temperature was the most important parameter to be controlled in order to minimize emissions of alkanes and alkenes. As for mono-aromatic compounds, benzene and toluene are the most thermally resistant compounds, and in some cases, an after-burning temperature of 1100 °C was not enough to get the complete destruction of benzene leading to a residual emission of 18 mg/kgsludge. PAHs showed an opposite trend with respect to aliphatic and mono-aromatic hydrocarbons being the thermal failure mode the main responsible of PIC emissions. A proper oxygen concentration is more important than elevated temperature thus reflecting the high thermal stability of PAHs. Overall, obtained results, even though obtained under flameless conditions that are different from those of the industrial plants, demonstrated that separation of primary and secondary sludge does not pose any drawbacks or concern regarding primary sludge being disposed of by incineration even though it is more contaminated than mixed digested sludge in terms of organic pollutants.

  19. Incidence and prevalence of primary hyperparathyroidism in a racially mixed population.

    PubMed

    Yeh, Michael W; Ituarte, Philip H G; Zhou, Hui Cynthia; Nishimoto, Stacie; Liu, In-Lu Amy; Harari, Avital; Haigh, Philip I; Adams, Annette L

    2013-03-01

    The epidemiology of primary hyperparathyroidism (PHPT) has generally been studied in Caucasian populations. The aim was to examine the incidence and prevalence of PHPT within a racially mixed population. A descriptive epidemiologic study was performed. The study population included 3.5 million enrollees within Kaiser Permanente Southern California. All patients with at least one elevated serum calcium level (>10.5 mg/dL, 2.6 mmol/L) between 1995 and 2010 were included. Cases of PHPT were identified by electronic query of laboratory values using biochemical criteria, after exclusion of secondary or renal and tertiary hyperparathyroidism cases. The incidence and prevalence rates of PHPT were calculated according to sex, race, age group by decade, and year. Initial case finding identified 15,234 patients with chronic hypercalcemia, 13,327 (87%) of which had PHPT as defined by elevated or inappropriately normal parathyroid hormone levels. The incidence of PHPT fluctuated from 34 to 120 per 100,000 person-years (mean 66) among women, and from 13 to 36 (mean 25) among men. With advancing age, incidence increased and sex differences became pronounced (incidence 12-24 per 100,000 for both sexes younger than 50 y; 80 and 36 per 100,000 for women and men aged 50-59 y, respectively; and 196 and 95 for women and men aged 70-79 y, respectively). The incidence of PHPT was highest among blacks (92 women; 46 men, P < .0001), followed by whites (81 women; 29 men), with rates for Asians (52 women, 28 men), Hispanics (49 women, 17 men), and other races (25 women, 6 men) being lower than that for whites (P < .0001). The prevalence of PHPT tripled during the study period, increasing from 76 to 233 per 100,000 women and from 30 to 85 per 100 000 men. Racial differences in prevalence mirrored those found in incidence. PHPT is the predominant cause of hypercalcemia and is increasingly prevalent. Substantial differences are found in the incidence and prevalence of PHPT between races.

  20. The effect of haemagglutinating factor from boar seminal vesicle fluid on leucocytes.

    PubMed

    Veselský, L; Sedláková, E; Dostál, J; Hruban, V; Pazdera, J

    1981-01-01

    Complete agglutination of porcine, bovine, ovine, and rabbit leucocytes and of porcine and bovine thrombocytes was observed after their exposure to a 1% solution of the haemagglutinating protein isolated from boar seminal fluid. Bull seminal vesicle fluid had the same agglutinating effect on leucocytes, but did not agglutinate thrombocytes. Ram seminal plasma and other fluids from the reproductive tract of boar and bull did not agglutinate either leucocytes or thrombocytes. A viability test showed that the agglutinin of boar seminal vesicle fluid, bull seminal vesicle fluid, and boar prostate fluid were all lethal for leucocytes.

  1. Profiling leucocyte subsets in tuberculosis-diabetes co-morbidity.

    PubMed

    Kumar, Nathella Pavan; Moideen, Kadar; Dhakshinraj, Sharmila D; Banurekha, Vaithilingam V; Nair, Dina; Dolla, Chandrakumar; Kumaran, Paul; Babu, Subash

    2015-10-01

    The immune system plays an important role in the pathogenesis of pulmonary tuberculosis-type 2 diabetes mellitus (PTB-DM) co-morbidity. However, the phenotypic profile of leucocyte subsets at homeostasis in individuals with active or latent tuberculosis (LTB) with coincident diabetes is not known. To characterize the influence of diabetes on leucocyte phenotypes in PTB or LTB, we examined the frequency (Fo ) of leucocyte subsets in individuals with TB with (PTB-DM) or without (PTB) diabetes; individuals with latent TB with (LTB-DM) or without (LTB) diabetes and non-TB-infected individuals with (NTB-DM) or without (NTB) diabetes. Coincident DM is characterized by significantly lower Fo of effector memory CD4(+) T cells in LTB individuals. In contrast, DM is characterized by significantly lower Fo of effector memory CD8(+) T cells and significantly higher Fo of central memory CD8(+) T cells in PTB individuals. Coincident DM resulted in significantly higher Fo of classical memory B cells in PTB and significantly higher Fo of activated memory and atypical B cells in LTB individuals. Coincident DM resulted in significantly lower Fo of classical and intermediate monocytes in PTB, LTB and NTB individuals. Finally, DM resulted in significantly lower Fo of myeloid and plasmacytoid dendritic cells in PTB, LTB and NTB individuals. Our data reveal that coincident diabetes alters the cellular subset distribution of T cells, B cells, dendritic cells and monocytes in both individuals with active TB and those with latent TB, thus potentially impacting the pathogenesis of this co-morbid condition. © 2015 John Wiley & Sons Ltd.

  2. Polymorphonuclear leucocyte motility in men with ankylosing spondylitis.

    PubMed Central

    Pease, C T; Fennell, M; Brewerton, D A

    1989-01-01

    The polymorphonuclear leucocyte (PMN) response to a chemotactic or chemokinetic stimulus is enhanced in men with ankylosing spondylitis (AS). This effect does not parallel the severity of disease activity or the size of the acute phase response, and it is independent of non-steroidal anti-inflammatory drug treatment. Polymorph function is normal in HLA-B27 positive brothers of probands with AS and in other HLA-B27 positive individuals in the absence of disease. Polymorph motility is also normal in patients with psoriasis vulgaris or Crohn's disease, indicating that enhanced PMN motility is not a non-specific consequence of all inflammatory disorders. PMID:2784306

  3. Griseofulvin inhibition of polymorphonuclear leucocyte chemotaxis in Boyden chambers.

    PubMed

    Bandmann, U; Norberg, B; Simmingsköld, G

    1975-09-01

    Griseofulvin inhibited the chemotaxis of polymorphonuclear leucocytes (PMN's) in vitro in the concentration range 0.1-1.0 mug/ml, i.e. at concentrations comparable to those obtained in serum during peroral treatment with griseofulvin. It is suggested that PMN chemotaxis is inhibited by griseofulvin interference with the redistribution of cytoplasmic microtubules, which is thought to be essential in the direction-finding of PMNs during chemotaxis. Furthermore, it is suggested that the griseofulvin inhibition of PMN chemotaxis - together with the previously known pharmacodynamic properties of griseofulvin - may provide the rationale for griseofulvin therapy in PMN-mediated tissue injury of the gut.

  4. Multidrug punch cards in primary care: a mixed methods study on patients' preferences and impact on adherence

    PubMed Central

    Boeni, Fabienne; Hersberger, Kurt E.; Arnet, Isabelle

    2014-01-01

    Background: Multidrug punch cards are frame cards with 28 plastic cavities filled with a patient's oral solid medication. They are used in primary care to facilitate medication management and to enhance adherence. Main criticism concerned handling difficulties and fading knowledge about medication of patients using them. This study aimed at exploring daily use, preferences, and adherence of primary care patients using multidrug punch cards. Methods: Community pharmacies in Switzerland recruited primary care patients using multidrug punch cards. A mixed methods approach was applied with quantitative interviews performed by telephone and qualitative interviews face-to-face. Results: Of 149 eligible patients from 21 community pharmacies, 22 participated 2011 in the quantitative and 11 participated 2013/14 in the qualitative interview. Patients were very satisfied with the multidrug punch cards and stated increased medication safety. All considered adherence as very important. Self-reported adherence was 10 (median) on a visual analog scale (0 = no intake, 10 = perfect adherence). The absence of package inserts and predefined handling difficulties e.g., tablets spiking at removal were not perceived as problems. Conclusions: Patients are satisfied with the multidrug punch cards, feel safe, mostly have no handling problems and adhere to their treatment. Trust in health-care professionals and patients' experiences emerged as key variables for initiating multidrug punch card use and for medication adherence. This mixed methods study invalidates previous concerns about disadvantages of multidrug punch cards. Health-care professionals should actively recommend them for primary care patients with polypharmacy and poor adherence. PMID:25324777

  5. Multidrug punch cards in primary care: a mixed methods study on patients' preferences and impact on adherence.

    PubMed

    Boeni, Fabienne; Hersberger, Kurt E; Arnet, Isabelle

    2014-01-01

    Multidrug punch cards are frame cards with 28 plastic cavities filled with a patient's oral solid medication. They are used in primary care to facilitate medication management and to enhance adherence. Main criticism concerned handling difficulties and fading knowledge about medication of patients using them. This study aimed at exploring daily use, preferences, and adherence of primary care patients using multidrug punch cards. Community pharmacies in Switzerland recruited primary care patients using multidrug punch cards. A mixed methods approach was applied with quantitative interviews performed by telephone and qualitative interviews face-to-face. Of 149 eligible patients from 21 community pharmacies, 22 participated 2011 in the quantitative and 11 participated 2013/14 in the qualitative interview. Patients were very satisfied with the multidrug punch cards and stated increased medication safety. All considered adherence as very important. Self-reported adherence was 10 (median) on a visual analog scale (0 = no intake, 10 = perfect adherence). The absence of package inserts and predefined handling difficulties e.g., tablets spiking at removal were not perceived as problems. Patients are satisfied with the multidrug punch cards, feel safe, mostly have no handling problems and adhere to their treatment. Trust in health-care professionals and patients' experiences emerged as key variables for initiating multidrug punch card use and for medication adherence. This mixed methods study invalidates previous concerns about disadvantages of multidrug punch cards. Health-care professionals should actively recommend them for primary care patients with polypharmacy and poor adherence.

  6. Influence of nutrients and mixing on the primary production and community respiration in the Gulf of Riga

    NASA Astrophysics Data System (ADS)

    Olesen, Michael; Lundsgaard, Claus; Andrushaitis, Andris

    1999-12-01

    Rates of plankton community production and respiration in relation to wind, solar radiation, biomass and nutrients were measured in the mixed layer during a late spring, a mid-summer and an early autumn situation in the Gulf of Riga. System metabolism was estimated by the in vitro oxygen method and from oxygen mass balance directly in the water. Gross production (GPP) remained fairly stable throughout the investigation periods with a mean of 1.8 g C m -2 day -1 (range: 0.5-3.9 g C m -2 day -1). Community respiration varied from 0.5 to 6.4 g C m -2 day -1 during the two summer studies (the high respiration rates ranging from 3.0 to 10.4 g C m -2 day -1 found during the autumn cruise were probably overestimates). Respiration rates generally exceeded GPP indicating that the system was apparently net-heterotrophic during the three investigation periods. The predominant limiting factor for phytoplankton growth were nutrients during spring and summer. In autumn the combination of low light levels and high vertical mixing due to wind lead to conditions of light limitation. However, growth conditions were affected by the stabilizing (solar heating) and destabilizing (wind) forces acting on the water column in all three situations. Depending on the relative strength of these forces, day-to-day primary production could vary considerably. Under conditions of nutrient limitation mixing may increase photosynthesis; this is a consequence of improved exploitation of incident light and of available nutrients in the mixed layer. The strong dependency of mixing on primary production implies that measurements based on in vitro incubations may underestimate "real" production.

  7. In vivo leucocyte migration in Behçet's syndrome.

    PubMed Central

    Efthimiou, J; Addison, I E; Johnson, B V

    1989-01-01

    Serial studies of leucocyte migration in vivo were carried out in 15 patients with Behçet's syndrome using a skin window technique. Where possible, patients with and without active disease were studied during and in the absence of treatment. In patients with active disease neutrophil migration was frequently greater than normal, particularly with respect to numbers of cells migrating. There was also an increased frequency of emigrating neutrophils with less or more nuclear lobes than normal. In three patients in whom function of skin window neutrophils was studied nitroblue tetrazolium reduction and phagocytosis and killing of Candida guilliermondiae were normal. The monocyte component of the skin window was more often reduced in patients than in normal controls. Corticosteroid treatment did not exert a major effect on leucocyte migration, though the doses involved were relatively small. Neutrophil abnormalities were common in patients and particularly those with active disease. These results suggest that neutrophil hyperactivity may have an important role in the pathogenesis of Behçet's syndrome. PMID:2649027

  8. The Genetic Origin of Leucocytic Mucopolysaccharides in Cancer Patients

    PubMed Central

    Riesco, A.; Coke, R. Cruz

    1973-01-01

    The presence or absence of lymphocytic mucopolysaccharides (MPS) is studied in 223 subjects: 100 normals (controls); 8 cancer patients cured for more than 6 years; 30 cancer patients at the start of their treatment; and 85 relatives of first degree consanguinity of these last patients. The data are studied by statistical and genetic analysis. The results confirm the findings reported earlier and show that the difference in the probability of a high frequency of leucocytic MPS between the relatives of cancer patients and the controls is highly significant. Furthermore, this probability in a relative of first degree of consanguinity of a cancer patient is more than three times greater than in an individual of the general population. Genetic segregation analysis shows that the high leucocytic MPS trait segregates in the families of cancer patients after a classic pattern of dominant autosomal inheritance. Applying Falconer's nomogram it is concluded that the whole of this phenotypic variation is of genetic origin. Its interrelationships with cancer are discussed and it is postulated that this disturbance of the lymphocytic MPS represents a subclinical variant, not known until now, of the clinical mucopolysaccaridoses. PMID:4270340

  9. Ethnicity and Ethnically "Mixed" Identity in Belize: A Study of Primary School-Age Children.

    ERIC Educational Resources Information Center

    Haug, Sarah Woodbury

    1998-01-01

    Nationalism, as taught in Belize schools, is panethnic and multiethnic, but because the increasingly widespread practice of ethnic mixing is not acknowledged, there is a discrepancy between what is taught and the daily life of students. Research results from 161 elementary school children show that the ethnic self-identification of children is…

  10. Experiences with primary healthcare in Fuzhou, urban China, in the context of health sector reform: a mixed methods study.

    PubMed

    McCollum, Rosalind; Chen, Lieping; ChenXiang, Tang; Liu, Xiaoyun; Starfield, Barbara; Jinhuan, Zheng; Tolhurst, Rachel

    2014-01-01

    China has recently placed increased emphasis on the provision of primary healthcare services through health sector reform, in response to inequitably distributed health services. With increasing funding for community level facilities, now is an opportune time to assess the quality of primary care delivery and identify areas in need of further improvement. A mixed methodology approach was adopted for this study. Quantitative data were collected using the Primary Care Assessment Tool-Chinese version (C-PCAT), a questionnaire previously adapted for use in China to assess the quality of care at each health facility, based on clients' experiences. In addition, qualitative data were gathered through eight semi-structured interviews exploring perceptions of primary care with health directors and a policy maker to place this issue in the context of health sector reform. The study found that patients attending community health and sub-community health centres are more likely to report better experiences with primary care attributes than patients attending hospital facilities. Generally low scores for community orientation, family centredness and coordination in all types of health facility indicate an urgent need for improvement in these areas. Healthcare directors and policy makers perceived the need for greater coordination between levels of health providers, better financial reimbursement, more formal government contracts and recognition/higher status for staff at the community level and more appropriate undergraduate and postgraduate training. Copyright © 2013 John Wiley & Sons, Ltd.

  11. A novel somatic MAPK1 mutation in primary ovarian mixed germ cell tumors.

    PubMed

    Zou, Yang; Deng, Wei; Wang, Feng; Yu, Xiao-Hong; Liu, Fa-Ying; Yang, Bi-Cheng; Huang, Mei-Zhen; Guo, Jiu-Bai; Xie, Qiu-Hua; He, Ming; Huang, Ou-Ping

    2016-02-01

    A recent exome-sequencing study revealed prevalent mitogen-activated protein kinase 1 (MAPK1) p.E322K mutation in cervical carcinoma. It remains largely unknown whether ovarian carcinomas also harbor MAPK1 mutations. As paralogous gene mutations co‑occur frequently in human malignancies, we analyzed here a total of 263 ovarian carcinomas for the presence of MAPK1 and paralogous MAPK3 mutations by DNA sequencing. A previously unreported MAPK1 p.D321N somatic mutation was identified in 2 out of 18 (11.1%) ovarian mixed germ cell tumors, while no other MAPK1 or MAPK3 mutation was detected in our samples. Of note, OCC‑115, the MAPK1‑mutated sample with bilateral cancerous ovaries affected, harbored MAPK1 mutation in the right ovary while retained the left ovary intact, implicating that the genetic alterations underlying ovarian mixed germ cell tumor may be different, even in patients with similar genetic backgrounds and tumor microenvironments. The results of evolutionary conservation and protein structure modeling analysis implicated that MAPK1 p.D321N mutation may be pathogenic. Additionally, mutations in protein phosphatase 2 regulatory subunit α (PPP2R1A), ring finger protein 43 (RNF43), DNA directed polymerase ε (POLE1), ribonuclease type III (DICER1), CCCTC‑binding factor (CTCF), ribosomal protein L22 (RPL22), DNA methyltransferase 3α (DNMT3A), transformation/transcription domain‑associated protein (TRRAP), isocitrate dehydrogenase (IDH)1 and IDH2 were not detected in ovarian mixed germ cell tumors, implicating these genetic alterations may be not associated with MAPK1 mutation in the development of this malignancy. The present study identified a previously unreported MAPK1 mutation in ovarian mixed germ cell tumors for the first time, and this mutation may be actively involved in the tumorigenesis of this disease.

  12. [Pathological features and origin of primary pineal mixed germ cell tumors].

    PubMed

    XIAO, Gang; FANG, Lu-xiong; QIU, Bing-hui; QI, Song-tao

    2011-03-01

    To investigate the origin of mixed germ cell tumors in the pineal region based on the image data, surgical findings and pathological examination of the tumor. The preoperative CT and magnetic resonance imaging (MRI) findings and tumor specimens were retrospectively analyzed in 15 cases of pineal mixed germ cell tumors confirmed by postoperative histological examination between January 2000 and September 2010. Radiographic examination of the tumor revealed calcification in 12 cases, cystic changes in 10 cases, and the presence of lipid in 5 cases. On the anteroposterior images, the tumors appeared round or elliptic with smooth edge in 6 cases, and showed irregular shape with multiple processes on the edge in 9 cases. Surgical exploration found all the tumors located in the the suprapineal recess enclosed by the arachnoidal envelope of the Galen vein. Pathologically, 13 specimens contained germinoma component, 9 contained teratoma component, 4 had embryonic carcinoma component, 3 had choriocarcinoma component, 7 showed yolk sac tumor component, and 3 showed rhabdomyoma component. Germinoma components were found on the tumor margin in 7 specimens, and intermingled germinoma and other components were found in 10 specimens. Pineal mixed germ cell tumor originates from the residue germ cells around the pineal gland, and most likely evolves from single primordial germ cells.

  13. Building Secure Attachments for Primary School Children: A Mixed Methods Study

    ERIC Educational Resources Information Center

    Ubha, Neerose; Cahill, Sharon

    2014-01-01

    Despite the wide implications of attachment theory there remains a lack of research exploring interventions which encapsulate the principles of an attachment-based framework in the school context. The aim of this research was to address this gap by implementing an intervention for a group of five primary-aged pupils with identified insecure…

  14. Mixed Messages: How Primary Agents of Socialization Influence Adolescent Females Who Identify as Multiracial-Bisexual

    ERIC Educational Resources Information Center

    King, Alissa R.

    2013-01-01

    The goals of this study were to highlight the often stigmatized and invisible identities of six female participants who identify as multiracial/biracial-bisexual/pansexual, focusing on the pre-college context. Findings, using in-depth interviews, indicated that the primary socializing agents within the pre-college context strongly influenced…

  15. Building Secure Attachments for Primary School Children: A Mixed Methods Study

    ERIC Educational Resources Information Center

    Ubha, Neerose; Cahill, Sharon

    2014-01-01

    Despite the wide implications of attachment theory there remains a lack of research exploring interventions which encapsulate the principles of an attachment-based framework in the school context. The aim of this research was to address this gap by implementing an intervention for a group of five primary-aged pupils with identified insecure…

  16. Safety in Numbers? Middle-Class Parents and Social Mix in London Primary Schools

    ERIC Educational Resources Information Center

    Vowden, Kim James

    2012-01-01

    Research into parents' secondary-school choices suggests that many middle-class parents are keen to secure a middle-class peer group for their children. This article reports the findings of a small-scale, qualitative study into whether a similar phenomenon exists at primary-school level and, if so, why. In-depth interviews were conducted with 56…

  17. Increased leucocyte apoptosis in transfused β-thalassaemia patients

    PubMed Central

    Walter, Patrick B.; Porter, John; Evans, Patricia; Kwiatkowski, Janet L.; Neufeld, Ellis J.; Coates, Thomas; Giardina, Patricia J.; Grady, Robert W.; Vichinsky, Elliott; Olivieri, Nancy; Trachtenberg, Felicia; Alberti, Daniele; Fung, Ellen; Ames, Bruce; Higa, Annie; Harmatz, Paul

    2015-01-01

    Summary This exploratory study assessed apoptosis in peripheral blood leucocytes (PBL) from β-thalassaemia patients receiving chronic transfusions and chelation therapy (deferasirox or deferoxamine) at baseline, 1, 6, and 12 months. At baseline, thalassaemic PBLs presented 50% greater levels of Bax (BAX), 75% higher caspase-3/7, 48% higher caspase-8 and 88% higher caspase-9 activities and 428% more nucleosomal DNA fragmentation than control subjects. Only neutrophils correlated significantly with apoptotic markers. Previously, we showed that over the treatment year, hepatic iron declined; we now show that the ratio of Bax/Bcl-2 (BCL2), (−27.3%/year), and caspase-9 activity (−13.3%/year) declined in both treatment groups, suggesting that chelation decreases body iron and indicators of PBL apoptosis. PMID:23216540

  18. Increased leucocyte apoptosis in transfused β-thalassaemia patients.

    PubMed

    Walter, Patrick B; Porter, John; Evans, Patricia; Kwiatkowski, Janet L; Neufeld, Ellis J; Coates, Thomas; Giardina, Patricia J; Grady, Robert W; Vichinsky, Elliott; Olivieri, Nancy; Trachtenberg, Felicia; Alberti, Daniele; Fung, Ellen; Ames, Bruce; Higa, Annie; Harmatz, Paul

    2013-02-01

    This exploratory study assessed apoptosis in peripheral blood leucocytes (PBL) from β-thalassaemia patients receiving chronic transfusions and chelation therapy (deferasirox or deferoxamine) at baseline, 1, 6, and 12 months. At baseline, thalassaemic PBLs presented 50% greater levels of Bax (BAX), 75% higher caspase-3/7, 48% higher caspase-8 and 88% higher caspase-9 activities and 428% more nucleosomal DNA fragmentation than control subjects. Only neutrophils correlated significantly with apoptotic markers. Previously, we showed that over the treatment year, hepatic iron declined; we now show that the ratio of Bax/Bcl-2 (BCL2), (-27·3%/year), and caspase-9 activity (-13·3%/year) declined in both treatment groups, suggesting that chelation decreases body iron and indicators of PBL apoptosis. © 2012 Blackwell Publishing Ltd.

  19. Novel distribution of the secretory leucocyte proteinase inhibitor in kidney.

    PubMed Central

    Ohlsson, S; Ljungkrantz, I; Ohlsson, K; Segelmark, M; Wieslander, J

    2001-01-01

    The secretory leucocyte proteinase inhibitor (SLPI) is a low molecular weight, tissue-specific inhibitor of, for example, elastase and cathepsin G, which also have antimicrobial capacity. SLPI has been localised to the respiratory, gastrointestinal and genital tracts, but so far not to the kidney. The presence of SLPI in renal tubuli cells was demonstrated using immunohistochemistry and, by means of in situ hybridisation on human renal biopsies, we were able to demonstrate SLPI production. In various inflammatory conditions in the kidneys, the protease-antiprotease balance is disturbed. For this reason, as well as the possible role in the defence against ascending urinary tract infections, it is interesting to establish a source of SLPI in renal tubuli cells. PMID:11817677

  20. Leucocyte function in Crohn's disease. Studies on mobilisation using a quantitative skin window technique and on the function of circulating polymorphonuclear leucocytes in vitro.

    PubMed Central

    Wandall, J H; Binder, V

    1982-01-01

    Leucocyte function was evaluated by mobilisation to skin windows with chambers and by the chemotactic, phagocytic, and nitro blue tetrazolium (NBT) reducing activity of circulating leucocytes in vitro in 20 patients with Crohn's disease, 21 healthy volunteers, and nine patients with sarcoidosis or tuberculosis. Leucocytes had been mobilised in significantly reduced numbers at 12, 24, 36, and 48 hours in Crohn's disease compared with healthy volunteers (P less than 0.01) and patients with sarcoidosis/tuberculosis (P less than 0.01). The leucocyte migration rate showed that mobilisation in Crohn's disease begins after a prolonged lag phase and is reduced compared with healthy volunteers (P less than 0.01) and patients with sarcoidosis/tuberculosis (P less than 0.02). The reduced mobilisation was not correlated with disease activity. In vitro random migration by leucocytes was slightly lower in Crohn's disease (P less than 0.05) than in healthy volunteers, but there was no difference after removal of the autologous plasma. Chemotactic response to casein did not differ between the groups studied. Serum independent and dependent phagocytosis did not differ from control groups. Serum independent phagocytosis was positively and significantly correlated to the disease activity (rho 0.4812, P less than 0.05). Resting leucocyte NBT reduction was increased in Crohn's disease and sarcoidosis/tuberculosis (P less than 0.01), but during phagocytosis a lower NBT reduction was found in Crohn's disease than in healthy volunteers (P less than 0.02). The inflammatory response in Crohn's disease, with reduced leucocyte accumulation, differs from patients with other granulomatous reactions and is independent of the disease activity. Our data suggest that the defect is not cellular. They support the hypothesis that a pathogenic factor in Crohn's disease may be foreign material that is normally eliminated remaining in the tissue and eliciting a chronic inflammatory response. PMID:7040174

  1. Sorption and Release of Organics by Primary, Anaerobic, and Aerobic Activated Sludge Mixed with Raw Municipal Wastewater

    PubMed Central

    Modin, Oskar; Saheb Alam, Soroush; Persson, Frank; Wilén, Britt-Marie

    2015-01-01

    New activated sludge processes that utilize sorption as a major mechanism for organics removal are being developed to maximize energy recovery from wastewater organics, or as enhanced primary treatment technologies. To model and optimize sorption-based activated sludge processes, further knowledge about sorption of organics onto sludge is needed. This study compared primary-, anaerobic-, and aerobic activated sludge as sorbents, determined sorption capacity and kinetics, and investigated some characteristics of the organics being sorbed. Batch sorption assays were carried out without aeration at a mixing velocity of 200 rpm. Only aerobic activated sludge showed net sorption of organics. Sorption of dissolved organics occurred by a near-instantaneous sorption event followed by a slower process that obeyed 1st order kinetics. Sorption of particulates also followed 1st order kinetics but there was no instantaneous sorption event; instead there was a release of particles upon mixing. The 5-min sorption capacity of activated sludge was 6.5±10.8 mg total organic carbon (TOC) per g volatile suspend solids (VSS) for particulate organics and 5.0±4.7 mgTOC/gVSS for dissolved organics. The observed instantaneous sorption appeared to be mainly due to organics larger than 20 kDa in size being sorbed, although molecules with a size of about 200 Da with strong UV absorbance at 215–230 nm were also rapidly removed. PMID:25768429

  2. Morphology of Mixed Primary and Secondary Organic Particles and the Adsorption of Spectator Organic Gases during Aerosol Formation

    SciTech Connect

    Vaden, Timothy D.; Song, Chen; Zaveri, Rahul A.; Imre, D.; Zelenyuk, Alla

    2010-04-13

    Traditional semi-empirical secondary organic aerosol (SOA) models assume that SOA mixes well with primary organic aerosols (POA), which significantly enhances the modeled SOA yields. These models further assume that the organic compounds in the gas phase do no condense on SOA as it forms. These assumptions were challenged through a detailed experimental investigation of the compositions and morphologies of SOA particles formed during ozonolysis of α-pinene in the presence of dioctyl phthalate (DOP) particles and DOP gas phase component using a single particle mass spectrometer. Ultraviolet (UV) laser depth-profiling experiments were used to characterize different types of mixed SOA/DOP particles: those formed by condensation of the oxidized α-pinene products on size-selected DOP particles and by condensation of DOP on size-selected α-pinene SOA particles. The results of these measurements conclusively show that the hydrophilic SOA and hydrophobic DOP do not mix, but instead form distinct phases. An examination of homogeneously-nucleated SOA particles formed in the presence of DOP shows them to be encapsulated by a thin DOP layer. Thus SOA can adsorb gas-phase DOP even though it has an extremely low vapor pressure (1.3×10-7 Torr), which has significant implications for SOA formation and fate in the atmosphere, where numerous organic compounds with various volatilities are present.

  3. Morphology, composition, and mixing state of primary particles from combustion sources - crop residue, wood, and solid waste.

    PubMed

    Liu, Lei; Kong, Shaofei; Zhang, Yinxiao; Wang, Yuanyuan; Xu, Liang; Yan, Qin; Lingaswamy, A P; Shi, Zongbo; Lv, Senlin; Niu, Hongya; Shao, Longyi; Hu, Min; Zhang, Daizhou; Chen, Jianmin; Zhang, Xiaoye; Li, Weijun

    2017-07-11

    Morphology, composition, and mixing state of individual particles emitted from crop residue, wood, and solid waste combustion in a residential stove were analyzed using transmission electron microscopy (TEM). Our study showed that particles from crop residue and apple wood combustion were mainly organic matter (OM) in smoldering phase, whereas soot-OM internally mixed with K in flaming phase. Wild grass combustion in flaming phase released some Cl-rich-OM/soot particles and cardboard combustion released OM and S-rich particles. Interestingly, particles from hardwood (pear wood and bamboo) and softwood (cypress and pine wood) combustion were mainly soot and OM in the flaming phase, respectively. The combustion of foam boxes, rubber tires, and plastic bottles/bags in the flaming phase released large amounts of soot internally mixed with a small amount of OM, whereas the combustion of printed circuit boards and copper-core cables emitted large amounts of OM with Br-rich inclusions. In addition, the printed circuit board combustion released toxic metals containing Pb, Zn, Sn, and Sb. The results are important to document properties of primary particles from combustion sources, which can be used to trace the sources of ambient particles and to know their potential impacts in human health and radiative forcing in the air.

  4. Effect of gonadotropin-releasing hormone on phagocytic leucocytes of rainbow trout.

    PubMed

    Yada, Takashi

    2012-03-01

    To clarify the role of gonadotropin-releasing hormone (GnRH) in the fish immune system, in vitro effect of GnRH was examined in phagocytic leucocytes of rainbow trout (Oncorhynchus mykiss). Gene expression of GnRH-receptor was detected by RT-PCR in leucocytes from head kidney. Administration of sGnRH increased proliferation and mRNA levels of a proinflammatory cytokine, tumor necrosis factor (TNF)-α, in trout leucocytes. Superoxide production in zymosan-stimulated phagocytic leucocytes was also increased by sGnRH in a dose-related manner from 0.01 to 100 nM. There was no significant effect of sGnRH on mRNA levels of growth hormone (GH) expressed in trout phagocytic leucocytes. Immunoneutralization of GH by addition of anti-salmon GH serum into the medium could not block the stimulatory effect of sGnRH on superoxide production. These results indicate that GnRH stimulates phagocytosis in fish leucocytes through a GnRH-receptor-dependent pathway, and that the effect of GnRH is not mediated through paracrine GH in leucocytes.

  5. The presence of eosinophil leucocytes in cervicovaginal smears with Actinomyces-like organisms: Light microscopic examination.

    PubMed

    Kaya, Dilek; Demirezen, Sayeste; Beksaç, Mehmet Sinan

    2012-10-01

    Actinomyces species are part of mucosal surfaces of oral cavity, gastrointestinal and genital tracts. When these mucosal surfaces disrupt, Actinomyces become pathogen and cause infection. Eosinophil leucocytes participate in host defense against helminthic infestation and they generally play a role in asthma and allergy. However, the role of eosinophil leucocytes in host defense against bacteria is conflicting. To determine whether there is a relationship between Actinomyces-like organisms (ALOs) and eosinophil leucocytes at light microscopic level. Cervicovaginal samples obtained from 200 patients were examined by both Pap smear microscopy and anaerobic culturing. Since the results obtained by these methods were not concordant for diagnosis of genital Actinomyces, 6 of 200 patients (3%) diagnosed with ALOs by Pap smear microscopy became the study group. Patients without any infectious agents (n=134) were the control group. Statistical analyses were conducted with χ(2) test using SPSS program. The study and control groups were compared statistically in view of the presence of eosinophil leucocytes and it was found that there was a significant correlation between the presence of ALOs and eosinophil leucocytes (P<0.05). Abundant polymorphonuclear leucocytes (PMNLs) and macrophages were also detected in the study group. This study implies that eosinophil leucocytes might have a role in host defense against Actinomyces in addition to PMNLs and macrophages.

  6. User-generated quality standards for youth mental health in primary care: a participatory research design using mixed methods

    PubMed Central

    Graham, Tanya; Rose, Diana; Murray, Joanna; Ashworth, Mark; Tylee, André

    2014-01-01

    Objectives To develop user-generated quality standards for young people with mental health problems in primary care using a participatory research model. Methods 50 young people aged 16–25 from community settings and primary care participated in focus groups and interviews about their views and experiences of seeking help for mental health problems in primary care, cofacilitated by young service users and repeated to ensure respondent validation. A second group of young people also aged 16–25 who had sought help for any mental health problem from primary care or secondary care within the last 5 years were trained as focus groups cofacilitators (n=12) developed the quality standards from the qualitative data and participated in four nominal groups (n=28). Results 46 quality standards were developed and ranked by young service users. Agreement was defined as 100% of scores within a two-point region. Group consensus existed for 16 quality standards representing the following aspects of primary care: better advertising and information (three); improved competence through mental health training and skill mix within the practice (two); alternatives to medication (three); improved referral protocol (three); and specific questions and reassurances (five). Alternatives to medication and specific questions and reassurances are aspects of quality which have not been previously reported. Conclusions We have demonstrated the feasibility of using participatory research methods in order to develop user-generated quality standards. The development of patient-generated quality standards may offer a more formal method of incorporating the views of service users into quality improvement initiatives. This method can be adapted for generating quality standards applicable to other patient groups. PMID:24920648

  7. Internet cognitive behavioural therapy for mixed anxiety and depression: a randomized controlled trial and evidence of effectiveness in primary care.

    PubMed

    Newby, J M; Mackenzie, A; Williams, A D; McIntyre, K; Watts, S; Wong, N; Andrews, G

    2013-12-01

    Major depressive disorder (MDD) and generalized anxiety disorder (GAD) have the highest co-morbidity rates within the internalizing disorders cluster, yet no Internet-based cognitive behavioural therapy (iCBT) programme exists for their combined treatment. We designed a six-lesson therapist-assisted iCBT programme for mixed anxiety and depression. Study 1 was a randomized controlled trial (RCT) comparing the iCBT programme (n = 46) versus wait-list control (WLC; n = 53) for patients diagnosed by structured clinical interview with MDD, GAD or co-morbid GAD/MDD. Primary outcome measures were the Patient Health Questionnaire nine-item scale (depression), Generalized Anxiety Disorder seven-item scale (generalized anxiety), Kessler 10-item Psychological Distress scale (distress) and 12-item World Health Organization Disability Assessment Schedule II (disability). The iCBT group was followed up at 3 months post-treatment. In study 2, we investigated the adherence to, and efficacy of the same programme in a primary care setting, where patients (n = 136) completed the programme under the supervision of primary care clinicians. The RCT showed that the iCBT programme was more effective than WLC, with large within- and between-groups effect sizes found (>0.8). Adherence was also high (89%), and gains were maintained at 3-month follow-up. In study 2 in primary care, adherence to the iCBT programme was low (41%), yet effect sizes were large (>0.8). Of the non-completers, 30% experienced benefit. Together, the results show that iCBT is effective and adherence is high in research settings, but there is a problem of adherence when translated into the 'real world'. Future efforts need to be placed on developing improved adherence to iCBT in primary care settings.

  8. Hypertension management in rural primary care facilities in Zambia: a mixed methods study.

    PubMed

    Yan, Lily D; Chirwa, Cindy; Chi, Benjamin H; Bosomprah, Samuel; Sindano, Ntazana; Mwanza, Moses; Musatwe, Dennis; Mulenga, Mary; Chilengi, Roma

    2017-02-03

    Improved primary health care is needed in developing countries to effectively manage the growing burden of hypertension. Our objective was to evaluate hypertension management in Zambian rural primary care clinics using process and outcome indicators to assess the screening, monitoring, treatment and control of high blood pressure. Better Health Outcomes through Mentoring and Assessment (BHOMA) is a 5-year, randomized stepped-wedge trial of improved clinical service delivery underway in 46 rural Zambian clinics. Clinical data were collected as part of routine patient care from an electronic medical record system, and reviewed for site performance over time according to hypertension related indicators: screening (blood pressure measurement), management (recorded diagnosis, physical exam or urinalysis), treatment (on medication), and control. Quantitative data was used to develop guides for qualitative in-depth interviews, conducted with health care providers at a proportional sample of half (20) of clinics. Qualitative data was iteratively analyzed for thematic content. From January 2011 to December 2014, 318,380 visits to 46 primary care clinics by adults aged ≥ 25 years with blood pressure measurements were included. Blood pressure measurement at vital sign screening was initially high at 89.1% overall (range: 70.1-100%), but decreased to 62.1% (range: 0-100%) by 48 months after intervention start. The majority of hypertensive patients made only one visit to the clinics (57.8%). Out of 9022 patients with at least two visits with an elevated blood pressure, only 49.3% had a chart recorded hypertension diagnosis. Process indicators for monitoring hypertension were <10% and did not improve with time. In in-depth interviews, antihypertensive medication shortages were common, with 15/20 clinics reporting hydrochlorothiazide-amiloride stockouts. Principal challenges in hypertension management included 1) equipment and personnel shortages, 2) provider belief that

  9. Accommodations for Patients with Disabilities in Primary Care: A Mixed Methods Study of Practice Administrators

    PubMed Central

    Pharr, Jennifer R

    2014-01-01

    Structural barriers that limit access to health care services for people with disabilities have been identified through qualitative studies; however, little is known about how patients with disabilities are accommodated in the clinical setting when a structural barrier is encountered. The purpose of this study was to identify how primary care medical practices in the United States accommodated people with disabilities when a barrier to service is encountered. Primary care practice administrators from the medical management organization were identified through the organization’s website. Sixty-three administrators from across the US participated in this study. Practice administrators reported that patients were examined in their wheelchairs (76%), that parts of the exam where skipped when a barrier was encountered (44%), that patients were asked to bring someone with them (52.4%) or that patients were refused treatment due to an inaccessible clinic (3.2%). These methods of accommodation would not be in compliance with requirements of the Americans with Disabilities Act. There was not a significant difference (p>0.05) in accommodations for patients with disabilities between administrators who could describe the application of the ADA to their clinic and those who could not. Practice administrators need a comprehensive understanding of the array of challenges encountered by patients with disabilities throughout the health care process and of how to best accommodate patients with disabilities in their practice. PMID:24373261

  10. Mixed primary squamous cell carcinoma, follicular carcinoma, and micropapillary carcinoma of the thyroid gland: A case report.

    PubMed

    Dong, Su; Song, Xue-Song; Chen, Guang; Liu, Jia

    2016-08-01

    Primary squamous cell carcinoma of the thyroid gland is rare, and mixed squamous cell and follicular carcinoma is even rarer still, with only a few cases reported in the literature. The simultaneous presentation of three primary cancers of the thyroid has not been reported previously. Here we report a case of primary squamous cell carcinoma of the thyroid, follicular thyroid carcinoma, and micropapillary thyroid carcinoma. A 62-year-old female patient presented with complaints of pain and a 2-month history of progressively increased swelling in the anterior region of the neck. Fine-needle-aspiration cytology of both lobes indicated the possibility of the presence of a follicular neoplasm. Total thyroidectomy with left-sided modified radical neck dissection was performed. Postoperative pathological examination confirmed the diagnosis of thyroid follicular carcinoma with squamous cell carcinoma and micropapillary carcinoma of the thyroid. Thyroid-stimulating hormone suppressive therapy with l-thyroxine was administered. Radioiodine and radiotherapy also were recommended, but the patient did not complete treatment as scheduled. The patient remained alive more than 9 months after operation. The present case report provides an example of the coexistence of multiple distinct malignancies in the thyroid.

  11. Food safety knowledge, practices and beliefs of primary food preparers in families with young children. A mixed methods study.

    PubMed

    Meysenburg, Rebecca; Albrecht, Julie A; Litchfield, Ruth; Ritter-Gooder, Paula K

    2014-02-01

    Food preparers in families with young children are responsible for safe food preparation and handling to prevent foodborne illness. To explore the food safety perceptions, beliefs, and practices of primary food preparers in families with children 10 years of age and younger, a mixed methods convergent parallel design and constructs of the Health Belief Model were used. A random sampling of 72 primary food handlers (36.2±8.6 years of age, 88% female) within young families in urban and rural areas of two Midwestern states completed a knowledge survey and participated in ten focus groups. Quantitative data were analyzed using SPSS. Transcribed interviews were analyzed for codes and common themes. Forty-four percent scored less than the average knowledge score of 73%. Participants believe children are susceptible to foodborne illness but perceive its severity to be low with gastrointestinal discomfort as the primary outcome. Using safe food handling practices and avoiding inconveniences were benefits of preventing foodborne illness. Childcare duties, time and knowledge were barriers to practicing food safety. Confidence in preventing foodborne illness was high, especially when personal control over food handling is present. The low knowledge scores and reported practices revealed a false sense of confidence despite parental concern to protect their child from harm. Food safety messages that emphasize the susceptibility and severity of foodborne illness in children are needed to reach this audience for adoption of safe food handling practices.

  12. Interprofessional mental health training in rural primary care: findings from a mixed methods study.

    PubMed

    Heath, Olga; Church, Elizabeth; Curran, Vernon; Hollett, Ann; Cornish, Peter; Callanan, Terrence; Bethune, Cheri; Younghusband, Lynda

    2015-05-01

    The benefits of interprofessional care in providing mental health services have been widely recognized, particularly in rural communities where access to health services is limited. There continues to be a need for more continuing interprofessional education in mental health intervention in rural areas. There have been few reports of rural programs in which mental health content has been combined with training in collaborative practice. The current study used a sequential mixed-method and quasi-experimental design to evaluate the impact of an interprofessional, intersectoral education program designed to enhance collaborative mental health capacity in six rural sites. Quantitative results reveal a significant increase in positive attitudes toward interprofessional mental health care teams and self-reported increases in knowledge and understanding about collaborative mental health care delivery. The analysis of qualitative data collected following completion of the program, reinforced the value of teaching mental health content within the context of collaborative practice and revealed practice changes, including more interprofessional and intersectoral collaboration. This study suggests that imbedding explicit training in collaborative care in content focused continuing professional education for more complex and chronic health issues may increase the likelihood that professionals will work together to effectively meet client needs.

  13. Investigating Primary Marine Aerosol Properties: CCN Activity of Sea Salt and Mixed Inorganic–Organic Particles

    PubMed Central

    2012-01-01

    Sea spray particles ejected as a result of bubbles bursting from artificial seawater containing salt and organic matter in a stainless steel tank were sampled for size distribution, morphology, and cloud condensation nucleus (CCN) activity. Bubbles were generated either by aeration through a diffuser or by water jet impingement on the seawater surface. Three objectives were addressed in this study. First, CCN activities of NaCl and two types of artificial sea salt containing only inorganic components were measured to establish a baseline for further measurements of mixed organic–inorganic particles. Second, the effect of varying bubble residence time in the bulk seawater solution on particle size and CCN activity was investigated and was found to be insignificant for the organic compounds studied. Finally, CCN activities of particles produced from jet impingement were compared with those produced from diffuser aeration. Analyses indicate a considerable amount of organic enrichment in the jet-produced particles relative to the bulk seawater composition when sodium laurate, an organic surfactant, is present in the seawater. In this case, the production of a thick foam layer during impingement may explain the difference in activation and supports hypotheses that particle production from the two methods of generating bubbles is not equal. PMID:22809370

  14. Testing of a Shrouded, Short Mixing Stack Gas Eductor Model Using High Temperature Primary Flow.

    DTIC Science & Technology

    1982-10-01

    are installed in conjuction with the gas turbine geerator sets 4 -- used to provide shipboard electrical power and aot with the 20 main proplusion...pumped head P - Po (Pa-Pot) for the tertiary flow to the driving head (U Z /2gc) ofp t thz primary flow t t p2 2gc a flow rate ratio, secondary to W_...44 CP 𔃾 E-1 0 N1- 99, 5-4 x 4 00 ’a.a w I- 100. i’-U" IS !.1.° -1 -- ,4 LA W2 % 4k 101 E-44 00 1-44 1020 141 I I £ Z 0 170 r’U 103 U ~ S. ri I 0 -Co

  15. Partial net primary production of a mixed dipterocarp forest: Spatial patterns and temporal dynamics

    NASA Astrophysics Data System (ADS)

    Tan, Zheng-Hong; Deng, Xiao-Bao; Hughes, Alice; Tang, Yong; Cao, Min; Zhang, Wen-Fu; Yang, Xiao-Fei; Sha, Li-Qing; Song, Liang; Zhao, Jun-Fu

    2015-03-01

    We examined how and why partial net primary production (NPPpart) varies across time and space in a Chinese dipterocarp forest. We hypothesize that (1) soil geochemistry explains the spatial pattern of NPPpart within the plot and (2) NPPpart can be used to measure the degree of drought resilience of a natural forest. Spatially, NPPpart was autocorrelated in the range of 75.3 m and homogenous. This spatial pattern could not be well explained by any of the soil properties individually or in combination. If drought sensitivity is defined by marked reduction in NPPpart, the studied forest is drought resilient even when a longer and drier than usual drought hit. Although annual NPPpart was unchanged (vary within 18.24 and 18.52 t ha-1 yr-1) after the drought, the allocation of NPPpart to short-lived litterfall increased, which has further effects on the ecosystem net carbon balance.

  16. National community pharmacy NHS influenza vaccination service in Wales: a primary care mixed methods study

    PubMed Central

    Evans, Andrew M; Wood, Fiona C; Carter, Ben

    2016-01-01

    Background Influenza is a significant cause of morbidity and excess mortality, yet vaccine coverage in the UK remains below target. Community pharmacies are increasingly being promoted as an alternative to vaccination by GPs. Aim To explore and verify the factors that influence the relative performance of pharmacies providing NHS influenza vaccinations. Design and setting A mixed methods study utilising qualitative, semi-structured interviews and quantitative analysis of predictors of vaccination numbers in community pharmacies in Wales. Method Interviews were conducted with 16 pharmacists who participated in the Welsh national pharmacy influenza service in 2013–2014. A purposive sampling strategy was used. Qualitative findings were analysed using framework analysis. Potential predictors of vaccination numbers were identified from interviews and a literature review, and included in a multivariable regression model. Results The contribution of community pharmacies towards vaccination in Wales is small. Findings suggest that community pharmacies reach younger at-risk individuals, in whom vaccine uptake is low, in greater proportion than influenza vaccination programmes as a whole. Extended opening hours and urban locations were positively associated with the number of vaccinations given, although pharmacists reported that workload, vaccine costs, unforeseen delays, lack of public awareness, and GPs’ views of the service limited their contribution. Pharmacists, aware of the potential for conflict with GPs, moderated their behaviour to mitigate such risk. Conclusion Before community pharmacies take greater responsibility for delivering healthcare services, obstacles including increasing pharmacist capacity, vaccine procurement, health service delays, managing GP–pharmacy relationships, and improving public awareness must be overcome. PMID:26965025

  17. Team dynamics, clinical work satisfaction, and patient care coordination between primary care providers: A mixed methods study.

    PubMed

    Song, Hummy; Ryan, Molly; Tendulkar, Shalini; Fisher, Josephine; Martin, Julia; Peters, Antoinette S; Frolkis, Joseph P; Rosenthal, Meredith B; Chien, Alyna T; Singer, Sara J

    Team-based care is essential for delivering high-quality, comprehensive, and coordinated care. Despite considerable research about the effects of team-based care on patient outcomes, few studies have examined how team dynamics relate to provider outcomes. The aim of this study was to examine relationships among team dynamics, primary care provider (PCP) clinical work satisfaction, and patient care coordination between PCPs in 18 Harvard-affiliated primary care practices participating in Harvard's Academic Innovations Collaborative. First, we administered a cross-sectional survey to all 548 PCPs (267 attending clinicians, 281 resident physicians) working at participating practices; 65% responded. We assessed the relationship of team dynamics with PCPs' clinical work satisfaction and perception of patient care coordination between PCPs, respectively, and the potential mediating effect of patient care coordination on the relationship between team dynamics and work satisfaction. In addition, we embedded a qualitative evaluation within the quantitative evaluation to achieve a convergent mixed methods design to help us better understand our findings and illuminate relationships among key variables. Better team dynamics were positively associated with clinical work satisfaction and quality of patient care coordination between PCPs. Coordination partially mediated the relationship between team dynamics and satisfaction for attending clinicians, suggesting that higher satisfaction depends, in part, on better teamwork, yielding more coordinated patient care. We found no mediating effects for resident physicians. Qualitative results suggest that sources of satisfaction from positive team dynamics for PCPs may be most relevant to attending clinicians. Improving primary care team dynamics could improve clinical work satisfaction among PCPs and patient care coordination between PCPs. In addition to improving outcomes that directly concern health care providers, efforts to

  18. Sources of unsafe primary care for older adults: a mixed-methods analysis of patient safety incident reports.

    PubMed

    Cooper, Alison; Edwards, Adrian; Williams, Huw; Evans, Huw P; Avery, Anthony; Hibbert, Peter; Makeham, Meredith; Sheikh, Aziz; J Donaldson, Liam; Carson-Stevens, Andrew

    2017-09-01

    older adults are frequent users of primary healthcare services, but are at increased risk of healthcare-related harm in this setting. to describe the factors associated with actual or potential harm to patients aged 65 years and older, treated in primary care, to identify action to produce safer care. a cross-sectional mixed-methods analysis of a national (England and Wales) database of patient safety incident reports from 2005 to 2013. 1,591 primary care patient safety incident reports regarding patients aged 65 years and older. we developed a classification system for the analysis of patient safety incident reports to describe: the incident and preceding chain of incidents; other contributory factors; and patient harm outcome. We combined findings from exploratory descriptive and thematic analyses to identify key sources of unsafe care. the main sources of unsafe care in our weighted sample were due to: medication-related incidents e.g. prescribing, dispensing and administering (n = 486, 31%; 15% serious patient harm); communication-related incidents e.g. incomplete or non-transfer of information across care boundaries (n = 390, 25%; 12% serious patient harm); and clinical decision-making incidents which led to the most serious patient harm outcomes (n = 203, 13%; 41% serious patient harm). priority areas for further research to determine the burden and preventability of unsafe primary care for older adults, include: the timely electronic tools for prescribing, dispensing and administering medication in the community; electronic transfer of information between healthcare settings; and, better clinical decision-making support and guidance.

  19. Effects of Silver Nanoparticles on Primary Mixed Neural Cell Cultures: Uptake, Oxidative Stress and Acute Calcium Responses

    PubMed Central

    Haase, Andrea; Rott, Stephanie; Mantion, Alexandre; Graf, Philipp; Plendl, Johanna; Thünemann, Andreas F.; Meier, Wolfgang P.; Taubert, Andreas; Luch, Andreas; Reiser, Georg

    2012-01-01

    In the body, nanoparticles can be systemically distributed and then may affect secondary target organs, such as the central nervous system (CNS). Putative adverse effects on the CNS are rarely investigated to date. Here, we used a mixed primary cell model consisting mainly of neurons and astrocytes and a minor proportion of oligodendrocytes to analyze the effects of well-characterized 20 and 40 nm silver nanoparticles (SNP). Similar gold nanoparticles served as control and proved inert for all endpoints tested. SNP induced a strong size-dependent cytotoxicity. Additionally, in the low concentration range (up to 10 μg/ml of SNP), the further differentiated cultures were more sensitive to SNP treatment. For detailed studies, we used low/medium dose concentrations (up to 20 μg/ml) and found strong oxidative stress responses. Reactive oxygen species (ROS) were detected along with the formation of protein carbonyls and the induction of heme oxygenase-1. We observed an acute calcium response, which clearly preceded oxidative stress responses. ROS formation was reduced by antioxidants, whereas the calcium response could not be alleviated by antioxidants. Finally, we looked into the responses of neurons and astrocytes separately. Astrocytes were much more vulnerable to SNP treatment compared with neurons. Consistently, SNP were mainly taken up by astrocytes and not by neurons. Immunofluorescence studies of mixed cell cultures indicated stronger effects on astrocyte morphology. Altogether, we can demonstrate strong effects of SNP associated with calcium dysregulation and ROS formation in primary neural cells, which were detectable already at moderate dosages. PMID:22240980

  20. Cultural responses to pain in UK children of primary school age: a mixed-methods study.

    PubMed

    Azize, Pary M; Endacott, Ruth; Cattani, Allegra; Humphreys, Ann

    2014-06-01

    Pain-measurement tools are often criticized for not addressing the influence of culture and ethnicity on pain. This study examined how children who speak English as a primary or additional language discuss pain. Two methods were used in six focus group interviews with 34 children aged 4-7 years: (i) use of drawings from the Pediatric Pain Inventory to capture the language used by children to describe pain; and (ii) observation of the children's placing of pain drawings on red/amber/green paper to denote perceived severity of pain. The findings demonstrated that children with English as an additional language used less elaborate language when talking about pain, but tended to talk about the pictures prior to deciding where they should be placed. For these children, there was a positive significant relationship between language, age, and length of stay in the UK. The children's placement of pain drawings varied according to language background, sex, and age. The findings emphasize the need for sufficient time to assess pain adequately in children who do not speak English as a first language.

  1. Initial examination of microwave pretreatment on primary, secondary and mixed sludges before and after anaerobic digestion.

    PubMed

    Eskicioglu, C; Kennedy, K J; Droste, R L

    2008-01-01

    The effects of microwave pretreatment on disintegration and mesophilic digestion of waste activated sludge (WAS), primary sludge (PS), combined (PS + WAS) sequencing batch reactor (SBR) sludge and anaerobically digested biocake were investigated by both household and bench scale industrial types microwaves at temperatures below and above boiling point. Pretreatment variables, temperature, intensity (cooking rate) and sludge concentration had statistically significant effects on solubilization. The microwave pretreatment also increased the bioavailability of sludge components under batch anaerobic digestion and enhanced the dewaterability of pretreated sludges after digestion. However, the level of improvements in solubilization and biodegradation from different waste sludges were different. While the largest improvement in ultimate biodegradation was observed in WAS, microwave irradiation only affected the rate of biodegradation of pretreated PS samples. Similarly, relatively lower solubilization ratios achieved for combined - SBR sludge was attributed to high sludge age of extended aeration SBR unit. It is possible that initial sludge characteristics may influence final pretreatment outcomes so that general statements of performance cannot always be made.

  2. Expression of inducible nitric oxide synthase causes delayed neurotoxicity in primary mixed neuronal-glial cortical cultures.

    PubMed

    Dawson, V L; Brahmbhatt, H P; Mong, J A; Dawson, T M

    1994-11-01

    Nitric oxide (NO) is a potent biological messenger molecule in the central nervous system (CNS). There are several potential sources of NO production in the CNS, including neurons and endothelial cells which express NO synthase (NOS) constitutively. Astrocytes and microglia can be induced by cytokines to express a NOS isoform similar to macrophage NOS (mNOS). Primary mixed glial cultures exposed to lipopolysaccharide (LPS) or a combination of LPS and gamma-interferon (INF-gamma) produce nitrite, a breakdown product of NO formation, in a dose-dependent manner. Nitrite production is detectable at 12 hr, peaks at 48 hr and is sustained for at least 96 hr. The NOS inhibitor, nitro-L-arginine (NArg), inhibits nitrite formation, but the immunosuppressant agent, FK506, does not. In mixed glial-neuronal cultures exposed to 50 ng LPS or 5 ng LPS and 1 microgram INF-gamma, neurons begin to die at 48 hr, approx. 24-36 hr after detectable nitrite production. Neurotoxicity is attenuated by 100 microM NArg. These data indicate that expression of inducible mNOS causes delayed neurotoxicity.

  3. Case management for dementia in primary health care: a systematic mixed studies review based on the diffusion of innovation model

    PubMed Central

    Khanassov, Vladimir; Vedel, Isabelle; Pluye, Pierre

    2014-01-01

    Background The purpose of this study was to examine factors associated with the implementation of case management (CM) interventions in primary health care (PHC) and to develop strategies to enhance its adoption by PHC practices. Methods This study was designed as a systematic mixed studies review (including quantitative and qualitative studies) with synthesis based on the diffusion of innovation model. A literature search was performed using MEDLINE, PsycInfo, EMBASE, and the Cochrane Database (1995 to August 2012) to identify quantitative (randomized controlled and nonrandomized) and qualitative studies describing the conditions limiting and facilitating successful CM implementation in PHC. The methodological quality of each included study was assessed using the validated Mixed Methods Appraisal Tool. Results Twenty-three studies (eleven quantitative and 12 qualitative) were included. The characteristics of CM that negatively influence implementation are low CM intensity (eg, infrequent follow-up), large caseload (more than 60 patients per full-time case manager), and approach, ie, reactive rather than proactive. Case managers need specific skills to perform their role (eg, good communication skills) and their responsibilities in PHC need to be clearly delineated. Conclusion Our systematic review supports a better understanding of factors that can explain inconsistent evidence with regard to the outcomes of dementia CM in PHC. Lastly, strategies are proposed to enhance implementation of dementia CM in PHC. PMID:24959072

  4. Case management for dementia in primary health care: a systematic mixed studies review based on the diffusion of innovation model.

    PubMed

    Khanassov, Vladimir; Vedel, Isabelle; Pluye, Pierre

    2014-01-01

    The purpose of this study was to examine factors associated with the implementation of case management (CM) interventions in primary health care (PHC) and to develop strategies to enhance its adoption by PHC practices. This study was designed as a systematic mixed studies review (including quantitative and qualitative studies) with synthesis based on the diffusion of innovation model. A literature search was performed using MEDLINE, PsycInfo, EMBASE, and the Cochrane Database (1995 to August 2012) to identify quantitative (randomized controlled and nonrandomized) and qualitative studies describing the conditions limiting and facilitating successful CM implementation in PHC. The methodological quality of each included study was assessed using the validated Mixed Methods Appraisal Tool. Twenty-three studies (eleven quantitative and 12 qualitative) were included. The characteristics of CM that negatively influence implementation are low CM intensity (eg, infrequent follow-up), large caseload (more than 60 patients per full-time case manager), and approach, ie, reactive rather than proactive. Case managers need specific skills to perform their role (eg, good communication skills) and their responsibilities in PHC need to be clearly delineated. Our systematic review supports a better understanding of factors that can explain inconsistent evidence with regard to the outcomes of dementia CM in PHC. Lastly, strategies are proposed to enhance implementation of dementia CM in PHC.

  5. Differences in leucocyte-endothelium interactions between normal and adenocarcinoma bearing tissues in response to radiation.

    PubMed Central

    Wu, N. Z.; Ross, B. A.; Gulledge, C.; Klitzman, B.; Dodge, R.; Dewhirst, M. W.

    1994-01-01

    Previously, we demonstrated that the interaction between leucocytes and endothelial cells in tumour tissues is greatly diminished compared with normal tissues under several induced inflammatory conditions. Radiation has been reported to cause release of inflammatory mediators and to promote neutrophil adhesions to cultured endothelial monolayers. In this study, we tested the hypothesis that radiation would cause increased leucocyte rolling and adhesion in both tumour and normal tissues. We examined these two parameters in response to 6 Gy of gamma-radiation in mammary adenocarcinomas implanted into rat skinfold window chambers as well as normal (i.e. non-tumour-bearing) preparations. Leucocyte rolling and adhesion were measured in terms of flux of rolling leucocytes (F(rolling)) and density of adhering leucocytes (D(adhering)) in microvessels. F(rolling) and D(adhering) were measured in two groups of preparations: irradiated and control. In normal preparations, F(rolling) and D(adhering) were both increased significantly by radiation. In contrast, in adenocarcinoma-bearing preparations, F(rolling) and D(adhering) were either unchanged (in the tumour centre) or reduced (in tumour periphery and the normal tissue surrounding the tumour) by radiation. Radiation did not cause changes in haemodynamics in these preparations, thus the observed changes in leucocyte rolling and adhesion could not be accounted for by haemodynamic factors. These results indicate that: (1) in normal preparations, radiation could cause inflammation as manifested by increased leucocyte rolling and adhesion; and (2) in tumour-bearing preparations, radiation caused changes in the vascular surface properties such that they became less adhesive to leucocytes. Such differences in radiation response may have important implications for radiation therapy and provide new insights into the unique features of tumours. Images Figure 2 PMID:8180019

  6. Selective inhibition of mitochondrial respiration and glycolysis in human leukaemic leucocytes by methylglyoxal.

    PubMed Central

    Biswas, S; Ray, M; Misra, S; Dutta, D P; Ray, S

    1997-01-01

    The effect of methylglyoxal on the oxygen consumption of mitochondria of both normal and leukaemic leucocytes was tested by using different respiratory substrates and complex specific artificial electron donors and inhibitors. The results indicate that methylglyoxal strongly inhibits mitochondrial respiration in leukaemic leucocytes, whereas, at a much higher concentration, methylglyoxal fails to inhibit mitochondrial respiration in normal leucocytes. Methylglyoxal strongly inhibits ADP-stimulated alpha-oxoglutarate and malate plus NAD+-dependent respiration, whereas, at a higher concentration, methylglyoxal fails to inhibit succinate and alpha-glycerophosphate-dependent respiration. Methylglyoxal also fails to inhibit respiration which is initiated by duroquinone and cannot inhibit oxygen consumption when the N,N,N', N'-tetramethyl-p-phenylenediamine by-pass is used. NADH oxidation by sub-mitochondrial particles of leukaemic leucocytes is also inhibited by methylglyoxal. Lactaldehyde, a catabolite of methylglyoxal, can exert a protective effect on the inhibition of leukaemic leucocyte mitochondrial respiration by methylglyoxal. Methylglyoxal also inhibits l-lactic acid formation by intact leukaemic leucocytes and critically reduces the ATP level of these cells, whereas methylglyoxal has no effect on normal leucocytes. We conclude that methylglyoxal inhibits glycolysis and the electron flow through mitochondrial complex I of leukaemic leucocytes. This is strikingly similar to our previous studies on mitochondrial respiration, glycolysis and ATP levels in Ehrlich ascites carcinoma cells [Ray, Dutta, Halder and Ray (1994) Biochem. J. 303, 69-72; Halder, Ray and Ray (1993) Int. J. Cancer 54, 443-449], which strongly suggests that the inhibition of electron flow through complex I of the mitochondrial respiratory chain and inhibition of glycolysis by methylglyoxal may be common characteristics of all malignant cells. PMID:9163322

  7. Selective inhibition of mitochondrial respiration and glycolysis in human leukaemic leucocytes by methylglyoxal.

    PubMed

    Biswas, S; Ray, M; Misra, S; Dutta, D P; Ray, S

    1997-04-15

    The effect of methylglyoxal on the oxygen consumption of mitochondria of both normal and leukaemic leucocytes was tested by using different respiratory substrates and complex specific artificial electron donors and inhibitors. The results indicate that methylglyoxal strongly inhibits mitochondrial respiration in leukaemic leucocytes, whereas, at a much higher concentration, methylglyoxal fails to inhibit mitochondrial respiration in normal leucocytes. Methylglyoxal strongly inhibits ADP-stimulated alpha-oxoglutarate and malate plus NAD+-dependent respiration, whereas, at a higher concentration, methylglyoxal fails to inhibit succinate and alpha-glycerophosphate-dependent respiration. Methylglyoxal also fails to inhibit respiration which is initiated by duroquinone and cannot inhibit oxygen consumption when the N,N,N', N'-tetramethyl-p-phenylenediamine by-pass is used. NADH oxidation by sub-mitochondrial particles of leukaemic leucocytes is also inhibited by methylglyoxal. Lactaldehyde, a catabolite of methylglyoxal, can exert a protective effect on the inhibition of leukaemic leucocyte mitochondrial respiration by methylglyoxal. Methylglyoxal also inhibits l-lactic acid formation by intact leukaemic leucocytes and critically reduces the ATP level of these cells, whereas methylglyoxal has no effect on normal leucocytes. We conclude that methylglyoxal inhibits glycolysis and the electron flow through mitochondrial complex I of leukaemic leucocytes. This is strikingly similar to our previous studies on mitochondrial respiration, glycolysis and ATP levels in Ehrlich ascites carcinoma cells [Ray, Dutta, Halder and Ray (1994) Biochem. J. 303, 69-72; Halder, Ray and Ray (1993) Int. J. Cancer 54, 443-449], which strongly suggests that the inhibition of electron flow through complex I of the mitochondrial respiratory chain and inhibition of glycolysis by methylglyoxal may be common characteristics of all malignant cells.

  8. Effects of photodynamic therapy on leucocyte-endothelium interaction: differences between normal and tumour tissue.

    PubMed Central

    Dellian, M.; Abels, C.; Kuhnle, G. E.; Goetz, A. E.

    1995-01-01

    An inflammatory reaction is regularly noticed in irradiated tissues following photodynamic therapy (PDT). This observation is potentially associated with leucocyte-mediated tissue damage, which might further contribute to the tumoricidal effect of this therapy. The objective of our study was to investigate the effects of PDT on leucocyte-endothelium interaction in the microvasculature of tumours and normal tissue. Experiments were performed in the dorsal skinfold chamber preparation of Syrian golden hamsters bearing amelanotic melanoma A-Mel-3. The photosensitiser. Photofrin (5 mg kg-1 i.v.) was injected 24 h before laser irradiation (630 nm, 100 mW cm-2, 10 J cm-2 or 100 J cm-2). Post-capillary confluent venules (diameter 15-40 microns) of subcutaneous (s.c.) tissue or the amelanotic melanoma A-Mel-3 were observed by intravital microscopy before, 5, 30, 60 and 180 min after laser irradiation and recorded for off-line analysis. Before treatment, the number of adherent leucocytes in tumour vessels was only 22% of the number observed in vessels of s.c. tissue (P < 0.01). The maximum increase in adhering leucocytes was observed in post-capillary venules of s.c. tissue 1 h after PDT (P < 0.01). In contrast, enhanced leucocyte-endothelium interaction was missing in tumour vessels and in control groups. These results indicate that the tumour destruction observed after PDT is not mediated by leucocyte-endothelium interaction in the tumour. Induction of leucocyte adhesion in the PDT-treated normal tissue suggests a contribution to the peritumoral inflammatory response. Different maturational status or biochemical properties of tumour microvascular endothelium may explain the lack of leucocyte adherence upon PDT. Images Figure 1 PMID:7577457

  9. Cortisol stimulates growth hormone gene expression in rainbow trout leucocytes in vitro.

    PubMed

    Yada, Takashi; Muto, Kohji; Azuma, Teruo; Hyodo, Susumu; Schreck, Carl B

    2005-05-15

    Extrapituitary expression of the growth hormone (GH) gene has been reported for the immune system of various vertebrates. In the rainbow trout (Oncorhynchus mykiss), GH mRNA could be detected in several lymphoid organs and leucocytes by reverse transcriptase-polymerase chain reaction (RT-PCR). To understand the control of GH expression in the fish immune system, mRNA levels for two distinct GH genes (GH1 and GH2) in trout leucocytes isolated from peripheral blood were quantified using a real-time PCR method. Both GH mRNAs could be detected in trout leucocytes, although their levels were extremely low compared to those in pituitary cells. The levels of GH2 mRNA in leucocytes were several times higher than those of GH1, while no difference was observed between GH1 and GH2 mRNA levels in the pituitary. Administration of dibutyryl cyclic AMP and cortisol produced a significant elevation of GH mRNA levels in trout leucocytes, although the levels were unchanged by T3. GH1 and GH2 mRNA levels showed similarities in responses to those factors. The effect of cortisol on GH mRNA appears biphasic; a dose-depending elevation of GH gene expression was observed in leucocytes treated with cortisol at below 200 nM, however, cortisol had no effect at 2000 nM. Cortisol-treated leucocytes showed no significant change in the mRNA level of beta-actin or proliferative activity during the experiments. Our results thus show that, at the low levels, GH gene expression in trout leucocytes is regulated by cortisol, which has been known as a regulatory factor of GH gene expression in pituitary cells, and suggest a physiological significance of paracrine GH produced in the fish immune system.

  10. Pediatric primary care providers' perspectives regarding hospital discharge communication: a mixed methods analysis.

    PubMed

    Leyenaar, JoAnna K; Bergert, Lora; Mallory, Leah A; Engel, Richard; Rassbach, Caroline; Shen, Mark; Woehrlen, Tess; Cooperberg, David; Coghlin, Daniel

    2015-01-01

    Effective communication between inpatient and outpatient providers may mitigate risks of adverse events associated with hospital discharge. However, there is an absence of pediatric literature defining effective discharge communication strategies at both freestanding children's hospitals and general hospitals. The objectives of this study were to assess associations between pediatric primary care providers' (PCPs) reported receipt of discharge communication and referral hospital type, and to describe PCPs' perspectives regarding effective discharge communication and areas for improvement. We administered a questionnaire to PCPs referring to 16 pediatric hospital medicine programs nationally. Multivariable models were developed to assess associations between referral hospital type and receipt and completeness of discharge communication. Open-ended questions asked respondents to describe effective strategies and areas requiring improvement regarding discharge communication. Conventional qualitative content analysis was performed to identify emergent themes. Responses were received from 201 PCPs, for a response rate of 63%. Although there were no differences between referral hospital type and PCP-reported receipt of discharge communication (relative risk 1.61, 95% confidence interval 0.97-2.67), PCPs referring to general hospitals more frequently reported completeness of discharge communication relative to those referring to freestanding children's hospitals (relative risk 1.78, 95% confidence interval 1.26-2.51). Analysis of free text responses yielded 4 major themes: 1) structured discharge communication, 2) direct personal communication, 3) reliability and timeliness of communication, and 4) communication for effective postdischarge care. This study highlights potential differences in the experiences of PCPs referring to general hospitals and freestanding children's hospitals, and presents valuable contextual data for future quality improvement initiatives

  11. Specificity of leucocyte migration inhibition test in coeliac disease. A reassessment using different gluten subfractions.

    PubMed Central

    Corazza, G R; Rawcliffe, P M; Frisoni, M; Sarchielli, P; Londei, M; Campieri, M; Lazzari, R; Gasbarrini, G

    1985-01-01

    Production of leucocyte migration inhibition factor by peripheral blood leucocytes in response to challenge with gluten fractions has been proposed as a reliable in vitro test for the diagnosis of coeliac disease. We have performed the leucocyte migration inhibition test with two different gluten fractions, GFIII and B2, in untreated and treated coeliac patients, patients with other intestinal diseases (abnormal controls) and healthy controls, and evaluated the sensitivity, specificity and positive and negative predictability of the test for the diagnosis of coeliac disease. Using GFIII as antigen leucocyte migration was significantly inhibited, compared to healthy controls, not only in treated and untreated coeliacs but also in abnormal controls. Using B2 gluten subfraction as antigen only treated coeliacs and abnormal controls differed significantly from healthy controls. The elevated number of abnormal controls showing migration inhibition consistently affected the diagnostic value of the test, which did not vary using B2 subfraction instead of GFIII as antigen. Our study confirms previous observations of gluten sensitization, as detected by leucocyte migration inhibition, in coeliac patients but strongly questions the claim that coeliac disease can be diagnosed on the basis of a positive leucocyte migration inhibition test without the need for intestinal biopsy. PMID:4006297

  12. An evaluation of leucocyte analysis on the Coulter STKS.

    PubMed

    Robertson, E P; Lai, H W; Wei, D C

    1992-01-01

    The performance of leucocyte analysis on the Coulter STKS (Coulter, Hialeah, FL, USA) was evaluated for accuracy, precision and reliability. The results were compared with those obtained from visual examination of a Romanowsky stained blood film together with the automated WBC-diff. from the Technicon H*1 (Technicon, Tarrytown, NY, USA). The relationship between the number of cells counted per WBC-diff. and the WBC count of the sample was established. Precision of the STKS WBC-diff. was acceptable on blood samples with normal and low WBC counts. Correlation with an 800 cell manual WBC-diff. (n = 104) was excellent (r = 0.97, 0.97, 0.83, 0.98 and 0.53 for neutrophils, lymphocytes, monocytes, eosinophils and basophils respectively). Blood specimens, collected into dipotassium EDTA, could be stored at 20-25 degrees C for at least 8 h with no significant effect on the STKS WBC-diff. In a study of 513 patient samples, the BLASTS suspect flag gave 5.4% false positives and zero false negatives, the VARIANT LYMPHS flag gave 1.5% false positives and 0.4% false negatives, and the IMM GRANS/BANDS flag gave 30.8% false positives and 2.3% false negatives. Several instrument and sample related problems were encountered during this study. Despite these limitations, the STKS can provide efficient 5 part WBC-diffs. and effective screening for WBC abnormalities.

  13. Labile disulfide bonds are common at the leucocyte cell surface

    PubMed Central

    Metcalfe, Clive; Cresswell, Peter; Ciaccia, Laura; Thomas, Benjamin; Barclay, A. Neil

    2011-01-01

    Redox conditions change in events such as immune and platelet activation, and during viral infection, but the biochemical consequences are not well characterized. There is evidence that some disulfide bonds in membrane proteins are labile while others that are probably structurally important are not exposed at the protein surface. We have developed a proteomic/mass spectrometry method to screen for and identify non-structural, redox-labile disulfide bonds in leucocyte cell-surface proteins. These labile disulfide bonds are common, with several classes of proteins being identified and around 30 membrane proteins regularly identified under different reducing conditions including using enzymes such as thioredoxin. The proteins identified include integrins, receptors, transporters and cell–cell recognition proteins. In many cases, at least one cysteine residue was identified by mass spectrometry as being modified by the reduction process. In some cases, functional changes are predicted (e.g. in integrins and cytokine receptors) but the scale of molecular changes in membrane proteins observed suggests that widespread effects are likely on many different types of proteins including enzymes, adhesion proteins and transporters. The results imply that membrane protein activity is being modulated by a ‘redox regulator’ mechanism. PMID:22645650

  14. Leucocyte protein Trojan, a possible regulator of apoptosis.

    PubMed

    Petrov, Petar; Syrjänen, Riikka; Uchida, Tatsuya; Vainio, Olli

    2017-02-01

    Trojan is a leucocyte-specific protein, cloned from chicken embryonic thymocyte cDNA library. The molecule is a type I transmembrane protein with an extracellular CCP domain, followed by two FN3 domains. Its cytoplasmic tail is predicted to possess a MAPK docking and a PKA phosphorylation sites. Trojan has been proposed to have an anti-apoptotic role based on its differential expression on developing thymocyte subpopulations. Using a chicken cell line, our in vitro studies showed that upon apoptosis induction, Trojan expression rises dramatically on the surface of surviving cells and gradually decreases towards its normal levels as cells recover. When sorted based on their expression levels of Trojan, cells with high expression appeared less susceptible to apoptotic induction than those bearing no or low levels of Trojan on their surface. The mechanism by which the molecule exerts its function is yet to be discovered. We found that cells overexpressing Trojan from a cDNA plasmid show elevated steady-state levels of intracellular calcium, suggesting the molecule is able to transmit cytoplasmic signals. The mechanistic nature of Trojan-induced signalling is a target of future investigation. In this article, we conducted a series of experiments that suggest Trojan as an anti-apoptotic regulator.

  15. Uterus human leucocyte antigen expression in the perspective of transplantation.

    PubMed

    Gauthier, Tristan; Filloux, Matthieu; Guillaudeau, Angelique; Essig, Marie; Bibes, Romain; Pacha, Adam Fodil; Piver, Pascal; Aubard, Yves; Marquet, Pierre; Drouet, Mireille

    2016-12-01

    To describe class I and II human leucocyte antigen (HLA) expression using different uterine tissues in the perspective of uterus transplantation. Human uterine tissues were obtained from 12 women who had undergone hysterectomy for the treatment of benign disease. HLA class I and HLA-antigen D related (DR) expression were assessed via immunochemistry. HLA class I expression in the uterus was compared with expression in other organs and tissues, including kidney and myocardium samples. HLA class I expression was strong in the endometrial glands and mild in the myometrium. Staining of endometrial glands was similar to glomerular staining in the kidney. The myometrium seems to express HLA class I similarly to hepatocytes and myocardial cells. HLA class I expression in the uterus did not differ in younger or post-menopausal women. HLA-DR was expressed in the endometrial glands, but not in the myometrium. A lack of HLA-DR expression seemed to be correlated with cell proliferation. HLA expression in the endometrium and myometrium is different. The endometrium should be the major target of alloreactive response. As for other transplanted organs, assessment of HLA unacceptable antigens and multiple immunosuppressive treatments is necessary in uterus transplantation. © 2016 Japan Society of Obstetrics and Gynecology.

  16. Exploring the actual and potential role of the primary care nurse in the prevention of cancer: a mixed methods study.

    PubMed

    McIlfatrick, S; Keeney, S; McKenna, H; McCarley, N; McIlwee, G

    2014-05-01

    The aim of this study was to investigate the actual and the potential role of the primary care nurse (PCN) in the prevention of cancer. International studies have indicated that a range of strategies can have an impact on the incidence of cancer. Due to their frequent front-line contact with the public, PCNs can play an important role in the primary prevention of cancer. Nonetheless, there is a lack of information on their actual and potential role in cancer prevention. A sequential confirmatory mixed methods approach was used. Postal questionnaires were administered to PCNs [n = 500; 225 returns (response rate 45%)] followed by semi-structured interviews (n = 15). PCNs provided high levels of cancer prevention activities, specifically focusing on smoking cessation, obesity and cervical screening. They considered that their cancer prevention role could be improved through additional practice-based training and more collaborative inter-professional working. They also identified the need for a better understanding of how to change people's attitudes and behaviours regarding cancer prevention. Evidence from this study provide important insights into the potential of the PCN to empower individuals to take responsibility for their own health and make more informed lifestyle choices. © 2013 John Wiley & Sons Ltd.

  17. The implementation and sustainability of a combined lifestyle intervention in primary care: mixed method process evaluation.

    PubMed

    Berendsen, Brenda A J; Kremers, Stef P J; Savelberg, Hans H C M; Schaper, Nicolaas C; Hendriks, Marike R C

    2015-03-17

    The impact of physical inactivity and unhealthy diet on health is increasingly profound. Lifestyle interventions targeting both behaviors simultaneously might decrease the prevalence of overweight and comorbidities. The Dutch 'BeweegKuur' is a combined lifestyle intervention (CLI) in primary care, to improve physical activity and dietary behavior in overweight people. In a cluster randomized controlled trial, the (cost-) effectiveness of an intensively guided program has been compared to a less intensively guided program. This process evaluation aimed to assess protocol adherence and potential differences between clusters. In addition, sustainability (i.e. continuation of the CLI in practice after study termination) was evaluated. Existing frameworks were combined to design the process evaluation for our intervention and setting specifically. We assessed reach, fidelity, dose delivered and received, context and implementation strategy. Both qualitative and quantitative data were used for a comprehensive evaluation. Data were collected in semi-structured interviews with health care providers (HCPs, n = 25), drop-out registration by HCPs, regular questionnaires among participants (n = 411) and logbooks kept by researchers during the trial. Protocol adherence by professionals and participants varied between the programs and clusters. In both programs the number of meetings with all HCPs was lower than planned in the protocol. Participants of the supervised program attended, compared to participants of the start-up program, more meetings with physiotherapists, but fewer with lifestyle advisors and dieticians. The 'BeweegKuur' was not sustained, but intervention aspects, networks and experiences were still utilized after finalization of the project. Whether clusters continued to offer a CLI seemed dependent on funding opportunities and collaborations. Protocol adherence in a CLI was problematic in both HCPs and participants. Mainly the amount of dietary guidance was

  18. Physical activity assessment in practice: a mixed methods study of GPPAQ use in primary care

    PubMed Central

    2014-01-01

    Background Insufficient physical activity (PA) levels which increase the risk of chronic disease are reported by almost two-thirds of the population. More evidence is needed about how PA promotion can be effectively implemented in general practice (GP), particularly in socio-economically disadvantaged communities. One tool recommended for the assessment of PA in GP and supported by NICE (National Institute for Health and Care Excellence) is The General Practice Physical Activity Questionnaire (GPPAQ) but details of how it may be used and of its acceptability to practitioners and patients are limited. This study aims to examine aspects of GPPAQ administration in non-urgent patient contacts using different primary care electronic recording systems and to explore the views of health professionals regarding its use. Methods Four general practices, selected because of their location within socio-economically disadvantaged areas, were invited to administer GPPAQs to patients, aged 35-75 years, attending non-urgent consultations, over two-week periods. They used different methods of administration and different electronic medical record systems (EMIS, Premiere, Vision). Participants’ (general practitioners (GPs), nurses and receptionists) views regarding GPPAQ use were explored via questionnaires and focus groups. Results Of 2,154 eligible consultations, 192 (8.9%) completed GPPAQs; of these 83 (43%) were categorised as inactive. All practices were located within areas ranked as being in the tertile of greatest socio-economic deprivation in Northern Ireland. GPs/nurses in two practices invited completion of the GPPAQ, receptionists did so in two. One practice used an electronic template; three used paper copies of the questionnaires. End-of-study questionnaires, completed by 11 GPs, 3 nurses and 2 receptionists and two focus groups, with GPs (n = 8) and nurses (n = 4) indicated that practitioners considered the GPPAQ easy to use but not in every consultation

  19. Physical activity assessment in practice: a mixed methods study of GPPAQ use in primary care.

    PubMed

    Heron, Neil; Tully, Mark A; McKinley, Michelle C; Cupples, Margaret E

    2014-01-15

    Insufficient physical activity (PA) levels which increase the risk of chronic disease are reported by almost two-thirds of the population. More evidence is needed about how PA promotion can be effectively implemented in general practice (GP), particularly in socio-economically disadvantaged communities. One tool recommended for the assessment of PA in GP and supported by NICE (National Institute for Health and Care Excellence) is The General Practice Physical Activity Questionnaire (GPPAQ) but details of how it may be used and of its acceptability to practitioners and patients are limited. This study aims to examine aspects of GPPAQ administration in non-urgent patient contacts using different primary care electronic recording systems and to explore the views of health professionals regarding its use. Four general practices, selected because of their location within socio-economically disadvantaged areas, were invited to administer GPPAQs to patients, aged 35-75 years, attending non-urgent consultations, over two-week periods. They used different methods of administration and different electronic medical record systems (EMIS, Premiere, Vision). Participants' (general practitioners (GPs), nurses and receptionists) views regarding GPPAQ use were explored via questionnaires and focus groups. Of 2,154 eligible consultations, 192 (8.9%) completed GPPAQs; of these 83 (43%) were categorised as inactive. All practices were located within areas ranked as being in the tertile of greatest socio-economic deprivation in Northern Ireland. GPs/nurses in two practices invited completion of the GPPAQ, receptionists did so in two. One practice used an electronic template; three used paper copies of the questionnaires.End-of-study questionnaires, completed by 11 GPs, 3 nurses and 2 receptionists and two focus groups, with GPs (n = 8) and nurses (n = 4) indicated that practitioners considered the GPPAQ easy to use but not in every consultation. Its use extended consultation time

  20. Morphology of mixed primary and secondary organic particles and the adsorption of spectator organic gases during aerosol formation

    PubMed Central

    Vaden, Timothy D.; Song, Chen; Zaveri, Rahul A.; Imre, Dan; Zelenyuk, Alla

    2010-01-01

    Primary organic aerosol (POA) and associated vapors can play an important role in determining the formation and properties of secondary organic aerosol (SOA). If SOA and POA are miscible, POA will significantly enhance SOA formation and some POA vapor will incorporate into SOA particles. When the two are not miscible, condensation of SOA on POA particles forms particles with complex morphology. In addition, POA vapor can adsorb to the surface of SOA particles increasing their mass and affecting their evaporation rates. To gain insight into SOA/POA interactions we present a detailed experimental investigation of the morphologies of SOA particles formed during ozonolysis of α-pinene in the presence of dioctyl phthalate (DOP) particles, serving as a simplified model of hydrophobic POA, using a single-particle mass spectrometer. Ultraviolet laser depth-profiling experiments were used to characterize two different types of mixed SOA/DOP particles: those formed by condensation of the oxidized α-pinene products on size-selected DOP particles and by condensation of DOP on size-selected α-pinene SOA particles. The results show that the hydrophilic SOA and hydrophobic DOP do not mix but instead form layered phases. In addition, an examination of homogeneously nucleated SOA particles formed in the presence of DOP vapor shows them to have an adsorbed DOP coating layer that is ∼4 nm thick and carries 12% of the particles mass. These results may have implications for SOA formation and behavior in the atmosphere, where numerous organic compounds with various volatilities and different polarities are present. PMID:20194795

  1. Adoption of a Portal for the Primary Care Management of Pediatric Asthma: A Mixed-Methods Implementation Study.

    PubMed

    Fiks, Alexander G; DuRivage, Nathalie; Mayne, Stephanie L; Finch, Stacia; Ross, Michelle E; Giacomini, Kelli; Suh, Andrew; McCarn, Banita; Brandt, Elias; Karavite, Dean; Staton, Elizabeth W; Shone, Laura P; McGoldrick, Valerie; Noonan, Kathleen; Miller, Dorothy; Lehmann, Christoph U; Pace, Wilson D; Grundmeier, Robert W

    2016-06-29

    Patient portals may improve communication between families of children with asthma and their primary care providers and improve outcomes. However, the feasibility of using portals to collect patient-reported outcomes from families and the barriers and facilitators of portal implementation across diverse pediatric primary care settings have not been established. We evaluated the feasibility of using a patient portal for pediatric asthma in primary care, its impact on management, and barriers and facilitators of implementation success. We conducted a mixed-methods implementation study in 20 practices (11 states). Using the portal, parents of children with asthma aged 6-12 years completed monthly surveys to communicate treatment concerns, treatment goals, symptom control, medication use, and side effects. We used logistic regression to evaluate the association of portal use with child characteristics and changes to asthma management. Ten clinician focus groups and 22 semistructured parent interviews explored barriers and facilitators of use in the context of an evidence-based implementation framework. We invited 9133 families to enroll and 237 (2.59%) used the portal (range by practice, 0.6%-13.6%). Children of parents or guardians who used the portal were significantly more likely than nonusers to be aged 6-9 years (vs 10-12, P=.02), have mild or moderate/severe persistent asthma (P=.009 and P=.04), have a prescription of a controller medication (P<.001), and have private insurance (P=.002). Portal users with uncontrolled asthma had significantly more medication changes and primary care asthma visits after using the portal relative to the year earlier (increases of 14% and 16%, respectively). Qualitative results revealed the importance of practice organization (coordinated workflows) as well as family (asthma severity) and innovation (facilitated communication and ease of use) characteristics for implementation success. Although use was associated with higher

  2. Adoption of a Portal for the Primary Care Management of Pediatric Asthma: A Mixed-Methods Implementation Study

    PubMed Central

    DuRivage, Nathalie; Mayne, Stephanie L; Finch, Stacia; Ross, Michelle E; Giacomini, Kelli; Suh, Andrew; McCarn, Banita; Brandt, Elias; Karavite, Dean; Staton, Elizabeth W; Shone, Laura P; McGoldrick, Valerie; Noonan, Kathleen; Miller, Dorothy; Lehmann, Christoph U; Pace, Wilson D; Grundmeier, Robert W

    2016-01-01

    Background Patient portals may improve communication between families of children with asthma and their primary care providers and improve outcomes. However, the feasibility of using portals to collect patient-reported outcomes from families and the barriers and facilitators of portal implementation across diverse pediatric primary care settings have not been established. Objective We evaluated the feasibility of using a patient portal for pediatric asthma in primary care, its impact on management, and barriers and facilitators of implementation success. Methods We conducted a mixed-methods implementation study in 20 practices (11 states). Using the portal, parents of children with asthma aged 6-12 years completed monthly surveys to communicate treatment concerns, treatment goals, symptom control, medication use, and side effects. We used logistic regression to evaluate the association of portal use with child characteristics and changes to asthma management. Ten clinician focus groups and 22 semistructured parent interviews explored barriers and facilitators of use in the context of an evidence-based implementation framework. Results We invited 9133 families to enroll and 237 (2.59%) used the portal (range by practice, 0.6%-13.6%). Children of parents or guardians who used the portal were significantly more likely than nonusers to be aged 6-9 years (vs 10-12, P=.02), have mild or moderate/severe persistent asthma (P=.009 and P=.04), have a prescription of a controller medication (P<.001), and have private insurance (P=.002). Portal users with uncontrolled asthma had significantly more medication changes and primary care asthma visits after using the portal relative to the year earlier (increases of 14% and 16%, respectively). Qualitative results revealed the importance of practice organization (coordinated workflows) as well as family (asthma severity) and innovation (facilitated communication and ease of use) characteristics for implementation success

  3. Patient Safety Incidents Involving Sick Children in Primary Care in England and Wales: A Mixed Methods Analysis

    PubMed Central

    Rees, Philippa; Edwards, Adrian; Powell, Colin; Williams, Huw; Carter, Ben; Luff, Donna; Parry, Gareth; Avery, Anthony; Sheikh, Aziz; Donaldson, Liam; Carson-Stevens, Andrew

    2017-01-01

    Background The UK performs poorly relative to other economically developed countries on numerous indicators of care quality for children. The contribution of iatrogenic harm to these outcomes is unclear. As primary care is the first point of healthcare contact for most children, we sought to investigate the safety of care provided to children in this setting. Methods and Findings We undertook a mixed methods investigation of reports of primary care patient safety incidents involving sick children from England and Wales’ National Reporting and Learning System between 1 January 2005 and 1 December 2013. Two reviewers independently selected relevant incident reports meeting prespecified criteria, and then descriptively analyzed these reports to identify the most frequent and harmful incident types. This was followed by an in-depth thematic analysis of a purposive sample of reports to understand the reasons underpinning incidents. Key candidate areas for strengthening primary care provision and reducing the risks of systems failures were then identified through multidisciplinary discussions. Of 2,191 safety incidents identified from 2,178 reports, 30% (n = 658) were harmful, including 12 deaths and 41 cases of severe harm. The children involved in these incidents had respiratory conditions (n = 387; 18%), injuries (n = 289; 13%), nonspecific signs and symptoms, e.g., fever (n = 281; 13%), and gastrointestinal or genitourinary conditions (n = 268; 12%), among others. Priority areas for improvement included safer systems for medication provision in community pharmacies; triage processes to enable effective and timely assessment, diagnosis, and referral of acutely sick children attending out-of-hours services; and enhanced communication for robust safety netting between professionals and parents. The main limitations of this study result from underreporting of safety incidents and variable data quality. Our findings therefore require further exploration in longitudinal

  4. Development and implementation of an online clinical pathway for adult chronic kidney disease in primary care: a mixed methods study.

    PubMed

    Donald, Maoliosa; McBrien, Kerry; Jackson, Wes; Manns, Braden J; Tonelli, Marcello; King-Shier, Kathryn; Jindal, Kailash; Lewanczuk, Richard Z; Scott-Douglas, Nairne; Braun, Ted; Straus, Sharon E; Naugler, Christopher; Elliott, Meghan J; Jun, Min; Hemmelgarn, Brenda R

    2016-08-17

    Primary care physicians and other primary health care professionals from Alberta, Canada identified a clinical pathway as a potential tool to facilitate uptake of clinical practice guidelines for the diagnosis, management and referral of adults with chronic kidney disease. We describe the development and implementation of a chronic kidney disease clinical pathway (CKD-CP; www.ckdpathway.ca ). The CKD-CP was developed and implemented based on the principles of the Knowledge-To-Action Cycle framework. We used a mixed methods approach to identify the usability and feasibility of the CKD-CP. This included individual interviews, an online survey and website analytics, to gather data on barriers and facilitators to use, perceived usefulness and characteristics of users. Results are reported using conventional qualitative content analysis and descriptive statistics. Eighteen individual interviews were conducted with primary care physicians, nephrologists, pharmacists and nurse practitioners to identify themes reflecting both barriers and facilitators to integrating the CKD-CP into clinical practice. Themes identified included: communication, work efficiency and confidence. Of the 159 participants that completed the online survey, the majority (52 %) were first time CKD-CP users. Among those who had previously used the CKD-CP, 94 % agreed or strongly agreed that the pathway was user friendly, provided useful information and increased their knowledge and confidence in the care of patients with CKD. Between November 2014 and July 2015, the CKD-CP website had 10,710 visits, 67 % of which were new visitors. The 3 most frequently visited web pages were home, diagnose and medical management. Canada, Indonesia and the United States were the top 3 countries accessing the website during the 9 month period. An interactive, online, point-of-care tool for primary care providers can be developed and implemented to assist in the care of patients with CKD. Our findings are important

  5. Primary production within the sea-ice zone west of the Antarctic Peninsula: I—Sea ice, summer mixed layer, and irradiance

    NASA Astrophysics Data System (ADS)

    Vernet, Maria; Martinson, Douglas; Iannuzzi, Richard; Stammerjohn, Sharon; Kozlowski, Wendy; Sines, Karie; Smith, Ray; Garibotti, Irene

    2008-09-01

    In shelf waters of the western Antarctic Peninsula (wAP), with abundant macro- and micronutrients, water-column stability has been suggested as the main factor controlling primary production; freshwater input from sea-ice melting stabilizes the upper water column by forming a shallow summer mixed layer. Retreating sea ice in the spring and summer thus defines the area of influence, the sea-ice zone (SIZ) and the marginal ice zone (MIZ). A 12-year time series (1995-2006) was analyzed to address two main questions: (1) what are the spatial and temporal patterns in primary production; and (2) to what extent and in what ways is primary production related to sea-ice dynamics. Data were collected on cruises performed during January of each year, at the height of the growth season, within the region bounded by 64°S and 64°W to the north and 68°S and 66°W to the south. Average daily integrated primary production varied by an order of magnitude, from ˜250 to ˜1100 mg C m -2 d -1, with an average cruise primary production of 745 mg C m -2 d -1. A strong onshore-offshore gradient was evident along the shelf with higher production observed inshore. Inter-annual regional production varied by a factor of 7: maximum rates were measured in 2006 (1788 mg C m -2 d -1) and minimum in 1999 (248 mg C m -2 d -1). The results support the hypothesis that primary production in the wAP shelf is related to sea-ice dynamics. To first order, shallower summer mixed-layer depths in the shelf correlated with late sea retreat and primary production. Principal component analysis showed that high primary production in January was associated with enhanced shelf production toward the coast and in the south, explaining 63% of the variability in space and time. This first mode captured the inter-annual variability in regional production. Temporal variability in primary production (time series of anomalies defined for each location) showed spatial dependence: higher primary production correlated

  6. Epigenetic changes in peripheral leucocytes as biomarkers in intrauterine growth retardation rat

    PubMed Central

    Xu, Xue-Feng; Xu, Shan-Shan; Fu, Lin-Cheng; Hu, Qiong-Yao; Lv, Ying; Du, Li-Zhong

    2016-01-01

    Epigenetics plays an important role in the fetal origins of adult disease. Intrauterine growth retardation (IUGR) can cause increased histone acetylation of the endothelin-1 (ET-1) gene from pulmonary vascular endothelial cells or the whole lung tissue and persist into later life, likely resulting in increased risk of pulmonary hypertension or asthma later in life. However, little is known regarding the correlation of epigenetic changes between specific tissue and peripheral leucocytes. In the present study, an IUGR rat model was established by maternal nutrient restriction. Peripheral blood leucocytes were isolated to detect the ET-1 expression level. Chromatin immunoprecipitation was used to analyze histone modification of the ET-1 gene promoter. The ET-1 protein expression of leucocytes from the 1-week IUGR group was similar to that from the 1-week control group. ET-1 protein expression of leucocytes from 10-week IUGR rats was obviously higher than that of the other groups (P<0.05). The levels of acetylated histone H3 in the ET-1 promoter of leucocytes from the 1-week IUGR rats were significantly higher than those from the age-matched control group (P=0.004). Furthermore, the trends continued ≤10 weeks after birth. In conclusion, epigenetic modifications of leucocytes can in part reflect the epigenetic changes of lung tissue in IUGR rats. Epigenetics of peripheral leucocytes may be used as a biomarker for predicting the risk of the development of disease, and may be used as a surrogate to investigate the subsequent development of pulmonary vascular disease or asthma. PMID:27882215

  7. Epigenetic changes in peripheral leucocytes as biomarkers in intrauterine growth retardation rat.

    PubMed

    Xu, Xue-Feng; Xu, Shan-Shan; Fu, Lin-Cheng; Hu, Qiong-Yao; Lv, Ying; Du, Li-Zhong

    2016-11-01

    Epigenetics plays an important role in the fetal origins of adult disease. Intrauterine growth retardation (IUGR) can cause increased histone acetylation of the endothelin-1 (ET-1) gene from pulmonary vascular endothelial cells or the whole lung tissue and persist into later life, likely resulting in increased risk of pulmonary hypertension or asthma later in life. However, little is known regarding the correlation of epigenetic changes between specific tissue and peripheral leucocytes. In the present study, an IUGR rat model was established by maternal nutrient restriction. Peripheral blood leucocytes were isolated to detect the ET-1 expression level. Chromatin immunoprecipitation was used to analyze histone modification of the ET-1 gene promoter. The ET-1 protein expression of leucocytes from the 1-week IUGR group was similar to that from the 1-week control group. ET-1 protein expression of leucocytes from 10-week IUGR rats was obviously higher than that of the other groups (P<0.05). The levels of acetylated histone H3 in the ET-1 promoter of leucocytes from the 1-week IUGR rats were significantly higher than those from the age-matched control group (P=0.004). Furthermore, the trends continued ≤10 weeks after birth. In conclusion, epigenetic modifications of leucocytes can in part reflect the epigenetic changes of lung tissue in IUGR rats. Epigenetics of peripheral leucocytes may be used as a biomarker for predicting the risk of the development of disease, and may be used as a surrogate to investigate the subsequent development of pulmonary vascular disease or asthma.

  8. Bactericidal activity of rat leucocytic extracts. I. Antibacterial spectrum and the subcellular localization of the bactericidal activity.

    PubMed

    FISHMAN, M; SILVERMAN, M S

    1957-06-01

    All extract of polymorphonuclear leucocytes of the rat, prepared by means of ultrasonic vibration, was found to be bactericidal against M. aureus. The bactericidal activity was primarily confined to the mitochondrial fraction of the leucocytes. The rat leucocyte mitochondrial extract was bactericidal against both Gram-positive (M. aureus, beta-Streptococci, Diplococcus pneumoniae, Corynebacterium diphtheriae) and Gram-negative (Shigella dysenteriae, Salmonella enteritidis, Pasteurella pestis) bacteria. The Gram-positive organisms were more susceptible to the bactericidal activity of the mitochondrial extracts.

  9. The compatibility of prescribing guidelines and the doctor-patient partnership: a primary care mixed-methods study

    PubMed Central

    Solomon, Josie; Raynor, David K; Knapp, Peter; Atkin, Karl

    2012-01-01

    Background UK policy expects health professionals to involve patients in decisions about their care (including medicines use) and, at the same time, to follow prescribing guidelines. The compatibility of these approaches is unclear. Aim To explore the relationship between prescribing guidelines and patient-partnership by exploring the attitudes of patients, GPs and primary care trust (PCT) prescribing advisors. Design and setting A mixed-methods study using qualitative, semi-structured interviews followed by a quantitative, questionnaire survey in primary care in Northern England. Method Interviews were conducted with 14 patients taking a statin or a proton pump inhibitor, eight GPs and two prescribing advisors. A multi-variate sampling strategy was used. Qualitative findings were analysed using framework analysis. Questionnaires based on themes derived from the interviews were distributed to 533 patients and 305 GPs of whom 286 (54%) and 142 (43%) responded. Results Areas of tension between guidelines and patient partnership were identified, including potential damage to trust in the doctor and reduced patient choice, through the introduction of the policy maker as a third stakeholder in prescribing decisions. Other areas of tension related to applying single condition guidelines to patients with multiple illnesses, competition for doctors' time and the perception of cost containment. Many GPs coped with these tensions by adopting a flexible approach or prioritising the doctor–patient relationship over guidelines. Conclusion Rigidly applied guidelines can limit patient choice and may damage the doctor–patient relationship. GPs need flexibility in order to optimise the implementation of prescribing guidelines, while responding to individuals' needs and preferences. PMID:22520915

  10. The primary care provider (PCP)-cancer specialist relationship: A systematic review and mixed-methods meta-synthesis.

    PubMed

    Dossett, Lesly A; Hudson, Janella N; Morris, Arden M; Lee, M Catherine; Roetzheim, Richard G; Fetters, Michael D; Quinn, Gwendolyn P

    2017-03-01

    Although they are critical to models of coordinated care, the relationship and communication between primary care providers (PCPs) and cancer specialists throughout the cancer continuum are poorly understood. By using predefined search terms, the authors conducted a systematic review of the literature in 3 databases to examine the relationship and communication between PCPs and cancer specialists. Among 301 articles identified, 35 met all inclusion criteria and were reviewed in-depth. Findings from qualitative, quantitative, and disaggregated mixed-methods studies were integrated using meta-synthesis. Six themes were identified and incorporated into a preliminary conceptual model of the PCP-cancer specialist relationship: 1) poor and delayed communication between PCPs and cancer specialists, 2) cancer specialists' endorsement of a specialist-based model of care, 3) PCPs' belief that they play an important role in the cancer continuum, 4) PCPs' willingness to participate in the cancer continuum, 5) cancer specialists' and PCPs' uncertainty regarding the PCP's oncology knowledge/experience, and 6) discrepancies between PCPs and cancer specialists regarding roles. These data indicate a pervasive need for improved communication, delineation, and coordination of responsibilities between PCPs and cancer specialists. Future interventions aimed at these deficiencies may improve patient and physician satisfaction and cancer care coordination. CA Cancer J Clin 2017;67:156-169. © 2016 American Cancer Society.

  11. Evidence for a human leucocyte antigen-DM-induced structural change in human leucocyte antigen-DObeta.

    PubMed

    Deshaies, Francis; Diallo, Djibril A; Fortin, Jean-Simon; O'Rourke, Helen M; Pezeshki, Abdul Mohammad; Bellemare-Pelletier, Angélique; Raby, Nicola; Bédard, Nathalie; Brunet, Alexandre; Denzin, Lisa K; Thibodeau, Jacques

    2009-07-01

    Human leucocyte antigen (HLA)-DO is a non-classical major histocompatibility complex class II molecule which modulates the function of HLA-DM and the loading of antigenic peptides on molecules such as HLA-DR. The bulk of HLA-DO associates with HLA-DM and this interaction is critical for HLA-DO egress from the endoplasmic reticulum. HLA-DM assists the early steps of HLA-DO maturation presumably through the stabilization of the interactions between the N-terminal regions of the alpha and beta chains. To evaluate a possible role for HLA-DM in influencing the conformation of HLA-DO, we made use of a monoclonal antibody, Mags.DO5, that was raised against HLA-DO/DM complexes. Using transfected cells expressing mismatched heterodimers between HLA-DR and -DO chains, we found that the epitope for Mags.DO5 is located on the DObeta chain and that Mags.DO5 reactivity was increased upon cotransfection with HLA-DM. Our results suggest that HLA-DM influences the folding of HLA-DO in the endoplasmic reticulum. A mutant HLA-DO showing reduced capacity for endoplasmic reticulum egress was better recognized by Mags.DO5 in the presence of HLA-DM. On the other hand, an HLA-DO mutant capable of endoplasmic reticulum egress on its own was efficiently recognized by Mags.DO5, irrespective of the presence of HLA-DM. Taken together, our results suggest that HLA-DM acts as a private chaperone, directly assisting the folding of HLA-DO to promote egress from the endoplasmic reticulum.

  12. Evaluation of silver nanospheres on viability and innate cellular parameters of gilthead seabream (Sparus aurata L.) head-kidney leucocytes.

    PubMed

    Guardiola, F A; Logothetis, P; Meseguer, J; Esteban, M A

    2017-08-16

    The increasing use of nanomaterials, e.g. nanosilver, has lead to concerns about environmental contamination and possible toxic effects on aquatic organisms. Here, we present evidence for the impact of silver nanospheres (AgNSs) on fish innate immune cells after in vitro exposure. AgNSs of 20, 50 or 100 nm in diameter were tested with the smallest ones (20 nm) clearly having the most deleterious effects, after an exposure period of 30 min, followed by the medium-sized ones; the NSs of 100 nm had no impact. The effective concentration was determined at 10 μg ml(-1) while lower concentrations (1, 2.5 or 5 μg ml(-1)) were ineffective. Head-kidney mixed leucocyte population showed significant viability reduction which was attributable to diminished viability of macrophages/monocytes and lymphocytes only whereas granulocytes' viability was not affected at the above exposure regime. Furthermore, cellular respiratory burst activity, phagocytic capacity and phagocytic ability were all reduced, with the first two parameters exhibiting the sharper reductions. Finally, transmission electron microscopy revealed that the AgNSs' internalization was brought about via phagocytosis, pinocytosis, receptor-mediated endocytosis and macropinocytosis; also, that cell death could be effected in either an apoptotic or a necrotic manner. It is concluded that AgNSs are potentially very noxious for the teleost fish immune system as they can adversely affect the function and viability of the head-kidney leucocytes. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Expression of natriuretic peptide receptor mRNA and functional response to atrial natriuretic peptide and C-type natriuretic peptide in rainbow trout (Oncorhynchus mykiss) head kidney leucocytes.

    PubMed

    Powell, M D; McWilliam, H; McLeod, J; Nankervis, S; Butler, R; Toop, T

    2008-04-01

    The stimulatory effect of vasomodulatory natriuretic peptide hormones on macrophages and peripheral blood leucocytes in mammals is well-established. However, the relationship in lower vertebrates has not been characterised. Expression of atrial natriuretic peptide, ventricular natriuretic peptide and C-type natriuretic peptide-1, and the guanylyl cyclase-linked (GC) natriuretic peptide receptor-A and -B-type receptors (NPR-A and NPR-B, respectively) was determined by PCR from the mRNA of rainbow trout head kidney leucocytes yielding gene fragments with 100% homology to the same respective natriuretic peptide and NPR-A and -B sequences obtained from other rainbow trout tissues. A mixed population of isolated rainbow trout head kidney leucocytes was stimulated in vitro with trout atrial natriuretic peptide (specific NPR-A agonist) and trout C-type natriuretic peptide (NPR-A and -B agonist) as well as the cGMP agonist 8-bromo-cGMP or the GC inhibitor 8-bromo-phenyl-eutheno-cGMP. Respiratory burst was stimulated by trout atrial natriuretic peptide, trout C-type natriuretic peptide-1 and 8-bromo-cGMP in a dose dependant manner with the highest activity as a result of stimulation with trout C-type natriuretic peptide-1 in excess of that achieved by phorbol myristate acetate (PMA). Equimolar concentrations of the inhibitor, inhibited the respiratory burst caused by the natriuretic peptides and 8-bromo-cGMP. The natriuretic peptide receptors on rainbow trout head kidney leucocytes appear to have a stimulatory function with regard to respiratory burst that is activated through a cGMP second messenger pathway and the natriuretic peptides expressed in the head kidney leucocytes may well act in a paracrine/autocrine manner.

  14. Phagocytosis and Respiratory Burst Activity in Lumpsucker (Cyclopterus lumpus L.) Leucocytes Analysed by Flow Cytometry

    PubMed Central

    Haugland, Gyri T.; Jakobsen, Ragnhild Aakre; Vestvik, Nils; Ulven, Kristian; Stokka, Lene; Wergeland, Heidrun I.

    2012-01-01

    In the present study, we have isolated leucocytes from peripheral blood, head kidney and spleen from lumpsucker (Cyclopterus lumpus L.), and performed functional studies like phagocytosis and respiratory burst, as well as morphological and cytochemical analyses. Different leucocytes were identified, such as lymphocytes, monocytes/macrophages and polymorphonuclear cells with bean shaped or bilobed nuclei. In addition, cells with similar morphology as described for dendritic cells in trout were abundant among the isolated leucocytes. Flow cytometry was successfully used for measuring phagocytosis and respiratory burst activity. The phagocytic capacity and ability were very high, and cells with different morphology in all three leucocyte preparations phagocytised beads rapidly. Due to lack of available cell markers, the identity of the phagocytic cells could not be determined. The potent non-specific phagocytosis was in accordance with a high number of cells positive for myeloperoxidase, an enzyme involved in oxygen-dependent killing mechanism present in phagocytic cells. Further, high respiratory burst activity was present in the leucocytes samples, verifying a potent oxygen- dependent degradation. At present, the specific antibody immune response could not be measured, as immunoglobulin or B-cells have not yet been isolated. Therefore, analyses of the specific immune response in this fish species await further clarification. The present study presents the first analyses of lumpsucker immunity and also the first within the order Scopaeniformes. PMID:23112870

  15. Protein nitration is predominantly mediated by a peroxynitrite-dependent pathway in cultured human leucocytes.

    PubMed Central

    Galiñanes, Manuel; Matata, Bashir M

    2002-01-01

    Protein nitration is a common characteristic of oxidative injury caused by the invasion of leucocytes into inflammatory lesions. Two distinct pathways of nitration of protein tyrosine residues, namely the peroxynitrite (ONOO(-))-mediated pathway and another catalysed by the haem-containing peroxidases, have been reported under experimental conditions. However, the contribution of these two pathways in human leucocytes is still controversial. The present study demonstrates that the process of phenolic nitration of proteins in cultured human leucocytes is mainly ONOO(-)-mediated and that it differs between granulocytes and mononuclear cells, depending on the cell compartment and the stimuli. We have also shown that NO induces protein nitration via a ONOO(-)-dependent pathway, whereas NO(2)(-), the NO metabolite, does not increase but decreases nitration in PMA-stimulated leucocytes. The inhibition of myeloperoxidase activity did not reduce protein nitration; on the other hand, the myeloperoxidase inhibitor aminobenzoic hydrazide caused increased nitration, which was mediated by ONOO(-). These results suggest that protein nitration is predominantly mediated by a ONOO(-)-dependent pathway in cultured human leucocytes and that the myeloperoxidase-catalysed pathway does not play a significant role in protein nitration. PMID:12099887

  16. Fatty acid binding protein 4 in circulating leucocytes reflects atherosclerotic lesion progression in Apoe(-/-) mice.

    PubMed

    Agardh, Hanna E; Gertow, Karl; Salvado, Dolores M; Hermansson, Andreas; van Puijvelde, Gijs H; Hansson, Göran K; n-Berne, Gabrielle Paulsso; Gabrielsen, Anders

    2013-02-01

    Discovery of novel biomarkers for atherosclerosis is important to aid in early diagnosis of pre-symptomatic patients at high risk of cardiovascular events. The aim of the present study was therefore to identify potential biomarkers in circulating cells reflecting atherosclerotic lesion progression in the vessel wall. We performed gene arrays on circulating leucocytes from atherosclerosis prone Apoe(-/-) mice with increasing ages, using C57BL/6 mice as healthy controls. We identified fatty acid binding protein 4 (FABP4) mRNA to be augmented in mice with established disease compared with young Apoe(-/-) or controls. Interestingly, the transcript FABP4 correlated significantly with lesion size, further supporting a disease associated increase. In addition, validation of our finding on protein level showed augmented FABP4 in circulating leucocytes whereas, importantly, no change could be observed in plasma. Immunofluorescence analysis demonstrated FABP4 to be present mainly in circulating neutrophils and to some extent in monocytes. Moreover, FABP4-positive neutrophils and macrophages could be identified in the subintimal space in the plaque. Using human circulating leucocytes, we confirmed the presence of FABP4 protein in neutrophils and monocytes. In conclusion, we have showed that cellular levels of FABP4 in circulating leucocytes associate with lesion development in the experimental Apoe(-/-) model. The increased expression is primarily localized to neutrophils, but also in monocytes. We have identified FABP4 in leucocytes as a potential and easy accessible biomarker of atherosclerosis which could be of future clinical relevance.

  17. Phagocytosis and respiratory burst activity in lumpsucker (Cyclopterus lumpus L.) leucocytes analysed by flow cytometry.

    PubMed

    Haugland, Gyri T; Jakobsen, Ragnhild Aakre; Vestvik, Nils; Ulven, Kristian; Stokka, Lene; Wergeland, Heidrun I

    2012-01-01

    In the present study, we have isolated leucocytes from peripheral blood, head kidney and spleen from lumpsucker (Cyclopterus lumpus L.), and performed functional studies like phagocytosis and respiratory burst, as well as morphological and cytochemical analyses. Different leucocytes were identified, such as lymphocytes, monocytes/macrophages and polymorphonuclear cells with bean shaped or bilobed nuclei. In addition, cells with similar morphology as described for dendritic cells in trout were abundant among the isolated leucocytes. Flow cytometry was successfully used for measuring phagocytosis and respiratory burst activity. The phagocytic capacity and ability were very high, and cells with different morphology in all three leucocyte preparations phagocytised beads rapidly. Due to lack of available cell markers, the identity of the phagocytic cells could not be determined. The potent non-specific phagocytosis was in accordance with a high number of cells positive for myeloperoxidase, an enzyme involved in oxygen-dependent killing mechanism present in phagocytic cells. Further, high respiratory burst activity was present in the leucocytes samples, verifying a potent oxygen- dependent degradation. At present, the specific antibody immune response could not be measured, as immunoglobulin or B-cells have not yet been isolated. Therefore, analyses of the specific immune response in this fish species await further clarification. The present study presents the first analyses of lumpsucker immunity and also the first within the order Scopaeniformes.

  18. Paracrine effects of uterine leucocytes on gene expression of human uterine stromal fibroblasts.

    PubMed

    Germeyer, Ariane; Sharkey, Andrew Mark; Prasadajudio, Mirari; Sherwin, Robert; Moffett, Ashley; Bieback, Karen; Clausmeyer, Susanne; Masters, Leanne; Popovici, Roxana Maria; Hess, Alexandra Petra; Strowitzki, Thomas; von Wolff, Michael

    2009-01-01

    The endometrium contains a distinct population of immune cells that undergo cyclic changes during the menstrual cycle and implantation. The majority of these leucocytes are uterine NK (uNK) cells, however how these cells interact with uterine stromal fibroblasts remains unclear. We therefore investigated the paracrine effect of medium conditioned by uterine decidual leucocytes (which are enriched for uNK cells) on the gene expression profile of endometrial stromal fibroblasts in vitro using a cDNA microarray. Our results, verified by real-time PCR, ELISA and FACS analysis, reveal that soluble factors from uterine leucocytes substantially alter endometrial stromal fibroblast gene expression. The largest group of up-regulated genes found was chemokines and cytokines. These include IL-8, CCL8 and CXCL1, which have also been shown to be stimulated by contact of stromal fibroblasts with trophoblast, suggesting that uNK cells work synergistically to support trophoblast migration during implantation. The decidual leucocytes also up-regulated IL-15 and IL-15Ralpha in stromal fibroblasts which could produce a niche for uNK cells allowing proliferation within and recruitment into the uterus, as seen in bone marrow. Overall this study demonstrates, for the first time, the paracrine communication between uterine leucocytes and uterine stromal fibroblasts, and adds to the understanding of how the uterine immune system contributes to the changes seen within the cycling endometrium.

  19. Resolution of leucocyte-mediated mucosal diseases. A novel in vivo paradigm for drug development

    PubMed Central

    Persson, Carl; Uller, Lena

    2012-01-01

    Removal of disease-driving inflammatory leucocytes is central to resolution of inflammation. The current pharmacological dogma teaches leucocyte elimination through apoptosis followed by phagocytosis. However, actual resolving roles of apoptotic–phagocytic processes have been difficult to demonstrate in the major diseases that are characterized by mucosal tissue inflammation. Many current in vivo observations rather demonstrate that leucocyte elimination occurs by transepithelial locomotion. Findings in diseased gut and bladder mucosae support this notion. Respiratory disease data are particularly compelling. Eosinophils and neutrophils abound in sputum and tracheal aspirates during treatment-induced recovery from severe asthma. Prolonged sputum neutrophilia, along with clinical improvement, follows upon smoking cessation in COPD. Eosinophils, neutrophils, lymphocytes, mast cells and dendritic cells also move in large numbers into the bronchial lumen at spontaneous inflammation resolution following allergen challenge in allergic rhinitis and asthma. A corresponding reduction of infiltrated cells in the bronchial mucosal tissue demonstrates efficiency of the transepithelial elimination pathway. Underscoring its operational role, drugs impeding transepithelial elimination of leucocytes aggravate mucosal/parenchymal inflammation. Hence, relying on lumen cell data alone can lead to paradoxical conclusions regarding anti-inflammatory drug efficacy. Conversely, drugs promoting non-injurious transepithelial elimination of leucocytes could resolve mucosal inflammatory diseases. PMID:22053825

  20. Leucocyte subset-specific type 1 interferon signatures in SLE and other immune-mediated diseases

    PubMed Central

    Jovanovic, Vojislav; Teo, Boon Wee; Mak, Anselm; Thumboo, Julian; McKinney, Eoin F; Lee, James C; MacAry, Paul; Kemeny, David M; Jayne, David RW; Fong, Kok Yong; Lyons, Paul A; Smith, Kenneth GC

    2016-01-01

    Objectives Type 1 interferons (IFN-1) are implicated in the pathogenesis of systemic lupus erythematosus (SLE), but most studies have only reported the effect of IFN-1 on mixed cell populations. We aimed to define modules of IFN-1-associated genes in purified leucocyte populations and use these as a basis for a detailed comparative analysis. Methods CD4+ and CD8+ T cells, monocytes and neutrophils were purified from patients with SLE, other immune-mediated diseases and healthy volunteers and gene expression then determined by microarray. Modules of IFN-1-associated genes were defined using weighted gene coexpression network analysis. The composition and expression of these modules was analysed. Results 1150 of 1288 IFN-1-associated genes were specific to myeloid subsets, compared with 11 genes unique to T cells. IFN-1 genes were more highly expressed in myeloid subsets compared with T cells. A subset of neutrophil samples from healthy volunteers (HV) and conditions not classically associated with IFN-1 signatures displayed increased IFN-1 gene expression, whereas upregulation of IFN-1-associated genes in T cells was restricted to SLE. Conclusions Given the broad upregulation of IFN-1 genes in neutrophils including in some HV, investigators reporting IFN-1 signatures on the basis of whole blood samples should be cautious about interpreting this as evidence of bona fide IFN-1-mediated pathology. Instead, specific upregulation of IFN-1-associated genes in T cells may be a useful biomarker and a further mechanism by which elevated IFN-1 contributes to autoimmunity in SLE. PMID:27252891

  1. Heparan sulfate on activated glomerular endothelial cells and exogenous heparinoids influence the rolling and adhesion of leucocytes.

    PubMed

    Rops, Angelique L; Jacobs, Cor W; Linssen, Peter C; Boezeman, Jan B; Lensen, Joost F; Wijnhoven, Tessa J; van den Heuvel, Lambert P; van Kuppevelt, Toin H; van der Vlag, Johan; Berden, Jo H

    2007-04-01

    Proliferative glomerulonephritides are characterized by the influx of leucocytes. Heparan sulfate (HS) plays an important role in the recruitment, rolling and firm adhesion of leucocytes to activated endothelium. Recently, we have shown the importance of HS on activated mouse glomerular endothelial cells (mGEnC-1) for the firm adhesion of leucocytes in a static adhesion assay. In the present study, we evaluated the role of HS on glomerular endothelial cells and the effect of adding heparinoids on the leucocyte-glomerular endothelium interaction under dynamic flow conditions. The number of rolling and firmly adhering leucocytes, and the rolling velocity of leucocytes was determined on a monolayer of unactivated or TNF-alpha-activated mGEnC-1 under dynamic flow conditions using physiological relevant shear stress rates in a flow chamber system. Furthermore, the effects of removal of HS on TNF-alpha-activated mGEnC-1 by heparinase III treatment, and of different concentrations of heparin, tinzaparin and HS, on the rolling and adhesion of leucocytes were evaluated. At the calculated physiological shear stress rate of 0.8 dynes/cm2 the number of rolling and firmly adhering leucocytes to mGEnC-1 increased 2-fold after activation with TNF-alpha, whereas the rolling velocity of the leucocytes decreased 2-fold. Addition of heparin, tinzaparin or HS, and the removal of HS on mGEnC-1 reduced the number of leucocytes rolling and adhering to activated mGEnC-1 about 2-3-fold, while the rolling velocity increased more than 2-fold. HS on activated glomerular endothelial cells is important for the interaction with leucocytes under flow conditions, while exogenous heparinoids interfere with this interaction. These results suggest that supplementary treatment of proliferative glomerulonephritides with heparinoids is an interesting option to pursue.

  2. Rat leucocyte response to the bites of rat fleas (Siphonaptera: Pulicidae).

    PubMed

    Vaughan, J A; Jerse, A E; Azad, A F

    1989-09-01

    The host response to bites of the oriental rat flea, Xenopyslla cheopis Rothschild, was investigated by examining rat blood leucocyte kinetics, histopathology, and the effect that the host response had upon subsequent flea feeding and longevity. Test rats were subjected to controlled exposures of fleas, and leucocyte data from test rats were compared to those of unexposed controls. Of the five leucocyte types examined, only the basophil appeared to play a role in the host blood response to flea bites. Significant increases in blood basophil levels occurred 2-3 d after exposure but subsided to control levels within a week. However, flea feeding did not produce histopathology at the flea feeding sites nor did the basophilic blood response of rats affect subsequent feeding or longevity of the fleas.

  3. Isolation of two polypeptides comprising the neutrophil-immobilizing factor of human leucocytes.

    PubMed Central

    Watt, K W; Brightman, I L; Goetzl, E J

    1983-01-01

    Human leucocyte lysosomal polypeptides of mol. wt 4000-5000, which constitute the neutrophil-immobolizing factor (NIF), were isolated from the 22,000 g supernate of sonicates of human neutrophils by filtration on Sephadex G-75. The larger (NIF-1) and smaller (NIF-2) of the polypeptides were resolved by filtration on Bio-Gel P6 and purified to homogeneity by sequential reverse-phase high performance liquid chromatography and paper electrophoresis. The results of analyses of amino acid composition indicated that NIF-1 and NIF-2 are distinct polypeptides composed of an apparent total of 41 and 38 amino acids, respectively. Both NIF polypeptides contain one cysteine and one methionine, lack isoleucine, tyrosine and phenylalanine, and are rich in histidine and proline. The sequence of 20 of the amino-terminal amino acids of both NIF polypeptides is identical, but NIF-2 possesses an additional alanine at the amino-terminus. Highly purified NIF-1 and NIF-2 inhibited human neutrophil random migration and chemotaxis to diverse stimuli in a concentration-dependent manner, with 50% inhibition of chemotaxis by 0.31-1 x 10(-8) M NIF-1 and 1-3 x 10(-7) M NIF-2. Neither NIF polypeptide was cytotoxic for neutrophils, altered neutrophil phagocytosis or release of lysosomal enzymes, or inhibited mononuclear leucocyte chemotaxis. The leucocyte and functional specificity of the NIF polypeptides and the quantitites released upon stimulation of the human leucocytes suggest that the transition to a mononuclear leucocyte population in chronic inflammation may be attributable in part to the NIF derived from the leucocyte infiltrates of acute responses. PMID:6848456

  4. Flow cytometric detection of alpha-1-acid glycoprotein on feline circulating leucocytes.

    PubMed

    Paltrinieri, S; Marchini, I; Gelain, M E

    2012-08-01

    To assess whether alpha-1-acid glycoprotein (AGP) can be detected on the membrane of feline circulating leucocytes. The presence of AGP on circulating leucocytes was investigated in both clinically healthy cats and cats with different diseases. A group of feline coronavirus (FCoV)-positive cats, comprising cats with feline infectious peritonitis (FIP) and cats not affected by FIP but seropositive for FCoV, were included in this study because the serum concentration of AGP increases during FCoV infection. Flow cytometry (using an anti-feline AGP antibody), serum protein electrophoresis, routine haematology and measurement of the serum AGP concentration were performed using blood samples from 32 healthy cats (19 FCoV-seropositive), 13 cats with FIP and 12 with other diseases (6 FCoV-seropositive). The proportion of cats with AGP-positive leucocytes in the different groups (e.g. controls vs sick; FIP vs other diseases, etc.) or in cats with different intensities of inflammatory response was compared using a Chi-square test. AGP-positive leucocytes were found in 23% of cats. Compared with controls, the proportion of patients with positive granulocytes and monocytes was higher among sick cats (especially cats with diseases other than FIP) and cats with high serum AGP concentration, but not in cats with leucocytosis or that were FCoV-seropositive. AGP-positive leucocytes can be found in feline blood, especially during inflammation. Conversely, no association between AGP-positive leucocytes and FIP was found. Further studies are needed to elucidate the mechanism responsible for this finding and its diagnostic role in cats with inflammation. © 2012 The Authors. Australian Veterinary Journal © 2012 Australian Veterinary Association.

  5. Access to primary health care for acute vascular events in rural low income settings: a mixed methods study.

    PubMed

    Ahmed, Shyfuddin; Chowdhury, Muhammad Ashique Haider; Khan, Md Alfazal; Huq, Nafisa Lira; Naheed, Aliya

    2017-01-18

    Cardiovascular diseases (CVDs) are the leading cause of global mortality. Among the CVDs, acute vascular events (AVE) mainly ischemic heart diseases and stroke are the largest contributors. To achieve 25% reduction in preventable deaths from CVDs by 2025, health systems need to be equipped with extended service coverage in order to provide person-centered care. The overall goal of this proposed study is to assess access to health care in-terms of service availability, care seeking patterns and barriers to access care after AVE in rural Bangladesh. We will consider myocardial infarction (MI) and stroke as acute vascular events. We will conduct a mixed methods study in rural Matlab, Bangladesh. This study will comprise of a) health facility survey, b) structured questionnaire interview and c) qualitative study. We will assess service availabilities by creating an inventory of public and private health facilities. Readiness of the facilities to deliver services for AVE will be assessed through a health facility survey using 'service availability and readiness assessment' (SARA) tools of the World Health Organization (WHO). We will interview survivors of AVE and caregivers (present and accompanied the person during the event) of person who died from AVE for exploring patterns of care seeking during an AVE. For exploring barriers to access care for AVE, we will conduct in-depth interview with survivors of AVE and caregivers of the person who died from AVE. We will also conduct key informant interviews with the service providers at primary health care (PHC) facilities and government high level officials at central health administration of Bangladesh. This study will provide a comprehensive picture of access to primary health care services during acute cardiovascular events as stroke & MI in rural context of Bangladesh. It will explore available service facilities in rural area for management, utilization of services and barriers to access care during an acute emergency

  6. Characterization of binding specificities of bovine leucocyte class I molecules: impacts for rational epitope discovery

    PubMed Central

    Hansen, Andreas M.; Rasmussen, Michael; Svitek, Nicholas; Harndahl, Mikkel; Golde, William T.; Barlow, John; Nene, Vishvanath; Buus, Søren

    2014-01-01

    The binding of peptides to classical major histocompatibility complex (MHC) class I proteins is the single most selective step in antigen presentation. However, the peptide-binding specificity of cattle MHC (bovine leucocyte antigen, BoLA) class I (BoLA-I) molecules remains poorly characterized. Here, we demonstrate how a combination of high-throughput assays using positional scanning combinatorial peptide libraries, peptide dissociation, and peptide-binding affinity binding measurements can be combined with bioinformatics to effectively characterize the functionality of BoLA-I molecules. Using this strategy, we characterized eight BoLA-I molecules, and found the peptide specificity to resemble that of human MHC-I molecules with primary anchors most often at P2 and P9, and occasional auxiliary P1/P3/P5/P6 anchors. We analyzed nine reported CTL epitopes from Theileria parva, and in eight cases, stable and high affinity binding was confirmed. A set of peptides were tested for binding affinity to the eight BoLA proteins and used to refine the predictors of peptide–MHC binding NetMHC and NetMHCpan. The inclusion of BoLA-specific peptide-binding data led to a significant improvement in prediction accuracy for reported T. parva CTL epitopes. For reported CTL epitopes with weak or no predicted binding, these refined prediction methods suggested presence of nested minimal epitopes with high-predicted binding affinity. The enhanced affinity of the alternative peptides was in all cases confirmed experimentally. This study demonstrates how biochemical high-throughput assays combined with immunoinformatics can be used to characterize the peptide-binding motifs of BoLA-I molecules, boosting performance of MHC peptide-binding prediction methods, and empowering rational epitope discovery in cattle. PMID:25186069

  7. Enhancement of mite antigen-induced histamine release by deuterium oxide from leucocytes of chronic urticarial patients

    SciTech Connect

    Numata, T.; Yamamoto, S.; Yamura, T.

    1981-09-01

    The mite antigen-induced histamine release from leucocytes of chronic urticarial patients was enhanced in the presence of deuterium oxide, which stabilizes microtubules. This enhancing effect of deuterium oxide on the histamine release from leucocytes may provide a useful means for the detection of allergens in vitro in chronic urticaria.

  8. Measurement tools and process indicators of patient safety culture in primary care. A mixed methods study by the LINNEAUS collaboration on patient safety in primary care

    PubMed Central

    Parker, Dianne; Wensing, Michel; Esmail, Aneez; Valderas, Jose M

    2015-01-01

    ABSTRACT Background: There is little guidance available to healthcare practitioners about what tools they might use to assess the patient safety culture. Objective: To identify useful tools for assessing patient safety culture in primary care organizations in Europe; to identify those aspects of performance that should be assessed when investigating the relationship between safety culture and performance in primary care. Methods: Two consensus-based studies were carried out, in which subject matter experts and primary healthcare professionals from several EU states rated (a) the applicability to their healthcare system of several existing safety culture assessment tools and (b) the appropriateness and usefulness of a range of potential indicators of a positive patient safety culture to primary care settings. The safety culture tools were field-tested in four countries to ascertain any challenges and issues arising when used in primary care. Results: The two existing tools that received the most favourable ratings were the Manchester patient safety framework (MaPsAF primary care version) and the Agency for healthcare research and quality survey (medical office version). Several potential safety culture process indicators were identified. The one that emerged as offering the best combination of appropriateness and usefulness related to the collection of data on adverse patient events. Conclusion: Two tools, one quantitative and one qualitative, were identified as applicable and useful in assessing patient safety culture in primary care settings in Europe. Safety culture indicators in primary care should focus on the processes rather than the outcomes of care. PMID:26339832

  9. Efficacy of a cognitive and behavioural psychotherapy applied by primary care psychologists in patients with mixed anxiety-depressive disorder: a research protocol.

    PubMed

    Jauregui, Amale; Ponte, Joaquín; Salgueiro, Monika; Unanue, Saloa; Donaire, Carmen; Gómez, Maria Cruz; Burgos-Alonso, Natalia; Grandes, Gonzalo

    2015-03-20

    In contrast with the recommendations of clinical practice guidelines, the most common treatment for anxiety and depressive disorders in primary care is pharmacological. The aim of this study is to assess the efficacy of a cognitive-behavioural psychological intervention, delivered by primary care psychologists in patients with mixed anxiety-depressive disorder compared to usual care. This is an open-label, multicentre, randomized, and controlled study with two parallel groups. A random sample of 246 patients will be recruited with mild-to-moderate mixed anxiety-depressive disorder, from the target population on the lists of 41 primary care doctors. Patients will be randomly assigned to the intervention group, who will receive standardised cognitive-behavioural therapy delivered by psychologists together with usual care, or to a control group, who will receive usual care alone. The cognitive-behavioural therapy intervention is composed of eight individual 60-minute face-to face sessions conducted in eight consecutive weeks. A follow-up session will be conducted over the telephone, for reinforcement or referral as appropriate, 6 months after the intervention, as required. The primary outcome variable will be the change in scores on the Short Form-36 General Health Survey. We will also measure the change in the frequency and intensity of anxiety symptoms (State-Trait Anxiety Inventory) and depression (Beck Depression Inventory) at baseline, and 3, 6 and 12 months later. Additionally, we will collect information on the use of drugs and health care services. The aim of this study is to assess the efficacy of a primary care-based cognitive-behavioural psychological intervention in patients with mixed anxiety-depressive disorder. The international scientific evidence has demonstrated the need for psychologists in primary care. However, given the differences between health policies and health services, it is important to test the effect of these psychological interventions

  10. The Fps/Fes kinase regulates leucocyte recruitment and extravasation during inflammation.

    PubMed

    Parsons, Sean A; Mewburn, Jeffrey D; Truesdell, Peter; Greer, Peter A

    2007-12-01

    Fps/Fes and Fer comprise a distinct subfamily of cytoplasmic protein-tyrosine kinases, and have both been implicated in the regulation of innate immunity. Previous studies showed that Fps/Fes-knockout mice were hypersensitive to systemic lipopolysaccharide (LPS) challenge, and Fer-deficient mice displayed enhanced recruitment of leucocytes in response to localized LPS challenge. We show here for the first time, a role for Fps in the regulation of leucocyte recruitment to areas of inflammation. Using the cremaster muscle intravital microscopy model, we observed increased leucocyte adherence to venules, and increased rates and degrees of transendothelial migration in Fps/Fes-knockout mice relative to wild-type animals subsequent to localized LPS challenge. There was also a decreased vessel wall shear rate in the post-capillary venules of LPS-challenged Fps/Fes-knockout mice, and an increase in neutrophil migration into the peritoneal cavity subsequent to thioglycollate challenge. Using flow cytometry to quantify the expression of surface molecules, we observed prolonged expression of the selectin ligand PSGL-1 on peripheral blood neutrophils from Fps/Fes-knockout mice stimulated ex vivo with LPS. These observations provide important insights into the observed in vivo behaviour of leucocytes in LPS-challenged Fps/Fes-knockout mice and provide evidence that the Fps/Fes kinase plays an important role in the innate immune response.

  11. VLA-4 blockade by natalizumab inhibits sickle reticulocyte and leucocyte adhesion during simulated blood flow.

    PubMed

    White, Jennell; Krishnamoorthy, Sriram; Gupta, Dipti; Lancelot, Moira; Moore, Nancy; Sarnaik, Sharada; Hobbs, William E; Light, David R; Hines, Patrick

    2016-09-01

    Very Late Antigen-4 (VLA-4, α4β1-integrin, ITGA4) orchestrates cell-cell and cell-endothelium adhesion. Given the proposed role of VLA-4 in sickle cell disease (SCD) pathophysiology, we evaluated the ability of the VLA-4 blocking antibody natalizumab to inhibit SCD blood cell adhesion. Natalizumab recognized surface VLA-4 on leucocytes and reticulocytes in whole blood from SCD subjects. SCD reticulocytes were positive for VLA-4, while VLA-4 staining of non-SCD reticulocytes was undetectable. Titrations with natalizumab revealed the presence of saturable levels of VLA-4 on both SCD reticulocytes and leucocytes similar to healthy subject leucocytes. Under physiological flow conditions, the adhesion of SCD whole blood cells and isolated SCD leucocytes to immobilized vascular cell adhesion molecule 1 (VCAM-1) was blocked by natalizumab in a dose-dependent manner, which correlated with cell surface receptor binding. Natalizumab also inhibited >50% of whole blood cell binding to TNF-α activated human umbilical vein endothelial cell monolayers under physiological flow at clinically relevant concentrations (10 to 100 μg/ml). This indicates that VLA-4 is the dominant receptor that drives SCD reticulocyte and mononuclear cell adhesion to VCAM-1 and that the VLA-4 adhesion to VCAM-1 is a significant contributor to SCD blood cell adhesion to endothelium. Thus, VLA-4 blockade may be beneficial in sickle cell disease.

  12. Characterization of binding specificities of Bovine Leucocyte class I molecules: Impacts for rational epitope discovery

    USDA-ARS?s Scientific Manuscript database

    The binding of peptides to classical major histocompatibility complex (MHC) class-I proteins is the single most selective step in antigen presentation. However, the peptide binding specificity of cattle MHC (bovine leucocyte antigen, BoLA) class I (BoLA-I) molecules remains poorly characterized. Her...

  13. The haematology of hyperthyroidism: abnormalities of erythrocytes, leucocytes, thrombocytes and haemostasis.

    PubMed Central

    Ford, H. C.; Carter, J. M.

    1988-01-01

    The abnormalities of erythrocytes, leucocytes, thrombocytes and coagulation that have been reported, particularly in more recent years, to be associated with hyperthyroidism are surveyed. Several areas are highlighted where further investigations could lead to clinically useful insights, improved information about the haematological processes involved or to a better understanding of thyroid hormone action. PMID:3076660

  14. Discriminative protection against hydroxyl and superoxide anion radicals by quercetin in human leucocytes in vitro.

    PubMed

    Wilms, Lonneke C; Kleinjans, Jos C S; Moonen, Edwin J C; Briedé, Jacob J

    2008-03-01

    Antioxidants play a vital role in the cellular protection against oxidative damage. Quercetin is a well-investigated antioxidant and known to be able to protect against cellular oxidative DNA damage. In this study, we tried to relate the protection by quercetin pre-treatment against oxidative DNA damage in human leucocytes in vitro to the interaction of quercetin in solution with hydroxyl and superoxide anion radicals as measured by electron spin resonance (ESR) spectrometry, using DMPO as a spin trap. Further, scavenging capacity of quercetin-treated leucocytes in vitro was evaluated by ESR spectrometry. Quercetin appears capable of protecting human leucocytes against oxidative DNA damage caused by hydrogen peroxide in a dose-dependent manner. The protection of leucocytes against superoxides is ambiguous. Incubation concentrations of quercetin (1, 10, and 50 microM) reduced levels of superoxide-induced oxidative DNA damage, while at 100 microM the amount of damage was increased. These results are supported by ESR-findings on quercetin in solution, also showing a prooxidant effect at 100 microM. ESR spectroscopy showed rate constant values for the reaction kinetics of quercetin in lowering iron-dependent hydroxyl radical formation and NADH-dependent superoxide anion formation of respectively 3.2 x 10(12)M(-1)s(-1) and 1.1 x 10(4)M(-1)s(-1). This shows that quercetin is a more potent inhibitor of hydroxyl radical formation than a scavenger of superoxide anions.

  15. Interaction between leucocytes and human spermatozoa influencing reactive oxygen intermediates release.

    PubMed

    Fraczek, Monika; Sanocka, Dorota; Kurpisz, Maciej

    2004-04-01

    The relationship between the presence of white blood cells (WBCs) and the fertilizing potential of human semen is still an open question. It is well known that the presence of leucocytes in human semen can be related to the production of reactive oxygen intermediates (ROI). Semen samples were obtained from 15 normozoospermic men and leucocytes were isolated from heparinized blood drawn from 15 volunteers. Lucigenin and luminol-mediated chemiluminescence assays were used to determine reactive oxygen species (ROS) generation by non-activated or activated leucocytes through 12-myristate-13-acetate or N-formyl-methionyl-leucyl-phenyalanine (FMLP) before the addition of spermatozoa isolated by swim-up or Percoll procedures. All spermatozoal fractions used in this study were characterized by defining their motility, morphology and viability. The levels of ROS formation by non-activated as well as stimulated leucocytes were significantly decreased after addition of swim-up separated spermatozoa (p < 0.01). The ability to inhibit the basal chemiluminescence was of lower degree for spermatozoa isolated from 90% Percoll fractions than for swim-up sperm. However, addition of sperm cells from 47% Percoll fraction was found to increase both lucigenin and luminol signals. Moreover, the determined ROI levels changed depending on the type of inducing factor used for oxidative burst. Then, spermatozoa selected by swim-up procedure although with only slightly higher viability and morphology than sperm obtained from 90% Percoll fraction clearly exhibited much higher capacity to inhibit ROI secretion by receptor-stimulated leucocytes (FMLP-activation) than Percoll fractionated sperm. Such results may indicate that within normal semen may exist sperm subpopulations with different biochemical mechanisms controlling the interaction between spermatozoa and contaminating leucocytes. When ROI levels contained in normozoospermic semen are dependent on the WBCs activation, it seems that

  16. Report of the First International Workshop on Equine Leucocyte Antigens, Cambridge, UK, July 1991.

    PubMed

    Kydd, J; Antczak, D F; Allen, W R; Barbis, D; Butcher, G; Davis, W; Duffus, W P; Edington, N; Grünig, G; Holmes, M A

    1994-07-01

    The First International Workshop on Equine Leucocyte Antigens was organized and convened for the purposes of identifying immunologically relevant cell surface molecules of equine leucocytes and establishing a system of nomenclature for those molecules. Participating members of the workshop represented the majority of laboratories world-wide engaged in the tasks of production and characterization of equine leucocyte and lymphocyte markers using monoclonal antibodies. The workshop confirmed the identification of several equine CD molecules described previously by individual laboratories, and in addition recognized antibodies identifying new CD molecules. The workshop also succeeded in fostering co-operation between laboratories around the world which study equine immunobiology. Equine CD molecules identified by the current battery of monoclonal antibodies include EqCD2, EqCD4, EqCD5, EqCD8, EqCD11a/18, EqCD13 and EqCD44. Other antibodies are markers for MHC class I and class II molecules, for B cells, granulocytes, macrophages, T cell subsets distinct from those defined by CD4 and CD8, and other sub-populations of horse leucocytes that do not have obvious counterparts in humans, rodents, or other species. Despite the progress made in the first workshop, there are still substantial gaps in the armory of reagents available to study equine leucocyte biology, and further definition of the structure, function, and genetics of the antigens identified by the workshop clusters (WC1, WC2 etc.) and other molecules of immunological importance will be a goal of future workshops. The study of equine immunobiology and resistance to disease also urgently requires the development of tools to study equine immunoglobulins and cytokines, and these needs will provide ample scope for future studies.

  17. Differential leucocyte count for ewe milk with low and high somatic cell count.

    PubMed

    Albenzio, Marzia; Caroprese, Mariangela

    2011-02-01

    This study was undertaken to compare flow cytometry (FC) and direct microscopic leucocyte count (MDLC) for the differentiation of macrophages, lymphocytes and polymorphonuclear leucocyte (PMN) and to evaluate leucocyte distribution in ewe milk with low and high somatic cell count (SCC). Milk samples were grouped for somatic cell count in low SCC (LSCC) when the content was lower than 5·00 × 10(5)/ml and high SCC (HSCC) when the content was higher than 1·00 × 10(6)/ml. No differences were found between the two methods tested suggesting that FC could be used as a routine test for rapid discrimination of leucocytes. Percentages of lymphocytes in ewe milk were higher in LSCC (50%) than in HSCC (39%) and count ranged from 273·91 ± 56·62 × 10(3) cells/ml (LSCC) to 308·90 ± 46·15 × 10(3) cells/ml (HSCC). PMN number was lower in LSCC than in HSCC (248·83 ± 46·87 × 10(3) cells/ml v. 444·38 ± 58·62 × 10(3) cells/ml); accordingly the percentage was lower in LSCC (40%) than in HSCC (57%). No differences were found for macrophages which were 36·36 ± 5·51 × 10(3) cells/ml and 39·32 ± 6·83 × 10(3) cells/ml in LSCC and HSCC, respectively. Lymphocytes in ewe milk did not vary with increased number of somatic cells and were the predominant cell type in LSCC. PMN represented the main population detected in HSCC and the correlation with SCC evidenced that this leucocyte class could be useful in differentiating ewe milk cell count, being strictly responsible for the SCC increase.

  18. Clock gene expression in peripheral leucocytes of patients with type 2 diabetes.

    PubMed

    Ando, H; Takamura, T; Matsuzawa-Nagata, N; Shima, K R; Eto, T; Misu, H; Shiramoto, M; Tsuru, T; Irie, S; Fujimura, A; Kaneko, S

    2009-02-01

    Recent studies have demonstrated relationships between circadian clock function and the development of metabolic diseases such as type 2 diabetes. We investigated whether the peripheral circadian clock is impaired in patients with type 2 diabetes. Peripheral leucocytes were obtained from eight patients with diabetes and six comparatively young non-diabetic volunteers at 09:00, 15:00, 21:00 and 03:00 hours (study 1) and from 12 male patients with diabetes and 14 age-matched men at 09:00 hours (study 2). Transcript levels of clock genes (CLOCK, BMAL1 [also known as ARNTL], PER1, PER2, PER3 and CRY1) were determined by real-time quantitative PCR. In study 1, mRNA expression patterns of BMAL1, PER1, PER2 and PER3 exhibited 24 h rhythmicity in the leucocytes of all 14 individuals. The expression levels of these mRNAs were significantly (p < 0.05) lower in patients with diabetes than in non-diabetic individuals at one or more time points. Moreover, the amplitudes of mRNA expression rhythms of PER1 and PER3 genes tended to diminish in patients with diabetes. In study 2, leucocytes obtained from patients with diabetes expressed significantly (p < 0.05) lower transcript levels of BMAL1, PER1 and PER3 compared with leucocytes from control individuals, and transcript expression was inversely correlated with HbA(1c) levels (rho = -0.47 to -0.55, p < 0.05). These results suggest that rhythmic mRNA expression of clock genes is dampened in peripheral leucocytes of patients with type 2 diabetes. The impairment of the circadian clock appears to be closely associated with the pathophysiology of type 2 diabetes in humans.

  19. Signalling profiles of circulating leucocytes in patients recovered from reactive arthritis.

    PubMed

    Alanärä, T; Aittomäki, S; Kuuliala, K; Kuuliala, A; Siitonen, S; Leirisalo-Repo, M; Repo, H

    2012-08-01

    Reactive arthritis (ReA) is a sterile joint inflammation triggered by a remote infection and associated with human leucocyte antigen (HLA)-B27. Its pathogenesis is unknown, but abnormal response to microbial structures or endogenous inflammatory mediators may be involved. We studied responses in leucocyte signalling profiles in patients with previous ReA after a full recovery. The study comprised 10 HLA-B27-positive healthy subjects with a history of Yersinia enterocolitica-triggered ReA (B27+ReA+) and 20 healthy reference subjects, of whom 10 carried HLA-B27 (B27+ReA-) and 10 did not (B27-ReA-). Phosphospecific fluorescent monoclonal antibodies and flow cytometry were used to determine activation of nuclear factor kappa B (NF-κB), signal transducers and activators of transcription (STATs) 1, 3, 5, and 6, and two mitogen-activated protein (MAP) kinases, p38 and extracellular signal-regulated kinase (ERK)1/2, in monocytes, lymphocytes, lymphocyte subsets, and neutrophils. B27+ReA+ and B27-ReA- whole-blood samples were incubated with Yersinia with or without infliximab to study the role of tumour necrosis factor (TNF) in lymphocyte subset activation. Samples of the three subject groups were studied using soluble bacterial or endogenous stimuli. Fluorescence levels were determined as relative fluorescence units (RFU) and the proportion of positively fluorescing cells. The intracellular activation of circulating leucocytes in response to soluble stimuli was consistently comparable in B27+ReA+, B27+ReA-, and B27-ReA- subjects. Infliximab inhibited Yersinia-induced lymphocyte NF-κB phosphorylation similarly in B27+ReA+ and B27-ReA- groups. ReA susceptibility is not reflected in leucocyte signalling profiles elicited by phlogistic stimuli. However, the possibility remains that aberrations occur in response to combinations of stimuli, such as those associated with leucocyte adhesion.

  20. Decreased gene expression of LC3 in peripheral leucocytes of patients with coronary artery disease.

    PubMed

    Wu, Guanghua; Wei, Guanghe; Huang, Jian; Pang, Shuchao; Liu, Lixin; Yan, Bo

    2011-09-01

    Coronary artery disease (CAD) is a common and multifactorial arterial disease that is mainly caused by atherosclerosis. Macrophages, lymphocytes and neutrophils have been implicated in atherosclerotic plaque development. Autophagy, a highly conserved cellular process for the removal of long-lived protein and organelles, plays a variety of pathophysiological roles. However, the roles of autophagy in peripheral leucocytes in atherosclerosis and CAD have not been explored. LC3 is a marker gene for autophagy, and LC3-II, a conjugated form of LC3 protein, is a membrane marker for autophagosome and autophagolysosomes. In this study, LC3 gene expression levels and LC3-II protein levels in peripheral leucocytes were measured in patients with CAD (n = 146) and healthy controls (n = 87). In patients with CAD, LC3 gene expression levels in the peripheral leucocytes were significantly decreased compared with age- and sex-matched healthy controls (P < 0·01). LC3-II protein levels were also significantly decreased in patients with CAD (P < 0·01). Multivariate logistic analyses showed that decreased LC3 gene expression levels were strongly associated with CAD. There were no differences in LC3 transcripts and LC3-II protein levels between subgroups of patients with CAD. LC3 gene expression in the peripheral leucocytes was significantly decreased in patients with CAD, indicating that autophagosome formation is decreased. These data suggest that autophagy in circulating leucocytes may be involved in the pathogenesis of atherosclerosis and CAD. © 2011 The Authors. European Journal of Clinical Investigation © 2011 Stichting European Society for Clinical Investigation Journal Foundation.

  1. Perceptions of health managers and professionals about mental health and primary care integration in Rio de Janeiro: a mixed methods study.

    PubMed

    Athié, Karen; Menezes, Alice Lopes do Amaral; da Silva, Angela Machado; Campos, Monica; Delgado, Pedro Gabriel; Fortes, Sandra; Dowrick, Christopher

    2016-09-30

    Community-based primary mental health care is recommended in low and middle-income countries. The Brazilian Health System has been restructuring primary care by expanding its Family Health Strategy. Due to mental health problems, psychosocial vulnerability and accessibility, Matrix Support teams are being set up to broaden the professional scope of primary care. This paper aims to analyse the perceptions of health professionals and managers about the integration of primary care and mental health. In this mixed-method study 18 health managers and 24 professionals were interviewed from different primary and mental health care services in Rio de Janeiro. A semi-structured survey was conducted with 185 closed questions ranging from 1 to 5 and one open-ended question, to evaluate: access, gateway, trust, family focus, primary mental health interventions, mental health records, mental health problems, team collaboration, integration with community resources and primary mental health education. Two comparisons were made: health managers and professionals' (Mann-Whitney non-parametric test) and health managers' perceptions (Kruskall-Wallis non parametric-test) in 4 service designs (General Traditional Outpatients, Mental Health Specialised Outpatients, Psychosocial Community Centre and Family Health Strategy)(SPSS version 17.0). Qualitative data were subjected to Framework Analysis. Firstly, health managers and professionals' perceptions converged in all components, except the health record system. Secondly, managers' perceptions in traditional services contrasted with managers' perceptions in community-based services in components such as mental health interventions and team collaboration, and converged in gateway, trust, record system and primary mental health education. Qualitative data revealed an acceptance of mental health and primary care integration, but a lack of communication between institutions. The Mixed Method demonstrated that interviewees consider mental

  2. Connexin36 identified at morphologically mixed chemical/electrical synapses on trigeminal motoneurons and at primary afferent terminals on spinal cord neurons in adult mouse and rat.

    PubMed

    Bautista, W; McCrea, D A; Nagy, J I

    2014-03-28

    Morphologically mixed chemical/electrical synapses at axon terminals, with the electrical component formed by gap junctions, is common in the CNS of lower vertebrates. In mammalian CNS, evidence for morphologically mixed synapses has been obtained in only a few locations. Here, we used immunofluorescence approaches to examine the localization of the neuronally expressed gap junction forming protein connexin36 (Cx36) in relation to the axon terminal marker vesicular glutamate transporter-1 (vglut1) in the spinal cord and the trigeminal motor nucleus (Mo5) of rat and mouse. In adult rodents, immunolabeling for Cx36 appeared exclusively as Cx36-puncta, and was widely distributed at all rostro-caudal levels in most spinal cord laminae and in the Mo5. A high proportion of Cx36-puncta was co-localized with vglut1, forming morphologically mixed synapses on motoneurons, in intermediate spinal cord lamina, and in regions of medial lamina VII, where vglut1-containing terminals associated with Cx36 converged on neurons adjacent to the central canal. Unilateral transection of lumbar dorsal roots reduced immunolabeling of both vglut1 and Cx36 in intermediate laminae and lamina IX. Further, vglut1-terminals displaying Cx36-puncta were contacted by terminals labeled for glutamic acid decarboxylase65, which is known to be contained in presynaptic terminals on large-diameter primary afferents. Developmentally, mixed synapses begin to emerge in the spinal cord only after the second to third postnatal week and thereafter increase to adult levels. Our findings demonstrate that axon terminals of primary afferent origin form morphologically mixed synapses containing Cx36 in broadly distributed areas of adult rodent spinal cord and Mo5.

  3. The effects of ambient temperature and mixing time of glass ionomer cement material on the survival rate of proximal ART restorations in primary molars

    PubMed Central

    Kemoli, Arthur M

    2014-01-01

    Objective: Temperature fluctuations and material mixing times are likely to affect the consistency and integrity of the material mixture, and hence the restoration made out of it. The purpose of the present study was to determine the influence of the ambient temperature and the mixing time of glass ionomer cement (GIC) restorative material on the survival rate of proximal atraumatic restorative treatment (ART) restorations placed in primary molars. Materials and Methods: A total of 804 restorations were placed in the primary molars of 6-8-year-olds using the ART approach. The restorations were then followed for a period of 2 years and evaluated at given intervals. The data collected were analyzed using SPSS computer statistical program, and the results tested and compared using the Chi-square, Kaplan Meier survival analysis and Cox Proportional hazard statistical tests. Results: The cumulative survival rate of the restorations dropped from the initial 94.4% to 30.8% at the end of 2 years. The higher survival rate of the restorations was associated with the experienced operators and assistants when using the rubber dam isolation method. However, there was no statistically significant difference in the survival rate of the restorations when related to the room temperature and the mixing time of the GIC materials used in spite of the variations in the temperature recoded and the methods used in mixing the materials. Conclusion: The ambient temperature and mixing time of GIC did not have a significant effect on the survival of the proximal ART restorations. PMID:24808692

  4. Leg ischaemia before circulatory arrest alters brain leucocyte count and respiratory chain redox state.

    PubMed

    Yannopoulos, Fredrik S; Arvola, Oiva; Haapanen, Henri; Herajärvi, Johanna; Miinalainen, Ilkka; Jensen, Hanna; Kiviluoma, Kai; Juvonen, Tatu

    2014-03-01

    Remote ischaemic preconditioning and its neuroprotective abilities are currently under investigation and the method has shown significant effects in several small and large animal studies. In our previous studies, leucocyte filtration during cardiopulmonary bypass reduced cerebrocortical adherent leucocyte count and mitigated cerebral damage after hypothermic circulatory arrest (HCA) in piglets. This study aimed to obtain and assess direct visual data of leucocyte behaviour in cerebral vessels after hypothermic circulatory arrest following remote ischaemic preconditioning. Twelve native stock piglets were randomized into a remote ischaemic preconditioning group (n = 6) and a control group (n = 6). The intervention group underwent hind-leg ischaemia, whereas the control group received a sham-treatment before a 60-min period of hypothermic circulatory arrest. An intravital microscope was used to obtain measurements from the cerebrocortical vessel in vivo. It included three sets of filters: a violet filter to visualize microvascular perfusion and vessel diameter, a green filter for visualization of rhodamine-labelled leucocytes and an ultraviolet filter for reduced nicotinamide adenine dinucleotide (NADH) analysis. The final magnification on the microscope was 400. After the experiment, cerebral and cerebellar biopsies were collected and analysed with transmission electron microscope by a blinded analyst. In the transmission electron microscope analysis, the entire intervention group had normal, unaffected rough endoplasmic reticulum's in their cerebellar tissue, whereas the control group had a mean score of 1.06 (standard deviation 0.41) (P = 0.026). The measured amount of adherent leucocytes was lower in the remote ischaemic preconditioning group. The difference was statistically significant at 5, 15 and 45 min after circulatory arrest. Statistically significant differences were seen also in the recovery phase at 90 and 120 min after reperfusion. Nicotinamide

  5. Leg ischaemia before circulatory arrest alters brain leucocyte count and respiratory chain redox state

    PubMed Central

    Yannopoulos, Fredrik S.; Arvola, Oiva; Haapanen, Henri; Herajärvi, Johanna; Miinalainen, Ilkka; Jensen, Hanna; Kiviluoma, Kai; Juvonen, Tatu

    2014-01-01

    OBJECTIVES Remote ischaemic preconditioning and its neuroprotective abilities are currently under investigation and the method has shown significant effects in several small and large animal studies. In our previous studies, leucocyte filtration during cardiopulmonary bypass reduced cerebrocortical adherent leucocyte count and mitigated cerebral damage after hypothermic circulatory arrest (HCA) in piglets. This study aimed to obtain and assess direct visual data of leucocyte behaviour in cerebral vessels after hypothermic circulatory arrest following remote ischaemic preconditioning. METHODS Twelve native stock piglets were randomized into a remote ischaemic preconditioning group (n = 6) and a control group (n = 6). The intervention group underwent hind-leg ischaemia, whereas the control group received a sham-treatment before a 60-min period of hypothermic circulatory arrest. An intravital microscope was used to obtain measurements from the cerebrocortical vessel in vivo. It included three sets of filters: a violet filter to visualize microvascular perfusion and vessel diameter, a green filter for visualization of rhodamine-labelled leucocytes and an ultraviolet filter for reduced nicotinamide adenine dinucleotide (NADH) analysis. The final magnification on the microscope was 400. After the experiment, cerebral and cerebellar biopsies were collected and analysed with transmission electron microscope by a blinded analyst. RESULTS In the transmission electron microscope analysis, the entire intervention group had normal, unaffected rough endoplasmic reticulum's in their cerebellar tissue, whereas the control group had a mean score of 1.06 (standard deviation 0.41) (P = 0.026). The measured amount of adherent leucocytes was lower in the remote ischaemic preconditioning group. The difference was statistically significant at 5, 15 and 45 min after circulatory arrest. Statistically significant differences were seen also in the recovery phase at 90 and 120 min after

  6. Comparison of the antibacterial effect of modified 3-mix paste versus Ultrapex over anaerobic microorganisms from infected root canals of primary teeth: an in vitro study.

    PubMed

    Velasco-Loera, N; De Alba-Vazquez, Y; Garrocho-Rangel, A; Gonzalez-Amaro, A M; Flores-Reyes, H; Pozos-Guillen, A J

    2012-01-01

    The aim of this study was to evaluate in vitro the antimicrobial efficacy of a modified 3-mix paste and to compare it with an iodoform paste (Ultrapex) against anaerobic microorganisms isolated from root canals of infected or necrotic primary teeth. An in vitro experimental assay was performed over isolated and identified anaerobic microorganisms of 21 samples, in order to compare the antimicrobial ability of both root canal filling materials, using a disc-diffusion method. A total of 21 microbial samples (15 polymicrobial and 6 monomicrobial) were obtained, from which 19 different strains were identified. Modified 3-mix paste showed an excellent antimicrobial effect against most of both kinds of microbial samples, although some of them exhibited resistance; on the other hand, Ultrapex showed only minimal antimicrobial ability (null or low categories). Clostridium ramosum exhibited the most resistance to both materials. The bactericidal effect of the modified 3-mix paste was superior to Ultrapex, with a statistically significant difference, against anaerobic microorganisms isolated from infected root canals of primary teeth.

  7. Allergic women show reduced T helper type 1 alloresponses to fetal human leucocyte antigen mismatch during pregnancy

    PubMed Central

    Prescott, S L; Breckler, L A; Witt, C S; Smith, L; Dunstan, J A; Christiansen, F T

    2010-01-01

    Low-level alloreactivity between mother and fetus may provide stimulation for fetal T helper type 1 (Th1) cell immune maturation. This study explored the effects of human leucocyte antigen (HLA) mismatch on materno–fetal interactions detected as cytokine responses and lymphoproliferation in mixed lymphocyte reactions, and whether this was altered in allergic women (n = 62) who have a Th2 propensity compared with non-allergic women (n = 65). HLA-DRβ1 mismatch was associated with significantly increased Th1 interferon (IFN)-γ, Th2 interleukin (IL)-13 and lymphoproliferative responses by both mothers and fetuses. Allergic women showed significantly lower IFN-γ Th1 production in response to HLA-DRβ1 mismatch. The infants of these women also showed significantly lower IL-10 and lower IFN-γ production relative to IL-13. Both HLA-DRβ1 mismatch and maternal allergy had significant independent effects on maternal IFN-γ Th1 responses. Maternal allergy modifies HLA-mediated alloreactivity between the mother and the fetus, reducing Th1 activation. This may affect the cytokine milieu at the materno–fetal interface and could be implicated in the attenuated Th1 responses observed commonly in infants of atopic mothers. PMID:19860744

  8. The Assessment of Math Learning Difficulties in a Primary Grade-4 Child with High Support Needs: Mixed Methods Approach

    ERIC Educational Resources Information Center

    Mundia, Lawrence

    2012-01-01

    This mixed-methods study incorporated elements of survey, case study and action research approaches in investigating an at-risk child. Using an in-take interview, a diagnostic test, an error analysis, and a think-aloud clinical interview, the study identified the child's major presenting difficulties. These included: inability to use the four…

  9. Factors associated with the utilization of primary care emergency centers in a Spanish region with high population dispersion: a mixed-methods study.

    PubMed

    Sanz-Barbero, Belén; Otero-García, Laura; Blasco-Hernández, Teresa; San Sebastián, Miguel

    2014-09-03

    Adequate access to primary care emergency centers is particularly important in rural areas isolated from urban centers. However, variability in utilization of emergency services located in primary care centers among inhabitants of nearby geographical areas is understudied. The objectives of this study are twofold: 1) to analyze the association between the availability of municipal emergency care centers and utilization of primary care emergency centers (PCEC), in a Spanish region with high population dispersion; and 2) to determine healthcare providers' perceptions regarding PCEC utilization. A mixed-methods study was conducted. Quantitative phase: multilevel logistic regression modeling using merged data from the 2003 Regional Health Survey of Castile and Leon and the 2001 census data (Spain). Qualitative phase:14 in-depth- interviews of rural-based PCEC providers. Having PCEC as the only emergency center in the municipality was directly associated with its utilization (p < 0.001). Healthcare providers perceived that distance to hospital increased PCEC utilization, and distance to PCEC decrease its use. PCEC users were considered to be predominantly workers and students with scheduling conflicts with rural primary care opening hours. The location of emergency care centers is associated with PCEC utilization. Increasing access to primary care by extending hours may be an important step toward optimal PCEC utilization. Further research would determine whether lower PCEC use by certain groups is associated with disparities in access to care.

  10. /sup 111/Indium leucocyte scanning in ampicillin-associated right-sided hemorrhagic colitis

    SciTech Connect

    Keshavarzian, A.; Saverymuttu, S.H.; Chadwick, V.S.

    1984-07-01

    Hemorrhagic colitis is a rare but well-recognized complication with ampicillin or penicillin derivative treatment. Early colonoscopy has been advocated in establishing the diagnosis by demonstrating the characteristic pattern of only right-sided involvement and so distinguishing it from other colitides. The authors report a patient who developed colitis after amoxycillin therapy in whom /sup 111/Indium leucocyte scan demonstrated right-sided colitis which alerted them to the diagnosis. Discontinuation of the antibiotic resulting in rapid improvement, and return of the /sup 111/Indium leucocyte scan to normal in this patient suggests that ampicillin-associated colitis should not be considered purely as a hemorrhagic disease but may in some cases have an inflammatory component.

  11. Characterization of swine leucocyte antigen alleles in a crossbred pig to be used in xenotransplant studies.

    PubMed

    Reyes, L M; Blosser, R J; Smith, R F; Miner, A C; Paris, L L; Blankenship, R L; Tector, M F; Tector, A J

    2014-11-01

    We have characterized swine leucocyte antigen (SLA) classes I and II molecules of a domestic pig as a model for use in our xenotransplant program. Molecular characterization of the SLA classes I and II genes is critical to understanding the adaptive immune responses between swine and humans in the event of xenotransplantation. Seven swine leucocyte antigen genes (SLA-1, SLA-2, SLA-3, DQB1, DRB1, DQA and DRA) were analyzed and 15 alleles were identified. A novel DRA*w04re01 is reported for this limited polymorphic class II gene. The heterozygous haplotypes, Hp-32.0/35.0 and Hp-0.13/0.23 were deduced for our IU-pig model, for SLA classes I and II regions, respectively. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  12. Effect of hydrocortisone on the mobilisation of leucocytes in corneal wounds.

    PubMed Central

    Basu, P K; Avaria, M; Jankie, R

    1981-01-01

    We have studied in rabbits the effect of subconjunctivally injected hydrocortisone on the polymorphonuclear leucocyte invasion of corneal wounds at different times after an injury. One group of rabbits was treated with the steroid (hydrocortisone group) and the other not (control group). After making nonpenetrating trephine incisions on the cornea we obtained cellular samples by the impression technique at a given postoperative period (2, 4, or 6 hours), and then the animal was killed. The cornea was processed for histological study of the infiltrating cells. At any postoperative period the number of polymorphonuclear leucocytes in the corneal wounds of the hydrocortisone group was significantly less than the number in the identical wounds of the control group (p less than 0.01 to 0.001). Images PMID:7317321

  13. Initial experience with indium-111 autologous leucocyte imaging in patients with acute pancreatitis.

    PubMed Central

    Anderson, J R; Spence, R A; Laird, J D; Ferguson, W R; Kennedy, T L

    1983-01-01

    Indium-111 labelled autologous leucocyte imaging was used to assess severity in 13 patients with acute pancreatitis. All three patients with severe disease as judged by a prognostic factor grading system had a positive result on imaging. A fourth patient with mild disease as judged by prognostic factors had a positive imaging result and 14 days later developed a pseudocyst. There were no false positive or false negative scans. Evidence from three patients suggested that a positive 111In-leucocyte imaging result implies substantial fat necrosis. In this study imaging was as accurate as prognostic factor grading. The technique may be a useful method of separating mild from severe acute pancreatitis. Images FIG 1 FIG 2 PMID:6411262

  14. Hepatitis B virus (HBV) DNA in leucocytes in acquired immune deficiency syndrome (AIDS).

    PubMed

    Lie-Injo, L E; Volberding, P; Golden, J A; Herrera, A R

    1985-01-01

    Earlier reported findings of hepatitis B virus (HBV) DNA in white blood cells of patients with hepatoma, and in a patient with autoimmune haemolytic anaemia, led to the examination of HBV DNA in a series of twenty three patients with acquired immune deficiency syndrome (AIDS), including nine with opportunistic infections and fourteen with Kaposi's sarcoma, by Southern blot hybridization method, using 32P labelled HBV DNA specific probe obtained by nick translation of HBV DNA cloned into plasmid pBR325. Four of the patients were found to be positive for HBV DNA or HBV related DNA in their leucocytes. The HBV DNA was found free or integrated in the leucocytes of the patients.

  15. The role of PMN-leucocyte lysosomes in tissue injury, inflammation and hypersensitivity

    PubMed Central

    Movat, H. Z.; Uriuhara, T.; Taichman, N. S.; Rowsell, H. C.; Mustard, J. F.

    1968-01-01

    Anaphylaxis due to intravascular interaction of hyperimmune antibody with antigen was studied in rabbits, swine and rats. Obstruction of the pulmonary vessels by the immune precipitates was found to initiate the process. This is followed by aggregation of PMN-leucocytes and platelets in pulmonary vessels and phagocytosis of the precipitates by these blood elements. During this process degranulation of the cells takes place with release of lysosomal contents. As a concomitant a rise in plasma acid protease and other hydrolases was demonstrated, presumably derived from the degranulating PMN-leucocytes and platelets. Unlike leukopaenic animals, normal ones showed a more marked hypotension, a greater tendency to protracted shock and developed focal and confluent haemorrhagic pulmonary lesions. It is suggested that anaphylaxis due to intravascular antigen—antibody interaction or aggregate anaphylaxis is a systemic or pulmonary Arthus reaction, rather than a `true' anaphylaxis. ImagesFIG. 5FIG. 6FIG. 3FIG. 4 PMID:4231971

  16. Leucocyte migration inhibition response to tissue antigens in asymptomatic individuals infected with Trypanosoma cruzi.

    PubMed Central

    Peralta, J M; Gill, K; Cordeiro Lima, M F; Coura, J R

    1981-01-01

    The direct leucocyte migration inhibition test was used to study 31 asymptomatic humans chronically infected with Trypanosoma cruzi and 23 normal uninfected controls. The antigenic preparations used were made from mouse and guinea-pig heart, skeletal muscle, kidney, liver and brain. Positive responses were found in the parasite-infected individuals to kidney, liver and brain antigen but not to antigen prepared from heart of skeletal muscle tissue. No correlation was found between T. cruzi antibody titres and migration index values to these various antigens. On the other hand, a positive correlation was only noted between the titres of tissue-reacting immunoglobulins and the migration indices induced by brain antigens: when titres of tissue-reacting immunoglobulins were elevated, less leucocyte migration inhibition was detected. PMID:6802533

  17. Mechanisms controlling primary and new production in a global ecosystem model Part I: The role of the large-scale upper mixed layer variability

    NASA Astrophysics Data System (ADS)

    Popova, E. E.; Coward, A. C.; Nurser, G. A.; de Cuevas, B.; Fasham, M. J. R.; Anderson, T. R.

    2006-07-01

    A global general circulation model coupled to a simple six-compartment ecosystem model is used to study the extent to which global variability in primary and export production can be realistically predicted on the basis of advanced parameterizations of upper mixed layer physics, without recourse to introducing extra complexity in model biology. The ''K profile parameterization'' (KPP) scheme employed, combined with 6-hourly external forcing, is able to capture short-term periodic and episodic events such as diurnal cycling and storm-induced deepening. The model realistically reproduces various features of global ecosystem dynamics that have been problematic in previous global modelling studies, using a single generic parameter set. The realistic simulation of deep convection in the North Atlantic, and lack of it in the North Pacific and Southern Oceans, leads to good predictions of chlorophyll and primary production in these contrasting areas. Realistic levels of primary production are predicted in the oligotrophic gyres due to high frequency external forcing of the upper mixed layer (accompanying paper Popova et al., 2006) and novel parameterizations of zooplankton excretion. Good agreement is shown between model and observations at various JFOFS time series sites: BATS, KERFIX, Papa and station India. One exception is that the high zooplankton grazing rates required to maintain low chlorophyll in high-nutrient low-chlorophyll and oligotrophic systems lessened agreement between model and data in the northern North Atlantic, where mesozooplankton with lower grazing rates may be dominant. The model is therefore not globally robust in the sense that additional parameterizations were needed to realistically simulate ecosystem dynamics in the North Atlantic. Nevertheless, the work emphasises the need to pay particular attention to the parameterization of mixed layer physics in global ocean ecosystem modelling as a prerequisite to increasing the complexity of ecosystem

  18. Opsonic activity of anti-flagellar serum against Clostridium chauvoei by mouse polymorphonuclear leucocytes.

    PubMed

    Tamura, Y; Tanaka, M

    1987-05-01

    The role of anti-flagellar serum against Clostridium chauvoei in phagocytosis by mouse polymorphonuclear leucocytes was examined. Anti-flagellar serum markedly increased phagocytic rate against the flagellated strain Okinawa but not against a non-flagellated mutant (NFM) derived from the same strain, while anti-NFM serum increased the phagocytic rate against both strains. These results indicate that anti-flagellar serum exerts its protective effect by opsonic activity.

  19. ILK mediates LPS-induced vascular adhesion receptor expression and subsequent leucocyte trans-endothelial migration.

    PubMed

    Hortelano, Sonsoles; López-Fontal, Raquel; Través, Paqui G; Villa, Natividad; Grashoff, Carsten; Boscá, Lisardo; Luque, Alfonso

    2010-05-01

    The inflammatory response to injurious agents is tightly regulated to avoid adverse consequences of inappropriate leucocyte accumulation or failed resolution. Lipopolysaccharide (LPS)-activated endothelium recruits leucocytes to the inflamed tissue through controlled expression of membrane-associated adhesion molecules. LPS responses in macrophages are known to be regulated by integrin-linked kinase (ILK); in this study, we investigated the role of ILK in the regulation of the LPS-elicited inflammatory response in endothelium. This study was performed on immortalized mouse endothelial cells (EC) isolated from lung and coronary vasculature. Cells were thoroughly characterized and the role of ILK in the regulation of the LPS response was investigated by suppressing ILK expression using siRNA and shRNA technologies. Phenotypic and functional analyses confirmed that the immortalized cells behaved as true EC. LPS induced the expression of the inflammatory genes E-selectin, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). ILK knockdown impaired LPS-mediated endothelial activation by preventing the induction of ICAM-1 and VCAM-1. Blockade of the LPS-induced response inhibited the inflammatory-related processes of firm adhesion and trans-endothelial migration of leucocytes. ILK is involved in the expression of cell adhesion molecules by EC activated with the inflammatory stimulus LPS. This reduced expression modulates leucocyte adhesion to the endothelium and the extravasation process. This finding suggests ILK as a potential anti-inflammatory target for the development of vascular-specific treatments for inflammation-related diseases.

  20. Increased collagenase and dipeptidyl peptidase I activity in leucocytes from healthy elderly people

    PubMed Central

    Llorente, L; Richaud-Patin, Y; Díaz-Borjón, A; Jakez-Ocampo, J; Alvarado-De La Barrera, C

    1999-01-01

    The incidence of infectious diseases increases with ageing. The enzymatic activity of leucocytes may have a relevant role in the morbidity and mortality due to infections in the elderly. In this study we have compared the activity of enzymes involved in the inflammatory response in leucocytes from young and elderly women. A total of 35 healthy females was studied, 20 volunteers aged 78–98 years (mean 89.1 years) and 15 young controls aged 19–34 years (mean 26 years). All of them were in good clinical condition, without any acute or chronic disease. Intracellular enzyme activity was analysed by flow cytometry in leucocytes from young and elderly women. The enzyme substrates employed were for oxidative burst, l-aminopeptidase, collagenase, cathepsin B, C, D and, G and dipeptidyl peptidase I. The intracellular enzyme activity assessed by flow cytometry in leucocytes from young and elderly women was similar, as far as oxidative burst, l-aminopeptidase, cathepsin B, C, D and G are concerned. An increased collagenase activity was detected in granulocytes from elders. The mean fluorescence channels for this enzyme corresponded to 86 ± 23 and 60 ± 15 in cells from elders and controls, respectively (P = 0.01224). An increased dipeptidyl peptidase I activity was detected in lymphocytes from elderly women. The corresponding values for this enzyme in elders and the young were 65.9 ± 43.3 and 17.3 ± 5, respectively (P = 0.0036). The proper functional activity of intracellular enzymes involved in inflammatory responses is likely to be determinant for successful ageing. PMID:10361229

  1. Specific characteristics of peritoneal leucocyte populations during sterile peritonitis associated with icodextrin CAPD fluids.

    PubMed

    Glorieux, Griet; Lameire, Norbert; Van Biesen, Wim; Dequidt, Clement; Vanholder, Raymond

    2003-08-01

    Icodextrin dialysate used for peritoneal dialysis contains an iso-molar glucose polymer solution, which provides sustained ultrafiltration over long dwell times and is considered a valuable approach to reduce intraperitoneal glucose exposure. However, several side effects have been described, including abdominal pain and allergic and hypersensitivity reactions. Also, reactions compatible with chemical peritonitis have been reported. Over the period of a few months (January 2002-May 2002), a remarkable increase in the number of continuous ambulatory peritoneal dialysis (CAPD) patients using icodextrin dialysate diagnosed with sterile peritonitis was observed in our unit. Five of the CAPD patients using icodextrin dialysate in our unit and diagnosed with sterile peritonitis were screened for leucocyte count and leucocyte differentiation during a follow-up period of 77 +/- 23 days. In addition, expression of CD14, a receptor for lipopolysaccharide (LPS), on the peripheral and peritoneal monocyte population was analysed. These results were compared to CAPD patients suffering from bacterial peritonitis. The peritoneal leucocyte count of CAPD patients using icodextrin dialysate and diagnosed with sterile peritonitis did not decrease significantly before treatment with icodextrin dialysate was interrupted, whereas it currently disappeared within 2-4 days in proven bacterial peritonitis. The sterile, cloudy icodextrin effluent contained an excess of macrophages on the day of diagnosis, whereas in bacterial peritonitis essentially an increase in the granulocyte population was observed. No elevation in the eosinophil population was observed. In contrast to bacterial peritonitis, we observed no increase in CD14 expression on the peripheral and peritoneal macrophages on the day of presentation and during the follow-up period. Specific batches of the icodextrin CAPD fluids contain a macrophage chemotactic agent, which causes a sustained inflammatory state in the peritoneal

  2. Association of proinflammatory cytokines and islet resident leucocytes with islet dysfunction in type 2 diabetes.

    PubMed

    Butcher, Matthew J; Hallinger, Daniel; Garcia, Eden; Machida, Yui; Chakrabarti, Swarup; Nadler, Jerry; Galkina, Elena V; Imai, Yumi

    2014-03-01

    Chronic inflammation in type 2 diabetes is proposed to affect islets as well as insulin target organs. However, the nature of islet inflammation and its effects on islet function in type 2 diabetes remain unclear. Moreover, the immune cell profiles of human islets in healthy and type 2 diabetic conditions are undefined. We aimed to investigate the correlation between proinflammatory cytokine expression, islet leucocyte composition and insulin secretion in type 2 diabetic human islets. Human islets from organ donors with or without type 2 diabetes were studied. First and second phases of glucose-stimulated insulin secretion were determined by perifusion. The expression of inflammatory markers was obtained by quantitative PCR. Immune cells within human islets were analysed by FACS. Type 2 diabetic islets, especially those without first-phase insulin secretion, displayed higher CCL2 and TNFa expression than healthy islets. CD45(+) leucocytes were elevated in type 2 diabetic islets, to a greater extent in moderately functional type 2 diabetic islets compared with poorly functional ones, and corresponded with elevated ALOX12 but not with CCL2 or TNFa expression. T and B lymphocytes and CD11c(+) cells were detectable within both non-diabetic and type 2 diabetic islet leucocytes. Importantly, the proportion of B cells was significantly elevated within type 2 diabetic islets. Elevated total islet leucocyte content and proinflammatory mediators correlated with islet dysfunction, suggesting that heterogeneous insulitis occurs during the development of islet dysfunction in type 2 diabetes. In addition, the altered B cell content highlights a potential role for the adaptive immune response in islet dysfunction.

  3. Association of proinflammatory cytokines and islet resident leucocytes with islet dysfunction in type 2 diabetes

    PubMed Central

    Butcher, Matthew J.; Hallinger, Daniel; Garcia, Eden; Machida, Yui; Chakrabarti, Swarup; Nadler, Jerry; Galkina, Elena V.; Imai, Yumi

    2014-01-01

    Aims/hypothesis Chronic inflammation in type 2 diabetes is proposed to affect islets as well as insulin target organs. However, the nature of islet inflammation and its effects on islet function in type 2 diabetes remain unclear. Moreover, the immune cell profiles of human islets in healthy and type 2 diabetic conditions are undefined. We aimed to investigate the correlation between proinflammatory cytokine expression, islet leucocyte composition and insulin secretion in type 2 diabetic human islets. Methods Human islets from organ donors with or without type 2 diabetes were studied. First and second phases of glucose-stimulated insulin secretion were determined by perifusion. The expression of inflammatory markers was obtained by quantitative PCR. Immune cells within human islets were analysed by FACS. Results Type 2 diabetic islets, especially those without first-phase insulin secretion, displayed higher CCL2 and TNFa expression than healthy islets. CD45+ leucocytes were elevated in type 2 diabetic islets, to a greater extent in moderately functional type 2 diabetic islets compared with poorly functional ones, and corresponded with elevated ALOX12 but not with CCL2 or TNFa expression. T and B lymphocytes and CD11c+ cells were detectable within both non-diabetic and type 2 diabetic islet leucocytes. Importantly, the proportion of B cells was significantly elevated within type 2 diabetic islets. Conclusions/interpretation Elevated total islet leucocyte content and proinflammatory mediators correlated with islet dysfunction, suggesting that heterogeneous insulitis occurs during the development of islet dysfunction in type 2 diabetes. In addition, the altered B cell content highlights a potential role for the adaptive immune response in islet dysfunction. PMID:24429578

  4. Social and environmental factors modulate leucocyte profiles in free-living Greylag geese (Anser anser)

    PubMed Central

    Ludwig, Sonja C.; Hemetsberger, Josef; Kotrschal, Kurt; Wascher, Claudia A.F.

    2017-01-01

    Background Blood parameters such as haematocrit or leucocyte counts are indicators of immune status and health, which can be affected, in a complex way, by exogenous as well as endogenous factors. Additionally, social context is known to be among the most potent stressors in group living individuals, therefore potentially influencing haematological parameters. However, with few exceptions, this potential causal relationship received only moderate scientific attention. Methods In a free-living and individually marked population of the highly social and long-lived Greylag goose, Anser anser, we relate variation in haematocrit (HCT), heterophils to lymphocytes ratio (H/L) and blood leucocyte counts to the following factors: intrinsic (sex, age, raising condition, i.e. goose- or hand-raised), social (pair-bond status, pair-bond duration and parental experience) and environmental (biologically relevant periods, ambient temperature) factors. Blood samples were collected repeatedly from a total of 105 focal birds during three biologically relevant seasons (winter flock, mating season, summer). Results We found significant relationships between haematological parameters and social as well as environmental factors. During the mating season, unpaired individuals had higher HCT compared to paired and family individuals and this pattern reversed in fall. Similarly, H/L ratio was positively related to pair-bond status in a seasonally dependent way, with highest values during mating and successful pairs had higher H/L ratio than unsuccessful ones. Also, absolute number of leucocytes tended to vary depending on raising condition in a seasonally dependent way. Discussion Haematology bears a great potential in ecological and behavioural studies on wild vertebrates. In sum, we found that HTC, H/L ratio and absolute number of leucocytes are modulated by social factors and conclude that they may be considered valid indicators of individual stress load. PMID:28070455

  5. Chemotactic activity of Helicobacter pylori sonicate for human polymorphonuclear leucocytes and monocytes.

    PubMed Central

    Nielsen, H; Andersen, L P

    1992-01-01

    The immunopathology of Helicobacter pylori associated active chronic gastritis, which is characterised by predominance of polymorphonuclear leucocyte infiltration, is largely unknown. To evaluate the role of bacterial components as inflammatory mediators ultracentrifuged sonicated preparations were made of clinical isolates of Helicobacter pylori. The crude sonicates were shown to exhibit chemotactic activity for human polymorphonuclear leucocytes and blood monocytes in a concentration dependent fashion. The potency was comparable with previously described bacterial derived cytotaxins. The cytotaxin(s) was non-dialysable and completely destroyed by proteinase. Heat treatment did not decrease the chemotactic activity, but in sonicate subjected to 100 degrees C for 15 minutes all activity disappeared after dialysis suggesting the breakdown of a larger protein to small fragments that are still biological active. By ammonium sulphate precipitation at increasing concentrations the cytotaxin(s) was selectively found in 10% ammonium sulphate saturation, and by further molecular gel separation the chemotactic activity was found in the molecular size range from 25 to 35 kDa. The demonstration of a polymorphonuclear leucocyte and monocyte cytotaxin from Helicobacter pylori sonicate may help in understanding the mucosal immune response in gastric inflammatory diseases. PMID:1624151

  6. Leucocyte profiles of Arctic marine birds: correlates of migration and breeding phenology

    PubMed Central

    Mallory, Mark L.; Little, Catherine M.; Boyd, Ellen S.; Ballard, Jennifer; Elliott, Kyle H.; Gilchrist, H. Grant; Hipfner, J. Mark; Petersen, Aevar; Shutler, Dave

    2015-01-01

    Most Arctic marine birds are migratory, wintering south of the limit of annual pack ice and returning north each year for the physiologically stressful breeding season. The Arctic environment is changing rapidly due to global warming and anthropogenic activities, which may influence the timing of breeding in relation to arrival times following migration, as well as providing additional stressors (e.g. disturbance from ships) to which birds may respond. During stressful parts of their annual cycle, such as breeding, birds may reallocate resources so that they have increased heterophil-to-lymphocyte ratios in their white blood cell (leucocyte) profiles. We analysed leucocyte profiles of nine species of marine birds to establish reference ranges for these species in advance of future Arctic change. Leucocyte profiles tended to cluster among taxonomic groups across studies, suggesting that reference values for a particular group can be established, and within species there was evidence that birds from colonies that had to migrate farther had higher heterophil-to-lymphocyte ratios during incubation than those that did not have to travel as far, particularly for species with high wing loading. PMID:27293713

  7. Exposure to anthrax toxin alters human leucocyte expression of anthrax toxin receptor 1.

    PubMed

    Ingram, R J; Harris, A; Ascough, S; Metan, G; Doganay, M; Ballie, L; Williamson, E D; Dyson, H; Robinson, J H; Sriskandan, S; Altmann, D M

    2013-07-01

    Anthrax is a toxin-mediated disease, the lethal effects of which are initiated by the binding of protective antigen (PA) with one of three reported cell surface toxin receptors (ANTXR). Receptor binding has been shown to influence host susceptibility to the toxins. Despite this crucial role for ANTXR in the outcome of disease, and the reported immunomodulatory consequence of the anthrax toxins during infection, little is known about ANTXR expression on human leucocytes. We characterized the expression levels of ANTXR1 (TEM8) on human leucocytes using flow cytometry. In order to assess the effect of prior toxin exposure on ANTXR1 expression levels, leucocytes from individuals with no known exposure, those exposed to toxin through vaccination and convalescent individuals were analysed. Donors could be defined as either 'low' or 'high' expressers based on the percentage of ANTXR1-positive monocytes detected. Previous exposure to toxins appears to modulate ANTXR1 expression, exposure through active infection being associated with lower receptor expression. A significant correlation between low receptor expression and high anthrax toxin-specific interferon (IFN)-γ responses was observed in previously infected individuals. We propose that there is an attenuation of ANTXR1 expression post-infection which may be a protective mechanism that has evolved to prevent reinfection.

  8. Leucocyte profiles of Arctic marine birds: correlates of migration and breeding phenology.

    PubMed

    Mallory, Mark L; Little, Catherine M; Boyd, Ellen S; Ballard, Jennifer; Elliott, Kyle H; Gilchrist, H Grant; Hipfner, J Mark; Petersen, Aevar; Shutler, Dave

    2015-01-01

    Most Arctic marine birds are migratory, wintering south of the limit of annual pack ice and returning north each year for the physiologically stressful breeding season. The Arctic environment is changing rapidly due to global warming and anthropogenic activities, which may influence the timing of breeding in relation to arrival times following migration, as well as providing additional stressors (e.g. disturbance from ships) to which birds may respond. During stressful parts of their annual cycle, such as breeding, birds may reallocate resources so that they have increased heterophil-to-lymphocyte ratios in their white blood cell (leucocyte) profiles. We analysed leucocyte profiles of nine species of marine birds to establish reference ranges for these species in advance of future Arctic change. Leucocyte profiles tended to cluster among taxonomic groups across studies, suggesting that reference values for a particular group can be established, and within species there was evidence that birds from colonies that had to migrate farther had higher heterophil-to-lymphocyte ratios during incubation than those that did not have to travel as far, particularly for species with high wing loading.

  9. Leucocyte populations in semen and male accessory gland function: relationship with antisperm antibodies and seminal quality.

    PubMed

    Kortebani, G; Gonzales, G F; Barrera, C; Mazzolli, A B

    1992-01-01

    Semen samples from 279 infertility patients attending an Immunological Centre were analysed to evaluate the relationship between the populations of leucocytes, seminal quality, antisperm antibodies, and seminal vesicle function. The most frequent finding between leucocytospermic samples was asthenozoospermia (57%), whereas in non-leucocytospermic samples normozoospermia was the most frequent finding (47%). In the samples with asthenozoospermia, granulocytes predominated, whereas in those with oligozoospermia and azoospermia a reduction in the number of macrophages and lymphocytes was observed, suggesting an obstructive process at the level of epididymis and/or vas deferens where these leucocytes are mostly produced. In the case of hypofunction of the seminal vesicles there was a predominance in granulocytes. The increased levels of each type of leucocytes affected seminal quality only when seminal vesicles were affected. Only the elevated granulocytes count was related to a decrease in sperm motility. In those samples with leucocytospermia, positive antisperm antibodies (ASA) were associated with low sperm motility, low sperm normal morphology, and low value of seminal corrected fructose, whereas, in the absence of leucocytospermia, ASA, were more related to low sperm counts. These data suggest that granulocytes were more related to seminal vesicles dysfunction and sperm motility changes, and that ASA may be observed in the presence or absence of leucocytospermia.

  10. Microcystin-LR modulates selected immune parameters and induces necrosis/apoptosis of carp leucocytes.

    PubMed

    Rymuszka, Anna; Sierosławska, Anna; Bownik, Adam; Skowroński, Tadeusz

    2010-03-01

    Microcystins (MCs) are potent hepatotoxins acting by the inhibition of protein phosphatase 1 and 2A, and may promote liver tumors. Moreover, studies also suggest they are nephrotoxic. The aim of the present study was to assess possible in vitro effects of microcystin-LR (which contains the amino acids leucine and arginine, the most widely studied and distributed variant of all microcystins) on the selected immune functions of the cells isolated from the head kidney of carp. In the experiments, pure microcystin-LR (MC-LR), was used at concentrations of 0.01, 0.1, 0.5, and 1 microg/ml RPMI-1640 medium. Leucocytes (lymphocytes and phagocytes) were isolated by centrifugation on a density gradient. Lymphocyte proliferation, intracellular production of reactive oxygen species by phagocytes, and the presence of apoptotic and/or necrotic cells were assessed. The respiratory burst activity of phagocytic cells was increased at the lowest toxin concentration used in the study, but it was decreased at higher concentrations. Using a sensitive luminescent immunoassay, MC-LR was observed to have no influence on the T-cell proliferation but decreased the proliferation of B lymphocytes. Moreover, it was noted that MC-LR induced necrosis to a higher degree than apoptosis in fish leucocytes. The results of the present study suggest the modulatory potency of microcystin-LR on fish leucocytes.

  11. Role of Siglecs on the leucocytes during the process of the joint's inflammation in rheumatoid arthritis.

    PubMed

    Yang, Xiao; Wang, Yue

    2011-12-01

    Rheumatoid arthritis (RA) is considered as an autoimmune disease that intermittently causes the chronic and acute inflammation of the patient's small joints which can destroy the tissues around the joints resulting in the limitation of the joint's function. In the synovium and synovia of the joints, the infiltration and/or phagocytosis of the different kinds of leucocytes were demonstrated according to the phases of the acute and chronic inflammation. Also, Siglecs (sialic acid binding Ig-like lectins) were reported on the leucocytes which can induce the active and inhibitory immune response by the specific binding with sialic acid on the conjugates including the sialylation of the immunoglobulin which has been reported there was striking increasing in the synovium and synovia of the small joints, also in the sera on RA cases. This hypothesis proposed Siglecs on the leucocytes which infiltrate into the joint's cavity and the increasing sialic acid conjugates might play a role during the acute and chronic inflammation on RA disease. It might be helpful to explain the mechanism of the different inflammation in different circumstances in the RA.

  12. 111-Indium labelled autologous leucocytes in diagnosis of inflammatory bowel disease

    SciTech Connect

    Wandall, J.H.; Edeling, C.J.; Jensen, J.T.; Lund, J.O.; Bonnevie, O.; Haxholdt, H.; Jensen, H.C.; Matzen, P.; Myschetsky, P.S.; Nielsen, A.M.

    1984-01-01

    111-Indium labelled leucocytes have been used to visualize inflammatory lesions in ulcerative colitis (CU) and in Crohn's disease (CD). The aim of this study was to compare findings by scintigraphy, radiology and endoscopy. Material: Twelve patients with CU and 15 patients with CD were studied. All patients were non-febrile. Two patients received prednisolone 5 mg/daily, 8 sulphasalazine. Methods: Autologous leucocytes were labelled with 111-In-Oxine and given i.v. Scintigrams were obtained 3 and 24 hrs. p.i. Double contrast x-ray studies were done of the colon and small intestine after 2 and 14 days respectively. Colonscopy with biopsy was done after 4 days. Results: Active lesions were found in 24 and 27 patients. Scintigrams 24 hrs.p.i. did not give and additional information compared with scintigrams 3 hrs.p.i. Intraluminal activity masked the location and extension of lesions after 24 hrs. Excretion in the stool was 2.4-25.8% of administered activity. Compared with scintigraphy a corresponding extension and location was found by colonscopy. In 4 patients x-ray of the colon was normal but scintigraphy and colonscopy showed active inflammation. Conclusion: Scintigraphy after injection of 111-In labelled leucocytes is a atraumatic method for visualization of inflammatory lesions in UC and CD. Furthermore, it appears to be more sensitive than conventional x-ray studies.

  13. Leucocyte interactions with the mouse cremaster muscle microcirculation in vivo in response to tumour-conditioned medium.

    PubMed Central

    Brown, N. J.; Reed, M. W.

    1997-01-01

    Leucocyte interactions with the cremaster muscle microcirculation in vivo were investigated in response to culture medium conditioned with different cell types in 25 adult male Swiss mice. Animals were divided into five groups. Three groups received ex vivo fluorescently labelled lymphokine activated killer (LAK) cells systemically and had either tumour (murine melanoma K1735)-conditioned medium (TCM), fibroblast (murine 3T3)-conditioned medium (FCM) or fresh culture medium administered topically to the cremaster muscle. In the two remaining groups, the host leucocytes were labelled fluorescently by systemic administration of acridine red, and either TCM or FCM was applied topically to the cremaster muscle. There was an immediate but transient increase in the frequency of rolling and adherent LAK cells, and a subsequent (90-120 min later) increase in rolling and adherent host leucocytes, demonstrating temporal differences in the response to topical administration of TCM. These increases in contact with the vascular endothelium occurred in all vessel types, venules, arterioles and capillaries, with the greatest response observed in the venules. The FCM and normal culture medium did not affect the distribution and localization of either LAK cells or host leucocytes. These data suggest that there are one or more soluble tumour-specific chemoattractants for leucocytes present in the conditioned medium. The mouse cremaster muscle microcirculation is therefore a useful model to investigate the mechanism of leucocyte-endothelium interactions in tumour biology. PMID:9083334

  14. Flow cytometry analyses of phagocytic and respiratory burst activities and cytochemical characterization of leucocytes isolated from wrasse (Labrus bergylta A.).

    PubMed

    Haugland, Gyri T; Rønneseth, Anita; Wergeland, Heidrun I

    2014-07-01

    We have isolated leucocytes from peripheral blood (PBL), head kidney (HKL) and spleen (SL) of wrasse (Labrus bergylta A.) and studied the innate immune responses phagocytosis and respiratory burst using flow cytometry. Further, we have characterized the phenotypic properties of the leucocytes by cytochemical staining. We could differentiate between several subsets of leucocytes; lymphocytes, monocytes/macrophages, neutrophils, eosinophils, basophils and small leucocytes that might be precursor or immature cells. One striking observation was the eosinophils which were present among HKL, PBL and SL. The neutrophils had rounded, bean shaped or bi-lobed nuclei and resembled neutrophils in Atlantic cod (Gadus morhua L.) and lumpsucker (Cyclopterus lumpus L.), but were different from the polymorphonucleated neutrophils in Atlantic salmon (Salmo salar L.) and humans. Basophils were observed, but they were rare. Phagocytosis and respiratory burst activities were detected among different cell types. Highest phagocytic activity was observed among monocytes/macrophages and small leucocytes. Several different subtypes had ability to perform an oxygen-dependent degradation of microbes, measured as respiratory burst activity. Knowledge of the basic properties of wrasse's leucocytes and innate immunology can benefit further studies on its adaptive immune responses.

  15. [Functional state of endothelium and oxidant activity of leucocytes in patients with ischemic heart disease after coronary bypass surgery].

    PubMed

    Panov, A V; Abesadze, I T; Korzhenevskaia, K V; Nil'k, R Ia; Kozulin, V Iu; Gordeev, M L; Shliakhto, E V

    2007-01-01

    Relationship between disorders of endothelial function, proinflammatory activity of leucocytes and effects of therapy with simvastatin or its combination with ezetimibe was studied in 72 patients with ischemic heart disease subjected to coronary artery bypass grafting (CABG). Vascular endothelial function was assessed by ultrasound detection of brachial artery response to its compression, oxidant activity of leucocytes - by chemiluminescent microscopy, severity of coronary artery atherosclerosis - by invasive coronary angiography. Twenty two healthy individuals comprised control group. Endothelial function, activity of leucocytes, and lipid levels were evaluated before and in 12 months after CABG. Patients with multivessel involvement revealed more complex disorders of endothelial function and higher levels of leucocytes activity compared to patients with single-vessel disease. CABG resulted in disappearance of anginal attacks and negative stress echo test during 1-year observation in 80.6% of patients. Patients with recurrent angina after CABG had more severe endothelial and leucocytes disorders. Combined lipid lowering therapy (simvastatin plus ezetimibe) compared to simvastatin alone demonstrated higher efficacy in terms of achievement of target lipid levels, improvement of endothelial function and leucocytes disorders.

  16. Disturbed flow promotes deposition of leucocytes from flowing whole blood in a model of a damaged vessel wall.

    PubMed

    Skilbeck, Christopher A; Walker, Peter G; David, Tim; Nash, Gerard B

    2004-08-01

    Departure from simple laminar flow in arteries may promote the local attachment of leucocytes either to intact endothelium or platelet thrombi. We perfused blood through a chamber with a backward facing step, to observe whether adhesion from whole blood to P-selectin was indeed localized to a region of recirculating flow, and whether platelets binding to collagen in such a region could capture leucocytes. Blood flowing over the step established a stable vortex, a reattachment point where forward and backward flow separated, and a simple laminar flow with wall shear rate c. 400/s further downstream. Fluorescently labelled leucocytes were observed to attach to P-selectin immediately upstream or downstream of the reattachment point, and to roll back towards the step or away from it, respectively. There was negligible adhesion further downstream. When a P-selectin-Fc chimaera was used to coat the chamber, stable attachment occurred, again preferentially in the disturbed flow region. Numerous platelets adhered to a collagen coating throughout the chamber, although there were local maxima either side of the reattachment point. The adherent platelets captured flowing leucocytes in these regions alone. Leucocytes may adhere from flowing blood in vessels with high shear rate if the flow is disturbed. While platelets can adhere over a wider range of shear rates, their ability to capture leucocytes may be restricted to regions of disturbed flow. Copyright 2004 Blackwell Publishing Ltd

  17. Evaluation of C3a receptor expression on human leucocytes by the use of novel monoclonal antibodies

    PubMed Central

    ZWIRNER, J; GÖTZE, O; BEGEMANN, G; KAPP, A; KIRCHHOFF, K; WERFEL, T

    1999-01-01

    Varying results have been published in the past regarding the reactivity of different leucocyte subpopulations, including neutrophils, monocytes and B lymphocytes, to the anaphylatoxin C3a and its degradation product C3a(desArg). To better characterize the cellular distribution of C3a receptor (C3aR) expression, monoclonal antibodies against two different epitopes on the third extracellular domain of the human C3aR were generated. Quantification of C3aR as compared with C5aR densities was performed on peripheral blood leucocytes by quantitative indirect immunofluorescence. Eosinophils and basophils expressed similar numbers of C3aR and C5aR molecules/cell. On eosinophils 10 700±4500 (mean±SD) C3aR and 14 700±4100 C5aR were found, whereas basophils carried 8100±2100 C3aR and 13 500±3800 C5aR. Monocytes expressed approximately six times more C5aR than C3aR molecules on their surface (6000±2500 C3aR versus 34 100±9300 C5aR molecules) whereas on neutrophils, the expression of C5aR was more than 20 times higher than the expression of C3aR (3100±1000 C3aR versus 63 500±12 200 C5aR). No C3aR expression was detectable on peripheral blood-derived B lymphocytes and on tonsillar B cells before and after stimulation with interleukin-2/Staphylococcus aureus Cowan strain I. Our findings correspond well with the paucity of data on C3a-induced functional activities in monocytes and neutrophils and suggest that eosinophilic and basophilic granulocytes represent the primary effector cells in the peripheral blood which can be stimulated by C3a. PMID:10447728

  18. [A role of the adhesive properties of leucocytes and blood serum in the pathogenesis of multiple sclerosis].

    PubMed

    Zhirnova, I G; Larina, I V; Komel'kova, L V; Tsareva, M I; Gannushkina, I V; Zavalishin, I A

    2008-01-01

    Adhesive properties of leucocytes were studied using an original technique based on the leukocyte adherence inhibition reaction and measuring the values of spontaneous adhesion index (SAI) and adhesion-strengthening effect (ASE) under the influence of autoserum. One hundred patients with multiple sclerosis (MS) and 53 controls were included in the study. Immunophenotyping of lymphocytes (LP) with monoclonal antibodies--markers CD3, CD4, CD8, CD16, CD20, CD25, HLA-DR and CD95--and determination of IgG, IgA, IgM and content of immune complexes were carried out in parallel. The increase of adhesion parameters (ASE and SAI) was found in MS. It was most significant in patients with primary progressive course and in disease exacerbation. The greatest changes of phenotypic LP content were associated with debut and exacerbation-remission periods. Significant positive correlations between higher SAI values and phenotypes CD20, CD95, HLA-DR and amount of natural killer cells were revealed in patients with MS in contrast to the negative correlations of SAI with CD3 and CD4 in the control group. A role of membrane and soluble forms of adhesion molecules in the initiation and progression of immunopathological process in MS is discussed.

  19. Differential response of planktonic primary, bacterial, and dimethylsulfide production rates to static vs. dynamic light exposure in upper mixed-layer summer sea waters

    NASA Astrophysics Data System (ADS)

    Galí, M.; Simó, R.; Pérez, G. L.; Ruiz-González, C.; Sarmento, H.; Royer, S.-J.; Fuentes-Lema, A.; Gasol, J. M.

    2013-12-01

    Microbial plankton experience short-term fluctuations in total solar irradiance and in its spectral composition as they are vertically moved by turbulence in the oceanic upper mixed layer (UML). The fact that the light exposure is not static but dynamic may have important consequences for biogeochemical processes and ocean-atmosphere fluxes. However, most biogeochemical processes other than primary production, like bacterial production or dimethylsulfide (DMS) production, are seldom measured in sunlight and even less often in dynamic light fields. We conducted four experiments in oligotrophic summer stratified Mediterranean waters, where a sample from the UML was incubated in ultraviolet (UV)-transparent bottles at three fixed depths within the UML and on a vertically moving basket across the same depth range. We assessed the response of the phyto- and bacterioplankton community with physiological indicators based on flow cytometry singe-cell measurements, fast repetition rate fluorometry (FRRf), phytoplankton pigment concentrations and particulate light absorption. Dynamic light exposure caused a subtle disruption of the photoinhibition and photoacclimation processes associated with ultraviolet radiation (UVR), which slightly alleviated bacterial photoinhibition but did not favor primary production. Gross DMS production (GPDMS) decreased sharply with depth in parallel to shortwave UVR, and displayed a dose-dependent response that mixing did not significantly disrupt. To our knowledge, we provide the first measurements of GPDMS under in situ UV-inclusive optical conditions.

  20. Differential response of planktonic primary, bacterial, and dimethylsulfide production rates to vertically-moving and static incubations in upper mixed-layer summer sea waters

    NASA Astrophysics Data System (ADS)

    Galí, M.; Simó, R.; Pérez, G. L.; Ruiz-González, C.; Sarmento, H.; Royer, S.-J.; Fuentes-Lema, A.; Gasol, J. M.

    2013-05-01

    Microbial plankton experience fluctuations in total solar irradiance and in its spectral composition as they are vertically moved by turbulence in the oceanic upper mixed layer (UML). The fact that the light exposure is not static but dynamic may have important consequences for biogeochemical processes and ocean-atmosphere fluxes. However, most biogeochemical processes other than primary production, like bacterial production or dimethylsulfide (DMS) production, are seldom measured in sunlight and even less often in dynamic light fields. We conducted four experiments in oligotrophic summer stratified Mediterranean waters, where a sample from the UML was incubated in ultraviolet (UV)-transparent bottles at three fixed depths within the UML and on a vertically-moving basket across the same depth range. We assessed the response of the phyto- and bacterioplankton community with physiological indicators based on flow cytometry singe-cell measurements, Fast Repetition Rate fluorometry (FRRf), phytoplankton pigment concentrations and particulate light absorption. Dynamic light exposure caused a disruption of the photoinhibition and photoacclimation processes associated to ultraviolet radiation (UVR), which slightly alleviated bacterial photoinhibition but did not favor primary production. Gross DMS production (GPDMS) decreased sharply with depth in parallel to shortwave UVR, and displayed a dose-dependent response that mixing did not significantly disrupt. To our knowledge, we provide the first measurements of GPDMS under in situ UV-inclusive optical conditions.

  1. Primary care physicians' perceived barriers and facilitators to conservative care for older adults with chronic kidney disease: design of a mixed methods study.

    PubMed

    Tam-Tham, Helen; Hemmelgarn, Brenda; Campbell, David; Thomas, Chandra; Quinn, Robert; Fruetel, Karen; King-Shier, Kathryn

    2016-01-01

    Guideline committees have identified the need for research to inform the provision of conservative care for older adults with stage 5 chronic kidney disease (CKD) who have a high burden of comorbidity or functional impairment. We will use both qualitative and quantitative methodologies to provide a comprehensive understanding of barriers and facilitators to care for these patients in primary care. Our objectives are to (1) interview primary care physicians to determine their perspectives of conservative care for older adults with stage 5 CKD and (2) survey primary care physicians to determine the prevalence of key barriers and facilitators to provision of conservative care for older adults with stage 5 CKD. A sequential exploratory mixed methods design was adopted for this study. The first phase of the study will involve fundamental qualitative description and the second phase will be a cross-sectional population-based survey. The research is conducted in Alberta, Canada. The participants are primary care physicians with experience in providing care for older adults with stage 5 CKD not planning on initiating dialysis. The first objective will be achieved by undertaking interviews with primary care physicians from southern Alberta. Participants will be selected purposively to include physicians with a range of characteristics (e.g., age, gender, and location of clinical practice). Interviews will be recorded, transcribed verbatim, and analyzed using conventional content analysis to generate themes. The second objective will be achieved by undertaking a population-based survey of primary care physicians in Alberta. The questionnaire will be developed based on the findings from the qualitative interviews and pilot tested for face and content validity. Physicians will be provided multiple options to complete the questionnaire including mail, fax, and online methods. Descriptive statistics and associations between demographic factors and barriers and facilitators to

  2. Evaluating process and clinical outcomes of a primary care mental health integration project in rural Rwanda: a prospective mixed-methods protocol

    PubMed Central

    Smith, Stephanie L; Misago, Claire Nancy; Osrow, Robyn A; Franke, Molly F; Iyamuremye, Jean Damascene; Dusabeyezu, Jeanne D'Arc; Mohand, Achour A; Anatole, Manzi; Kayiteshonga, Yvonne; Raviola, Giuseppe J

    2017-01-01

    Introduction Integrating mental healthcare into primary care can reduce the global burden of mental disorders. Yet data on the effective implementation of real-world task-shared mental health programmes are limited. In 2012, the Rwandan Ministry of Health and the international healthcare organisation Partners in Health collaboratively adapted the Mentoring and Enhanced Supervision at Health Centers (MESH) programme, a successful programme of supported supervision based on task-sharing for HIV/AIDS care, to include care of neuropsychiatric disorders within primary care settings (MESH Mental Health). We propose 1 of the first studies in a rural low-income country to assess the implementation and clinical outcomes of a programme integrating neuropsychiatric care into a public primary care system. Methods and analysis A mixed-methods evaluation will be conducted. First, we will conduct a quantitative outcomes evaluation using a pretest and post-test design at 4 purposively selected MESH MH participating health centres. At least 112 consecutive adults with schizophrenia, bipolar disorder, depression or epilepsy will be enrolled. Primary outcomes are symptoms and functioning measured at baseline, 8 weeks and 6 months using clinician-administered scales: the General Health Questionnaire and the brief WHO Disability Assessment Scale. We hypothesise that service users will experience at least a 25% improvement in symptoms and functioning from baseline after MESH MH programme participation. To understand any outcome improvements under the intervention, we will evaluate programme processes using (1) quantitative analyses of routine service utilisation data and supervision checklist data and (2) qualitative semistructured interviews with primary care nurses, service users and family members. Ethics and dissemination This evaluation was approved by the Rwanda National Ethics Committee (Protocol #736/RNEC/2016) and deemed exempt by the Harvard University Institutional Review

  3. Adhesion of leucocytes onto polytetrafluoroethylene (PTFE) vascular grafts and the effect of low molecular weight dextran (LMWD).

    PubMed

    al-Huneidi, W; Owunwanne, A; Christenson, J T

    1990-01-01

    Platelets are known to interact with the surface of synthetic grafts. In the present study we have investigated another blood constituent, the white blood cell, to evaluate the contribution of leucocyte adhesion onto synthetic vascular graft surfaces. Furthermore the effect of low molecular weight dextran (LMWD) was evaluated in vivo. Polytetrafluoroethylene (PTFE) grafts were interpositioned in the femoral circulation in 12 healthy adult sheep. Autologous leucocytes were labelled with Indium-111-oxine and re-injected i.v. after purity and functional evaluation. Graft uptake of 111In-labelled leucocytes were studied continuously for 2.5 hours and two consecutive days using a gamma camera. Six animals received LMWD, 15 ml/h, i.v. infusion during the first day while the other six animals were infused with normal saline in the same amount and rate (controls). The labelled leucocytes showed normal phagocytosis of bacterias and the leucocyte purity was 74 +/- 6%. There was a rapid increase in graft activity initially. In control animals a continuous increase of graft activity was observed throughout the experiment. In the LMWD-treated animals graft activity remained on a steady level after the initial built up of activity and after 2.5 hours there was a highly significant difference between the groups, p less than 0.001. These differences were confirmed by in vitro activity measurement, autoradiography and histological examination of the grafts at the end of the experiment. It was concluded that leucocyte adhesion onto the surface of PTFE grafts occur during the early period after implantation and could therefore be a contributing factor in the thrombogenesis. Administration of LMWD seems to have a beneficial effect since less leucocyte adhesion occurred in those animals treated with LMWD.

  4. Similar change in platelets and leucocytes 24 h after injury is associated with septic shock a week later.

    PubMed

    Jol, Saskia; Hietbrink, Falco; Leenen, Luke P H; Koenderman, Leo; van Wessem, Karlijn J P

    2017-03-01

    Septic shock is a severe complication in polytrauma patients. Early identification of patients at risk can guide future prevention strategies. Platelets (PLTs) and leucocytes presumably play an important role in the post-injury inflammatory response. The role of early changes in PLT and leucocyte counts was investigated in search for the aetiology of the development of septic complications. Polytrauma patients (aged 16-80 years) admitted to the intensive care unit with an expected stay of at least 3 days were included. PLT and leucocyte counts were measured on a daily basis for 14 days. A total of 41 patients were included, of whom nine (22%) developed septic shock. There was no difference in (New) Injury Severity Score or Acute Physiology and Chronic Health Evaluation II scores between patients who developed septic shock and patients who did not. Three patients died, one of them in septic shock. Patients who developed septic shock during hospital stay had lower PLTs and a slower recovery to normal PLT counts than patients without septic shock. Patients who developed either a decrease in both PLTs and leucocytes or an increase in PLTs and leucocytes in the first 24 h after trauma were more likely to develop septic shock. This correlation was not found in patients who did not develop septic shock. A similar change in PLT and leucocyte counts in the first 24 h after trauma is associated with the development of septic shock after a week. This indicates an early interaction between PLTs and leucocytes, which needs further investigation to gain more insight in the aetiology of post-injury septic complications. © 2017 Royal Australasian College of Surgeons.

  5. [The comparative evaluation of information value of traditionally analyzed indicators of total blood test and leucocytes index of intoxication in women with physiological and complicated course of pregnancy].

    PubMed

    Skriabina, V V

    2013-12-01

    The article demonstrates that on the basis of retrospective analysis of 34 cases of physiological and 218 cases of complicated course of pregnancy the evaluation of leucocytes index of intoxication is more informative than the standard analysis of leukogram. The increase of leucocytes index of intoxication and tendency of increase of percentage of concentration of leucocytes at early stage of pregnancy are detected mainly in women with complicated course of pregnancy.

  6. Patient safety initiatives in Central and Eastern Europe: A mixed methods approach by the LINNEAUS collaboration on patient safety in primary care

    PubMed Central

    Godycki-Cwirko, Maciek; Esmail, Aneez; Dovey, Susan; Wensing, Michel; Parker, Dianne; Kowalczyk, Anna; Błaszczyk, Honorata; Kosiek, Katarzyna

    2015-01-01

    ABSTRACT Background: Despite patient safety being recognized as an important healthcare issue in the European Union, there has been variable implementation of patient safety initiatives in Central and Eastern Europe (CEE). Objective: To assess the status of patient safety initiatives in countries in CEE; to describe a process of engagement in Poland, which can serve as a template for the implementation of patient safety initiatives in primary care. Methods: A mixed methods design was used. We conducted a review of literature focusing on publications from CEE, an inventory of patient safety initiatives in CEE countries, interviews with key informants, international survey, review of national reporting systems, and pilot demonstrator project in Poland with implementation of patient safety toolkits assessment. Results: There was no published patient safety research from Albania, Belarus, Greece, Latvia, Lithuania, Romania, or Russia. Nine papers were found from Bulgaria, Croatia, the Czech Republic, Poland, Serbia, and Slovenia. In most of the CEE countries, patient safety had been addressed at the policy level although the focus was mainly in hospital care. There was a dearth of activity in primary care. The use of patient improvement strategies was low. Conclusion: International cooperation as exemplified in the demonstrator project can help in the development and implementation of patient safety initiatives in primary care in changing the emphasis away from a blame culture to one where greater emphasis is placed on improvement and learning. PMID:26339839

  7. Ovarian cavity fluid of the viviparous surfperch Neoditrema ransonnetii suppresses the spontaneous cytotoxic activity of head-kidney leucocytes against xenogeneic targets.

    PubMed

    Yokozawa, N; Nakamura, O; Saito, E; Tsutsui, S

    2015-01-01

    In this study, the effect of ovarian cavity fluid (OCF) from a surfperch Neoditrema ransonnetii on the cytotoxic activity of leucocytes was investigated. In an assay targeting RTG-2, a cell line derived from rainbow trout Oncorhynchus mykiss gonads, leucocytes from both the spleen and head kidney showed spontaneous killing activity. Pre-incubation with OCF significantly suppressed the cytotoxic activity of head-kidney leucocytes towards RTG-2. This suppressive activity was due to the presence of low molecular-mass materials. These results suggest that OCF plays significant roles in pregnancy by its ability to modulate cytotoxicity with maternal leucocytes.

  8. Science Self-Efficacy in the Primary Classroom: Using Mixed Methods to Investigate Sources of Self-Efficacy

    NASA Astrophysics Data System (ADS)

    Webb-Williams, Jane

    2017-04-01

    Self-efficacy has been shown to influence student engagement, effort and performance as well as course selection and future career choice. Extending our knowledge regarding the development of self-efficacy has important implications for educators and for those concerned about the international uptake of science careers. Previous research has identified four sources that may contribute towards self-efficacy: mastery experiences, vicarious experiences, verbal persuasion and physiological/affective states. Very little research has been conducted within the school environment that looks at the formation of these sources and yet early school experiences have been posited to be a key factor in girls' lack of engagement in post compulsory science education. This paper investigates children's self-efficacy beliefs in science and reports on findings from mixed method research conducted with 182 children aged between 10 and 12 years. Classroom data were collected through focus groups, individual interviews and surveys. Findings revealed that although girls and boys held similar levels of academic performance in science, many girls underestimated their capability. The four sources of self-efficacy identified by Bandura (1997) plus self-regulation as an additional source, were evident in the children's descriptions, with boys being more influenced by mastery experience and girls by a combination of vicarious experience and physiological/affective states. Girl's appraisal of information appeared to operate through a heuristic process whereby girls disregarded salient information such as teacher feedback in favour of reliance on social comparison. Contextual factors were identified. Implications for science teachers are discussed.

  9. Primary care physicians' perspectives on computer-based health risk assessment tools for chronic diseases: a mixed methods study.

    PubMed

    Voruganti, Teja R; O'Brien, Mary Ann; Straus, Sharon E; McLaughlin, John R; Grunfeld, Eva

    2015-09-24

    Health risk assessment tools compute an individual's risk of developing a disease. Routine use of such tools by primary care physicians (PCPs) is potentially useful in chronic disease prevention. We sought physicians' awareness and perceptions of the usefulness, usability and feasibility of performing assessments with computer-based risk assessment tools in primary care settings. Focus groups and usability testing with a computer-based risk assessment tool were conducted with PCPs from both university-affiliated and community-based practices. Analysis was derived from grounded theory methodology. PCPs (n = 30) were aware of several risk assessment tools although only select tools were used routinely. The decision to use a tool depended on how use impacted practice workflow and whether the tool had credibility. Participants felt that embedding tools in the electronic medical records (EMRs) system might allow for health information from the medical record to auto-populate into the tool. User comprehension of risk could also be improved with computer-based interfaces that present risk in different formats. In this study, PCPs chose to use certain tools more regularly because of usability and credibility. Despite there being differences in the particular tools a clinical practice used, there was general appreciation for the usefulness of tools for different clinical situations. Participants characterised particular features of an ideal tool, feeling strongly that embedding risk assessment tools in the EMR would maximise accessibility and use of the tool for chronic disease management. However, appropriate practice workflow integration and features that facilitate patient understanding at point-of-care are also essential.

  10. A(1c) control in a primary care setting: self-titrating an insulin analog pre-mix (INITIATEplus trial).

    PubMed

    Oyer, David S; Shepherd, Mark D; Coulter, Franklin C; Bhargava, Anuj; Brett, Jason; Chu, Pei-Ling; Trippe, Bruce S

    2009-11-01

    To study glycemic control and hypoglycemia development upon initiation of insulin through a self-titration schedule in a 24-week trial, conducted with 4875 insulin-naïve patients with poorly controlled type 2 diabetes, predominantly in a primary care setting. Subjects initiated twice-daily biphasic insulin aspart 70/30 with 6 units prebreakfast and 6 units presupper, self-titrating according to self-measured blood glucose values. Subjects were randomized (1:1:1) to telephone counseling provided by a registered dietician: no counseling (NC), 1 counseling session (1C), or 3 sessions (3C). Mean baseline HbA(1c) (9.9% across groups) decreased approximately 2.5% to 7.49% + or - 1.48, 7.48% + or - 1.50, and 7.44% + or - 1.46 in the NC, 1C, and 3C groups, respectively. Within these groups, a hemoglobin A(1c) (HbA(1c)) value <7% was achieved by 40.2%, 41.6%, and 41.8% of subjects, respectively. Eight-point blood glucose profiles were substantially improved from baseline for all groups. Hypoglycemia was experienced by 10.2%-11.4% of the subjects in each group. Rates of minor and major hypoglycemia were low but decreased as dietary counseling increased (minor hypoglycemia: 56 vs 50 vs 45 episodes per 100 patient-years; major hypoglycemia, 9 vs 6 vs 4 episodes per 100 patient-years, for the NC vs 1C vs 3C groups, respectively; P <.001, 3C vs NC). Weight increased by 3.13, 3.40, and 2.88 kg for the NC, 1C, and 3C groups, respectively. In the primary care setting, self-titration of biphasic insulin aspart 70/30 was effective in achieving recommended HbA(1c) goals even with minimal dietary counseling.

  11. The Influence of Cytomegalovirus on Expression of HLA-G and its Ligand KIR2DL4 by Human Peripheral Blood Leucocyte Subsets.

    PubMed

    Albayati, Z; Alyami, A; Alomar, S; Middleton, D; Bonnett, L; Aleem, S; Flanagan, B F; Christmas, S E

    2017-08-17

    HLA-G is a non-classical class I HLA antigen, normally expressed in high levels only on extravillous cytotrophoblast. It has immunosuppressive properties in pregnancy and has also been found to be upregulated on leucocytes in viral infection. In this study, proportions of all leucocyte subsets expressing HLA-G were found to be low in healthy subjects positive or negative for cytomegalovirus (CMV). Significantly greater proportions of CD4+ CD69+ and CD56+ T cells expressed HLA-G compared to other T cells. However, following stimulation with CMV antigens or intact CMV, proportions of CD4+, CD8+, CD69+ and CD56+ T cells, and also B cells expressing HLA-G, were significantly increased in CMV+ subjects. Despite some subjects having alleles of HLA-G associated with high levels of expression, no relationship was found between HLA-G genotype and expression levels. Purified B cells from CMV+ subjects stimulated in mixed culture with CMV antigens showed significantly increased HLA-G mRNA expression by real-time polymerase chain reaction. Serum levels of soluble HLA-G were similar in CMV- and CMV+ subjects but levels in culture supernatants were significantly higher in cells from CMV+ than from CMV- subjects stimulated with CMV antigens. The HLA-G ligand KIR2DL4 was mainly expressed on NK cells and CD56+ T cells with no differences between CMV+ and CMV- subjects. Following stimulation with IL-2, an increase in the proportion of CD56+ T cells positive for KIR2DL4 was found, together with a significant decrease in CD56dimCD16+ NK cells. The results show that CMV influences HLA-G expression in healthy subjects and may contribute to viral immune evasion. © 2017 The Foundation for the Scandinavian Journal of Immunology.

  12. Flow cytometry assays of respiratory burst in Atlantic salmon (Salmo salar L.) and in Atlantic cod (Gadus morhua L.) leucocytes.

    PubMed

    Kalgraff, Cathrine A K; Wergeland, Heidrun I; Pettersen, Eirin Fausa

    2011-09-01

    The oxidation of dihydrorhodamine 123 (DHR) to the fluorescent rhodamine 123 (RHO) was detected using flow cytometry. This assay for detection of respiratory burst activity was established in peripheral blood leucocytes (PBL) and head kidney leucocytes (HKL) of Atlantic salmon and Atlantic cod. The leucocytes were stimulated by phorbol 12-myristate 13-acetate (PMA). For cod cells 10 times lower concentration of PMA had to be used compared to salmon cells, as higher concentrations were toxic and resulted in considerable cell death. The cells found to be RHO-positive were monocytes/macrophages and neutrophils based on the scatter dot plots, but for salmon also some small cells were found to have high fluorescence intensity both in the flow cytometry analyses and by fluorescence microscopy of cytospin preparations. The nature of these cells is not known. For cod leucocytes, such cells were not obvious. The instrument settings are a bit more demanding for cod, as cod cells die more easily compared to salmon cells. In both assays the limit between negative and positive cells has to be carefully considered. The presented flow cytometry protocols for measurements of respiratory burst in salmon and cod leucocytes can be applied in various studies where respiratory burst functions are involved, such as to verify if it is activated or suppressed in connection with infections and immunostimulation.

  13. Fatty acid binding protein 4 in circulating leucocytes reflects atherosclerotic lesion progression in Apoe−/− mice

    PubMed Central

    Agardh, Hanna E; Gertow, Karl; Salvado, Dolores M; Hermansson, Andreas; Puijvelde, Gijs H; Hansson, Göran K; n-Berne, Gabrielle Paulsso; Gabrielsen, Anders

    2013-01-01

    Discovery of novel biomarkers for atherosclerosis is important to aid in early diagnosis of pre-symptomatic patients at high risk of cardiovascular events. The aim of the present study was therefore to identify potential biomarkers in circulating cells reflecting atherosclerotic lesion progression in the vessel wall. We performed gene arrays on circulating leucocytes from atherosclerosis prone Apoe−/− mice with increasing ages, using C57BL/6 mice as healthy controls. We identified fatty acid binding protein 4 (FABP4) mRNA to be augmented in mice with established disease compared with young Apoe−/− or controls. Interestingly, the transcript FABP4 correlated significantly with lesion size, further supporting a disease associated increase. In addition, validation of our finding on protein level showed augmented FABP4 in circulating leucocytes whereas, importantly, no change could be observed in plasma. Immunofluorescence analysis demonstrated FABP4 to be present mainly in circulating neutrophils and to some extent in monocytes. Moreover, FABP4-positive neutrophils and macrophages could be identified in the subintimal space in the plaque. Using human circulating leucocytes, we confirmed the presence of FABP4 protein in neutrophils and monocytes. In conclusion, we have showed that cellular levels of FABP4 in circulating leucocytes associate with lesion development in the experimental Apoe−/− model. The increased expression is primarily localized to neutrophils, but also in monocytes. We have identified FABP4 in leucocytes as a potential and easy accessible biomarker of atherosclerosis which could be of future clinical relevance. PMID:23387955

  14. Reprogramming Neutral Lipid Metabolism in Mouse Dendritic Leucocytes Hosting Live Leishmania amazonensis Amastigotes

    PubMed Central

    Lecoeur, Hervé; Giraud, Emilie; Prévost, Marie-Christine; Milon, Geneviève; Lang, Thierry

    2013-01-01

    Background After loading with live Leishmania (L) amazonensis amastigotes, mouse myeloid dendritic leucocytes/DLs are known to undergo reprogramming of their immune functions. In the study reported here, we investigated whether the presence of live L. amazonensis amastigotes in mouse bone marrow-derived DLs is able to trigger re-programming of DL lipid, and particularly neutral lipid metabolism. Methodology/Principal Findings Affymetrix-based transcriptional profiles were determined in C57BL/6 and DBA/2 mouse bone marrow-derived DLs that had been sorted from cultures exposed or not to live L. amazonensis amastigotes. This showed that live amastigote-hosting DLs exhibited a coordinated increase in: (i) long-chain fatty acids (LCFA) and cholesterol uptake/transport, (ii) LCFA and cholesterol (re)-esterification to triacyl-sn-glycerol (TAG) and cholesteryl esters (CE), respectively. As these neutral lipids are known to make up the lipid body (LB) core, oleic acid was added to DL cultures and LB accumulation was compared in live amastigote-hosting versus amastigote-free DLs by epi-fluorescence and transmission electron microscopy. This showed that LBs were both significantly larger and more numerous in live amastigote-hosting mouse dendritic leucocytes. Moreover, many of the larger LB showed intimate contact with the membrane of the parasitophorous vacuoles hosting the live L. amazonensis amastigotes. Conclusions/Significance As leucocyte LBs are known to be more than simple neutral lipid repositories, we set about addressing two related questions. Could LBs provide lipids to live amastigotes hosted within the DL parasitophorous vacuole and also deliver? Could LBs impact either directly or indirectly on the persistence of L. amazonensis amastigotes in rodent skin? PMID:23785538

  15. Reprogramming neutral lipid metabolism in mouse dendritic leucocytes hosting live Leishmania amazonensis amastigotes.

    PubMed

    Lecoeur, Hervé; Giraud, Emilie; Prévost, Marie-Christine; Milon, Geneviève; Lang, Thierry

    2013-01-01

    After loading with live Leishmania (L) amazonensis amastigotes, mouse myeloid dendritic leucocytes/DLs are known to undergo reprogramming of their immune functions. In the study reported here, we investigated whether the presence of live L. amazonensis amastigotes in mouse bone marrow-derived DLs is able to trigger re-programming of DL lipid, and particularly neutral lipid metabolism. Affymetrix-based transcriptional profiles were determined in C57BL/6 and DBA/2 mouse bone marrow-derived DLs that had been sorted from cultures exposed or not to live L. amazonensis amastigotes. This showed that live amastigote-hosting DLs exhibited a coordinated increase in: (i) long-chain fatty acids (LCFA) and cholesterol uptake/transport, (ii) LCFA and cholesterol (re)-esterification to triacyl-sn-glycerol (TAG) and cholesteryl esters (CE), respectively. As these neutral lipids are known to make up the lipid body (LB) core, oleic acid was added to DL cultures and LB accumulation was compared in live amastigote-hosting versus amastigote-free DLs by epi-fluorescence and transmission electron microscopy. This showed that LBs were both significantly larger and more numerous in live amastigote-hosting mouse dendritic leucocytes. Moreover, many of the larger LB showed intimate contact with the membrane of the parasitophorous vacuoles hosting the live L. amazonensis amastigotes. As leucocyte LBs are known to be more than simple neutral lipid repositories, we set about addressing two related questions. Could LBs provide lipids to live amastigotes hosted within the DL parasitophorous vacuole and also deliver? Could LBs impact either directly or indirectly on the persistence of L. amazonensis amastigotes in rodent skin?

  16. Increased glucocerebrosidase (GBA) 2 activity in GBA1 deficient mice brains and in Gaucher leucocytes.

    PubMed

    Burke, Derek G; Rahim, Ahad A; Waddington, Simon N; Karlsson, Stefan; Enquist, Ida; Bhatia, Kailash; Mehta, Atul; Vellodi, Ashok; Heales, Simon

    2013-09-01

    Lysosomal glucocerebrosidase (GBA1) deficiency is causative for Gaucher disease. Not all individuals with GBA1 mutations develop neurological involvement raising the possibility that other factors may provide compensatory protection. One factor may be the activity of the non-lysosomal β-glucosidase (GBA2) which exhibits catalytic activity towards glucosylceramide and is reported to be highly expressed in brain tissue. Here, we assessed brain GBA2 enzymatic activity in wild type, heterozygote and GBA1 deficient mice. Additionally, we determined activity in leucocytes obtained from 13 patients with Gaucher disease, 10 patients with enzymology consistent with heterozygote status and 19 controls. For wild type animals, GBA2 accounted for over 85 % of total brain GBA activity and was significantly elevated in GBA1 deficient mice when compared to heterozygote and wild types (GBA1 deficient; 92.4 ± 5.6, heterozygote; 71.5 ± 2.4, wild type 76.8 ± 5.1 nmol/h/mg protein). For the patient samples, five Gaucher patients had GBA2 leucocyte activities markedly greater than controls. No difference in GBA2 activity was apparent between the control and carrier groups. Undetectable GBA2 activity was identified in four leucocyte preparations; one in the control group, two in the carrier group and one from the Gaucher disease group. Work is now required to ascertain whether GBA2 activity is a disease modifying factor in Gaucher disease and to identify the mechanism(s) responsible for triggering increased GBA2 activity in GBA1 deficiency states.

  17. The melanocortin MC1 receptor agonist BMS-470539 inhibits leucocyte trafficking in the inflamed vasculature

    PubMed Central

    Leoni, G; Voisin, M-B; Carlson, K; Getting, SJ; Nourshargh, S; Perretti, M

    2010-01-01

    Background and purpose: Over three decades of research evaluating the biology of melanocortin (MC) hormones and synthetic peptides, activation of the MC type 1 (MC1) receptor has been identified as a viable target for the development of novel anti-inflammatory therapeutic agents. Here, we have tested a recently described selective agonist of MC1 receptors, BMS-470539, on leucocyte/post-capillary venule interactions in murine microvascular beds. Experimental approach: Intravital microscopy of two murine microcirculations were utilized, applying two distinct modes of promoting inflammation. The specificity of the effects of BMS-470539 was assessed using mice bearing mutant inactive MC1 receptors (the recessive yellow e/e colony). Key results: BMS-470539, given before an ischaemia–reperfusion protocol, inhibited cell adhesion and emigration with no effect on cell rolling, as assessed 90 min into the reperfusion phase. These properties were paralleled by inhibition of tissue expression of both CXCL1 and CCL2. Confocal investigations of inflamed post-capillary venules revealed immunostaining for MC1 receptors on adherent and emigrated leucocytes. Congruently, the anti-inflammatory properties of BMS-470539 were lost in mesenteries of mice bearing the inactive mutant MC1 receptors. Therapeutic administration of BMS-470539 stopped cell emigration, but did not affect cell adhesion in the cremasteric microcirculation inflamed by superfusion with platelet-activating factor. Conclusions and implications: Activation of MC1 receptors inhibited leucocyte adhesion and emigration. Development of new chemical entities directed at MC1 receptors could be a viable approach in the development of novel anti-inflammatory therapeutic agents with potential application to post-ischaemic conditions. PMID:20331604

  18. Barriers and facilitators influencing self-management among COPD patients: a mixed methods exploration in primary and affiliated specialist care.

    PubMed

    Hillebregt, Chantal F; Vlonk, Auke J; Bruijnzeels, Marc A; van Schayck, Onno Cp; Chavannes, Niels H

    2017-01-01

    Self-management is becoming increasingly important in COPD health care although it remains difficult to embed self-management into routine clinical care. The implementation of self-management is understood as a complex interaction at the level of patient, health care provider (HCP), and health system. Nonetheless there is still a poor understanding of the barriers and effective facilitators. Comprehension of these determinants can have significant implications in optimizing self-management implementation and give further directions for the development of self-management interventions. Data were collected among COPD patients (N=46) and their HCPs (N=11) in three general practices and their collaborating affiliated hospitals. Mixed methods exploration of the data was conducted and collected by interviews, video-recorded consultations (N=50), and questionnaires on consultation skills. Influencing determinants were monitored by 1) interaction and communication between the patient and HCP, 2) visible and invisible competencies of both the patient and the HCP, and 3) degree of embedding self-management into the health care system. Video observations showed little emphasis on effective behavioral change and follow-up of given lifestyle advice during consultation. A strong presence of COPD assessment and monitoring negatively affects the patient-centered communication. Both patients and HCPs experience difficulties in defining personalized goals. The satisfaction of both patients and HCPs concerning patient centeredness during consultation was measured by the patient feedback questionnaire on consultation skills. The patients scored high (84.3% maximum score) and differed from the HCPs (26.5% maximum score). Although the patient-centered approach accentuating self-management is one of the dominant paradigms in modern medicine, our observations show several influencing determinants causing difficulties in daily practice implementation. This research is a first step

  19. Mindfulness-based stress reduction for GPs: results of a controlled mixed methods pilot study in Dutch primary care

    PubMed Central

    Verweij, Hanne; Waumans, Ruth C; Smeijers, Danique; Lucassen, Peter LBJ; Donders, A Rogier T; van der Horst, Henriëtte E; Speckens, Anne EM

    2016-01-01

    Background Burnout is highly prevalent in GPs and can have a negative influence on their wellbeing, performance, and patient care. Mindfulness-based stress reduction (MBSR) may be an effective intervention to decrease burnout symptoms and increase wellbeing. Aim To gain insight into the feasibility and effectiveness of MBSR on burnout, empathy, and (work-related) wellbeing in GPs. Design and setting A mixed methods pilot study, including a waiting list-controlled pre-/post-study and a qualitative study of the experiences of participating GPs in the Netherlands. Method Participants were sent questionnaires assessing burnout, work engagement, empathy, and mindfulness skills, before and at the end of the MBSR training/waiting period. Qualitative data on how GPs experienced the training were collected during a plenary session and with evaluation forms at the end of the course. Results Fifty Dutch GPs participated in this study. The MBSR group reported a greater decrease in depersonalisation than the control group (adjusted difference −1.42, 95% confidence interval [CI] = −2.72 to −0.21, P = 0.03). Dedication increased more significantly in the MBSR group than in the control group (adjusted difference 2.17, 95% CI = 0.51 to 3.83, P = 0.01). Mindfulness skills increased significantly in the MBSR group compared with the control group (adjusted difference 6.90, 95% CI = 1.42 to 12.37, P = 0.01). There was no significant change in empathy. The qualitative data indicated that the MBSR course increased their wellbeing and compassion towards themselves and others, including their patients. Conclusion The study shows that MBSR for GPs is feasible and might result in fewer burnout symptoms and increased work engagement and wellbeing. However, an adequately powered randomised controlled trial is needed to confirm the study’s findings. PMID:26823271

  20. Physician self-disclosure in primary care: a mixed methods study of GPs’ attitudes, skills, and behaviour

    PubMed Central

    Allen, Emily-Charlotte Frances; Arroll, Bruce

    2015-01-01

    Background There is a debate in medicine about the use and value of self-disclosure by the physician as a communication tool. There is little empirical evidence about GPs and self-disclosure. Aim To explore what GPs’ attitudes, skills, and behaviour are with regard to self-disclosure during a clinical consultation and whether there is a need for the development of training resources. Design and setting Mixed methods using open-ended and semi-structured interviews in Auckland, New Zealand, and the surrounding districts. Method Sixteen GPs were interviewed on the issue of self-disclosure in clinical practice. A general inductive approach was used for data analysis. Results Self-disclosure was common in this group of GPs, contrary to training in some of the groups, and was seen as a potentially positive activity. Family and physical topics were most common, yet psychological and relationship issues were also discussed. Knowing patients made self-disclosure more likely, but a GP’s intuition played the main role in determining when to self-disclose, and to whom. GPs have developed their own guidelines, shaped by years of experience; however, there was a consensus that training would be helpful. Conclusion Self-disclosure is common and, in general, seen as positive. Major personal issues were acceptable for some GPs to self-disclose, especially to known patients. Although participants had developed their own guidelines, exposure of trainees to the issue of self-disclosure would be of value to prevent future mistakes and to protect both doctor and patient from any unintended harm, for example, developing a dependent relationship. PMID:26324497

  1. Barriers and facilitators influencing self-management among COPD patients: a mixed methods exploration in primary and affiliated specialist care

    PubMed Central

    Hillebregt, Chantal F; Vlonk, Auke J; Bruijnzeels, Marc A; van Schayck, Onno CP; Chavannes, Niels H

    2017-01-01

    Self-management is becoming increasingly important in COPD health care although it remains difficult to embed self-management into routine clinical care. The implementation of self-management is understood as a complex interaction at the level of patient, health care provider (HCP), and health system. Nonetheless there is still a poor understanding of the barriers and effective facilitators. Comprehension of these determinants can have significant implications in optimizing self-management implementation and give further directions for the development of self-management interventions. Data were collected among COPD patients (N=46) and their HCPs (N=11) in three general practices and their collaborating affiliated hospitals. Mixed methods exploration of the data was conducted and collected by interviews, video-recorded consultations (N=50), and questionnaires on consultation skills. Influencing determinants were monitored by 1) interaction and communication between the patient and HCP, 2) visible and invisible competencies of both the patient and the HCP, and 3) degree of embedding self-management into the health care system. Video observations showed little emphasis on effective behavioral change and follow-up of given lifestyle advice during consultation. A strong presence of COPD assessment and monitoring negatively affects the patient-centered communication. Both patients and HCPs experience difficulties in defining personalized goals. The satisfaction of both patients and HCPs concerning patient centeredness during consultation was measured by the patient feedback questionnaire on consultation skills. The patients scored high (84.3% maximum score) and differed from the HCPs (26.5% maximum score). Although the patient-centered approach accentuating self-management is one of the dominant paradigms in modern medicine, our observations show several influencing determinants causing difficulties in daily practice implementation. This research is a first step

  2. Physician self-disclosure in primary care: a mixed methods study of GPs' attitudes, skills, and behaviour.

    PubMed

    Allen, Emily-Charlotte Frances; Arroll, Bruce

    2015-09-01

    There is a debate in medicine about the use and value of self-disclosure by the physician as a communication tool. There is little empirical evidence about GPs and self-disclosure. To explore what GPs' attitudes, skills, and behaviour are with regard to self-disclosure during a clinical consultation and whether there is a need for the development of training resources. Mixed methods using open-ended and semi-structured interviews in Auckland, New Zealand, and the surrounding districts. Sixteen GPs were interviewed on the issue of self-disclosure in clinical practice. A general inductive approach was used for data analysis. Self-disclosure was common in this group of GPs, contrary to training in some of the groups, and was seen as a potentially positive activity. Family and physical topics were most common, yet psychological and relationship issues were also discussed. Knowing patients made self-disclosure more likely, but a GP's intuition played the main role in determining when to self-disclose, and to whom. GPs have developed their own guidelines, shaped by years of experience; however, there was a consensus that training would be helpful. Self-disclosure is common and, in general, seen as positive. Major personal issues were acceptable for some GPs to self-disclose, especially to known patients. Although participants had developed their own guidelines, exposure of trainees to the issue of self-disclosure would be of value to prevent future mistakes and to protect both doctor and patient from any unintended harm, for example, developing a dependent relationship. © British Journal of General Practice 2015.

  3. Changes in beta-adrenoceptors and leucocyte subpopulations after physical exercise in normal subjects.

    PubMed

    Landmann, R; Portenier, M; Staehelin, M; Wesp, M; Box, R

    1988-03-01

    Seven healthy volunteers were subjected to standardized bicycle ergometry. Before and at the end of exercise, leucocyte and lymphocyte subset distribution was assessed by immunofluorescence labelling with monoclonal antibodies and the cytofluorograf. Competition binding studies were performed on mononuclear leucocytes with a fixed amount of the radioligand 125I-(-)-cyanopindolol (125I-CYP) and increasing concentrations of the hydrophilic ligand (-)4-(3-tertiary-butylamino-2-hydroxypropoxy)-benzimidazol-2-one hydrochloride (CGP-12177). Total numbers of beta-adrenoceptors per cell and the receptor-ligand affinities were then derived by computer analysis. In separate experiments with blood obtained from resting subjects, beta-adrenoceptor numbers of lymphocyte subsets, which had been sorted by the fluorescence activated cell sorter, were determined by saturation binding of 125I-CYP. During exercise there was a twofold increase in total leucocyte numbers. The ratio between monocytes (4%) and lymphocytes (55%) remained constant, but the composition of lymphocyte subsets had changed. A twofold increase was observed for the lymphocyte population carrying the Leu-7 and the CD8 antigens or only the Leu-7 antigen, whereas the number of B cells and CD4 positive T cells increased only slightly. The lymphocyte phenotype changes appeared after 3 min, reached a maximum at the end of ergometry and had disappeared 30 min after exercise. Exercise led to a doubling of the number of beta-adrenoceptors in unseparated mononuclear leucocytes from 240 +/- 46 to 535 +/- 190 sites per cell (SD, n = 6). beta-Adrenoceptor numbers were higher on sorted Leu-7 positive cells (KD 14 +/- 6 pmol/l, Bmax 1174 +/- 233 sites/cell) and on CD8 positive cells (KD 145 +/- 79 pmol/l, Bmax 1577 +/- 670 sites/cell) than on monocytes (KD 39 +/- 31 pmol/l, 647 +/- 91 sites/cell). Very low specific 125I-CYP binding was found on Leu-7 and CD8 negative cells (less than 0.2 pmol/l). Before exercise the

  4. Rapid enrichment of leucocytes and genomic DNA from blood based on bifunctional core shell magnetic nanoparticles

    NASA Astrophysics Data System (ADS)

    Xie, Xin; Nie, Xiaorong; Yu, Bingbin; Zhang, Xu

    2007-04-01

    A series of protocols are proposed to extract genomic DNA from whole blood at different scales using carboxyl-functionalized magnetic nanoparticles as solid-phase absorbents. The enrichment of leucocytes and the adsorption of genomic DNA can be achieved with the same carboxyl-functionalized magnetic nanoparticles. The DNA bound to the bead surfaces can be used directly as PCR templates. By coupling cell separation and DNA purification, the whole operation can be accomplished in a few minutes. Our simplified protocols proved to be rapid, low cost, and biologically and chemically non-hazardous, and are therefore promising for microfabrication of a DNA-preparation chip and routine laboratory use.

  5. Indium 111 autologous leucocyte scanning in lobar pneumonia and lung abscesses.

    PubMed Central

    Saverymuttu, S H; Phillips, G; Peters, A M; Lavender, J P

    1985-01-01

    Indium 111 leucocyte scanning was used to examine the kinetics of granulocyte localisation in five patients with lobar pneumonia and four patients with lung abscess. Lung abscesses showed dense accumulation of labelled granulocytes within four hours of injection. In contrast, in all cases of lobar pneumonia granulocytes failed to accumulate over a period of up to 24 hours despite evidence of adequate perfusion and of uptake in inflammation elsewhere. These results suggest that the recruitment of granulocytes to lobar pneumonia may be terminated early in its clinical course. Images PMID:4095674

  6. Ocular microbiota and polymorphonuclear leucocyte recruitment during overnight contact lens wear.

    PubMed

    Stapleton, F; Willcox, M D; Sansey, N; Holden, B A

    1997-05-01

    Bacterial colonization of the ocular surface and contact lens (CL) and recruitment of polymorphonuclear leucocytes (PMN) during overnight CL wear was examined in 11 asymptomatic wearers. The ocular surface was more frequently colonized than the CL, with commensal bacteria (P < 0.05). Following sleep, more bacteria were recovered from the CL compared with daily use (P < 0.05), and fewer PMN were recruited compared to sleep without CL wear (P < 0.05). Overnight CL wear may inhibit physiological PMN recruitment to the cornea by preventing their access, by modifying the chemotactic signal or by altering the activation state of the recruited cells.

  7. Kinetics of the inhibition of human leucocyte elastase by eglin from the leech Hirudo medicinalis.

    PubMed Central

    Baici, A; Seemüller, U

    1984-01-01

    The rate constants for the inhibition of human leucocyte elastase by eglin from the leech Hirudo medicinalis were determined by using a pre-steady-state kinetic approach. kon and koff for complex-formation and dissociation were 1 X 10(6)M-1 X S-1 and 8 X 10(-4)S-1 respectively. Ki was calculated as the ratio koff/kon = 8 X 10(-10)M, the binding of eglin to elastase was reversible and the inhibition mechanism was of the fully competitive type. The mechanistic properties of the system and the biological significance of the rate constants are discussed. PMID:6562888

  8. Preventing childhood obesity during infancy in UK primary care: a mixed-methods study of HCPs' knowledge, beliefs and practice

    PubMed Central

    2011-01-01

    Background There is a strong rationale for intervening in early childhood to prevent obesity. Over a quarter of infants gain weight more rapidly than desirable during the first six months of life putting them at greater risk of obesity in childhood. However, little is known about UK healthcare professionals' (HCPs) approach to primary prevention. This study explored obesity-related knowledge of UK HCPs and the beliefs and current practice of general practitioners (GPs) and practice nurses in relation to identifying infants at risk of developing childhood obesity. Method Survey of UK HCPs (GPs, practice nurses, health visitors, nursery, community and children's nurses). HCPs (n = 116) rated their confidence in providing infant feeding advice and completed the Obesity Risk Knowledge Scale (ORK-10). Semi-structured interviews with a sub-set of 12 GPs and 6 practice nurses were audio recorded, taped and transcribed verbatim. Thematic analysis was applied using an interpretative, inductive approach. Results GPs were less confident about giving advice about infant feeding than health visitors (p = 0.001) and nursery nurses (p = 0.009) but more knowledgeable about the health risks of obesity (p < 0.001) than nurses (p = 0.009). HCPs who were consulted more often about feeding were less knowledgeable about the risks associated with obesity (r = -0.34, n = 114, p < 0.001). There was no relationship between HCPs' ratings of confidence in their advice and their knowledge of the obesity risk. Six main themes emerged from the interviews: 1) Attribution of childhood obesity to family environment, 2) Infant feeding advice as the health visitor's role, 3) Professional reliance on anecdotal or experiential knowledge about infant feeding, 4) Difficulties with recognition of, or lack of concern for, infants "at risk" of becoming obese, 5) Prioritising relationship with parent over best practice in infant feeding and 6) Lack of shared understanding for dealing with early years

  9. Quantitative effects of a nickel-titanium palatal expander on skeletal and dental structures in the primary and mixed dentition: a preliminary study.

    PubMed

    Ferrario, Virgilio F; Garattini, Giovanna; Colombo, Anna; Filippi, Vittorio; Pozzoli, Silvio; Sforza, Chiarella

    2003-08-01

    The present study analysed the six-month effects of a nickel-titanium (NiTi) palatal expander on the dental and palatal structures of four primary (mean age 5.8 years) and nine mixed dentition children (mean age 8.7 years), with a posterior unilateral crossbite. Standardized dental and palatal landmarks were digitized using a three-dimensional (3D) electromagnetic instrument. Collected data were analysed with geometric-mathematical models. During a six-month interval, the natural growth and development of the dental arches and hard tissue palate was negligible, as assessed in seven control children (two in the primary dentition, mean age 4.4 years; five in the mixed dentition, mean age 7.7 years). In all children the crossbite was completely corrected. Indeed, dental expansion was always more than or corresponded to the palatal expansion. A smoothing of the size-independent (shape) palatal curvature in the transverse plane was observed. No differences in maximum palatal height were noted. Symmetrical derotation of the anchorage teeth in a distal direction occurred in almost all children. The inclination of the facial axis of the clinical crown (FACC) in the anatomical transverse plane of those teeth with differences between dental and palatal expansion always showed significant modifications (vestibular inclination up to 16.7 degrees). The clinical crown height of anchorage teeth remained nearly the same in all patients. No significant modifications in mandibular arch size were observed. The increase in maxillary arch width, especially in younger children, was probably due to a combination of different effects: opening of the midpalatal suture, tipping of the alveolar process, and molar tipping.

  10. The impact of polio eradication on routine immunization and primary health care: a mixed-methods study.

    PubMed

    Closser, Svea; Cox, Kelly; Parris, Thomas M; Landis, R Matthew; Justice, Judith; Gopinath, Ranjani; Maes, Kenneth; Banteyerga Amaha, Hailom; Mohammed, Ismaila Zango; Dukku, Aminu Mohammed; Omidian, Patricia A; Varley, Emma; Tedoff, Pauley; Koon, Adam D; Nyirazinyoye, Laetitia; Luck, Matthew A; Pont, W Frank; Neergheen, Vanessa; Rosenthal, Anat; Nsubuga, Peter; Thacker, Naveen; Jooma, Rashid; Nuttall, Elizabeth

    2014-11-01

    After 2 decades of focused efforts to eradicate polio, the impact of eradication activities on health systems continues to be controversial. This study evaluated the impact of polio eradication activities on routine immunization (RI) and primary healthcare (PHC). Quantitative analysis assessed the effects of polio eradication campaigns on RI and maternal healthcare coverage. A systematic qualitative analysis in 7 countries in South Asia and sub-Saharan Africa assessed impacts of polio eradication activities on key health system functions, using data from interviews, participant observation, and document review. Our quantitative analysis did not find compelling evidence of widespread and significant effects of polio eradication campaigns, either positive or negative, on measures of RI and maternal healthcare. Our qualitative analysis revealed context-specific positive impacts of polio eradication activities in many of our case studies, particularly disease surveillance and cold chain strengthening. These impacts were dependent on the initiative of policy makers. Negative impacts, including service interruption and public dissatisfaction, were observed primarily in districts with many campaigns per year. Polio eradication activities can provide support for RI and PHC, but many opportunities to do so remain missed. Increased commitment to scaling up best practices could lead to significant positive impacts. © The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America.

  11. The Impact of Polio Eradication on Routine Immunization and Primary Health Care: A Mixed-Methods Study

    PubMed Central

    Closser, Svea; Cox, Kelly; Parris, Thomas M.; Landis, R. Matthew; Justice, Judith; Gopinath, Ranjani; Maes, Kenneth; Banteyerga Amaha, Hailom; Mohammed, Ismaila Zango; Dukku, Aminu Mohammed; Omidian, Patricia A.; Varley, Emma; Tedoff, Pauley; Koon, Adam D.; Nyirazinyoye, Laetitia; Luck, Matthew A.; Pont, W. Frank; Neergheen, Vanessa; Rosenthal, Anat; Nsubuga, Peter; Thacker, Naveen; Jooma, Rashid; Nuttall, Elizabeth

    2014-01-01

    Background. After 2 decades of focused efforts to eradicate polio, the impact of eradication activities on health systems continues to be controversial. This study evaluated the impact of polio eradication activities on routine immunization (RI) and primary healthcare (PHC). Methods. Quantitative analysis assessed the effects of polio eradication campaigns on RI and maternal healthcare coverage. A systematic qualitative analysis in 7 countries in South Asia and sub-Saharan Africa assessed impacts of polio eradication activities on key health system functions, using data from interviews, participant observation, and document review. Results. Our quantitative analysis did not find compelling evidence of widespread and significant effects of polio eradication campaigns, either positive or negative, on measures of RI and maternal healthcare. Our qualitative analysis revealed context-specific positive impacts of polio eradication activities in many of our case studies, particularly disease surveillance and cold chain strengthening. These impacts were dependent on the initiative of policy makers. Negative impacts, including service interruption and public dissatisfaction, were observed primarily in districts with many campaigns per year. Conclusions. Polio eradication activities can provide support for RI and PHC, but many opportunities to do so remain missed. Increased commitment to scaling up best practices could lead to significant positive impacts. PMID:24690667

  12. Morphologically mixed chemical-electrical synapses formed by primary afferents in rodent vestibular nuclei as revealed by immunofluorescence detection of connexin36 and vesicular glutamate transporter-1.

    PubMed

    Nagy, J I; Bautista, W; Blakley, B; Rash, J E

    2013-11-12

    Axon terminals forming mixed chemical/electrical synapses in the lateral vestibular nucleus of rat were described over 40 years ago. Because gap junctions formed by connexins are the morphological correlate of electrical synapses, and with demonstrations of widespread expression of the gap junction protein connexin36 (Cx36) in neurons, we investigated the distribution and cellular localization of electrical synapses in the adult and developing rodent vestibular nuclear complex, using immunofluorescence detection of Cx36 as a marker for these synapses. In addition, we examined Cx36 localization in relation to that of the nerve terminal marker vesicular glutamate transporter-1 (vglut-1). An abundance of immunolabeling for Cx36 in the form of Cx36-puncta was found in each of the four major vestibular nuclei of adult rat and mouse. Immunolabeling was associated with somata and initial dendrites of medium and large neurons, and was absent in vestibular nuclei of Cx36 knockout mice. Cx36-puncta were seen either dispersed or aggregated into clusters on the surface of neurons, and were never found to occur intracellularly. Nearly all Cx36-puncta were localized to large nerve terminals immunolabeled for vglut-1. These terminals and their associated Cx36-puncta were substantially depleted after labyrinthectomy. Developmentally, labeling for Cx36 was already present in the vestibular nuclei at postnatal day 5, where it was only partially co-localized with vglut-1, and did not become fully associated with vglut-1-positive terminals until postnatal day 20-25. The results show that vglut-1-positive primary afferent nerve terminals form mixed synapses throughout the vestibular nuclear complex, that the gap junction component of these synapses contains Cx36, that multiple Cx36-containing gap junctions are associated with individual vglut-1 terminals and that the development of these mixed synapses is protracted over several postnatal weeks.

  13. Methodological and practical viewpoints of qualitative-driven mixed method design: the case of decentralisation of primary healthcare services in Nepal.

    PubMed

    Regmi, Krishna

    2017-09-11

    Although considerable attention has been paid to the use of quantitative methods in health research, there has been limited focus on decentralisation research using a qualitative-driven mixed method design. Decentralisation presents both a problematic concept and methodological challenges, and is more context-specific and is often multi-dimensional. Researchers often consider using more than one method design when researching phenomena is complex in nature. Aim To explore the effects of decentralisation on the provision of primary healthcare services. Qualitative-driven mixed method design, employing three methods of data collections: focus group discussions (FGDs), semi-structured interviews (SSIs) and participant observations under two components, that is, core component and supplementary components were used. Four FGDs with health service practitioners, three FGDs with district stakeholders, 20 SSIs with health service users and 20 SSIs with national stakeholders were carried out. These were conducted sequentially. NVivo10, a data management program, was utilised to code the field data, employing a content analysis method for searching the underlying themes or concepts in the text material. Findings Both positive and negative experiences related to access, quality, planning, supplies, coordination and supervision were identified. This study suggests some evidence of the effects of decentralisation on health outcomes in general, as well as filling a gap of understanding and examining healthcare through a qualitative-driven mixed methods approach, in particular. Future research in the area of qualitative in-depth understanding of the problems (why decentralisation, why now and what for) would provoke an important data set that benefits the researchers and policy-makers for planning and implementing effective health services.

  14. Morphologically mixed chemical-electrical synapses formed by primary afferents in rodent vestibular nuclei as revealed by immunofluorescence detection of connexin36 and vesicular glutamate transporter-1

    PubMed Central

    Nagy, James I.; Bautista, Wendy; Blakley, Brian; Rash, John E.

    2013-01-01

    Axon terminals forming mixed chemical/electrical synapses in the lateral vestibular nucleus of rat were described over forty years ago. Because gap junctions formed by connexins are the morphological correlate of electrical synapses, and with demonstrations of widespread expression of the gap junction protein connexin36 (Cx36) in neurons, we investigated the distribution and cellular localization of electrical synapses in the adult and developing rodent vestibular nuclear complex, using immunofluorescence detection of Cx36 as a marker for these synapses. In addition, we examined Cx36 localization in relation to that of the nerve terminal marker vesicular glutamate transporter-1 (vglut-1). An abundance of immunolabelling for Cx36 in the form of Cx36-puncta was found in each of the four major vestibular nuclei of adult rat and mouse. Immunolabelling was associated with somata and initial dendrites of medium and large neurons, and was absent in vestibular nuclei of Cx36 knockout mice. Cx36-puncta were seen either dispersed or aggregated into clusters on the surface of neurons, and were never found to occur intracellularly. Nearly all Cx36-puncta were localized to large nerve terminals immunolabelled for vglut-1. These terminals and their associated Cx36-puncta were substantially depleted after labyrinthectomy. Developmentally, labelling for Cx36 was already present in the vestibular nuclei at postnatal day 5, where it was only partially co-localized with vglut-1, and did not become fully associated with vglut-1-positive terminals until postnatal day 20 to 25. The results show that vglut-1-positive primary afferent nerve terminals form mixed synapses throughout the vestibular nuclear complex, that the gap junction component of these synapses contain Cx36, that multiple Cx36-containing gap junctions are associated with individual vglut-1 terminals and that the development of these mixed synapses is protracted over several postnatal weeks. PMID:23912039

  15. Using theory to improve low back pain care in Australian Aboriginal primary care: a mixed method single cohort pilot study.

    PubMed

    Lin, Ivan B; Coffin, Juli; O'Sullivan, Peter B

    2016-04-12

    Low back pain (LBP) care is frequently discordant with research evidence. This pilot study evaluated changes in LBP care following a systematic, theory informed intervention in a rural Australian Aboriginal Health Service. We aimed to improve three aspects of care; reduce inappropriate LBP radiological imaging referrals, increase psychosocial oriented patient assessment and, increase the provision of LBP self-management information to patients. Three interventions to improve care were developed using a four-step systematic implementation approach. A mixed methods pre/post cohort design evaluated changes in the three behaviours using a clinical audit of LBP care in a six month period prior to the intervention and then following implementation. In-depth interviews elicited the perspectives of involved General Practitioners (GPs). Qualitative analysis was guided by the theoretical domains framework. The proportion of patients who received guideline inconsistent imaging referrals (GICI) improved from 4.1 GICI per 10 patients to 0.4 (95% CI for decrease in rate: 1.6 to 5.6) amongst GPs involved in the intervention. Amongst non-participating GPs (locum/part-time GPs who commenced post-interventions) the rate of GICI increased from 1.5 to 4.4 GICI per 10 patients (95 % CI for increase in rate: .5 to 5.3). There was a modest increase in the number of patients who received LBP self-management information from participating GPs and no substantial changes to psychosocial oriented patient assessments by any participants; however GPs qualitatively reported that their behaviours had changed. Knowledge and beliefs about consequences were important behavioural domains related to changes. Environmental and resource factors including protocols for locum staff and clinical tools embedded in patient management software were future strategies identified. A systematic intervention model resulted in partial improvements in LBP care. Determinants of practice change amongst GPs were

  16. Activity of some lysosomal enzymes in plasma and leucocytes of rabbits exposed to effect of retinol and hydrocortisone.

    PubMed

    Schmidt, M; Kołataj, A; Bulla, J; Król, T; Witek, B; Banasik, A

    1992-01-01

    Observing activity of some lysosomal enzymes in blood serum and leucocytes of rabbits subjected to injection of 200,000 units of retinol and 25 mg of hydrocortisone/kg of body weight it was found that: 1. In the effect of retinol administration there was an increase in the activity AP, BGAL, BGLU, AspAT and lipase in blood serum after 72 hours and NAGL after 168 hours while in leucocytes BGAL and NAGL after 72 hours and AGAL after 168 hours. 2. As a result of hydrocortisone injection the activity of all the enzymes examined (except Ala-Na) in blood serum increased markedly already after 24-48 hours. 3. In leucocytes hydrocortisone caused a significant increase in the activity of AP, BGRD, NAGL, BGAL, AGAL and cathepsin D. 4. The glucose level in blood plasma decreased after 48 hours and 120 hours after hydrocortisone injection and 168 hours after retinol injection.

  17. In vivo treatment with progestogens causes immunosuppression of carp Cyprinus carpio leucocytes by affecting nitric oxide production and arginase activity.

    PubMed

    Pietsch, C; Neumann, N; Preuer, T; Kloas, W

    2011-07-01

    In this study, carp Cyprinus carpio were injected with various steroid compounds, including synthetic and natural progestogens and the glucocorticoid cortisol, to investigate effects on leucocytes isolated from their kidneys. Injection of cortisol led to an increased spleeno-somatic index (I(S)) on day 21 post-injection (pi) and immunosuppressive effects measured as decreased nitric oxide (NO) production and increased arginase activity in isolated leucocytes on days 14 and 21 pi, respectively. Moreover, reduced NO production was also observed after injection of the synthetic progestogens, levonorgestrel (LEV) and medroxyprogesterone acetate. In addition, LEV influenced arginase activity in head kidney cells on day 14 and day 21 pi. This study is the first demonstration in fishes that the application of these steroid compounds in vivo affects NO production and arginase activity of isolated leucocytes.

  18. An automated image analysis system can be beneficial in preclassification of leucocytes in children with hematological disease.

    PubMed

    Portakal, Oytun; Tavil, Betul; Kuşkonmaz, Barış; Aytaç, Selin; Hasçelik, Gülşen

    2011-01-01

    This study was aimed to evaluate the analytical performance of an automated image analysis system (a pilot model of Diff Master(™) Octavia) for the preclassification of leucocytes in children with hematological disease. Manual microscopy performed by pediatric hematologists was used as the reference method. Five mature cell class and blasts were evaluated. Diff Master Octavia correctly preclassified 87.4% of all leucocytes with a high reproducibility. The overall accuracy was found to be 93.0%. Clinical sensitivity was 97.7% and specificity was 76.0%. The average time per slide for Diff Master(™) Octavia was 2.3  min lower than that of manual method. Our results indicated that the Diff Master(™) Octavia can detect and preclassify leucocytes accurately; therefore, it can be used as an efficient and fast method in pediatric hematology routine.

  19. Sustainability and scalability of a volunteer-based primary care intervention (Health TAPESTRY): a mixed-methods analysis.

    PubMed

    Kastner, Monika; Sayal, Radha; Oliver, Doug; Straus, Sharon E; Dolovich, Lisa

    2017-08-01

    Chronic diseases are a significant public health concern, particularly in older adults. To address the delivery of health care services to optimally meet the needs of older adults with multiple chronic diseases, Health TAPESTRY (Teams Advancing Patient Experience: Strengthening Quality) uses a novel approach that involves patient home visits by trained volunteers to collect and transmit relevant health information using e-health technology to inform appropriate care from an inter-professional healthcare team. Health TAPESTRY was implemented, pilot tested, and evaluated in a randomized controlled trial (analysis underway). Knowledge translation (KT) interventions such as Health TAPESTRY should involve an investigation of their sustainability and scalability determinants to inform further implementation. However, this is seldom considered in research or considered early enough, so the objectives of this study were to assess the sustainability and scalability potential of Health TAPESTRY from the perspective of the team who developed and pilot-tested it. Our objectives were addressed using a sequential mixed-methods approach involving the administration of a validated, sustainability survey developed by the National Health Service (NHS) to all members of the Health TAPESTRY team who were actively involved in the development, implementation and pilot evaluation of the intervention (Phase 1: n = 38). Mean sustainability scores were calculated to identify the best potential for improvement across sustainability factors. Phase 2 was a qualitative study of interviews with purposively selected Health TAPESTRY team members to gain a more in-depth understanding of the factors that influence the sustainability and scalability Health TAPESTRY. Two independent reviewers coded transcribed interviews and completed a multi-step thematic analysis. Outcomes were participant perceptions of the determinants influencing the sustainability and scalability of Health TAPESTRY. Twenty

  20. Flow cytometry assay for intracellular detection of Infectious Pancreatic Necrosis virus (IPNV) in Atlantic salmon (Salmo salar L.) leucocytes.

    PubMed

    Rønneseth, Anita; Pettersen, Eirin Fausa; Wergeland, Heidrun I

    2012-12-01

    Infectious Pancreatic Necrosis virus (IPNV) is traditionally detected in adherent leucocytes using immunofluorescence labelled specific antibodies, PCR or by further cultivation of infected cell material in cell lines. We present a flow cytometry (FCM) assay for detection of intracellular IPNV in salmon leucocytes, where each single cell is analysed for presence of virus. The method is established using in vitro challenge of salmon leucocytes and CHSE-214 cells. For detection of intracellular virus antigen the Cytofix/Cytoperm kit from BD is optimal compared with paraformaldehyde or acetone/methanol for cell permeabilisation. This is combined with labelling procedures allowing both internal virus antigen labelling and external antibody labelling of cell markers to identify B-cells and neutrophils. The secondary antibodies were Alexa Fluor 647 for the internal labelling and RPE for the external labelling of bound cell subtype specific antibodies. The presences of virus within cells are also demonstrated by confocal and light microscopy of infected cells. IPNV is successfully detected in blood and head kidney leucocyte samples. IPNV is found both in B-cells and neutrophils as well as in other types of leucocytes that could not be identified due to lack of cell-specific antibodies. Serial samples from cultivation of in vitro infected leucocytes and CHSE-214 cells analysed by flow cytometry showed that number of infected cells increased with increasing number of days. The flow cytometry protocol for detection of intracellular IPNV is verified using CHSE-214 cells persistently infected with IPNV. These analyses are compared with virus titre and virus infected naive CHSE-214 cells. The detection of IPNV in persistently infected cells indicates that carrier fish can be analysed, as such cells are considered to have virus titres similar to carriers.

  1. Effects of eprosartan on mitochondrial membrane potential and H2O2 levels in leucocytes in hypertension.

    PubMed

    Labiós, M; Martínez, M; Gabriel, F; Guiral, V; Ruiz-Aja, S; Beltrán, B; Muñoz, A

    2008-07-01

    We investigated whether circulating leucocytes from hypertensive patients exhibit more spontaneous, stimulated hydrogen peroxide (H2O2) production and greater mitochondrial membrane potential (Deltapsi) than those from normotensive individuals. We also investigated the effects of oral treatment with the angiotensin II (AT II) type 1 receptor blocker eprosartan (600 mg day(-1)) on these markers of oxidative stress. In 25 hypertensive patients and 28 healthy volunteers, spontaneous H2O2 formation was measured by flow cytometry after preincubation of buffy coat-leucocytes from fresh peripheral venous blood at 37 degrees C with 2',7' dichlorofluorescein. Stimulation of H2O2 formation by circulating leucocytes was elicited by the addition of tert-butylhydroperoxide (tBHP). Deltapsi was determined by flow cytometry after the addition of tetramethylrhodamine methyl ester (TMRM). Compared with healthy individuals, lymphocytes from hypertensive patients exhibited higher Deltapsi (12.28+/-3.20 vs 16.25+/-2.88 arbitrary fluorescence units (AFU), respectively; P<0.001) and greater spontaneous H2O2 production (4.75+/-5.15 vs 8.98+/-9.97 AFU, respectively; P<0.05). tBHP stimulation was associated with higher H2O2 levels in circulating leucocytes in patients with uncorrected hypertension than in normotensive individuals. H2O2 overproduction was corrected by eprosartan treatment. These results suggest that oxidative stress could be important in the pathogenesis of hypertension. Furthermore, measurement of leucocyte oxidant activities may be useful for the evaluation of oxidative stress, which may be reduced with the use of antihypertensive drugs. Our results demonstrate that treatment of hypertension with eprosartan normalizes blood pressure and corrects oxidative disturbances, suggesting that leucocytes could be a target for this drug.

  2. Melatonin counteracts alterations in oxidative metabolism and cell viability induced by intracellular calcium overload in human leucocytes: changes with age.

    PubMed

    Espino, Javier; Bejarano, Ignacio; Paredes, Sergio D; González, David; Barriga, Carmen; Reiter, Russel J; Pariente, José A; Rodríguez, Ana B

    2010-07-01

    Ageing is associated with an increased production of free radicals and alterations in the mechanisms of adaptation to oxidative stress. In fact, the free radical theory of ageing proposes that deleterious actions of free radicals are responsible for the functional deterioration associated with ageing. Moreover, a close relationship exists between calcium homeostasis and oxidative stress. The current work was aimed at proving that intracellular calcium overload induced by N-formyl-methionyl-leucyl-phenylalanine (FMLP) and/or thapsigargin leads to oxidative stress. We additionally examined the effect of melatonin on the levels of reactive oxygen species (ROS) and cell viability in human leucocytes collected from young (20-30-year-old) and elderly (65-75-year-old) individuals under both basal and oxidative stress-induced conditions. Treatments with 10 nM FMLP and/or 1 microM thapsigargin induced a transient increase in cytosolic free-calcium concentration ([Ca(2 + )](c)) in human leucocytes due to calcium release from internal stores, and led in turn to oxidative stress, as assessed by intracellular ROS measurement. Non-treated leucocytes from aged individuals exhibited higher ROS levels and lower rates of cell survival when compared to leucocytes from young individuals. Similar results were obtained in FMLP and/or thapsigargin-treated leucocytes from elderly individuals when compared to those from the young individuals. Melatonin treatment significantly reduced both hydrogen peroxide (H(2)O(2)) and superoxide anion levels, likely due to its free-radical scavenging properties, and enhanced leucocyte viability in both age groups. Therefore, melatonin may be a useful tool for the treatment of disease states and processes where an excessive production of oxidative damage occurs.

  3. Variability in prescription drug expenditures explained by adjusted clinical groups (ACG) case-mix: a cross-sectional study of patient electronic records in primary care.

    PubMed

    Aguado, Alba; Guinó, Elisabet; Mukherjee, Bhramar; Sicras, Antoni; Serrat, Josep; Acedo, Mateo; Ferro, Juan Jose; Moreno, Victor

    2008-03-04

    In view of rapidly increasing prescription costs, case-mix adjustment should be considered for effective control of costs. We have estimated the variability in pharmacy costs explained by ACG in centers using patient electronic records, profiled centers and physicians and analyzed the correlation between cost and quality of prescription. We analyzed 65,630 patient records attending five primary care centers in Spain during 2005. Variables explored were age, gender, registered diagnosed episodes of care during 2005, total cost of prescriptions, physician and center. One ACG was assigned to each patient with ACG case-mix software version 7.1. In a two-part model, logistic regression was used to explain the incurrence of drug expenditure at the first stage and a linear mixed model that considered the multilevel structure of data modeled the cost, conditional upon incurring any expense. Risk and efficiency indexes in pharmacy cost adjusted for ACG were obtained for centers and physicians. Spearman rank correlation between physician expenditure, adjusted for ACG, and a prescription quality index was also obtained. Pediatric and adult data were analyzed separately. No prescription was recorded for 13% of adults and 39.6% of children. The proportion of variance of the incurrence of expenditure explained by ACGs was 0.29 in adults and 0.21 in children. For adults with prescriptions, the variance of cost explained by ACGs was 35.4%, by physician-center was 1.8% and age 10.5% (residual 52.3%). For children, ACGs explained 22.4% of cost and physician-center 10.9% (residual 66.7%). Center efficiency index for adults ranged 0.58 to 1.22 and for children 0.32 to 2.36. Spearman correlation between expenditure and prescription quality index was -0.36 in family physicians (p = 0.019, N = 41) and -0.52 in pediatricians (p = 0.08, N = 12). In our setting, ACG is the variable studied that explains more variability in pharmacy cost in adults compared to physician and center. In

  4. Establishment of a cell line from leucocytes of a cow with chronic lymphocytic leukemia.

    PubMed

    Adomaitiene, D; Tamosiunas, V; Mauricas, M; Surovas, V; Markevicius, A

    1983-07-01

    A cell line was established from blood leucocytes of a cow with chronic lymphocytic leukemia. The leucocytes were cultured with conditioned medium (culture fluid of mouse cell line L). In vitro cell transformation was demonstrated by adaptation to permanent growth, modification of cell morphology, the alteration of cell surface phenotype, kinetic behaviour and the loss of the euploid stability of the cell karyotype. Ultrastructural studies showed rather a uniform cell pattern in a culture population heterogeneous for degree of cell vacuolization. A wide variation in the expression of surface markers in cells was demonstrated by E-, EA- and EAC-rosetting. In suspension culture the cell population was found to be sIg negative. Expression of leukemia-associated antigens by a fraction of the cultured cells was evidenced by a cytotoxic technique using complement and heterologous antisera against bovine leukemic lymphocytes, absorbed with normal lymphoid cells. Virus-like particles and BLV antigens were not identified. Culture cells failed to show spontaneous or antibody-dependent killer cytotoxicity. Comparison with blood lymphocytes of healthy and leukemic cattle was done. The established culture should be useful as a model for experimental immunology and oncology.

  5. Isolation of various canine leucocytes and their characterization by surface marker analysis.

    PubMed Central

    Ho, C K; Babiuk, L A

    1978-01-01

    Various techniques were used to separate canine peripheral blood leucocytes into populations enriched in lymphocytes, polymorphonuclear leucocytes, phagocytic mononuclear cells (monocytes) and macrophages. Surface markers on each cell population were determined by rosette formation. Fc receptors for IgG and complement receptors (C3b and C3d) were present on PMN, monocytes, macrophages as well as on a sub-population of lymphocytes. Purification of the lymphocytes into T-and B-cell-enriched populations revealed that these receptors were present only on the B lymphocytes and not on the T lymphocytes. In addition, a third lymphocyte population, which did not possess surface immunoglobulin, and Fc receptor but not the complement receptor. None of the cell populations exhibited C4 complement receptors or Fc receptors for IgM. When different cell populations were tested for their ability to form rosettes directly with human type 'O' red blood cells it was found that most populations could rosette, suggesting that this technique could not be used as a specific marker for canine T lymphocytes. PMID:309854

  6. Cytokine expression in leucocytes and gut cells of rainbow trout, Oncorhynchus mykiss Walbaum, induced by probiotics.

    PubMed

    Kim, Do-Hyung; Austin, Brian

    2006-12-15

    Understanding how the various host cells respond to probiotic bacteria in vitro may provide important insight into elaborate immune responses triggered by beneficial bacteria. The aim of this study was to investigate the detailed pattern of the mRNA expression of cytokines (IL-1beta, IL-8, TNF-alpha and TGF-beta) in head kidney (HK) leucocytes and gut cells isolated from rainbow trout (Oncorhynchus mykiss Walbaum) after co-culturing with live probiotics. HK leucocytes and gut cells adjusted to 5 x 10(6) and 2 x 10(6) ml(-1), respectively, in L-15 medium containing 25% decomplemented FCS and 300 mg l(-1) L-glutamine were co-cultured with Carnobacterium maltaromaticum B26 and C. divergens B33 at an multiplicity of infection of 25 for 6 and 12 h. Quantitative real-time reverse transcriptase polymerase chain reaction using SYBR Green I was employed to determine the mRNA expression of studied genes. Although neither probiotic strains significantly induced mRNA of the cytokines in gut cells, expression ratios of IL-1beta and TNF-alpha of HK cells were significantly higher, suggesting that these bacteria can stimulate innate immunity in rainbow trout.

  7. Somatotropic gene response to recombinant growth hormone treatment in buffalo leucocytes.

    PubMed

    Castigliego, Lorenzo; Li, Xiao-Ning; Armani, Andrea; Razzano, Maria; Mazzi, Marco; Rosati, Remo; Gianfaldoni, Daniela; Guidi, Alessandra

    2011-12-01

    The use of recombinant bovine growth hormone (rbGH) to increase milk yield in cows is banned in some countries. In others, where it is authorised, it has triggered harsh debates on labelling of dairy products. If many studies have been performed on bovines, there is a lack of information on buffaloes, which are sometimes treated with rbGH and re-present an important economical resource for dairy products in some countries. Analytical methods with legal value for surveillance of rbGH treatments do not yet exist. Research on gene expression biomarkers is one of the most promising approaches to this purpose. For this reason, we treated five buffaloes for 10 weeks with a sustained-release formulation of rbGH and analysed the response of 20 somatotropic axis genes in leucocytes by real-time polymerase chain reaction. Overall changes in gene expression levels were of low magnitude and sometimes affected by the 'time' factor. Only the IGFBP-1 gene showed a significant under-expression (about two-fold; p <0.001) in treated animals. Taken together, these results give evidence that expression analysis of the somatotropic axis genes in leucocytes is little helpful for discrimination of rbGH-treated buffaloes, but do not exclude that another array of genes could provide useful patterns of variation.

  8. Response of peripheral blood leucocytes to mitogenic factor(s) in porcine seminal plasma.

    PubMed

    Kovacs, D P; Tekpetey, F R; Armstrong, D T

    1994-04-01

    Porcine seminal plasma (PSP) contains a potent mitogenic substance capable of causing proliferation and extensive agglutination in cultured porcine peripheral blood leucocytes (PBL). In order to determine the specific leucocyte cell population affected by this mitogen, lymphocyte, monocyte and polymorphonuclear (PMN)-enriched cell fractions were separated from PBL and treated with increasing concentrations of PSP (1-8%, v/v). For monocyte cell populations enriched through elutriation and adherence to plastic, cell proliferation in response to PSP treatment was consistently higher than PSP-treated unfractionated PBL. Monocyte-depleted PBL and lymphocyte populations enriched through elutriation demonstrated a decrease in proliferation compared to PSP-treated unfractionated PBL. PMN populations separated from PBL by discontinuous Percoll gradient centrifugation were unresponsive to PSP. Agglutination was observed for the unfractionated PBL and each enriched cell population. These results demonstrate that PSP contains a potent mitogen which induces proliferation in monocyte-enriched cell populations and may reflect the potential of PSP to act as an immune regulator in the uterine environment during early embryo development and implantation.

  9. Sex differences in leucocyte telomere length in a free-living mammal.

    PubMed

    Watson, Rebecca L; Bird, Ellen J; Underwood, Sarah; Wilbourn, Rachael V; Fairlie, Jennifer; Watt, Kathryn; Salvo-Chirnside, Eliane; Pilkington, Jill G; Pemberton, Josephine M; McNeilly, Tom N; Froy, Hannah; Nussey, Daniel H

    2017-06-01

    Mounting evidence suggests that average telomere length reflects previous stress and predicts subsequent survival across vertebrate species. In humans, leucocyte telomere length (LTL) is consistently shorter during adulthood in males than in females, although the causes of this sex difference and its generality to other mammals remain unknown. Here, we measured LTL in a cross-sectional sample of free-living Soay sheep and found shorter telomeres in males than in females in later adulthood (>3 years of age), but not in early life. This observation was not related to sex differences in growth or parasite burden, but we did find evidence for reduced LTL associated with increased horn growth in early life in males. Variation in LTL was independent of variation in the proportions of different leucocyte cell types, which are known to differ in telomere length. Our results provide the first evidence of sex differences in LTL from a wild mammal, but longitudinal studies are now required to determine whether telomere attrition rates or selective disappearance are responsible for these observed differences. © 2016 The Authors. Molecular Ecology Published by John Wiley & Sons Ltd.

  10. Increased leucocyte Na-K ATPase in obesity: reversal following weight loss

    SciTech Connect

    Turaihi, K.; Baron, D.N.; Dandona, P.

    1987-09-01

    Ouabain-sensitive /sup 86/Rb influx and (/sup 3/H) ouabain binding capacity were investigated in the leucocytes of 17 obese patients and 15 control subjects. Both were significantly increased in the obese when compared with controls. Following dietary restriction and a 4% to 5% weight reduction in the obese over 2 weeks, (/sup 3/H) ouabain binding and ouabain-sensitive /sup 86/Rb influx (a model for K+ influx) decreased to levels similar to those in controls. This shows that the number of Na-K ATPase sites on leucocyte membranes of the obese are significantly increased and that this is associated with accelerated /sup 86/Rb transport. Since both of these indices decreased following 4% to 5% reduction in body weight while the patients were still obese, increased Na-K ATPase is neither a marker of nor cardinal to the pathogenesis of obesity. We conclude that (1) increase in Na-K ATPase units and /sup 86/Rb influx are not characteristic of obesity itself and (2) dietary restriction over the short-term with limited weight reduction restores Na-K ATPase units and /sup 86/Rb influx to normal.

  11. Effects of fluoxetine on the oxidative status of peripheral blood leucocytes of restraint-stressed mice.

    PubMed

    Novío, Silvia; Núñez, María Jesús; Amigo, Gonzalo; Freire-Garabal, Manuel

    2011-11-01

    Emotional stress can be viewed as a cause of adverse circumstances that induces a wide range of biochemical and behavioural changes. Oxidative stress is a critical route of damage in various psychological stress-induced disorders such as depression. Antidepressants are widely prescribed to treat these conditions; however, no animal study has investigated the effect of selective serotonin reuptake inhibitors (SSRIs) on the levels of intracellular reactive oxygen species in peripheral blood leucocytes of stressed mice. In this study, mice were immobilized for a period of 6 hr. Fluoxetine (5 mg/kg of body-weight) was administered 30 min. before subjecting the animals to acute stress. The level of intracellular reactive oxygen species in leucocytes of the peripheral blood of stressed mice was investigated using a 2',7'-dichlorofluorescein diacetate probe, and the antioxidant response of fluoxetine was evaluated by superoxide dismutase, diaphorase, catalase and reduced glutathione. Our results show that restraint stress significantly increases the generation of reactive oxygen species in the peripheral defence cells. Treatment with fluoxetine partially reverses the adverse effects of stress. The improvement in cellular oxidative status may be an important mechanism underlying the protective pharmacological effects of fluoxetine, which are clinically observed in the treatment of depressive disorders.

  12. Mice overexpressing p40 in lungs have reduced leucocyte influx and slightly impaired resistance during tuberculosis

    PubMed Central

    Leemans, Jaklien C; Wieland, Catharina W; Florquin, Sandrine; van der Poll, Tom; Vervoordeldonk, Margriet J B M

    2006-01-01

    Interleukin (IL)-12 (p70) is a heterodimeric cytokine composed of p40 and p35, that plays a major role in the protective immune response to Mycobacterium tuberculosis. To define the role of p40 in lungs during pulmonary M. tuberculosis infection we generated transgenic (Tg) mice overexpressing p40 under control of the surfactant protein C promoter. Tg mice expressed the transgene in their lungs, yet demonstrated elevated pulmonary p40 protein levels. After infection, Tg mice displayed higher pulmonary p40 and p70 levels than wild type mice. Interferon-γ concentrations were similar in uninfected and infected Tg and wild type mice, arguing against agonistic effects of p40. Tg mice demonstrated reduced recruitment of macrophages, lymphocytes and neutrophils to the lungs early after infection. This was accompanied by reduced pulmonary tumour necrosis factor-α, macrophage inflammatory protein (MIP)-2 and MIP-1 α levels. This suggests that elevated p40 concentrations inhibited the chemotactic effects of p70 on leucocytes. Furthermore, Tg mice displayed slightly higher pulmonary mycobacterial outgrowth late in the infection than wild type mice. Taken together, we demonstrate that constitutive overexpression of p40 in lungs negatively influences IL-12-mediated leucocyte migration and protection against lung tuberculosis. This suggests a novel antagonistic role for p40 homodimers in regulating the chemotactic bioactivity of IL-12 after pulmonary mycobacterial infection. PMID:16476061

  13. Relationship between somatic cell count, polymorphonuclear leucocyte count and quality parameters in bovine bulk tank milk.

    PubMed

    Wickström, Erik; Persson-Waller, Karin; Lindmark-Månsson, Helena; Ostensson, Karin; Sternesjö, Ase

    2009-05-01

    The somatic cell count (SCC) in bovine bulk tank milk is presently used as an indicator of raw milk quality, reflecting the udder health status of the herd. During mastitis, SCC increases, mostly owing to an influx of polymorphonuclear leucocytes (PMN) from blood into milk, with a concomitant change in milk composition. Bulk tank milk samples were categorized according to their SCC, as well as polymorphonuclear leucocyte count (PMNC), to study relationships between SCC, PMNC and various raw milk quality traits, i.e. contents of total protein, whey protein, casein, fat and lactose, casein number, proteolysis and rheological properties. The proportion of PMN, obtained by direct microscopy, was significantly higher in samples with high SCC compared with low SCC samples. SCC and PMNC were strongly correlated, yielding a correlation coefficient of 0.85. High SCC samples had lower lactose and casein contents, lower casein number and more proteolysis than low SCC samples. Samples with high PMNC had a lower casein number than low PMNC samples. Samples with high and low SCC or PMNC did not differ in respect to rheological properties. Our results do not indicate that PMNC is a better biomarker than SCC for raw bulk tank milk quality, as previously proposed.

  14. A comparison of the efficacy of organic and mixed-organic polymers with polyaluminium chloride in chemically assisted primary sedimentation (CAPS).

    PubMed

    De Feo, G; Galasso, M; Landi, R; Donnarumma, A; De Gisi, S

    2013-01-01

    CAPS is the acronym for chemically assisted primary sedimentation, which consists of adding chemicals to raw urban wastewater to increase the efficacy of coagulation, flocculation and sedimentation. The principal benefits of CAPS are: upgrading of urban wastewater treatment plants; increasing efficacy of primary sedimentation; and the major production of energy from the anaerobic digestion of primary sludge. Metal coagulants are usually used because they are both effective and cheap, but they can cause damage to the biological processes of anaerobic digestion. Generally, biodegradable compounds do not have these drawbacks, but they are comparatively more expensive. Both metal coagulants and biodegradable compounds have preferential and penalizing properties in terms of CAPS application. The problem can be solved by means of a multi-criteria analysis. For this purpose, a series of tests was performed in order to compare the efficacy of several organic and mixed-organic polymers with that of polyaluminium chloride (PACl) under specific conditions. The multi-criteria analysis was carried out coupling the simple additive weighting method with the paired comparison technique as a tool to evaluate the criteria priorities. Five criteria with the following priorities were used: chemical oxygen demand (COD) removal > turbidity, SV60 > coagulant dose, and coagulant cost. The PACl was the best alternative in 70% of the cases. The CAPS process using PACl made it possible to obtain an average COD removal of 68% compared with 38% obtained, on average, with natural sedimentation and 61% obtained, on average, with the best PACl alternatives (cationic polyacrylamide, natural cationic polymer, dicyandiamide resin).

  15. Adaptive capacity of the Adjusted Clinical Groups Case-Mix System to the cost of primary healthcare in Catalonia (Spain): a observational study

    PubMed Central

    Velasco-Velasco, Soledad; Navarro-Artieda, Ruth; Prados-Torres, Alexandra; Bolibar-Ribas, Buenaventura; Violan-Fors, Concepción

    2012-01-01

    Objectives To describe the adaptive capacity of the Adjusted Clinical Groups (ACG) system to the cost of care in primary healthcare centres in Catalonia (Spain). Design Retrospective study (multicentres) conducted using computerised medical records. Setting 13 primary care teams in 2008 were included. Participants All patients registered in the study centres who required care between 1 January and 31 December 2008 were finally studied. Patients not registered in the study centres during the study period were excluded. Outcome measures Demographic (age and sex), dependent (cost of care) and case-mix variables were studied. The cost model for each patient was established by differentiating the fixed and variable costs. To evaluate the adaptive capacity of the ACG system, Pearson's coefficient of variation and the percentage of outliers were calculated. To evaluate the explanatory power of the ACG system, the authors used the coefficient of determination (R2). Results The number of patients studied was 227 235 (frequency: 5.9 visits per person per year), with a mean of 4.5 (3.2) episodes and 8.1 (8.2) visits per patient per year. The mean total cost was €654.2. The explanatory power of the ACG system was 36.9% for costs (56.5% without outliers). 10 ACG categories accounted for 60.1% of all cases and 19 for 80.9%. 5 categories represented 71% of poor performance (N=78 887, 34.7%), particularly category 0300-Acute Minor, Age 6+ (N=26 909, 11.8%), which had a coefficient of variation =139% and 6.6% of outliers. Conclusions The ACG system is an appropriate manner of classifying patients in routine clinical practice in primary healthcare centres in Catalonia, although improvements to the adaptive capacity through disaggregation of some categories according to age groups and, especially, the number of acute episodes in paediatric patients would be necessary to reduce intra-group variation. PMID:22734115

  16. Type I allergic hypersensitivity reactions due to ethylene oxide sterilised leucocyte filters in patients with thalassaemia: report of four cases

    PubMed Central

    Belen, Burcu; Polat, Meltem

    2015-01-01

    Ethylene oxide (EO) is a highly reactive gas used in sterilisation of heat sensitive medical devices, such as infusion sets, cannulae, intubation materials, ventriculoperitoneal shunts, dialysis catheters and stents. Allergic reactions due to EO have been reported in haemodialysis patients, patients undergoing extracorporeal photopheresis and donors of plasmapheresis. Clinical manifestations vary considerably and generally do not allow differentiation between IgE-mediated anaphylaxis and anaphylactoid reactions. We report four patients with thalassaemia who experienced anaphylaxis during transfusion due to ethylene oxide sterilised leucocyte filters. The aim of this report is to highlight the fact that frequently transfused patients can have allergic reactions due to EO particles left in leucocyte filters. PMID:25725028

  17. Identification of cultural determinants of antibiotic use cited in primary care in Europe: a mixed research synthesis study of integrated design "Culture is all around us".

    PubMed

    Touboul-Lundgren, Pia; Jensen, Siri; Drai, Johann; Lindbæk, Morten

    2015-09-17

    Inappropriate antibiotic prescribing, particularly for respiratory tract infections (RTI) in ambulatory care, has become a worldwide public health threat due to resulting antibiotic resistance. In spite of various interventions and campaigns, wide variations in antibiotic use persist between European countries. Cultural determinants are often referred to as a potential cause, but are rarely defined. To our knowledge, so far no systematic literature review has focused on cultural determinants of antibiotic use. The aim of this study was to identify cultural determinants, on a country-specific level in ambulatory care in Europe, and to describe the influence of culture on antibiotic use, using a framework of cultural dimensions. A computer-based systematic literature review was conducted by two research teams, in France and in Norway. Eligible publications included studies exploring antibiotic use in primary care in at least two European countries based on primary study results, featuring a description of cultural determinants, and published between 1997 and 2015. Quality assessment was conducted independently by two researchers, one in each team, using appropriate checklists according to study design. Each included paper was characterized according to method, countries involved, sampling and main results, and cultural determinants mentioned in each selected paper were extracted, described and categorized. Finally, the influence of Hofstede's cultural dimensions associated with antibiotic consumption within a primary care setting was described. Among 24 eligible papers, 11 were rejected according to exclusion criteria. Overall, 13 papers meeting the quality assessment criteria were included, of which 11 used quantitative methods and two qualitative or mixed methods. The study participants were patients (nine studies) and general practitioners (two studies). This literature review identified various cultural determinants either patient-related (illness perception

  18. Endotoxin activation of endothelium for polymorphonuclear leucocyte transendothelial migration and modulation by interferon-gamma.

    PubMed Central

    Issekutz, A C; Lopes, N

    1993-01-01

    Endotoxin [lipopolysaccharide (LPS)] is a potent inflammatory stimulus and can activate human umbilical vein endothelium (HUVE) for leucocyte adhesiveness and transendothelial migration. Here we investigated the role of HUVE-secreted cytokines in this process. When HUVE monolayers were grown on filters and preincubated for 3 hr with LPS, 51Cr-labelled polymorphonuclear leucocytes (PMNL) migrated across the HUVE in a dose- and time-dependent manner. Maximal PMNL transmigration with LPS (1 ng/ml) was 26 +/- 3% of added PMNL in 75 min. Neutralizing antibodies to interleukin-1 alpha (IL-1 alpha) and IL-1 beta, tumour necrosis factor-alpha (TNF-alpha), IL-8 or recombinant IL-1 receptor antagonist had no effect on the activation by LPS of the HUVE for supporting migration of PMNL. The HUVE 'activated state' declined with prolonged (22 hr) exposure to LPS, as reflected by a decrease in PMNL transendothelial migration to 5.5 +/- 1% and in the expression of the endothelial cell adhesion molecule, E-selectin, as compared to stimulation with LPS for 3 hr. However, simultaneous exposure to interferon-gamma (IFN-gamma) (200 IU/ml) and LPS maintained maximal PMNL transendothelial migration (28 +/- 4%) for at least 24 hr, prolonged E-selectin expression by HUVE and superinduced intracellular adhesion molecule-1 (ICAM-1) expression. The PMNL transendothelial migration was blocked by > 90% by monoclonal antibody (mAb) to CD18 with either 3 hr of LPS or 22 hr LPS + IFN-gamma stimulation. Migration was partially inhibited by mAb to E-selectin (30-40%) or to ICAM-1 (35-45%) and by a combination of both reagents (50-60%) under both stimulation conditions. Thus, LPS activation of HUVE for PMNL transendothelial migration: (a) does not require secretion of IL-1, TNF-alpha or IL-8 by the endothelium, (b) IFN-gamma enhances and prolongs endothelial activation by LPS and may increase leucocyte infiltration in LPS or bacterial inflammatory reactions, and (c) CD18-dependent mechanisms are

  19. Improving the network management of integrated primary mental healthcare for older people in a rural Australian region: protocol for a mixed methods case study

    PubMed Central

    Fuller, Jeffrey; Oster, Candice; Dawson, Suzanne; O'Kane, Deb; Lawn, Sharon; Henderson, Julie; Gerace, Adam; Reed, Richard; Nosworthy, Ann; Galley, Philip; McPhail, Ruth; Cochrane, Eimear Muir

    2014-01-01

    Introduction An integrated approach to the mental healthcare of older people is advocated across health, aged care and social care sectors. It is not clear, however, how the management of integrated servicing should occur, although interorganisational relations theory suggests a reflective network approach using evaluation feedback. This research will test a network management approach to help regional primary healthcare organisations improve mental health service integration. Methods and analysis This mixed methods case study in rural South Australia will test facilitated reflection within a network of health and social care services to determine if this leads to improved integration. Engagement of services will occur through a governance group and a series of three 1-day service stakeholder workshops. Facilitated reflection and evaluation feedback will use information from a review of health sector and local operational policies, a network survey about current service links, gaps and enablers and interviews with older people and their carers about their help seeking journeys. Quantitative and qualitative analysis will describe the policy enablers and explore the current and ideal links between services. The facilitated reflection will be developed to maximise engagement of senior management in the governance group and the service staff at the operational level in the workshops. Benefit will be assessed through indicators of improved service coordination, collective ownership of service problems, strengthened partnerships, agreed local protocols and the use of feedback for accountability. Ethics, benefits and dissemination Ethics approval will deal with the sensitivities of organisational network research where data anonymity is not preserved. The benefit will be the tested utility of a facilitated reflective process for a network of health and social care services to manage linked primary mental healthcare for older people in a rural region. Dissemination will

  20. Measurement of in vitro leucocyte mitogenesis in fish: ELISA based detection of the thymidine analogue 5'-bromo-2'-deoxyuridine

    USGS Publications Warehouse

    Gauthier, David T.; Cartwright, Deborah D.; Densmore, Christine L.; Blazer, Vicki; Ottinger, Christopher A.

    2003-01-01

    In this study we present a method for the measurement of in vitro mitogenesis in fish leucocytes that is based on the incorporation of the thymidine analogue 5′-bromo-2′-deoxyuridine (BrdU) into the DNA of replicating cells, followed by ELISA-based detection. This technique, adapted from methods developed for mammalian cells, operates on a similar biological principle to 3H-thymidine incorporation, but circumvents the logistical and safety issues inherent with the radioactive label. Because it directly measures DNA proliferation, the assay has advantages over other colorimetric methods that may be strongly influenced by leucocyte metabolic status. Using BrdU incorporation followed by ELISA, we evaluate the responsiveness of rainbow trout (Oncorhynchus mykiss [Walbaum]) leucocytes to the mammalian T-cell mitogen Concanavalin A (Con A) as well as the differential response of white perch (Morone americana [Gmelin]) leucocytes to Con A and pokeweed mitogen. Specific considerations intrinsic to the assay system are discussed, including the implications of utilising enzyme-based detection.

  1. Carbohydrate-structure-dependent recognition of desialylated serum glycoproteins in the liver and leucocytes. Two complementary systems.

    PubMed Central

    Bezouska, K; Táborský, O; Kubrycht, J; Pospísil, M; Kocourek, J

    1985-01-01

    Oligosaccharides with four different types of branching were prepared from purified human transferrin, alpha 2-macroglobulin, caeruloplasmin and alpha 1-acid glycoprotein and labelled with NaBH3 3H. Binding of these oligosaccharides to rat liver plasma membrane, rat leucocytes, pig liver plasma membranes and pig leucocyte plasma membranes was investigated. A striking dependence of binding on oligosaccharide branching was observed. The values of apparent association constants Ka at 4 degrees C vary from 10(6) M-1 (biantennary structure) to 10(9) M-1 (tetra-antennary structure) in the liver, whereas in the leucocytes the Ka values were found to be of reversed order, from 1.8 X 10(9) M-1 for biantennary to 2.2 X 10(6) M-1 for tetra-antennary structures. The binding is completely inhibited by 150 mM-D-galactose, but 150 mM-D-mannose has almost no effect on binding. Leucocyte plasma membranes bind preferentially 125I-asialoglycoproteins with biantennary oligosaccharides, thus completing the specificity pattern of the hepatic recognition system for desialylated glycoproteins. Possible physiological roles of these two complementary recognition systems under normal and pathological conditions are discussed. Images Fig. 2. PMID:4004770

  2. Isolation and partial characterisation of a new antiproliferative substance from human leucocytes inhibiting growth of Candida albicans

    PubMed Central

    Naess-Andresen, C F; Ekeberg, D; Fagerhol, M K; Sandvik, K; Staahl, L

    2003-01-01

    Aim: To purify and partially characterise a fraction from human leucocytes containing a substance cytotoxic to Candida albicans. Methods: Leucocytes were isolated from the buffy coats of healthy blood donors. The cytotoxic factor (CF) was isolated from the soluble fraction of the cells. A cell lysate was passed through a filter with a cut off value of 3 kDa, and the filtrate was processed by anionic exchange chromatography and gel filtration. The purified CF was analysed for its chemical and biological properties. The cytotoxicity of CF was tested on C albicans grown on agar plates. Results: Mass spectrometry showed a molecular mass of 2.148 kDa. CF was found in polymorphonuclear neutrophilic cells only. No amino acids were detected, and a low ultraviolet absorbance at 260 nm and resistance to nuclease indicate the absence of nucleic acids. An anthrone test was positive for carbohydrate. The substance was soluble in water. CF showed a dose related cytotoxicity in the range of 0.1–1 mg/ml. The cytotoxic effect was abrogated by zinc ions. Preliminary testing indicated that CF also had cytotoxic effects against some bacteria. Conclusions: This report describes a factor from isolated human leucocytes that is cytotoxic to C albicans. The substance contains a carbohydrate moiety, whereas no amino acids were detected. The cytotoxicity can be abrogated by zinc ions in vitro. This substance is probably part of the repertoire by which leucocytes prevent infections. PMID:12890745

  3. Comparison of the effects of xenon and sevoflurane anaesthesia on leucocyte function in surgical patients: a randomized trial.

    PubMed

    Fahlenkamp, A V; Coburn, M; Rossaint, R; Stoppe, C; Haase, H

    2014-02-01

    While most anaesthetics are known to suppress immune reactions, data from experimental studies indicate the enhancement of reactivity to inflammatory stimulators under xenon treatment. We investigated the effect of xenon anaesthesia on leucocyte function in surgical patients. We performed a subgroup analysis of subjects undergoing xenon or sevoflurane anaesthesia in a randomized clinical trial. After oral premedication with midazolam, two separate blood samples were obtained from subjects undergoing elective abdominal surgery, directly before and 1 h after induction of anaesthesia. General anaesthesia was maintained with either 60% xenon or 2.0% sevoflurane in 30% O2. Leucocyte count, phagocytotic function, and pro-inflammatory cytokine release after ex vivo lipopolysaccharide (LPS) stimulation were determined. Except for lymphocyte numbers, leucocyte subpopulations did not differ between the groups. Phagocytosis and oxidative burst of granulocytes were reduced in both groups after 1 h of anaesthesia, whereas monocytes were not affected. Pro-inflammatory cytokine release in response to LPS was not affected. In vivo, xenon and sevoflurane anaesthesia did not have a pro-inflammatory effect, at least in combination with the types of surgery performed in this study. Notably, the impact of xenon anaesthesia did not differ significantly from sevoflurane anaesthesia with regard to leucocyte function. However, an underestimation of treatment effects due to limited sample sizes cannot be fully excluded.

  4. Toxicological in vitro effects of heavy metals on gilthead seabream (Sparus aurata L.) head-kidney leucocytes.

    PubMed

    Morcillo, Patricia; Cordero, Héctor; Meseguer, José; Esteban, María Á; Cuesta, Alberto

    2015-12-25

    Heavy metals provoke toxicological effects on aquatic animal species, including fish, though their effects on fish leucocytes and immunotoxicology are still limited. In the present work the effects of heavy metals (Cd, Hg, Pb or As) on viability, oxidative stress and innate immune parameters of isolated head-kidney leucocytes from gilthead seabream (Sparus aurata) are studied. Cytotoxicity results indicated that short exposures (30 min or 2h) to Hg promoted both apoptosis and necrosis cell death of leucocytes whilst Cd, Pb and As did only by apoptosis, in all cases in a concentration- and time-dependent manner. In addition, production of free oxygen radicals was induced by Cd, Hg and As heavy metals. Cd failed to change phagocytosis but Hg and As increased the percentage of phagocytic cells but decreased the number of ingested particles per cell whilst Pb increased both phagocytic parameters. On the other hand, respiratory burst activity was significantly reduced by incubation with Cd, Hg and As but increased with Pb. Furthermore, the gene expression profiles partly support the functional finding of this work. This study provides an in vitro approach for elucidating the heavy metals toxicity, and particularly the immunotoxicity, in fish leucocytes.

  5. Net primary productivity and rain-use efficiency as affected by warming, altered precipitation, and clipping in a mixed-grass prairie.

    PubMed

    Xu, Xia; Sherry, Rebecca A; Niu, Shuli; Li, Dejun; Luo, Yiqi

    2013-09-01

    Grassland productivity in response to climate change and land use is a global concern. In order to explore the effects of climate change and land use on net primary productivity (NPP), NPP partitioning [fBNPP , defined as the fraction of belowground NPP (BNPP) to NPP], and rain-use efficiency (RUE) of NPP, we conducted a field experiment with warming (+3 °C), altered precipitation (double and half), and annual clipping in a mixed-grass prairie in Oklahoma, USA since July, 2009. Across the years, warming significantly increased BNPP, fBNPP , and RUEBNPP by an average of 11.6%, 2.8%, and 6.6%, respectively. This indicates that BNPP was more sensitive to warming than aboveground NPP (ANPP) since warming did not change ANPP and RUEANPP much. Double precipitation stimulated ANPP, BNPP, and NPP but suppressed RUEANPP , RUEBNPP , and RUENPP while half precipitation decreased ANPP, BNPP, and NPP but increased RUEANPP , RUEBNPP , and RUENPP . Clipping interacted with altered precipitation in impacting RUEANPP , RUEBNPP , and RUENPP , suggesting land use could confound the effects of precipitation changes on ecosystem processes. Soil moisture was found to be a main factor in regulating variation in ANPP, BNPP, and NPP while soil temperature was the dominant factor influencing fBNPP . These findings suggest that BNPP is critical point to future research. Additionally, results from single-factor manipulative experiments should be treated with caution due to the non-additive interactive effects of warming with altered precipitation and land use (clipping).

  6. A comparison of the quality of life in patients with primary and secondary lower-limb lymphedema: A mixed-methods study

    PubMed Central

    Stolldorf, Deonni P; Dietrich, Mary S.; Ridner, Sheila H.

    2016-01-01

    Patients with lower-limb lymphedema experience symptoms that may differ in intensity and distress. This mixed-methods study compares symptom intensity and distress and the impact of lymphedema on patients’ quality of life by primary and secondary (cancer and non-cancer) lymphedema groups. Individuals completed an online questionnaire (i.e., demographic form, Lymphedema Symptom Intensity and Distress Survey-Leg, and an open-ended question). Analyses included descriptive and inferential statistics (quantitative data) and content analysis (qualitative data). Participants differed statistically significantly by gender, employment status, and lymphedema location. Groups differed significantly in lack of self-confidence (χ2(df=2) =9.19; p=.010). Cancer patients reported higher intensity and distress scores for some symptoms, but these differences were not statistically significant. Patients reported lacking psychosocial well-being and resources and experiencing physical and functional impairments and treatment and care challenges. Patients with lower-limb lymphedema experience psychosocial impairments and problems with quality of and access to care. PMID:27151079

  7. Effects of polymorphonuclear leucocyte depletion on the pathogenesis of experimental Legionnaires' disease.

    PubMed Central

    Fitzgeorge, R. B.; Featherstone, A. S.; Baskerville, A.

    1988-01-01

    Guinea-pigs were depleted of circulating polymorphonuclear leucocytes (PMN) by administration of anti-polymorph serum. Groups of animals were then infected by aerosols containing different doses of Legionella pneumophila and the effects compared with those in intact infected controls. Elimination of PMN lowered the dose of L. pneumophila necessary to establish infection, increased bacterial numbers in the lungs and caused much higher mortality. It did not change the nature or extent of pulmonary lesions. The findings confirm the importance of PMN in defence of the lung against L. pneumophila infection and indicate that PMN and their enzymes are not responsible for the pulmonary lesions, which are probably caused directly by the bacteria. PMID:3348954

  8. Influence of captopril on glucose and fatty acid oxidation in human thrombocytes and mononuclear leucocytes.

    PubMed

    Haeckel, R; Colic, D

    1991-01-01

    Captopril (CAS 62571-86-2) may be beneficial for the treatment of diabetes because of its activating effect on peripheral glucose consumption besides its well known blood pressure degradation. The glucose oxidation has been found to be activated by captopril in thrombocytes and mononuclear leucocytes, cell types which are usually considered to be independent from insulin. Because the oxidation of pyruvate labelled in position C-1 but not of 2-14C-pyruvate and of 1-14C-acetate was enhanced, captopril most probably stimulated the pyruvate decarboxylation reaction. The metabolism of glucose labelled in positions 1 and 6 was equally activated by captopril indicating another step which may be affected by captopril.

  9. Experimental transmission of bovine leukosis virus by leucocytes recovered from the stable fly Stomoxys calcitrans L.

    PubMed

    Freitas, T R; Romero, C H

    1991-01-01

    1. Wild stable flies (Stomoxys calcitrans) feeding on heifers infected with bovine leukosis virus (BLV) carried viable bovine leucocytes in the midgut and proboscis that, when inoculated by the subcutaneous route into lambs aged 5 to 60 days, elicited the development of antibodies to glycoprotein (gp51) and polypeptide 25 (p25). 2. Antibodies were detected as early as one month later and persisted for an experimental period of 24 or 36 months. Uninoculated control lambs reared together with the experimental animals did not acquire the infection, indicating the lack of horizontal transmission. 3. S. calcitrans reared in the laboratory were intermittently allowed to feed on the skin of BLV-infected heifers and on five lambs over a period of 3-10 months. Although some of these lambs were bitten about 500 times, none developed antibodies to BLV (gp51 or p25) over observation periods of 30 or 36 months.

  10. Promoting Help-Seeking in Response to Symptoms amongst Primary Care Patients at High Risk of Lung Cancer: A Mixed Method Study

    PubMed Central

    Brindle, Lucy; Ewings, Sean; James, Elizabeth; Moore, Mike; Rivas, Carol; Esqueda, Ana Ibanez; Corner, Jessica

    2016-01-01

    Background Lung cancer symptoms are vague and difficult to detect. Interventions are needed to promote early diagnosis, however health services are already pressurised. This study explored symptomology and help-seeking behaviours of primary care patients at ‘high-risk’ of lung cancer (≥50 years old, recent smoking history), to inform targeted interventions. Methods Mixed method study with patients at eight general practitioner (GP) practices across south England. Study incorporated: postal symptom questionnaire; clinical records review of participant consultation behaviour 12 months pre- and post-questionnaire; qualitative participant interviews (n = 38) with a purposive sample. Results A small, clinically relevant group (n = 61/908, 6.7%) of primary care patients was identified who, despite reporting potential symptoms of lung cancer in questionnaires, had not consulted a GP ≥12 months. Of nine symptoms associated with lung cancer, 53.4% (629/1172) of total respondents reported ≥1, and 35% (411/1172) reported ≥2. Most participants (77.3%, n = 686/908) had comorbid conditions; 47.8%, (n = 414/908) associated with chest and respiratory symptoms. Participant consulting behaviour significantly increased in the 3-month period following questionnaire completion compared with the previous 3-month period (p = .002), indicating questionnaires impacted upon consulting behaviour. Symptomatic non-consulters were predominantly younger, employed, with higher multiple deprivation scores than their GP practice mean. Of symptomatic non-consulters, 30% (18/61) consulted ≤1 month post-questionnaire, with comorbidities subsequently diagnosed for five participants. Interviews (n = 39) indicated three overarching differences between the views of consulting and non-consulting participants: concern over wasting their own as well as GP time; high tolerance threshold for symptoms; a greater tendency to self-manage symptoms. Conclusions This first study to examine symptoms and

  11. Experiences of nurse practitioners and medical practitioners working in collaborative practice models in primary healthcare in Australia - a multiple case study using mixed methods.

    PubMed

    Schadewaldt, Verena; McInnes, Elizabeth; Hiller, Janet E; Gardner, Anne

    2016-07-29

    In 2010 policy changes were introduced to the Australian healthcare system that granted nurse practitioners access to the public health insurance scheme (Medicare) subject to a collaborative arrangement with a medical practitioner. These changes facilitated nurse practitioner practice in primary healthcare settings. This study investigated the experiences and perceptions of nurse practitioners and medical practitioners who worked together under the new policies and aimed to identify enablers of collaborative practice models. A multiple case study of five primary healthcare sites was undertaken, applying mixed methods research. Six nurse practitioners, 13 medical practitioners and three practice managers participated in the study. Data were collected through direct observations, documents and semi-structured interviews as well as questionnaires including validated scales to measure the level of collaboration, satisfaction with collaboration and beliefs in the benefits of collaboration. Thematic analysis was undertaken for qualitative data from interviews, observations and documents, followed by deductive analysis whereby thematic categories were compared to two theoretical models of collaboration. Questionnaire responses were summarised using descriptive statistics. Using the scale measurements, nurse practitioners and medical practitioners reported high levels of collaboration, were highly satisfied with their collaborative relationship and strongly believed that collaboration benefited the patient. The three themes developed from qualitative data showed a more complex and nuanced picture: 1) Structures such as government policy requirements and local infrastructure disadvantaged nurse practitioners financially and professionally in collaborative practice models; 2) Participants experienced the influence and consequences of individual role enactment through the co-existence of overlapping, complementary, traditional and emerging roles, which blurred perceptions of

  12. Neutrophil leucocyte chemotaxis is not induced by a spatial gradient of chemoattractant.

    PubMed

    Vicker, M G; Lackie, J M; Schill, W

    1986-08-01

    Chemotaxis and directed locomotion of neutrophil leucocytes are generally thought to be determined by the directed response of the cell to stable, spatial gradients of chemoattractants. In most cases, however, cells are also exposed to characteristic temporal changes in the attractant concentration during the lifetime of the gradient, especially as it develops. We have attempted to test whether neutrophils can respond to a spatial gradient in which these temporal changes are essentially absent. Gradients of formyl-peptides were made across a narrow barrier of agarose gel that separated two fluid reservoirs, and the cells were observed cinematographically as they moved between gel and glass. In gradients predeveloped at low temperature, at which cell motion and responses to attractant were inhibited, neutrophils showed no tendency to accumulate up-gradient when warmed to 37 degrees C. Yet their speed and turning behaviour was related to the local concentration of formyl-peptide. However, gradients that developed at 37 degrees C, whilst the cells were responsive, elicited directed locomotion. We also tested populations that were either spreading into or already evenly distributed across micropore filters to see how cells might sense directional cues. We reasoned that evenly distributed populations could accumulate in a spatial gradient only if cells were able to 'read' it. However, no redistribution occurred without an applied impulse of attractant. It seems that the oriented, temporal component of an attractant signal is essential if a directed response (i.e. non-random turning) is to occur; a spatial gradient of soluble attractant alone does not induce neutrophil accumulation or taxis. This finding has implications for the termination of the acute inflammatory response, for clinical tests of leucocyte behaviour and for morphogen signal interpretation by cells in developing tissues.

  13. Microarray analysis of the inflammatory and immune responses in head kidney turbot leucocytes treated with resveratrol.

    PubMed

    Domínguez, Berta; Pardo, Belén G; Noia, Manuel; Millán, Adrián; Gómez-Tato, Antonio; Martínez, Paulino; Leiro, José; Lamas, Jesús

    2013-03-01

    A DNA oligo-microarray enriched in genes and involved in inflammatory and immune responses was used to evaluate the effects of resveratrol on gene expression in turbot head kidney leucocytes. Leucocytes were cultured for 3, 6 and 24 h, in the presence or absence of resveratrol, or were stimulated with the membrane fraction of the parasite Philasterides dicentrarchi or with the membrane plus resveratrol. Gene expression changed considerably in control cells, and several of the regulated genes were related to inflammatory and immune responses and to the cytoskeleton. Similar changes in gene expression occurred in control cells and in cells stimulated with P. dicentrarchi membrane fraction. Treatment with resveratrol induced changes in the expression (mostly down-regulation) of several genes involved in immune responses and inflammation. Thus, the down-regulation of the transcription factor PU.1, pentraxin-multidomain protein, heme oxygenase 1, S100 calcium-binding protein A-16 (S100A16) and the signal transducer and activator of transcription 4 was observed after all three incubation times. The down-regulation of the suppressor of cytokine signalling 3a, LPS-induced tumour necrosis alpha, hepcidin, metallothionein, TLR8 and the calcium dependent lectin A was observed after 3 and 6 h. Resveratrol also decreased the expression of CCL20, IL-8, apolipoprotein E and glutathione S-transferase after incubation for 6 and 24 h, and of TNF-α after incubation for 3 and 24 h. Resveratrol also induced strong regulation of several cytoskeleton-related genes. The use of the turbot oligo-microarray enabled us to discover genes whose expression was not previously suspected of being modulated by this polyphenol.

  14. Mitochondrial DNA 4977-base pair common deletion in blood leucocytes and melanoma risk

    PubMed Central

    Shen, Jie; Wan, Jie; Huff, Chad; Fang, Shenying; Lee, Jeffrey E.; Zhao, Hua

    2017-01-01

    The 4977-base pair common deletion DmtDNA4977 is the most frequently observed mitochondrial DNA mutation in human tissues. Because mitochondrial DNA mutations are mainly caused by reactive oxygen species (ROS), and given that oxidative stress plays an important role in melanoma carcinogenesis, the investigation of DmtDNA4977 may be particularly relevant to the development of melanoma. In the current study, we compared DmtDNA4977 levels in blood leucocytes from 206 melanoma patients and 219 healthy controls. Overall, melanoma cases had significantly higher levels of DmtDNA4977 than healthy controls (median: 0.60 vs 0.20, P=0.008). The difference was evident among individuals who were older than 47 years old, women, and had pigmentation risk factors (e.g. blond or red hair, blue eye, fair skin, light or none tanning ability after prolonged sun exposure, and freckling in the sun as a child). The difference was also evident among those who had at least one lifetime sunburn with blistering and had no reported use of a sunlamp. Interestingly, among controls, DmtDNA4977 levels differed by phenotypic index and reported use of a sunlamp. In the risk assessment, increased levels of DmtDNA4977 were associated with a 1.23-fold increased risk of melanoma (Odds ratio (OR): 1.23, 95% Confidence Interval (90% CI): 1.01, 1.50). A significant dose-response relationship was observed in quartile analysis (P=0.001). In summary, our study suggests that high levels of DmtDNA4977 in blood leucocytes are associated with increased risk of melanoma, and that association is affected by both pigmentation and personal history of sun exposure. PMID:26988264

  15. Clinical evaluation of a leucocyte-depleting blood cardioplegia filter (BC1B) for elective open-heart surgery.

    PubMed

    Suzuki, I; Ogoshi, N; Chiba, M; Komatsu, T; Moizumi, Y

    1998-05-01

    Activation of leucocytes during extracorporeal circulation has attracted attention in recent years as a cause of reperfusion injury in open-heart surgery patients. In the present study, 40 adult patients undergoing elective open-heart surgery were randomized into two groups: 20 using the Pall BC1B leucocyte-depleting filter for blood cardioplegia (group 1) and the other 20 without the filter (group 2). In order to determine if the filter was effective in protecting the myocardium, CPK-MB and troponin-T (TnT) were measured. In addition, efforts were also made to determine appropriate sites at which the BC1B blood cardioplegia filter should be positioned. There were no significant differences between the two groups in terms of conditions of perfusion. No adverse effects were seen in either group. The total leucocyte reduction rate through the filter was 98.1% with the passage of 2 liters of blood through the filter. A pressure drop of 4.4 +/- 3.2 mmHg was observed through the filter during use. Statistically significant differences were noted between the two groups in CPK-MB (p = 0.031) and TnT (p = 0.004). Findings obtained in previous studies demonstrate that the various advantages of leucocyte reduction, shown in experimental studies, can be translated into clinical advantages. In conclusion, based on the results in this clinical study showing significant difference in CPK-MB and TnT which are known effective indicators for myocardial injury, between leucoreduced and non-leucoreduced group, the Pall BC1B leucocyte-depleting filter for blood cardioplegia has been shown to be effective in alleviating reperfusion injury in open-heart surgery patients.

  16. 300 The Method of Antigen Specific Damage of Leucocytes by Food Additives in Patients with Bronchial Asthma

    PubMed Central

    Titova, Nadya

    2012-01-01

    Background Diagnosis of adverse reactions to food additives is difficult due to a variety of mechanisms involved and the lack of sufficiently reliable methods for their determination. The diagnosis of intolerance to food additives is still based only on placebo-controlled oral provocation. Methods The aim of this study was to determine the incidence of intolerance to ponceau 4R (E124), indigo carmine (E132), azorubine (E122), tartrazine (E102), sunset yellow (E110) and sodium benzoate (E211) among patients with bronchial asthma. We studied 114 patients with bronchial asthma using clinical and laboratory methods. Also we used the method of antigen specific damage of leucocytes by food additives. After the incubation of leucocytes with solutions of food additives to leucocytes was added 0.05 mL of trypan blue and counted the percentage of stained (damaged) granulocytes with the food dye and in control tests. If damaging leucocytes were more than 20% in comparison with controls - the test considered positive. Results It was found that positive to ponceau 4R were 6 of 114 patients, to indigo carmine–3 of 73, to sodium benzoate–4 of 73, to azorubine–11 of 114, to tartrazine–7 of 114 and to sunset yellow–9 of 114. There was a correlation between the results obtained and data history. Between experienced and control group (the patients without allergic diseases) were the reliable differences (P < 0.05). Conclusions 1. Under influence of the food additives leukocytes of patients with bronchial asthma are damaged and painted by trypan blue. 2. The method of antigen specific damage of leucocytes by food additives can be used for diagnostics of the allergies to food dyes, sodium benzoate and other gaptens.

  17. Human leucocyte antigens B*08, DRB1*03 and DRB1*13 are significantly associated with autoimmune liver and biliary diseases in Finnish children.

    PubMed

    Ylinen, E; Salmela, L; Peräsaari, J; Jaatinen, T; Tenca, A; Vapalahti, O; Färkkilä, M; Jalanko, H; Kolho, K-L

    2017-02-01

    The human leucocyte antigen (HLA) allele and haplotype frequencies of the Finnish population are unique because of the restricted and homogenous gene population. There are no published data on HLA genotype associations in paediatric autoimmune liver diseases in Scandinavia. This study characterised the HLA genotypes of children with autoimmune liver or biliary disease in Finland. The study cohort comprised 19 paediatric patients (13 female) aged three years to 15 years treated for autoimmune liver or biliary disease at the Children's Hospital, Helsinki University Hospital, between 2000 and 2011, and followed up for four years and three months to 14.6 years. We genotyped HLA-B and HLA-DRB1 in the children, and the HLA antigen frequencies were compared with 19 807 records from the Finnish Bone Marrow Donor Registry. All paediatric patients with autoimmune liver or biliary disease had either autoimmune HLA haplotype B*08;DRB1*03 or DRB1*13. These were significantly more common among patients with autoimmune hepatitis, primary sclerosing cholangitis and autoimmune hepatitis/primary sclerosing cholangitis overlap syndrome than the Finnish control population. HLA RB1*04 was not found in the study cohort. Our study found that B*08, DRB1*03 and DRB1*13 were significantly associated with autoimmune liver and biliary diseases in Finnish paediatric patients. ©2016 Foundation Acta Paediatrica. Published by John Wiley & Sons Ltd.

  18. Implementing an mHealth system for substance use disorders in primary care: a mixed methods study of clinicians' initial expectations and first year experiences.

    PubMed

    Mares, Marie-Louise; Gustafson, David H; Glass, Joseph E; Quanbeck, Andrew; McDowell, Helene; McTavish, Fiona; Atwood, Amy K; Marsch, Lisa A; Thomas, Chantelle; Shah, Dhavan; Brown, Randall; Isham, Andrew; Nealon, Mary Jane; Ward, Victoria

    2016-09-29

    Millions of Americans need but don't receive treatment for substance use, and evidence suggests that addiction-focused interventions on smart phones could support their recovery. There is little research on implementation of addiction-related interventions in primary care, particularly in Federally Qualified Health Centers (FQHCs) that provide primary care to underserved populations. We used mixed methods to examine three FQHCs' implementation of Seva, a smart-phone app that offers patients online support/discussion, health-tracking, and tools for coping with cravings, and offers clinicians information about patients' health tracking and relapses. We examined (a) clinicians' initial perspectives about implementing Seva, and (b) the first year of implementation at Site 1. Prior to staggered implementation at three FQHCs (Midwest city in WI vs. rural town in MT vs. metropolitan NY), interviews, meetings, and focus groups were conducted with 53 clinicians to identify core themes of initial expectations about implementation. One year into implementation at Site 1, clinicians there were re-interviewed. Their reports were supplemented by quantitative data on clinician and patient use of Seva. Clinicians anticipated that Seva could help patients and make behavioral health appointments more efficient, but they were skeptical that physicians would engage with Seva (given high caseloads), and they were uncertain whether patients would use Seva. They were concerned about legal obligations for monitoring patients' interactions online, including possible "cries for help" or inappropriate interactions. One year later at Site 1, behavioral health care providers, rather than physicians, had incorporated Seva into patient care, primarily by discussing it during appointments. Given workflow/load concerns, only a few key clinicians monitored health tracking/relapses and prompted outreach when needed; two researchers monitored the discussion board and alerted the clinic as needed

  19. Deciphering the Temporal and Spatial Complexity in Submarine Canyons in Antarctica: the Role of Mixed Layer Depth in Regulating Primary Production

    NASA Astrophysics Data System (ADS)

    Carvalho, F.; Kohut, J. T.; Schofield, O.; Oliver, M. J.; Gorbunov, M. Y.

    2016-02-01

    There is a high spatial and temporal variability in the biophysical processes regulating primary productivity in submarine canyons in the West Antarctic Peninsula (WAP). WAP canyon heads are considered biological "hotspots" by providing predictable food resource and driving penguin foraging locations. Because the physiology and composition of the phytoplankton blooms and the physical mechanisms driving them aren't well understood, we aim to characterize the dynamics of the spring phytoplankton bloom at the head of a canyon in the WAP. A 6-year record of Slocum glider deployments is analyzed, corresponding to over 16,000 water column profiles. The mixed layer depth (MLD), determined by the maximum of the buoyancy frequency criteria, was found to be the MLD definition with the highest ecological relevance. The same holds true for other regions in Antarctica such as the Ross and Amundsen Seas. A FIRe sensor on a glider was used to evaluate physiological responses of phytoplankton to canyon dynamics using fluorescence kinetics. Initial results show a spatial influence, with increased photosynthetic efficiencies found at the canyon head. The strongest signal was the seasonal cycle. The shoaling of the MLD in early January results in increased chlorophyll a concentrations and as MLD deepens in mid season due to wind forcing, phytoplankton concentrations decrease, likely due to decreased light availability. A consistent secondary peak in chlorophyll matches a shoaling in MLD later in the growth season. A steady warming and increase in salinity of the MLD is seen throughout the season. Spatial differences were recorded at the head of the canyon and result from the local circulation. Shallower MLD found on the northern region are consistent with a fresher surface ocean (coastal influence) and increased chlorophyll concentrations. The southern region is thought to be more oceanic influenced as intrusions of warm deep water (mUCDW) to the upper water column were recorded

  20. Homeless people's access to primary care physiotherapy services: an exploratory, mixed-method investigation using a follow-up qualitative extension to core quantitative research.

    PubMed

    Dawes, Jo; Deaton, Stuart; Greenwood, Nan

    2017-06-30

    The purpose of this study was to appraise referrals of homeless patients to physiotherapy services and explore perceptions of barriers to access. This exploratory mixed-method study used a follow-up qualitative extension to core quantitative research design. Over 9 months, quantitative data were gathered from the healthcare records of homeless patients referred to physiotherapy by a general practitioner (GP) practice, including the number of referrals and demographic data of all homeless patients referred. Corresponding physiotherapy records of those people referred to physiotherapy were searched for the outcome of their care. Qualitative semi-structured telephone interviews, based on the quantitative findings, were carried out with staff involved with patient care from the referring GP practice and were used to expand insight into the quantitative findings. Two primary care sites provided data for this study: a GP practice dedicated exclusively to homeless people and the physiotherapy department receiving their referrals. Quantitative data from the healthcare records of 34 homeless patient referrals to physiotherapy were collected and analysed. In addition, five staff involved in patient care were interviewed. 34 referrals of homeless people were made to physiotherapy in a 9-month period. It was possible to match 25 of these to records from the physiotherapy department. Nine (36%) patients did not attend their first appointment; seven (28%) attended an initial appointment, but did not attend a subsequent appointment and were discharged from the service; five (20%) completed treatment and four patients (16%) had ongoing treatment. Semi-structured interviews revealed potential barriers preventing homeless people from accessing physiotherapy services, the complex factors being faced by those making referrals and possible ways to improve physiotherapy access. Homeless people with musculoskeletal problems may fail to access physiotherapy treatment, but opportunities

  1. Developing a service user informed intervention to improve participation and ability to perform daily activities in primary Sjögren's syndrome: a mixed-methods study protocol.

    PubMed

    Hackett, Katie L; Newton, Julia L; Deane, Katherine H O; Rapley, Tim; Deary, Vincent; Kolehmainen, Niina; Lendrem, Dennis; Ng, Wan-Fai

    2014-08-21

    A significant proportion of patients with primary Sjögren's syndrome (PSS) is functionally impaired and experience difficulties participating in various aspects of everyday life. There is currently no evidence of efficacy for non-pharmacological interventions aimed specifically at supporting the patients with PSS to improve their participation and ability to perform daily activities. This paper describes a research protocol for a mixed-methods study to develop an intervention to improve these outcomes. The protocol follows the Medical Research Council framework for complex interventions. We will use group concept mapping with the patients, adults who live with them and healthcare professionals to identify factors which prevent people with PSS from participating in daily life and performing daily activities. The factors will be prioritised by participants for importance and feasibility and will inform an intervention to be delivered within a National Health Service (NHS) setting. Evidence-based intervention techniques will be identified for the prioritised factors and combined into a deliverable intervention package. Key stakeholders will comment on the intervention content and mode of delivery through focus groups, and the data will be used to refine the intervention. The acceptability and feasibility of the refined intervention will be evaluated in a future study. The study has been approved by an NHS Research Ethics Committee, REC Reference: 13/NI/0190. The findings of this study will be disseminated in peer-reviewed journals and through presentation at national and international conferences. UKCRN Study ID: 15939. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

  2. Quality improvement and person-centredness: a participatory mixed methods study to develop the ‘always event’ concept for primary care

    PubMed Central

    Bowie, Paul; McNab, Duncan; Ferguson, Julie; de Wet, Carl; Smith, Gregor; MacLeod, Marion; McKay, John; White, Craig

    2015-01-01

    Objectives (1) To ascertain from patients what really matters to them on a personal level of such high importance that it should ‘always happen’ when they interact with healthcare professionals and staff groups. (2) To critically review existing criteria for selecting ‘always events’ (AEs) and generate a candidate list of AE examples based on the patient feedback data. Design Mixed methods study informed by participatory design principles. Subjects and setting Convenience samples of patients with a long-term clinical condition in Scottish general practices. Results 195 patients from 13 general practices were interviewed (n=65) or completed questionnaires (n=130). 4 themes of high importance to patients were identified from which examples of potential ‘AEs’ (n=8) were generated: (1) emotional support, respect and kindness (eg, “I want all practice team members to show genuine concern for me at all times”); (2) clinical care management (eg, “I want the correct treatment for my problem”); (3) communication and information (eg, “I want the clinician who sees me to know my medical history”) and (4) access to, and continuity of, healthcare (eg, “I want to arrange appointments around my family and work commitments”). Each ‘AE’ was linked to a system process or professional behaviour that could be measured to facilitate improvements in the quality of patient care. Conclusions This study is the first known attempt to develop the AE concept as a person-centred approach to quality improvement in primary care. Practice managers were able to collect data from patients on what they ‘always want’ in terms of expectations related to care quality from which a list of AE examples was generated that could potentially be used as patient-driven quality improvement (QI) measures. There is strong implementation potential in the Scottish health service. However, further evaluation of the utility of the method is also necessary. PMID:25922095

  3. An open-label study of the tolerability of mixed amphetamine salts in adults with attention-deficit/hyperactivity disorder and treated primary essential hypertension.

    PubMed

    Wilens, Timothy E; Zusman, Randall M; Hammerness, Paul G; Podolski, Amy; Whitley, Julia; Spencer, Thomas J; Gignac, Martin; Biederman, Joseph

    2006-05-01

    To evaluate the short-term tolerability of an extended-release preparation of the stimulant medication mixed amphetamine salts (MAS XR) in adults with attention-deficit/hyperactivity disorder (ADHD) whose hypertension has been successfully treated with antihypertensive medications. An 8-week, 2-phase, open-label study design was implemented. All adults had ADHD (DSM-IV diagnosis) and essential hypertension and were required to be normotensive (blood pressure < 135/85 mm Hg, treated) for at least 4 weeks at entry into the study. MAS XR was given for a 6-week period, titrated once each week to a target maximum dose of 60 mg/day given once daily in the morning (phase 1), and then discontinued for 2 weeks at the end of the study (phase 2). At baseline, subjects underwent a comprehensive clinical assessment, medical history, vital signs assessment, and electrocardiogram (ECG). Rating scales were used throughout the study to assess response to treatment, and blood pressure was measured manually at each study visit. The primary outcome was the effect of MAS XR on blood pressure and the development of hypertension. Thirteen subjects receiving antihypertensive therapy were entered and placed on MAS XR treatment and completed the trial. There were no serious adverse events. No sustained elevated blood pressure (> 140/90 mm Hg at 2 consecutive visits) was observed in the subjects treated with MAS XR. Similar rates of single episodes of hypertension were observed in phases 1 and 2. Similarly, there was no group mean increase in systolic or diastolic blood pressure or pulse during treatment with MAS XR. No clinically significant changes in the ECG were observed. During the 6-week medication phase, significant improvement was found on rating scales assessing ADHD symptoms and severity that reversed with discontinuation of MAS XR. The results of this open study suggest that adults with ADHD and controlled hypertension can be safely treated with MAS XR.

  4. Comparison of capillary and venous blood in the analysis of concentration and function of leucocyte sub-populations.

    PubMed

    Canetti, Elisa F D; Keane, J; McLellan, C P; Gray, A B

    2016-08-01

    Compare capillary and venous blood in the analysis of concentration and function of leucocyte sub-populations. This study hypothesised that capillary samples may be used in a site-specific manner as an alternative source of blood samples for assays of leucocyte concentration and neutrophilic phagocytic function and reactive oxygen species (ROS) production, allowing acquisition of multiple samples to better monitor transient but significant post-exercise immune modulation. Resting blood samples were simultaneously obtained from vein, finger and earlobe of healthy subjects (n = 10, age: 25.1 ± 3.1 years). Leucocyte concentrations were measured using a five-part differential haematological analyser. Leucocyte sub-populations (CD3, CD4, CD8, CD19, CD56, CD14) and granulocytic functional-related (CD11b, CD18, CD16b, CD66b) surface antigen markers, neutrophil phagocytosis (FITC-labelled Escherichia coli) and stimulated ROS production (DHR) were quantified utilizing flow cytometry. A MANOVA (α < 0.05 significance) analysed the effects of the different sampling sites in the concentrations of leucocyte populations, their surface antigen expression and granulocytic functions. Leucocyte concentration and neutrophilic ROS production yielded non-significant differences between sampling sites. Expression of granulocytic surface antigens was increased in both capillary sites compared to venous site (p = 0.008), particularly for adhesion markers CD11b/CD18. The percentage of neutrophils performing phagocytosis was higher in venous samples compared to finger (p = 0.025). Increased number of E. coli ingested was observed in venous sample compared to finger (p = 0.001) and to earlobe (p = 0.006). Whilst attention must be paid for varying neutrophilic surface antigen expression and further studies are needed to establish appropriate reference ranges, this study supports the use of capillary blood samples in a site-specific manner to enhance sampling capabilities field

  5. Primary production at 47°N and 20°W in the North Atlantic Ocean: a comparison between the 14C incubation method and the mixed layer carbon budget

    NASA Astrophysics Data System (ADS)

    Chipman, David W.; Marra, John; Takahashi, Taro

    Primary production in the oceanshas been estimated mainly on the basis of in vitro incubation measurements. An implicit assumption is that the growth rate of phytoplankton observed in vitro represents that occurring in the freely circulating water of the euphotic zone. We have tested this assumption at 47°N-20°W in the eastern North Atlantic Ocean during the initial stages of a spring phytoplankton bloom. The daily primary production was measured by means of the 14C assimilation method, in which the incubation bottles were suspended in the ocean from dawn to dusk daily (about 14 h). The mean daily carbon assimilation rate in the photic zone and in the mixed layer was 107 ± 23 and 84 ± 18 mmol C m-2day∗-1 (where day∗ = 14 daylight hours), respectively, during the 12-day period between 26 April and 7 May 1989. The mixed layer carbon assimilation data are found to be consistent with the in situ CO 2 utilization rate of 82 ± 17 mmol C m-day∗-1 estimated on the basis of the thickness of surface mixed layer, the CO 2 concentration in it, and the air-sea CO 2 flux. We conclude that primary production in the open ocean appears to be well represented by the in vitro measurements, if the samples are incubated under the in situ light and temperature conditions. The mean daily reduction rate of the total CO 2 concentration observed in the mixed layer over the 12-day period is 2.3 μmol kg -1 day -1, about 75% of the rate, 3.1 μmol kg -1 day ∗-1, expected from the rate of primary production. About 8.5% of this difference is explained by the atmospheric CO 2 flux, and the remaining 16.5% may be attributed to the respiration and the influx of CO 2-rich waters from the mixed layer.

  6. Sequence variation of a novel heptahelical leucocyte receptor through alternative transcript formation.

    PubMed Central

    Barella, L; Loetscher, M; Tobler, A; Baggiolini, M; Moser, B

    1995-01-01

    Chemoattractants, including chemokines such as interleukin 8 (IL-8) and related proteins, activate leucocytes via seven-transmembrane-domain G-protein-coupled receptors. A cDNA for a novel receptor of this kind consisting of 327 amino acids was isolated from a human blood monocyte cDNA library. The polypeptide, termed monocyte-derived receptor 15 (MDR15), is an alternative form of the Burkitt's lymphoma receptor 1 (BLR1) encoded by a human Burkitt's lymphoma cDNA [Dobner, Wolf, Emrich and Lipp (1992) Eur. J. Immunol. 22, 2795-2799]. MDR15 and BLR1 cDNAs differ in the 5' region, where the open reading frame of MDR15 is shorter by 45 codons. Southern-blot analysis indicates that the two transcripts for MDR15 and BLR1 are encoded by the same gene. Northern-blot analysis using a probe that hybridizes with both mRNAs demonstrated high-level expression in chronic B-lymphoid leukaemia and non-Hodgkin's lymphoma cells and, to a lesser extent, peripheral blood monocytes and lymphocytes. Reverse transcription-PCR studies with MDR15- and BLR1-specific primers showed similar levels of transcripts for both receptors in RNA that was positive in Northern-blot analysis. MDR15 and BLR1 have high structural similarity to receptors for human IL-8 (about 40% amino acid identity) and other chemokines. However, none of a series of radiolabelled chemokines (IL-8, NAP-2, GRO alpha, PF4, IP10, MCP-1, MCP-2, MCP-3, I-309, RANTES and MIP-1 alpha) and other ligands (C3a and leukotriene B4) bound to Jurkat transfectants that stably expressed either MDR15 or BLR1 mRNA. The fact that MDR15 and BLR1 are expressed on leucocytes and show marked sequence similarity to chemokine receptors suggests the existence of as yet unidentified chemokines. Alternative transcript formation affecting the 5'-terminal part of the coding region may be a way to modify ligand-binding selectivity. Images Figure 2 Figure 3 Figure 5 Figure 6 Figure 7 PMID:7639692

  7. Accuracy of cerebrospinal leucocyte count, protein and culture for the diagnosis of acute bacterial meningitis: a comparative study using Bayesian latent class analysis.

    PubMed

    Manning, Laurens; Laman, Moses; Mare, Trevor; Hwaiwhanje, Ilomo; Siba, Peter; Davis, Timothy M E

    2014-12-01

    To examine the utility of laboratory methods other than bacterial culture in diagnosing acute bacterial meningitis (ABM). Bayesian latent class analysis was used to estimate diagnostic precision of cerebrospinal fluid (CSF) culture, leucocyte counts and protein concentrations for ABM in Melanesian children. With a cut-off of ≥20 leucocytes/mm(3) , the area under the receiver operating characteristic curve (AUC ROC) was >97.5% for leucocyte counts. A lower (93%) AUC ROC was observed for CSF protein concentrations ≥1 g/l. CSF culture had poor sensitivity and high specificity. Leucocyte counts provide sufficient diagnostic precision to aid clinical decision-making in ABM. © 2014 John Wiley & Sons Ltd.

  8. Synthesis, structure characterization and biological activity of selected metal complexes of sulfonamide Schiff base as a primary ligand and some mixed ligand complexes with glycine as a secondary ligand

    NASA Astrophysics Data System (ADS)

    Sharaby, Carmen M.; Amine, Mona F.; Hamed, Asmaa A.

    2017-04-01

    The current work reports synthesis of metal complexes and mixed ligand complexes of a novel sulfonamide Schiff base ligand (HL) resulted from the condensation of sulfametrole [N‧-(4-methoxy-1,2,5-thiadiazol-3-yl]sulfanilamide and acetyl-acetone as a primary ligand and glycine as a secondary ligand. The metal complexes and mixed ligand complexes of HL Schiff base ligand were synthesized and characterized using different physicochemical studies as elemental analyses, mass spectra, conductivity measurement, IR spectra, 1H NMR spectra, UV-vis Spectra, solid reflectance, magnetic susceptibility, thermal analyses (TGA and DTA) and their microbial and anticancer activities. The spectroscopic data of the complexes suggest their 1:2(L1:M) complex structures and 1:2:2(L1:L2:M) mixed ligand complex structures, where L1 = HL and L2 = glycine. Also, the spectroscopic studies suggested the octahedral structure for all complexes. The synthesized Schiff base, its metal and mixed ligand complexes were screened for their bacterial, antifungal and anticancer activity. The activity data show that the metal complexes and mixed ligand complexes exhibited promising microbial and anticancer activities than their parent HL Schiff base ligand, also the data show that the mixed ligand complexes more effective than the metal complexes.

  9. A pseudo-cryptococcal artefact derived from leucocytes in wet India ink mounts of centrifuged cerebrospinal fluid.

    PubMed

    Thiruchelvan, N; Wuu, K Y; Arseculeratne, S N; Ashraful-Haq, J

    1998-03-01

    Wet India ink mounts of cerebrospinal fluid (CSF) are useful in the laboratory diagnosis of cryptococcal meningitis. Pseudo-cryptococcal artefacts in such mounts have been attributed to leucocytes in CSF but their mode of formation has not been explained. This report describes the reproduction of such an artefact in cryptococcus free CSF-leucocyte mixtures that had been subjected to high speed centrifugation. The viscosity of DNA that could provide a morphological pseudo-capsule, and the yellow-green fluorescence of the pseudo-capsular material on staining with acridine-orange, suggest that lymphocytic nuclear DNA, which possibly leaked out after damage to the lymphocyte membrane by centrifugation, was responsible for this artefact.

  10. Type I allergic hypersensitivity reactions due to ethylene oxide sterilised leucocyte filters in patients with thalassaemia: report of four cases.

    PubMed

    Belen, Burcu; Polat, Meltem

    2015-02-27

    Ethylene oxide (EO) is a highly reactive gas used in sterilisation of heat sensitive medical devices, such as infusion sets, cannulae, intubation materials, ventriculoperitoneal shunts, dialysis catheters and stents. Allergic reactions due to EO have been reported in haemodialysis patients, patients undergoing extracorporeal photopheresis and donors of plasmapheresis. Clinical manifestations vary considerably and generally do not allow differentiation between IgE-mediated anaphylaxis and anaphylactoid reactions. We report four patients with thalassaemia who experienced anaphylaxis during transfusion due to ethylene oxide sterilised leucocyte filters. The aim of this report is to highlight the fact that frequently transfused patients can have allergic reactions due to EO particles left in leucocyte filters. 2015 BMJ Publishing Group Ltd.

  11. Francisella noatunensis subsp. noatunensis replicates within Atlantic cod (Gadus morhua L.) leucocytes and inhibits respiratory burst activity.

    PubMed

    Vestvik, Nils; Rønneseth, Anita; Kalgraff, Cathrine A K; Winther-Larsen, Hanne C; Wergeland, Heidrun I; Haugland, Gyri T

    2013-09-01

    Francisella noatunensis subsp. noatunensis, causing granulomatosis in cod, has been shown to reside within cod immune cells, mainly within monocytes and macrophages. In the present study, we analysed the ability of the bacterium to replicate within adherent cells isolated from head kidney by in vitro infection of leucocytes. Two different technical approaches for flow cytometry analyses were performed for detection of intracellular bacteria. The presence of the wild type was assessed after identification by intracellular binding of specific antibodies to the pathogen. The other way was to use green fluorescent protein (GFP) transformed bacterium for infection studies allowing direct measurements of fluorescence from infected cells. By both methods we found an increase in fluorescence in infected cells, verifying bacterial replication, both after 4 and 28 h post infection in leucocytes isolated from head kidney (HKL). The GFP transformed bacterium was similar to the wild type in growth and infectivity pattern, showing that it can be a valuable tool for further studies of infection routes and pathology. Further, F. noatunensis subsp. noatunensis was found to inhibit respiratory burst activity, a potent pathogen killing mechanism, in cod leucocytes, but not in such cells from salmon. Our findings may indicate that inhibition of respiratory burst during Francisella infection is a key to its intracellular existence. This strategy seems to be conserved through evolution as it is also observed during infections in higher vertebrates caused by bacteria within the Francisella genus. The results presented here, showing the intracellular existence of Francisella, its replication within leucocytes and the inhibitory effect on respiratory burst, strongly support that these factors contribute to disease and pathology in infected cod. The intracellular replication shown in the present study might contribute to explain the problems of obtaining protective vaccines against

  12. Preliminary studies on the chemotactic potential of dogfish (Scyliorhinus canicula) leucocytes using the bipolar shape formation assay.

    PubMed

    Hunt, T C; Rowley, A F

    1986-06-01

    The bipolar shape formation assay, previously used to determine the chemotactic potential of various factors for mammalian leucocytes, was tested in the present study with granulocytes of the lesser spotted dogfish, Scyliorhinus canicula. Bipolar shape formation was found to be a temperature dependent process with maximal formation observed at 30 degrees C. Addition of the formyl peptide, N-formyl-methionyl-phenylalanine failed to induce any bipolar forms at all temperatures and concentrations tested.

  13. In vitro immunotoxicological effects of heavy metals on European sea bass (Dicentrarchus labrax L.) head-kidney leucocytes.

    PubMed

    Morcillo, Patricia; Cordero, Héctor; Meseguer, José; Esteban, M Ángeles; Cuesta, Alberto

    2015-11-01

    The knowledge about the direct effects of heavy metals on fish leucocytes is still limited. We investigate the in vitro effects of heavy metals (Cd, Hg, Pb or As) on oxidative stress, viability and innate immune parameters of head-kidney leucocytes (HKLs) from European sea bass (Dicentrarchus labrax). Production of free oxygen radicals was induced by Cd, Hg and As, mainly after 30 min of exposure. Cd and Hg promoted both apoptosis and necrosis cell death while Pb and As did only apoptosis, in all cases in a concentration-dependent manner. Moreover, expression of genes related to oxidative stress and apoptosis was significantly induced by Hg and Pb but down-regulated by As. In addition, the expression of the metallothionein A gene was up-regulated by Cd and Pb exposure though this transcript, as well as the heat shock protein 70, was down-regulated by Hg. Cd, methylmercury (MeHg) and As reduced the phagocytic ability, whereas Hg and Pb increased it. Interestingly, all the heavy metals decreased the phagocytic capacity (the number of ingested particles per cell). Leucocyte respiratory burst changed depending on the metal exposure, usually in a time- and dose-manner. Interestingly, the expression of immune-related genes was slightly affected by Cd, MeHg, As or Pb being Hg the form producing the greatest alterations, which included down-regulation of immunoglobulin M and hepcidin, as well as the up-regulation of interleukin-1 beta mRNA levels. This study provides an in vitro approach for elucidating the heavy metals toxicity, and particularly the immunotoxicity, in fish leucocytes.

  14. Effects of polyamines on cellular innate immune response and the expression of immune-relevant genes in gilthead seabream leucocytes.

    PubMed

    Reyes-Becerril, Martha; Ascencio-Valle, Felipe; Tovar-Ramírez, Dariel; Meseguer, José; Esteban, María Ángeles

    2011-01-01

    It is well known that the polyamines spermidine and spermine, along with the diamine putrescine, are involved in many cellular processes and they are known to play an important role in the control of the innate immune response in higher vertebrates. However, to the best of our knowledge, no studies have focused on their immunological implications in other vertebrates, such as fish. For this reason, the effects of polyamines on the cellular innate immune response and immune-related gene expression were evaluated in vitro, using seabream head-kidney leucocytes (HKL). For this study, head-kidney leucocytes were incubated with the polyamines putrescine, spermine or spermidine (0.005 and 0.0025%) for 0.50, 1, 2 or 4 h. No significant effect was observed on either leucocyte viability or the innate cellular immune responses (peroxidase content and phagocytic and respiratory burst activities). The polyamines produced an increase in respiratory burst and phagocytic ability when leucocytes were incubated principally with putrescine (0.005 and 0.0025%) after 2 and 4 h of the experiment. Finally, the expression levels of immune-associated genes (IgM, MHCIα, MHCIIα, C3, IL-1β, CD8, Hep, NCCRP-1, CSF-1 and TLR) were quantified by real-time PCR and some of them (C3, MHCI, CD8, IgM and Hep) were up-regulated by the higher polyamine concentration. Further studies are needed to ascertain how polyamines control the immune system of seabream as well as which mechanisms are involved.

  15. In vitro immunotoxicity of untreated and treated urban wastewaters using various treatment processes to rainbow trout leucocytes.

    PubMed

    Gagné, François; Fortier, Marlène; Fournier, Michel; Smyth, Shirley-Anne

    2013-07-01

    Municipal effluents are known to impede the immune system of aquatic organisms. The purpose of this study was to examine the immunotoxicity of urban wastewaters before and after 6 treatment processes from 12 cities toward trout leucocytes. Freshly prepared trout leucocytes were exposed to increasing concentrations of solid phase (C18) extracts of wastewaters for 24 hr at 150C. Immunocompetence was determined by following changes in leucocyte viability and the proportion of cells able to ingest at least one (immunoactivity) and at least three (immunoefficiency) fluorescent beads. The influents were treated by six different treatment strategies consisting of facultative aerated lagoons, activated sludge, biological aerated filter, biological nutrient removal, chemically-assisted physical treatment and trickling filter/solid contact. Water quality parameters of the wastewaters revealed that the plants effectively removed total suspended solids and reduced the chemical oxygen demand. The results revealed that the effluents' immunotoxic properties were generally more influenced by the properties of the untreated wastewaters than by the treatment processes. About half of the incoming influents decreased leucocyte viability while 4 treatment plants were able to reduce toxicity. The influents readily increased phagocytosis activity for 8/12 influents while it was decreased in 4/12 influents. This increase was abolished for 4/12 of the effluents using treatments involving biological and oxidative processes. In conclusion, municipal effluents have the potential to alter the immune system in fish and more research will be needed to improve the treatments of wastewaters to better protect the quality of the aquatic environment.

  16. Up-regulation of leucocytes genes implicated in telomere dysfunction and cellular senescence correlates with depression and anxiety severity scores.

    PubMed

    Teyssier, Jean-Raymond; Chauvet-Gelinier, Jean-Christophe; Ragot, Sylviane; Bonin, Bernard

    2012-01-01

    Major depressive disorder (MDD) is frequently associated with chronic medical illness responsible of increased disability and mortality. Inflammation and oxidative stress are considered to be the major mediators of the allostatic load, and has been shown to correlate with telomere erosion in the leucocytes of MDD patients, leading to the model of accelerated aging. However, the significance of telomere length as an exclusive biomarker of aging has been questioned on both methodological and biological grounds. Furthermore, telomeres significantly shorten only in patients with long lasting MDD. Sensitive and dynamic functional biomarkers of aging would be clinically useful to evaluate the somatic impact of MDD. To address this issue we have measured in the blood leucocytes of MDD patients (N=17) and controls (N=16) the expression of two genes identified as robust biomarkers of human aging and telomere dysfunction: p16(INK4a) and STMN1. We have also quantified the transcripts of genes involved in the repair of oxidative DNA damage at telomeres (OGG1), telomere regulation and elongation (TERT), and in the response to biopsychological stress (FOS and DUSP1). The OGG1, p16(INK4a), and STMN1 gene were significantly up-regulated (25 to 100%) in the leucocytes of MDD patients. Expression of p16(INK4a) and STMN1 was directly correlated with anxiety scores in the depression group, and that of p16(INK4a), STMN and TERT with the depression and anxiety scores in the combined sample (MDD plus controls). Furthermore, we identified a unique correlative pattern of gene expression in the leucocytes of MDD subjects. Expression of p16(INK4) and STMN1 is a promising biomarker for future epidemiological assessment of the somatic impact of depressive and anxious symptoms, at both clinical and subclinical level in both depressive patients and general population.

  17. Efavirenz induces interactions between leucocytes and endothelium through the activation of Mac-1 and gp150,95.

    PubMed

    Orden, Samuel; De Pablo, Carmen; Rios-Navarro, Cesar; Martinez-Cuesta, Maria Angeles; Peris, Jose E; Barrachina, Maria D; Esplugues, Juan V; Alvarez, Angeles

    2014-04-01

    The potential cardiovascular (CV) toxicity associated with combined antiretroviral therapy (cART) has been attributed mainly to the nucleoside reverse transcriptase inhibitors abacavir and didanosine. However, the other two components of cART--non-nucleoside reverse transcriptase inhibitors (NNRTIs) and protease inhibitors (PIs)--may also be implicated, either directly or by influencing the action of the other drugs. This study evaluates the acute direct effects of the NNRTIs efavirenz and nevirapine and one of the most widely employed PIs, lopinavir, on leucocyte-endothelium interactions, a hallmark of CV disease. Drugs were analysed in vitro in human cells (interactions of peripheral blood polymorphonuclear or mononuclear cells with human umbilical vein endothelial cells) using a flow chamber system, and in vivo in rat mesenteric vessels by means of intravital microscopy. The expression of adhesion molecules in leucocytes and endothelial cells was studied by flow cytometry, and the role of these molecules in white cell recruitment was evaluated by pre-treating human cells or rats with blocking antibodies. Efavirenz and nevirapine, but not lopinavir, increased the rolling flux and adhesion of leucocytes in vitro and in vivo while inducing emigration in rat venules. Efavirenz, but not nevirapine, augmented the levels of CD11b, CD11c and CD18 in neutrophils and monocytes. The actions of efavirenz, but not of nevirapine, were reversed by antibodies against Mac-1 (CD11b/CD18), gp150,95 (CD11c/CD18) or ICAM-1 (CD54). NNRTIs, but not PIs, interfere with leucocyte-endothelial interactions. However, differences between efavirenz and nevirapine suggest a specific CV profile for each compound.

  18. Up-Regulation of leucocytes Genes Implicated in Telomere Dysfunction and Cellular Senescence Correlates with Depression and Anxiety Severity Scores

    PubMed Central

    Teyssier, Jean-Raymond; Chauvet-Gelinier, Jean-Christophe; Ragot, Sylviane; Bonin, Bernard

    2012-01-01

    Background Major depressive disorder (MDD) is frequently associated with chronic medical illness responsible of increased disability and mortality. Inflammation and oxidative stress are considered to be the major mediators of the allostatic load, and has been shown to correlate with telomere erosion in the leucocytes of MDD patients, leading to the model of accelerated aging. However, the significance of telomere length as an exclusive biomarker of aging has been questioned on both methodological and biological grounds. Furthermore, telomeres significantly shorten only in patients with long lasting MDD. Sensitive and dynamic functional biomarkers of aging would be clinically useful to evaluate the somatic impact of MDD. Methodology To address this issue we have measured in the blood leucocytes of MDD patients (N = 17) and controls (N = 16) the expression of two genes identified as robust biomarkers of human aging and telomere dysfunction: p16INK4a and STMN1. We have also quantified the transcripts of genes involved in the repair of oxidative DNA damage at telomeres (OGG1), telomere regulation and elongation (TERT), and in the response to biopsychological stress (FOS and DUSP1). Results The OGG1, p16INK4a, and STMN1 gene were significantly up-regulated (25 to 100%) in the leucocytes of MDD patients. Expression of p16INK4a and STMN1 was directly correlated with anxiety scores in the depression group, and that of p16INK4a, STMN and TERT with the depression and anxiety scores in the combined sample (MDD plus controls). Furthermore, we identified a unique correlative pattern of gene expression in the leucocytes of MDD subjects. Conclusions Expression of p16INK4 and STMN1 is a promising biomarker for future epidemiological assessment of the somatic impact of depressive and anxious symptoms, at both clinical and subclinical level in both depressive patients and general population. PMID:23185405

  19. [Primary orthostatic hypotension].

    PubMed

    Lehmann, M; Hirsch, F W; Auch-Schwelk, W; Alnor, J; Ochs, A; Gastmann, U; Keul, J

    1986-02-01

    We studied a 25-year-old man suffering from primary orthostatic hypotension whose blood pressure decreased to 65/45 mm Hg during orthostasis and to 95/70 mm Hg during ergometric exercise (50 and 100 watt), and whose heart rate responses were inadequate. Resting catecholamine levels were within the normal range and did not show any significant increase related to orthostasis or to ergometric exercise. Hypersensitivity was observed to low doses of intravenous noradrenaline and isoproterenol. Specific binding of 3H-Yohimbine to intact platelets revealed a normal number of alpha-2-adrenoreceptors in agreement with the adrenaline-induced platelet aggregation in vitro, which was, however, in contrast to hypersensitivity to noradrenaline. Specific 3H-Dihydroalprenolol binding to intact polymorphonuclear leucocytes revealed an increased beta-2-adrenoreceptor density in agreement with hypersensitivity to Isoproterenol. Prescription of Fludrocortison improved orthostatic hypotension.

  20. Bin2 Is a Membrane Sculpting N-BAR Protein That Influences Leucocyte Podosomes, Motility and Phagocytosis

    PubMed Central

    Sánchez-Barrena, María José; Vallis, Yvonne; Clatworthy, Menna R.; Doherty, Gary J.; Veprintsev, Dmitry B.; Evans, Philip R.; McMahon, Harvey T.

    2012-01-01

    Cell motility, adhesion and phagocytosis are controlled by actin and membrane remodelling processes. Bridging integrator-2 (Bin2) also called Breast cancer-associated protein 1 (BRAP1) is a predicted N-BAR domain containing protein with unknown function that is highly expressed in leucocytic cells. In the present study we solved the structure of Bin2 BAR domain and studied its membrane binding and bending properties in vitro and in vivo. Live-cell imaging experiments showed that Bin2 is associated with actin rich structures on the plasma membrane, where it was targeted through its N-BAR domain. Pull-down experiments and immunoprecipitations showed that Bin2 C-terminus bound SH3 domain containing proteins such as Endophilin A2 and α-PIX. siRNA of endogenous protein led to decreased cell migration, increased phagocytosis and reduced podosome density and dynamics. In contrast, overexpression of Bin2 led to decreased phagocytosis and increased podosome density and dynamics. We conclude that Bin2 is a membrane-sculpting protein that influences podosome formation, motility and phagocytosis in leucocytes. Further understanding of this protein may be key to understand the behaviour of leucocytes under physiological and pathological conditions. PMID:23285027

  1. Continuous infusion of macrophage inflammatory protein MIP-1alpha enhances leucocyte recovery and haemopoietic progenitor cell mobilization after cyclophosphamide.

    PubMed Central

    Marshall, E.; Woolford, L. B.; Lord, B. I.

    1997-01-01

    Macrophage inflammatory protein 1alpha (MIP-1alpha) inhibits haemopoietic stem cell proliferation. This property has been exploited in a murine chemotherapy model and has been shown to ameliorate cytotoxic-induced myelosuppression after S-phase-specific cytotoxic therapy. We have now shown that BB-10010, a stable mutant of MIP-1alpha, (a) is more effective when administered as a continuous infusion than when bolus injected and (b), when administered via a 7-day infusion during and after cyclophosphamide treatment, results in an earlier recovery of leucocyte numbers. This effect was accompanied by progenitor cell mobilization into the peripheral blood and included primitive cells with marrow-repopulating ability (MRA). Maximal mobilization and recovery of leucocytes occurred when MIP-1alpha was combined with granulocyte colony-stimulating factor (G-CSF) therapy. The findings suggest that MIP1-alpha used alone or in combination with G-CSF may allow delivery of a greater chemotherapy dose intensity as a consequence of both accelerated leucocyte recovery and maintenance of high-quality mobilized progenitor cells for harvesting and peripheral blood stem cell transplantation. PMID:9192972

  2. Response of in vivo protein synthesis in T lymphocytes and leucocytes to an endotoxin challenge in healthy volunteers.

    PubMed

    Januszkiewicz, A; Loré, K; Essén, P; Andersson, B; McNurlan, M A; Garlick, P J; Ringdén, O; Andersson, J; Wernerman, J

    2002-11-01

    In vivo determination of protein synthesis in immune cells reflects metabolic activity and immunological activation. An intravenous injection of endotoxin to healthy volunteers was used as a human sepsis model, and in vivo protein synthesis of T lymphocytes and leucocytes was measured. The results were related to plasma concentrations of selected cytokines, peripheral cell counts and subpopulations of immune cells. The subjects (n = 8 + 8) were randomized to an endotoxin (4 ng/kg) or a saline group. In vivo protein synthesis was determined twice: before and 1-2.5 h after the endotoxin/saline injection. Protein synthesis decreased in isolated T lymphocytes, but increased in leucocytes. Plasma levels of TNF-alpha, IL-8, IL-6, IL-1 ra and IL-10 were elevated, whereas IL-2 and IFN-gamma, produced predominantly by T lymphocytes, did not change in response to endotoxin. Neutrophils increased, whereas lymphocytes and monocytes decreased 2.5 h after the endotoxin injection. Flow cytometry revealed a drop in total CD3+ T lymphocytes and CD56+ natural killer cells, accompanied by an increase in CD15+ granulocytes. In summary, in vivo protein synthesis decreased in T lymphocytes, while the total leucocyte population showed a concomitant increase immediately after the endotoxin challenge. The changes in protein synthesis were accompanied by alterations in immune cell subpopulations and in plasma cytokine levels.

  3. Third component of complement, immunoglobulin deposition, and leucocyte attachment related to surface sulfate on larval Taenia taeniaeformis.

    PubMed

    Letonja, T; Hammerberg, B

    1983-08-01

    Cysticerci and strobilocerci of Taenia taeniaeformis were incubated with leucocytes from peritoneal washings of normal and T. taeniaeformis-infected rats in the presence of either normal sera or sera from infected rats. Leucocytes from infected and normal rats attached exclusively to the scolices but not the bladders of the larvae in the presence of serum from normal or infected rats. Heat inactivation at 56 C for 30 min destroyed the serum-mediated cell attachment. Histochemical staining of the larval taeniids with acid Alcian Blue demonstrated high concentrations of sulfated mucopolysaccharides on bladders that were not present on scolices. Immunofluorescent staining detected no difference in IgG deposition on the surfaces of bladders and scolices after incubation with rat sera in contrast to the markedly greater amounts of complement protein C3 found on scolices versus bladders. These results indicate that polysulfated substances on the bladder of this larval taeniid are associated with regional resistance to C3 deposition and leucocyte attachment.

  4. Response of in vivo protein synthesis in T lymphocytes and leucocytes to an endotoxin challenge in healthy volunteers

    PubMed Central

    Januszkiewicz, A; Loré, K; EsséN, P; Andersson, B; Mcnurlan, M A; Garlick, P J; RingdéN, O; Andersson, J; Wernerman, J

    2002-01-01

    In vivo determination of protein synthesis in immune cells reflects metabolic activity and immunological activation. An intravenous injection of endotoxin to healthy volunteers was used as a human sepsis model, and in vivo protein synthesis of T lymphocytes and leucocytes was measured. The results were related to plasma concentrations of selected cytokines, peripheral cell counts and subpopulations of immune cells. The subjects (n = 8 + 8) were randomized to an endotoxin (4 ng/kg) or a saline group. In vivo protein synthesis was determined twice: before and 1–2·5 h after the endotoxin/saline injection. Protein synthesis decreased in isolated T lymphocytes, but increased in leucocytes. Plasma levels of TNF-α, IL-8, IL-6, IL-1 ra and IL-10 were elevated, whereas IL-2 and IFN-γ, produced predominantly by T lymphocytes, did not change in response to endotoxin. Neutrophils increased, whereas lymphocytes and monocytes decreased 2·5 h after the endotoxin injection. Flow cytometry revealed a drop in total CD3+ T lymphocytes and CD56+ natural killer cells, accompanied by an increase in CD15+ granulocytes. In summary, in vivo protein synthesis decreased in T lymphocytes, while the total leucocyte population showed a concomitant increase immediately after the endotoxin challenge. The changes in protein synthesis were accompanied by alterations in immune cell subpopulations and in plasma cytokine levels. PMID:12390314

  5. Quantitative and volume, conductivity and scatter changes in leucocytes of patients with acute undifferentiated febrile illness: a pilot study.

    PubMed

    Kalra, Varun; Ahmad, Sohaib; Shrivastava, Vikas; Mittal, Garima

    2016-05-01

    A single diagnostic test for acute undifferentiated febrile illnesses (AUFI) is elusive. This pilot study was undertaken on the premise that leucocytes, being the main cells of defence, undergo quantitative, structural and functional changes in AUFI. We evaluated the potential of volume, conductivity and scatter (VCS) parameters of leucocytes, generated with the haemogram report by the Coulter auto-analyzer, in differentiating the common etiologies of AUFI. The haematological and VCS data obtained from 800 controls and 200 cases of AUFI (50 cases each of acute malaria, dengue, scrub typhus and enteric fever) were retrieved for analysis. The cases and controls differed significantly with respect to relative numbers and the VCS parameters of neutrophils, lymphocytes and monocytes (p<0.05). The neutrophil and lymphocyte were significantly voluminous in acute malaria and scrub typhus as compared to dengue and enteric fevers (p<0.05). Enteric fever significantly enhanced the conductivity of neutrophils as compared to other subgroups while lymphocyte conductivity significantly differed from dengue and scrub typhus. Lymphocyte and neutrophil scatter values in malaria and scrub typhus were comparable but differed significantly from that in enteric fever. Etiology-specific changes occur in leucocytes, both in numbers and their VCS properties which can be identified without additional cost. © The Author 2016. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  6. Factors influencing in vitro respiratory burst assays with head kidney leucocytes from rainbow trout, Oncorhynchus mykiss (Walbaum).

    PubMed

    Chettri, J K; Holten-Andersen, L; Buchmann, K

    2010-07-01

    Abstract Head kidney leucocytes are central elements in a number of in vivo and in vitro assays elucidating innate and adaptive immune mechanisms in teleosts following stimulation with various antigens. These systems are sensitive to several factors affecting the outcome of the assays. The present work describes the importance of temperature, cell concentration, exposure time and immune-modulatory molecules on the respiratory burst activity (RBA) of rainbow trout head kidney leucocytes in vitro. Some variation in RBA was observed among individual fish. However, use of cells pooled from four individuals produced satisfactory results following exposure to phorbol 12-myristate 13-acetate, zymosan and beta-glucan. Temperature was shown to have a significant effect on production of reactive radicals as illustrated by a high activity in cells maintained at 15-20 degrees C and a reduced activity at temperature extremes (1, 4 and 30 degrees C). Highest activity was found at a cell concentration of 1 x 10(7) cells mL(-1). Reactivity showed a clear decline when cells were exposed for more than 4 h. Moreover, incubation of cells with inhibitory substances viz., DiMePE2, cortisol and superoxide dismutase decreased the RBA. It is concluded that several biotic and abiotic factors should be taken into account when conducting RBA assays with head kidney leucocytes for elucidation of rainbow trout immune responses.

  7. Different impact of heat-inactivated and viable lactic acid bacteria of aquatic origin on turbot (Scophthalmus maximus L.) head-kidney leucocytes.

    PubMed

    Muñoz-Atienza, Estefanía; Araújo, Carlos; Lluch, Nuria; Hernández, Pablo E; Herranz, Carmen; Cintas, Luis M; Magadán, Susana

    2015-05-01

    In aquaculture, several criteria should be considered to select an appropriate probiotic, including the aquatic origin and safety of the strain and its ability to modulate the host immune response. The properties and effects of probiotics are strain-specific and some factors such as viability, dose and duration of diet supplementation may regulate their immunomodulatory activities. In this study, we assessed the in vitro effect of eight heat-inactivated and viable lactic acid bacteria (LAB) of aquatic origin belonging to the genera Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Pediococcus and Weissella on the viability and innate immune response of turbot (Scophthalmus maximus L.) leucocytes. Head-kidney leucocytes were incubated with viable and heat-inactivated LAB at different concentrations. After incubation, the viability of leucocytes was evaluated using colorimetric assays (MTT and LDH) and flow cytometry (annexin V/propidium iodide). Heat-inactivated LAB showed no cytotoxic effect while viable LAB exerted variable influence on apoptosis of turbot phagocytes and lymphocytes. Leucocyte respiratory burst activity and phagocytosis were also differentially activated, as viable LAB stimulated leucocytes more efficiently than the heat-inactivated LAB. Our results suggest diverse strain-specific mechanisms of interaction between the evaluated LAB and turbot leucocytes. Furthermore, our work sets up in vitro systems to evaluate the effect of LAB as potential probiotics, which will be useful to develop efficient screening.

  8. Experiences of registered nurses transitioning from employment in acute care to primary health care-quantitative findings from a mixed-methods study.

    PubMed

    Ashley, Christine; Halcomb, Elizabeth; Brown, Angela; Peters, Kath

    2017-06-15

    To describe the experiences of registered nurses who transition from acute to primary health care (PHC) employment. Internationally the provision of health care in PHC settings is increasing. Nurses are moving from acute care employment to meet the growing demand for a PHC workforce. However, little is known about the transition experiences of these nurses. A sequential mixed-methods study comprising a survey, and semi-structured interviews. This study reports on survey findings relating to the transition experience. Convenience and snowballing techniques were used to recruit 111 registered nurses who had transitioned from Australian acute settings to PHC employment within the last 5 years. An online survey gathered data relating to personal and professional demographics, type of PHC setting and transition experiences. Most respondents (n = 90, 81.1%) reported receiving some orientation, although the length and content varied considerably. Those working in metropolitan locations were more likely to report concerns associated with their orientation, with respondents from rural or remote locations more likely to have access to a preceptor than city/metropolitan respondents. Just under half of respondents found prioritising workload (n = 47; 42.7%) or organisational knowledge (n = 45; 40.9%) difficult or very difficult, and 47.7% (n = 53) felt isolated or unsupported. 49.5% (n = 55) reported being overwhelmed with the new role either sometimes or regularly. Barriers to transitioning successfully included limited employer support to attend professional development activities. Availability of specific support measures may assist in the transition process. Findings from our study should be considered by employers when recruiting nurses new to PHC, and when designing orientation and ongoing education programmes. This study highlights the challenges faced by nurses who transition from acute care into PHC employment. Understanding the barriers and facilitators to

  9. Why service users do not complain or have 'voice': a mixed-methods study from Nepal's rural primary health care system.

    PubMed

    Gurung, Gagan; Derrett, Sarah; Gauld, Robin; Hill, Philip C

    2017-01-25

    Despite abundant literature on the different aspects of health care complaint management systems in high-income countries, little is known about this area in less developed health care systems and most research to date has been conducted in hospital settings. This article seeks to address this gap by reporting on research into complaint systems in primary health care (PHC) settings in Nepal. Using a mixed-methods design, qualitative interviews were conducted with key informants (n = 39) and six community focus groups (n = 56), in the Dang District of Nepal. In addition, interviewer-administered structured questionnaire interviews were held with 400 service users, health facility operation and management committee (HFMC) members and service providers from 22 of the 39 public health facilities. Qualitative data were transcribed, organized and then analyzed using the framework method in QSR NVivo 10, while quantitative data were analyzed using IBM SPSS 22. Despite service users having grievances with the health system, they did not complain frequently: 9% (n = 20) reported ever making complaints about the PHC services. Complaints made were about medicines, health facility opening hours, health facility physical environment, and service providers, and were categorized into environment/equipment, accessibility/availability, level of empathy in the care process and care/safety. Generally, complaints were made verbally to health providers or to HFMC members or female community health volunteers. Use of formal channels such as suggestion boxes or written complaints was almost non-existent. Reasons reported for not complaining included: a lack of complaint channels; lack of knowledge of service entitlements; power asymmetry between service providers and service users; lack of opportunity to choose alternative providers, lack of an established culture of complaining, and a perceived lack of responsiveness to complaints. Very few service users made complaints to

  10. Patient-, organization-, and system-level barriers and facilitators to preventive oral health care: a convergent mixed-methods study in primary dental care.

    PubMed

    Templeton, Anna Rose; Young, Linda; Bish, Alison; Gnich, Wendy; Cassie, Heather; Treweek, Shaun; Bonetti, Debbie; Stirling, Douglas; Macpherson, Lorna; McCann, Sharon; Clarkson, Jan; Ramsay, Craig

    2016-01-12

    Dental caries is the most common chronic disease of adult and childhood, a largely preventable yet widespread, costly public health problem. This study identified patient-, organization-, and system-level factors influencing routine delivery of recommended care for prevention and management of caries in primary dental care. A convergent mixed-methods design assessed six guidance-recommended behaviours to prevent and manage caries (recording risk, risk-based recall intervals, applying fluoride varnish, placing preventive fissure sealants, demonstrating oral health maintenance, taking dental x-rays). A diagnostic questionnaire assessing current practice, beliefs, and practice characteristics was sent to a random sample of 651 dentists in National Health Service (NHS) Scotland. Eight in-depth case studies comprising observation of routine dental visits and dental team member interviews were conducted. Patient feedback was collected from adult patients with recent checkups at case study practices. Key informant interviews were conducted with decision makers in policy, funding, education, and regulation. The Theoretical Domains Framework within the Behaviour Change Wheel was used to identify and describe patient-, organization-, and system-level barriers and facilitators to care. Findings were merged into a matrix describing theoretical domains salient to each behaviour. The matrix and Behaviour Change Wheel were used to prioritize behaviours for change and plan relevant intervention strategies. Theoretical domains associated with best practice were identified from the questionnaire (N-196), case studies (N = 8 practices, 29 interviews), and patient feedback (N = 19). Using the study matrix, key stakeholders identified priority behaviours (use of preventive fissure sealants among 6-12-year-olds) and strategies (audit and feedback, patient informational campaign) to improve guidance implementation. Proposed strategies were assessed as appropriate for immediate

  11. Induction of apoptosis in a carp leucocyte cell line infected with turbot (Scophthalmus maximus L.) rhabdovirus.

    PubMed

    Du, Changsheng; Zhang, Qiya; Li, Chunliang; Miao, Dali; Gui, Jianfang

    2004-05-01

    A rhabdovirus was observed from the diseased turbot (Scophthalmus maximus L.) with lethal syndrome. In this study, a carp leucocyte (CLC) cell line was used to investigate the infection process and cell death mechanism occurring during the virus infection. Strong cytopathogenic effect (CPE) and the morphological changes, such as extreme chromatin condensation, nucleus fragmentation, and apoptotic body formation, were observed under fluorescence microscopy after DAPI staining in the infected CLC cells. Transmission electron microscopy analysis showed cell shrinkage, plasma membrane blebbing, cytoplasm vacuolization, chromatin condensation, nuclear breakdown and formation of discrete apoptotic bodies. The bullet-shaped nucleocapsids were measured and ranged in size from 110 to 150 nm in length and 40 to 60 nm in diameter. And therefore the virus is called Scophthalmus maximus rhabdovirus (SMRV). Agarose gel electrophoresis analysis of the DNA extracted from infected cells showed typical DNA ladder in the course of SMRV infection. Flow cytometry analysis of SMRV infected CLC cells detected apoptotic peak in the virus infected CLC cells. Virus titre analysis and electron microscopic observation revealed that the virus replication fastigium was earlier than that of the apoptosis occurrence. No apoptosis was observed in the CLC infected with UV-inactivated SMRV. All these supported that SMRV infected CLC cells undergo apoptosis and the virus replication is necessary for apoptosis induction of CLC cells.

  12. Paroxysmal nocturnal haemoglobinuria phenotype cells and leucocyte subset telomere length in childhood acquired aplastic anaemia.

    PubMed

    Tutelman, Perri R; Aubert, Geraldine; Milner, Ruth A; Dalal, Bakul I; Schultz, Kirk R; Deyell, Rebecca J

    2014-03-01

    The significance of paroxysmal nocturnal haemoglobinuria (PNH(pos) ) cells and leucocyte subset telomere lengths in paediatric aplastic anaemia (AA) is unknown. Among 22 children receiving immunosuppressive therapy (IST) for AA, 73% (16/22) were PNH(pos) , of whom 94% achieved at least a partial response (PR) to IST; 11/16 (69%) achieved complete response (CR). Only 2/6 (33%) PNH(neg) patients achieved PR. PNH(pos) patients were less likely to fail IST compared to PNH(neg) patients (odds ratio 0·033; 95% confidence interval 0·002-0·468; P = 0·012). Children with AA had short granulocyte (P = 7·8 × 10(-9) ), natural killer cell (P = 6·0 × 10(-4) ), naïve T lymphocyte (P = 0·002) and B lymphocyte (P = 0·005) telomeres compared to age-matched normative data.

  13. Distinct evolutionary strategies of human leucocyte antigen loci in pathogen-rich environments

    PubMed Central

    Sanchez-Mazas, Alicia; Lemaître, Jean-François; Currat, Mathias

    2012-01-01

    Human leucocyte antigen (HLA) loci have a complex evolution where both stochastic (e.g. genetic drift) and deterministic (natural selection) forces are involved. Owing to their extraordinary level of polymorphism, HLA genes are useful markers for reconstructing human settlement history. However, HLA variation often deviates significantly from neutral expectations towards an excess of genetic diversity. Because HLA molecules play a crucial role in immunity, this observation is generally explained by pathogen-driven-balancing selection (PDBS). In this study, we investigate the PDBS model by analysing HLA allelic diversity on a large database of 535 populations in relation to pathogen richness. Our results confirm that geographical distances are excellent predictors of HLA genetic differentiation worldwide. We also find a significant positive correlation between genetic diversity and pathogen richness at two HLA class I loci (HLA-A and -B), as predicted by PDBS, and a significant negative correlation at one HLA class II locus (HLA-DQB1). Although these effects are weak, as shown by a loss of significance when populations submitted to rapid genetic drift are removed from the analysis, the inverse relationship between genetic diversity and pathogen richness at different loci indicates that HLA genes have adopted distinct evolutionary strategies to provide immune protection in pathogen-rich environments. PMID:22312050

  14. Albumin inhibits human polymorphonuclear leucocyte luminol-dependent chemiluminescence: evidence for oxygen radical scavenging.

    PubMed Central

    Holt, M. E.; Ryall, M. E.; Campbell, A. K.

    1984-01-01

    Luminol-dependent chemiluminescence of normal human polymorphonuclear leucocytes (PMN) which were resting, or stimulated by unopsonized latex beads, opsonized zymosan or the chemotactic peptide N-formyl-met-leu-phe was decreased more than 80% in the presence of physiological concentrations of albumin (4%, w/v). This inhibition did not result from impairment of light transmission, cellular toxicity, luminol excited-state quenching or a dialysable contaminant in the albumin preparation, but was reduced by 30% when the fall induced by albumin in extracellular free Ca2+ concentration was corrected. The inhibition was most apparent in the larger second phase of the PMN chemiluminescent response to chemotactic peptide or opsonized zymosan stimulation. The smaller first phase of these responses was in fact enhanced by low concentrations of albumin (0.05-0.5%, w/v) and only inhibited up to 50% by 4% (w/v) albumin. Albumin in the range 0.1-4% (w/v) exerted a similar effect on chemiluminescence resulting from superoxide anion (O-2) and hydrogen peroxide (H2O2) production by xanthine oxidase catalysed oxidation of xanthine in the presence of luminol. We suggest that the effect of albumin on PMN luminol-dependent chemiluminescence is mediated by modification of the oxygen radical generating pathway, or oxygen radical scavenging. This previously undocumented property of the major extracellular protein requires further examination if oxygen radicals are to be established as important mediators of inflammation. PMID:6712882

  15. Natural killer cells, killer immunoglobulin-like receptors and human leucocyte antigen class I in disease

    PubMed Central

    Boyton, R J; Altmann, D M

    2007-01-01

    Natural killer cells constitute a potent, rapid part of the innate immune response to infection or transformation, and also generate a link to priming of adaptive immunity. Their function can encompass direct cytotoxicity as well as the release of cytokines and chemokines. In humans, a major component of natural killer (NK) cell target recognition depends mainly on the surveillance of human leucocyte antigen (HLA) class I molecules by killer immunoglobulin-like receptors (KIR). Different KIR can transmit inhibitory or activatory signals to the cell, and effector function is considered to result from the balance of these contributing signals. The regulation of NK cell responses depends on a number of variables: KIR genotype, HLA genotype, heterozygosity versus homozygosity for these, whether there is cognate recognition between the HLA and KIR products carried by an individual, clonal variation between individual NK cells in KIR expression, and the specific modulation of HLA expression by infection, transformation or peptide binding. Different HLA/KIR genotypes can impart different thresholds of activation to the NK cell repertoire and such genotypic variation has been found to confer altered risk in a number of diseases including human immunodeficiency virus (HIV) susceptibility and progression, hepatitis C virus clearance, idiopathic bronchiectasis, autoimmunity and cancer. PMID:17521317

  16. Composition and properties of a cell-membrane fraction from the polymorphonuclear leucocyte

    PubMed Central

    Woodin, A. M.; Wieneke, Antonnette A.

    1966-01-01

    1. The composition of a vesicular cell-membrane fraction from leucocytes has been studied. The bulk of the mass is accounted for as protein and lipid. A small amount of carbohydrate, including some N-acetylneuraminic acid, is present. The phospholipid/cholesterol molar ratio is 1·4 and differs from that for the whole cell. 2. Labile phosphorus groups are present in the membrane but the analysis is complicated by the presence of phosphorus occluded in the membrane vesicles. 3. Leucocidin does not change the gross composition of the membranes or alter the amount or reactivity of the phosphorus compounds. 4. The cell-membrane fraction has considerable avidity for an impurity present in commercial [32P]orthophosphate. When this is removed [32P]orthophosphate or [32P]ATP does not label the membrane. 5. The presence of an NADH2–cytochrome c oxidoreductase and an alkaline phosphatase is described. The adenosine-triphosphatase activity of the membrane has not been found to depend on the presence of Na+ or K+. PMID:4287773

  17. Trypanosoma cruzi: sequence of phagocytosis and cytotoxicity by human polymorphonuclear leucocytes.

    PubMed Central

    Rimoldi, M T; Cardoni, R L; Olabuenaga, S E; de Bracco, M M

    1981-01-01

    We have studied the relationship between phagocytosis and cytotoxicity of human polymorphonuclear leucocytes (PMN) to sensitized Trypanosoma cruzi. Assays were done simultaneously using [3H]-uridine labelled epimastigotes as target cells. Phagocytosis was evaluated by the uptake and cytotoxicity by the release of parasite associated [3H]-uridine. Both reactions reached maximum levels at the same effector- to target-cell ratio and antibody concentration. Uptake of epimastigotes by PMN was highest at 30 min and intracellular disruption and release of parasite debris took place later. In conditions that precluded repeated uptake of sensitized radiolabelled T. cruzi, the release profile of [3H]-uridine from PMN that contained intracellular parasites was similar to that of the standard cytotoxic assay. However, as the ingestion phase was separated from the release step, no lag in the onset of the reaction was observed. Although we cannot rule out extracellular killing, the results of this study demonstrate that the bulk of damaged T. cruzi epimastigotes had been previously internalized by the PMN. PMID:7016743

  18. Allelic distribution of human leucocyte antigen in historical and recently diagnosed tuberculosis patients in Southern Italy

    PubMed Central

    Ruggiero, Giuseppina; Cosentini, Elena; Zanzi, Delia; Sanna, Veronica; Terrazzano, Giuseppe; Matarese, Giuseppe; Sanduzzi, Alessandro; Perna, Francesco; Zappacosta, Serafino

    2004-01-01

    This study addresses the analysis of the human leucocyte antigen (HLA) allele distribution in 54 historical and in 68 recently diagnosed tuberculosis (TB) patients. The historical cohort was characterized by the presence of large fibrocavernous lesions effectively treated with therapeutic pneumothorax during the period 1950–55. Patients and healthy controls enrolled in the study were from the Campania region of southern Italy. No significant association between HLA alleles and TB in the population of recently diagnosed TB patients was observed. On the contrary, among the historical TB patients there was a strong association with an increased frequency of the HLA-DR4 allele alone and/or in the presence of the HLA-B14 allele (P = 0·000004; Pc = 0·0008), as well as with a decreased frequency of the HLA-A2+,-B14−,DR4− allele association (P = 0·00005; Pc = 0·01). In order to exclude any interference from age-related factors, these results were confirmed by comparing the historical cohort of TB patients with an age-matched healthy control population of the same ethnic origin (P = 0·00004; Pc = 0·008; and P = 0·0001; and Pc = 0·02, respectively). PMID:15009432

  19. The effects of some antirheumatic drugs on an in vitro model of human polymorphonuclear leucocyte chemokinesis.

    PubMed Central

    Smith, M. J.; Walker, J. R.

    1980-01-01

    1 A rapid, reproducible in vitro assay for studying the chemokinetic movement of human polymorphonuclear leucocytes (PMNs) is described. Two synthetic peptides, formyl methionyl-leucyl-phenylalanine (FMLP) and formyl methionyl-phenylalanine (FMP), were used as a standard chemokinesins. 2 Maximal chemokinetic movement was observed with peptide concentrations of 2.5 nM (FMLP) and 100 muM (FMP). EC50 values of 650.0 +/- 60.0 pM and 27.0 +/- 3.5 muM respectively are similar to those reported for chemotactic activity of the peptides in micropore filter assays. 3 The PMN chemokinetic response to FMLP was enhanced by histamine (100 nM) and vitamin C (2.5 muM). 4 Human serum albumin was shown to induce chemokinesis but to antagonize the response to FMLP in a dose-related fashion. Fibrinogen similarly antagonized the cell response to peptide. 5 Levamisole (250 nM to 2.5 muM) significantly potentiated the chemokinetic responses to FMLP and FMP in a dose-related manner. The chemokinetic response to FMLP was unaffected by D-penicillamine (250 muM to 10 mM) while alclofenac (500 muM to 1 mM), salicylic acid (250 muM to 10 mM) and indomethacin (100 muM to 1 mM) caused dose-related inhibition. PMID:7397456

  20. Calcium ionophore A23187 induces release of chemokinetic and aggregating factors from polymorphonuclear leucocytes.

    PubMed Central

    Bray, M. A.; Ford-Hutchinson, A. W.; Shipley, M. E.; Smith, M. J.

    1980-01-01

    1. Rat and human polymorphonuclear leucocytes (PMNs) when exposed to calcium ionophore A23187 10 microM release products which cause aggregation of rat PMNs and chemokinesis of human PMNs. 2. Aggregating and chemokinetic activities are rapidly generated; maximal release occurs after 4 min, and can be detected in dilutions of the supernatant of up to 1:1000. 3. Generation of aggregating and chemokinetic activities is inhibited by nordihydroguaiaretic acid 10(-4) to 10(0-7) M, 5,8,11,14-eicosatetraynoic acid 10(-4) and 10(-5) M, BW 755C 10(-4) M and benoxaprofen 10(-4) M, all compounds known to inhibit lipoxygenase pathways of arachidonic acid (AA) metabolism. 4. Conventional non-steroidal anti-inflammatory agents, such as aspirin and indomethacin, inhibited little or not at all the generation of these activities. 5. We conclude that the aggregating and chemokinetic activities induced by A23187 represent generation of biologically active products of lipoxygenase pathways of AA metabolism. PMID:6781577

  1. [Inhibitory effect of nafamostat mesilate (FUT-175) on O2- production in rat polymorphonuclear leucocytes].

    PubMed

    Oda, M; Ogihara, M; Sato, T; Kurumi, M; Iwaki, M

    1986-05-01

    Effect of nafamostat mesilate (FUT-175), a serine protease inhibitor, having anti-inflammatory effects was studied on superoxide (O2-) production in rat polymorphonuclear leucocytes (PMN) and compared with those of other serine protease inhibitors and typical anti-inflammatory agents. 1) O2- productions in rat PMN stimulated with concanavalin A (Con A) and cytochalasin B (Cyt B) were too weak to observe. With NADH, however, strong O2- production was induced by Con A and Cyt B. 2) FUT-175 at 10(-6) and 10(-5) M inhibited O2- production in rat PMN induced by Con A and Cyt B with NADH in a concentration-dependent manner. 3) The serine protease inhibitor L-tosylamido-2-phenylethyl-chloromethyl ketone (TPCK) and soybean trypsin inhibitor (SBTI) inhibited O2- production at 10(-5) M and 10(-4) M, respectively, while aprotinin, chymostatin and leupeptin did not. 4) Neither indomethacin nor dexamethasone, typical anti-inflammatory agents, inhibited O2- production. Mepacrine, a phospholipase A2 inhibitor, strongly inhibited it. 5) O2- production in PMN prepared from the rat administered FUT-175, 200 mg/kg, p.o., was significantly decreased in comparison with that of the control rat. 6) FUT-175 had no effect on O2- production by hypoxanthine-xanthine oxidase. These results showed FUT-175 had a strong inhibitory effect on O2- production in rat PMN which other typical anti-inflammatory agents did not have.

  2. Reduced expression of galectin-1 and galectin-9 by leucocytes in asthma patients.

    PubMed

    Sanchez-Cuellar, S; de la Fuente, H; Cruz-Adalia, A; Lamana, A; Cibrian, D; Giron, R M; Vara, A; Sanchez-Madrid, F; Ancochea, J

    2012-12-01

    Accumulating evidence shows that galectins play roles in the initiation and resolution phases of inflammatory responses by promoting anti- or proinflammatory effects. This study investigated the presence of three members of the galectin family (galectin-1, -3 and -9) in induced sputum samples of asthma patients, as well as their possible implication in the immunopathogenesis of human asthma. Levels of interleukin (IL)-5, IL-13, and galectins were determined in leucocytes isolated from induced sputum samples by reverse transcription-polymerase chain reaction (RT-PCR) immunofluorescence and flow cytometry. High levels of IL-5 and IL-13 mRNA were detected in sputum cells from asthma patients. In parallel, immunoregulatory proteins galectin-1 and galectin-9 showed a reduced expression on macrophages from sputum samples compared with cells from healthy donors. In-vitro immunoassays showed that galectin-1 and galectin-9, but not galectin-3, are able to induce the production of IL-10 by peripheral blood mononuclear cells from healthy donors. These findings indicate that macrophages from sputum samples of asthma patients express low levels of galectin-1 and galectin-9, favouring the exacerbated immune response observed in this disease.

  3. Antibiotic uptake by cultured Atlantic cod leucocytes and effect on intracellular Francisella noatunensis subsp. noatunensis replication.

    PubMed

    Kaldestad, Marte; Haugland, Gyri T; Rønneseth, Anita; Wergeland, Heidrun I; Samuelsen, Ole Bent

    2014-02-04

    The granuloma disease caused by Francisella noatunensis subsp. noatunensis in farmed Atlantic cod has not been successfully treated by use of antibacterials, even when antibacterial resistance testing indicates a sufficient effect. The reason for this treatment failure may be the intracellular existence of the bacteria within immune cells, mainly macrophages. To investigate the effect of antibacterials on intracellular Francisella replication, we established a protocol for the detection of drugs within Atlantic cod immune cells using high-performance liquid chromatography (HPLC). When the uptake and intracellular concentrations of oxolinic acid and flumequine were analysed in isolated adherent head kidney leucocytes (HKLs) by HPLC, we found that uptake was rapid and the intracellular concentrations reflected the extracellular exposure concentrations. To investigate the effect of the antibacterial compounds on intracellular bacterial replication, adherent HKLs experimentally infected with the bacteria were analysed using flow cytometry and intracellular labelling of bacteria by specific antibodies. We found that flumequine did not inhibit intracellular bacterial replication. Unexpectedly, the results indicated that the intracellularly effiacy of the drug was reduced. The HPLC method used proved to be highly applicable for accurate determination of intracellular drug concentrations. When combined with sensitive and specific flow cytometry analyses for identification and measurement of intracellular bacterial replication, we suggest that this approach can be very valuable for the design of antibacterial treatments of intracellular pathogens.

  4. Influence of fucoidans and their derivatives on antitumor and phagocytic activity of human blood leucocytes.

    PubMed

    Anisimova, N Yu; Ustyuzhanina, N E; Donenko, F V; Bilan, M I; Ushakova, N A; Usov, A I; Nifantiev, N E; Kiselevskiy, M V

    2015-07-01

    The immunotropic activity of structurally different fucoidans and their derivatives towards isolated immune blood cells, effectors of innate immune system, was studied. The most potent effect was observed for high molecular weight fucoidan CF from the alga Chordaria flagelliformis, whose backbone is built of (1→3)-linked units of α-L-fucopyranose, and branches included residues of α-D-glucuronic acid and α-L-fucofuranose. This compound at the concentration of 0.05 mg/ml potentiated phagocytosis of Saccharomyces cerevisiae and Lactobacillus acidophilus by neutrophils, increasing relative quantity of phagocytes as well as their effectiveness. Along with this, 14% increase in the concentration of membrane-bound integrin CD11c molecules was observed. The systemic effect of CF at the dose of 0.01 mg/mouse i.p. led to potentiation of cytotoxic activity of spleen mononuclear leucocytes towards melanoma cells of line B16 by 1.9-fold and towards chronic myelogenous leukemia cells of line K-562 by 1.7-fold. These results indicate that fucoidan CF can stimulate anti-infective and antitumor activity of effectors of the innate immune system via CD11c integrins.

  5. Nitric oxide production by nurse shark (Ginglymostoma cirratum) and clearnose skate (Raja eglanteria) peripheral blood leucocytes.

    PubMed

    Walsh, Cathy J; Toranto, Jason D; Gilliland, C Taylor; Noyes, David R; Bodine, Ashby B; Luer, Carl A

    2006-01-01

    Reactive nitrogen intermediates, such as nitric oxide (NO), are important immunomodulators in vertebrate immune systems, but have yet to be identified as mediators of host defence in any member of class Chondrichthyes, the cartilaginous fishes. In the present study, production of NO by nurse shark (Ginglymostoma cirratum) peripheral blood leucocytes (PBL) stimulated with bacterial cell wall lipopolysaccharide (LPS) was investigated. PBL were cultured for 24 to 96 h following stimulation with LPS at concentrations ranging from 0 to 25 microg ml(-1), in both serum-supplemented and serum-free culture conditions. Production of NO was measured indirectly using the Griess reaction, with maximal NO production occurring after 72 h using 10% FBS and 10 microg LPS ml(-1). Application of these culture conditions to PBL from another cartilaginous fish (clearnose skate, Raja eglanteria) resulted in a similar NO response. Addition of a specific inhibitor of inducible nitric oxide synthase (iNOS), L-N(6)-(1-iminoethyl)lysine (L-NIL), resulted in a significant decrease in the production of NO by PBL from both species.

  6. Characterization of a neutral protease from lysosomes of rabbit polymorphonuclear leucocytes

    PubMed Central

    Davies, Philip; Rita, Giuseppe A.; Krakauer, Kathrin; Weissmann, Gerald

    1971-01-01

    1. The subcellular distribution has been investigated of a protease from rabbit polymorphonuclear leucocytes, obtained from peritoneal exudates. The enzyme, optimally active between pH7.0 and 7.5, hydrolyses histone but not haemoglobin, sediments almost exclusively with a granule fraction rich in other lysosomal enzymes, and is latent until the granules are disrupted by various means. 2. Enzymic analysis of specific and azurophilic granules separated by zonal centrifugation showed that neutral protease activity was confined to fractions rich in enzymes characteristic of azurophile granules. 3. Recovery of neutral protease activity from subcellular fractions was several times greater than that found in whole cells. This finding was explained by the presence of a potent inhibitor of the enzyme activity in the cytoplasm. 4. The effect of the inhibitor was reversed by increasing ionic strength (up to 2.5m-potassium chloride) and by polyanions such as heparin and dextran sulphate, but not by an uncharged polymer, dextran. 5. The enzyme was also inhibited, to a lesser extent, by 1-chloro-4-phenyl-3-l-toluene-p-sulphonamidobutan-2-one, soya-bean trypsin inhibitor and ∈-aminohexanoate (∈-aminocaproate). 6. The granule fractions failed to hydrolyse artificial substrates for trypsin and chymotrypsin. 7. Partial separation of the enzyme was achieved by Sephadex gel filtration at high ionic strength and by isoelectric focusing. The partially separated, activated enzyme showed an approximately 300-fold increase in specific activity over that in whole cells. PMID:5126908

  7. Diagnosing tumours on routine surgical sections by immunohistochemistry: use of cytokeratin, common leucocyte, and other markers.

    PubMed Central

    Poston, R N; Sidhu, Y S

    1986-01-01

    Tumours of uncertain tissue of origin were investigated by immunohistochemistry on formalin fixed paraffin embedded sections. Two antibodies--PD7/26, an anti common leucocyte antigen, and CAM5.2, an anticytokeratin--recognised most lymphomas and carcinomas, respectively: 88% of these tumours were identified by the two antibodies alone. These antibodies permitted the separation of the cases into groups: positive with CAM5.2, positive with PD7/26, and a third comprising those negative with both. The negative group contained other tumours and a small number of carcinomas and lymphomas; many of the lymphomas were, apparently, of histiocytic origin. Comparison of CAM5.2 with other epithelial markers showed that it was the most effective. Some further classification of the tumours was carried out with a panel of organ and cell specific antibodies: mesotheliomas were recognised by their pattern of reactivity with epithelial markers. Overall, the tumour type was determined in 90% of cases. Immunohistochemistry performed as described can be a potent aid to the diagnostic histopathology of tumours. Images PMID:2424934

  8. A soluble recombinant form of human leucocyte antigen-G 6 (srHLA-G6).

    PubMed

    Pelá, Flávia Porto; Rustiguel, Joane Kathelen; Rodrigues, Lilian Cataldi; Mendonça, Jacqueline Nakau; Andrade, Camillo Del Cistia; Lopes, Norberto Peporine; Rosa, José Cesar; Nonato, Maria Cristina; Favier, Benoit; Donadi, Eduardo Antônio; Dias-Baruffi, Marcelo

    2017-05-20

    Human Leucocyte Antigen-G (HLA-G) is a non classical major histocompatibility complex (MHC) molecule that through RNA splicing can encode seven isoforms which are membrane bound (-G1, -G2, -G3 and -G4) and soluble (-G5, -G6 and -G7). HLA-G is described as important immune suppressor endogenous molecule to favor maternal-fetal tolerance, transplant survival and tumor immune scape. HLA-G shows low protein variability and a unique structural complexity that is related with the expression of different isoforms followed by biochemical processes, such as, proteolytic cleavage, molecular interactions, and protein ubiquitination. Studies with HLA-G have shown difficult to assess the role of the individual isoforms. Thus, the aim of this work was to obtain a HLA-G6 recombinant form. The results indicated the production of high homogeneous preparations of soluble recombinant HLA-G6 (srHLA-G6) with molecular mass 23,603.76 Da, determined by MALD-TOF/TOF. In addition, native and denatured srHLA-G6 were detected by ELISA, using commercial monoclonal antibodies. Finally, we developed a suitable methodology to express srHLA-G6 that could contribute in structural and functional studies involving specific isoforms. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. Cytosolic Ca2+ content and membrane fluidity of platelets and polymorphonuclear leucocytes in diabetes mellitus.

    PubMed

    Caimi, G; Lo Presti, R; Canino, B; Montana, M; Ventimiglia, G; Catania, A; Sarno, A

    1995-08-01

    Considering the role played by platelets and leucocytes in diabetic disease and keeping in mind the strong correlation between functional and metabolic aspects that characterizes this clinical condition, we evaluated, in two groups of diabetics, respectively the platelet and polymorphonuclear (PMN) cytosolic Ca2+ content (employing the fluorescent probe Fura 2-AM) and membrane fluidity (using the fluorescent probe TMA-DPH and considering the fluorescence polarization degree, inversely related to the membrane fluidity). From the obtained results, it is evident that the platelet cytosolic Ca2+ content does not distinguish normals from diabetics of type 1 and 2; the platelet membrane fluidity instead does not discriminate normals from diabetics, but differentiates diabetics of type 1 and 2 (type 1 = 0.284 +/- 0.015; type 2 = 0.314 +/- 0.018; p < 0.001). PMN cytosolic Ca2+ content and membrane fluidity do not discriminate normals from diabetics. In the two groups of diabetics none of the platelet and PMN parameters (cytosolic Ca2+ content and membrane fluidity) are related to the glycometabolic pattern.

  10. [Effect of 3-oxypyridine and succinic acid derivatives on endometrial leucocyte infiltration and lipid peroxidation in recrudescence of inflammatory diseases of the uterus and its appendages].

    PubMed

    Volchegorskiĭ, I A; Pravdin, E V; Uzlova, T V

    2013-01-01

    The effect of domestic 3-oxypyridine and succinic acid derivatives (emoxipin, reamberin and mexidol) included in the complex treatment of recrudescence of inflammatory diseases of the uterus and its appendages, as manifested in endometrial leucocyte infiltration (ELI) compared to lipid peroxidation products concentration in blood serum, was assessed in a short-term, prospective placebo-controlled single-blind randomized trial. It has been found that two-week infusions of emoxipin (single dose, 150 mg), reamberin (single dose, 400 ml), and mexidol (single dose, 300 mg) favor a decrease in ELI and influence ambiguously on lipid peroxidation - antioxidant (LPA) system condition in blood serum of patients. It has been found that 3-oxypyridine derivative (emoxipin) decreases the intensity of ELI, but does not affect LPA system condition. Succinic acid derivative (reamberin) is inferior to emoxipin in the degree of decreasing ELI, but reduces the concentration of the antioxidant protein ceruloplasmin. Mexidol, being 3-oxipyridine and succinic acid derivative simultaneously, exceeds reamberin in decreasing ELI, increases concentration of blood antioxidant components (alpha-tocopherol and ceruloplasmin), and decreases the level of primary isopropanol-soluble products of lipid peroxidation.

  11. [The changes of peripheral blood leucocytes subpopulation after challenge with CSFV virulent strain Shimen].

    PubMed

    Zhou, Yuan-Cheng; Wang, Qin; Fan, Xue-Zheng; Xu, Lu; Xu, Zhi-Wen; Guo, Wan-Zhu; Liu, Jun; Chen, Lei; Tang, Bo

    2009-07-01

    To investigate the effects of classical swine fever virus (CSFV) virulent strain Shimen (SM) infection on piglets peripheral blood leucocytes, the 60-days weanling piglets were infected with the shinen strain and the peripheral blood samples of the piglets were collected to analyze the kinetics of the CSEV nucleic acid, the peripheral blood leucocytes subpopulation and SLA molecule expression on the peripheral blood leukocytes. The results showed that the piglets rectal temperature increased 48 hours after intramuscular injection of CSFV SM strain, the CSFV nucleic acid was detected in the peripheral blood at 2DPI, the content of CSFV nucleic acid increased and up-regulated to a peak at 6DPI as 10 (4.84 +/- 0.98 times as 2DPI. The amount of WBC, LYM and PLT significantly decreased, where in the amount of WBC decreased to 65.87% at 1DPI and 50% at 2DPI respectively; the amount of LYM decreased to 70.68%, 47.88% and 23.29% at 1DPI, 2DPI, and 3DPI, respectively; the amount of PLT decreased day by day and to 34.59% at 6DPI; the amount of NK, gammadeltaT, Tc, Th, CD3+ CD4+ CD8+ and CD3- CD4- CD8- cells decreased after infection; 78.49% of NK cells decreased at 1DPI and then there was no significant change from 2DPI to 6DPI. The amount of gammadeltaT, Tc, CD4- CD8- CD3-,CD4+ CD8+ CD3+ cells decreased to 41.74%, 43.83%, 15.87%, and 32.96% at 3DPI, respectively, However, the amount of T helper cells decreased continually to 42.95% at 6DPI; the amount of SLA I positive lymphocytes decreased significantly and the amount of SLA I positive CD3 cells decreased to 23.07% and 15.38% at 1DPI and 2DPI respectively; the SLA I positive granulocytes increased continually from 92.20% at 1DPI to 98.30% at 3DPI; the amount of CD3 SLA II + cells in lymphocytes decreased from 1.38% at 1DPI to 0.22% at 2DPI, while the SLA II + granulocytes increased continually to a peak at 3DPI and 53.76% of granulocytes expressed the SLA II molecule, but the percentage of the granulocytes expressing SLA

  12. Mixing method and apparatus

    DOEpatents

    Green, Norman W.

    1982-06-15

    Method of mixing particulate materials comprising contacting a primary source and a secondary source thereof whereby resulting mixture ensues; preferably at least one of the two sources has enough motion to insure good mixing and the particulate materials may be heat treated if desired. Apparatus for such mixing comprising an inlet for a primary source, a reactor communicating therewith, a feeding means for supplying a secondary source to the reactor, and an inlet for the secondary source. Feeding means is preferably adapted to supply fluidized materials.

  13. Nitric oxide-generating system as an autocrine mechanism in human polymorphonuclear leucocytes.

    PubMed Central

    Riesco, A; Caramelo, C; Blum, G; Montón, M; Gallego, M J; Casado, S; López Farré, A

    1993-01-01

    Recent data [Lopéz-Farré, Riesco, Moliz, Egido, Casado, Hernando and Caramelo (1991) Biochem. Biophys. Res. Commun. 178, 884-891] revealed that endothelin 1 (ET-1) increases intracellular free [Ca2+] in polymorphonuclear leucocytes (PMN) by a mechanism that can be inhibited by L-arginine. The aim of the present study was to clarify the mechanisms of the interaction between the effects of ET-1 and L-arginine in human PMN. The experimental findings showed that in human PMN: (a) ET-1 and the chemoattractant peptide N-formylmethionyl- leucyl-phenylalanine (fMLP) induce both the metabolism of L-arginine to L-citrulline and cyclic GMP (cGMP) formation; (b) the ET-1-induced cGMP production is inhibitable by the L-arginine antagonist NG-monomethyl-L-arginine, therefore suggesting the involvement of NO; (c) the ET-1- or fMLP-induced NO/cGMP stimulation is critically dependent on the availability of L-arginine; (d) human PMN possess a L-arginine transport system with both Na(+)-dependent and -independent components; (e) the L-arginine transport system in PMN appears to be feedback-regulated by NO/cGMP in ET-1-stimulated conditions, but not under baseline conditions; (f) the L-arginine transport system in PMN is independent of the gamma-glutamyl cycle and is not modified by either ET-1 or fMLP. The L-arginine/NO/cGMP-dependent mechanisms characterized in the present study may be relevant in the regulation of PMN activation in pathophysiological conditions in vivo. PMID:7686367

  14. Leucocyte expression of genes implicated in the plasminogen activation cascade is modulated by yoghurt peptides.

    PubMed

    Theodorou, Georgios; Politis, Ioannis

    2016-08-01

    The urokinase-plasminogen activator (u-PA), its receptor (u-PAR) and the inhibitors of u-PA (PAI-1 and PAI-2) provide a multi-molecular system in leucocytes that exerts pleiotropic functions influencing the development of inflammatory and immune responses. The objective of the present study was to examine the ability of water soluble extracts (WSE) obtained from traditional Greek yoghurt made from bovine or ovine milk to modulate the expression of u-PA, u-PAR, PAI-1 and PAI-2 in ovine monocytes and neutrophils. WSE were obtained from 8 commercial traditional type Greek yoghurts made from ovine or bovine milk. WSE upregulated the expression of all 4 u-PA related genes in monocytes but the upregulation was much higher in the PAI-1 (10-fold) than in u-PA and u-PAR (3-4 fold) thus, shifting the system towards inhibition. In line with this observation, WSE reduced total and membrane-bound u-PA activity in monocytes. In neutrophils, WSE caused small (50-60%) but significant (P < 0·05) reductions in expression of u-PAR and PAI-2 but had no effect on expression of u-PA, PAI-1 and on total cell-associated and membrane-bound u-PA activity. WSE from yoghurts made from bovine or ovine milk were essentially equally effective in affecting the u-PA system except for the u-PAR gene in ovine neutrophils that was affected (reduced) by the ovine and not the bovine WSE. In conclusion, peptides present in WSE modulated the expression of u-PA related genes but the effect was much more prominent in monocytes than in neutrophils.

  15. Soluble human leucocyte antigen-G and interleukin-10 levels in isocyanate-induced asthma.

    PubMed

    Mapp, C E; Ferrazzoni, S; Rizzo, R; Miotto, D; Stignani, M; Boschetto, P; Maestrelli, P; Baricordi, O R

    2009-06-01

    We previously reported that in moderate-to-severe asthma there is a deficit of IL-10 secretion that could prevent the production of soluble HLA-G (sHLA-G), a non-classical human leucocyte antigen class I molecule with tissue-protective properties in inflammatory responses. Our objective was to investigate the production of sHLA-G and the secretion of IL-10 by peripheral blood mononuclear cells (PBMCs) in asthma induced by isocyanates and to compare the results with those obtained in non-occupational allergic asthma. sHLA-G and IL-10 were measured by ELISA in the culture supernatants of unstimulated or lipopolysaccharide (LPS)-stimulated PBMCs obtained from 20 subjects with isocyanate asthma, 16 asymptomatic subjects exposed to isocyanates, 18 subjects with non-occupational allergic asthma, and 26 healthy control subjects. Occupational exposure to isocyanates was associated with high baseline levels of secretion of IL-10 by PBMCs, whether or not the exposed subjects had asthmatic symptoms. However, spontaneous production of sHLA-G by PBMC was significantly higher in subjects with isocyanate asthma compared with asymptomatic-exposed controls. In contrast, PBMCs from subjects with non-occupational allergic asthma produced sHLA-G only after LPS stimulation. sHLA-G production and IL-10 secretion are influenced by workplace exposure to isocyanates and by development of asthma. The different behaviour of both sHLA-G and IL-10 in asthma induced by isocyanates compared with non-occupational allergic asthma suggests a heterogeneous biological role for HLA-G molecules and for IL-10, a key cytokine of immune and inflammatory responses.

  16. Inheritance of mineralocorticoid effector abnormalities of human mononuclear leucocytes in families with pseudohypoaldosteronism.

    PubMed

    Wehling, M; Kuhnle, U; Daumer, C; Armanini, D

    1989-11-01

    In-vitro effects of aldosterone on intracellular sodium and potassium concentrations have been described for normal human mononuclear leucocytes (HML). After incubation for 1 h at 37 degrees C, intracellular sodium and potassium in HML are significantly higher in the presence of 1.4 nM aldosterone than after incubation without aldosterone. As published earlier, these effects were absent in patients with pseudohypoaldosteronism. In the present paper, the families of seven patients with pseudohypoaldosteronism (index cases) were studied. In the first family, two siblings were affected by the disease and had a reduced number of mineralocorticoid (MC) receptors on HML. Intracellular sodium and potassium in HML from these patients did not show a response to 1.4 nM aldosterone. The parents, who were first cousins, had no history of disease and normal receptor data, but in the mother, the response of HML electrolytes to aldosterone was abnormal. In the second family, the mother of a child with pseudohypoaldosteronism, the mother's sister, and her son, had low numbers of MC receptors. Only the aunt of the index case had an uncertain history of the disease. The MC effector mechanism was abnormal in both children and both mothers studied. In a third family, the effector defect was present only in HML of the father. In three further families the abnormality of the effector mechanism was detected in HML of the patient's mother. These data suggest an autosomal dominant inheritance of pseudohypoaldosteronism with variable expression of the gene.(ABSTRACT TRUNCATED AT 250 WORDS)

  17. Molecular characterization of swine leucocyte antigen class I genes in outbred pig populations.

    PubMed

    Ho, C-S; Lunney, J K; Franzo-Romain, M H; Martens, G W; Lee, Y-J; Lee, J-H; Wysocki, M; Rowland, R R R; Smith, D M

    2009-08-01

    The highly polymorphic swine leucocyte antigen (SLA) genes are one of the most important determinants in swine immune responses to infectious diseases, vaccines, and in transplantation success. Study of SLA influence requires accurate and effective typing methods. We developed a simple and rapid method to type alleles at the three classical SLA class I loci (SLA-1, SLA-3 and SLA-2) using the PCR-sequence-specific primer (PCR-SSP) strategy. This typing system relies on 47 discriminatory PCR primer pairs designed to amplify the SLA class I alleles by groups that have similar sequence motifs. We applied this low-resolution group-specific typing method to characterize the SLA class I alleles present in three outbred pig populations (n = 202). Alleles from 24 class I allele groups corresponding to 56 class I genotypes were detected. We also identified 23 low-resolution SLA class I haplotypes in these pigs and found haplotypes Lr-1.0 (SLA-1*01XX-SLA-3*01XX-SLA-2*01XX) and Lr-4.0 (SLA-1*04XX-SLA-3*04XX-SLA-2*04XX) in all three pig populations with a high prevalence. Over 80% of the pigs examined (n = 162) were found to bear at least one of these haplotypes, resulting in a combined haplotype frequency of nearly 50%. This PCR-SSP-based typing system demonstrates a reliable and unambiguous detection of SLA class I alleles, and can be used to effectively investigate the SLA diversity in outbred pig populations. It will help to identify the role of SLA antigens in disease-resistant pigs and may facilitate the development of effective vaccines.

  18. SRY gene transferred by extracellular vesicles accelerates atherosclerosis by promotion of leucocyte adherence to endothelial cells.

    PubMed

    Cai, Jin; Guan, Weiwei; Tan, Xiaorong; Chen, Caiyu; Li, Liangpeng; Wang, Na; Zou, Xue; Zhou, Faying; Wang, Jialiang; Pei, Fang; Chen, Xinjian; Luo, Hao; Wang, Xinquan; He, Duofen; Zhou, Lin; Jose, Pedro A; Zeng, Chunyu

    2015-08-01

    We set out to investigate whether and how SRY (sex-determining region, Y) DNAs in plasma EVs (extracellular vesicles) is involved in the pathogenesis of atherosclerosis. PCR and gene sequencing found the SRY gene fragment in plasma EVs from male, but not female, patients; EVs from male patients with CAD (coronary artery disease) had a higher SRY GCN (gene copy number) than healthy subjects. Additional studies found that leucocytes, the major source of plasma EVs, had higher SRY GCN and mRNA and protein expression in male CAD patients than controls. After incubation with EVs from SRY-transfected HEK (human embryonic kidney)-293 cells, monocytes (THP-1) and HUVECs (human umbilical vein endothelial cells), which do not endogenously express SRY protein, were found to express newly synthesized SRY protein. This resulted in an increase in the adherence factors CD11-a in THP-1 cells and ICAM-1 (intercellular adhesion molecule 1) in HUVECs. EMSA showed that SRY protein increased the promoter activity of CD11-a in THP-1 cells and ICAM-1 in HUVECs. There was an increase in THP-1 cells adherent to HUVECs after incubation with SRY-EVs. SRY DNAs transferred from EVs have pathophysiological significance in vivo; injection of SRY EVs into ApoE-/- (apolipoprotein-knockout) mice accelerated atherosclerosis. The SRY gene in plasma EVs transferred to vascular endothelial cells may play an important role in the pathogenesis of atherosclerosis; this mechanism provides a new approach to the understanding of inheritable CAD in men.

  19. Phenotypic study of peripheral blood leucocytes in HTLV-I-infected individuals from Minas Gerais, Brazil.

    PubMed

    Brito-Melo, G E A; Martins-Filho, O A; Carneiro-Proietti, A B F; Catalan-Soares, B; Ribas, J G; Thorum, G W; Barbosa-Stancioli, E F

    2002-06-01

    The human T-cell lymphotropic virus type I (HTLV-I)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) associated with the HTLV-I is a well-defined clinical-pathological entity in which the virus and host immune responses contribute to the pathological mechanism. In this study, flow cytometric analysis of whole peripheral blood leucocytes (PBL) was performed to evaluate the immunological status of HTLV-I-infected individuals in an effort to better understand the role of the immune system in the development of HAM/TSP. We have evaluated three groups of infected patients including asymptomatic (AS = 18), ambulatory/oligosymptomatic (AM = 14) and hospitalized HAM/TSP individuals (HO = 42). Noninfected healthy blood donors were used for the control group (NI = 32). Our results demonstrated that the HO group presents an increased percentage of circulating T cells and a decreased percentage of B and natural killer (NK) cells, leading to the highest T/B-cell ratio in comparison with the other groups. Interestingly, while an increased percentage of activated CD4+HLA-DR+ T lymphocytes was observed in both AM and HO, only HO presented higher percentage of activated CD8+HLA-DR+ in combination with the highest CD18 surface expression. This was true for all cell populations analysed, including T lymphocytes, monocytes and neutrophils. Moreover, the HO group was distinguished by a dramatic decrease in the percentage of CD8+CD28+ lymphocytes. Taken together, these findings demonstrate a potent cellular immune activation response involving primarily CD8+ T cells that is concomitant with disease progression in HAM/TSP. We also show that an upregulation of CD18 expression, a hallmark for increased cell migratory potential, might play a critical role in the development/maintenance of HAM/TSP.

  20. Reduction of leucocyte telomere length in radiographic hand osteoarthritis: a population‐based study

    PubMed Central

    Zhai, G; Aviv, A; Hunter, D J; Hart, D J; Gardner, J P; Kimura, M; Lu, X; Valdes, A M; Spector, T D

    2006-01-01

    Background Although age is the strongest predictor of osteoarthritis, the exact mechanism underlying this disorder remains elusive. Objective To examine the association between leucocyte telomere length (LTL), a bio‐indicator of ageing, and radiographic hand osteoarthritis. Methods An unselected, predominantly female sample from the TwinsUK Adult Twin Registry (Twin Research and Genetic Epidemiology Unit, St Thomas Hospital, London, UK) was studied. Radiographs of both hands were obtained with a standard posteroanterior view and assessed for radiographic osteoarthritis according to the Kellgren/Lawrence (K/L) score. Individual radiographic features including osteophytes and joint space narrowing (JSN) were also assessed on a four‐point scale using a standard atlas. Hand osteoarthritis was defined radiographically as having ⩾3 osteoarthritis‐affected joints of both hands (K/L score⩾2). Severity of hand osteoarthritis was indicated semiquantitatively by total K/L scores, osteophytes, JSN scores and proportion of joints affected. Mean LTL was measured by the terminal restriction fragment length using the Southern blot. Results A total of 1086 Caucasian subjects (mean (SD) age 55 (8.0) years) were studied. LTL was 6.95 (0.64) kb and was inversely correlated with age. After adjustment for age, sex, body mass index and smoking, LTL was significantly shorter by 178 bp in subjects with hand osteoarthritis (n = 160) than in those without (n = 926; p = 0.04). LTL was also significantly associated with semicontinuous measures of osteoarthritis (eg, total K/L score, JSN score, osteophyte score and proportion of joints affected) after adjustment (all p⩽0.02) in a dose–response fashion. Conclusion Shorter LTL equivalent to around 11 years of annual loss in normal people is associated with radiographic hand osteoarthritis and disease severity, suggesting potential shared mechanisms between osteoarthritis and ageing, and implicating oxidative

  1. The interaction of leucocidin with the cell membrane of the polymorphonuclear leucocyte

    PubMed Central

    Woodin, A. M.; Wieneke, Antonnette A.

    1966-01-01

    1. When leucocidin is incubated with leucocytes it is inactivated in solution and only a little adsorption takes place. This reaction has been used to purify the cell membrane. 2. The interaction of the membrane with leucocidin is very complex and at least three phenomena occur: (a) An inactivation of leucocidin in solution by large amounts of membrane which is synergistic between the two components of leucocidin, is thermolabile and is not inhibited by electrolyte. (b) An adsorption of leucocidin which is synergistic between the two components of leucocidin, does not proceed to the same extent as the inactivation in solution and is a function of the phospholipid components. Phospholipids isolated from the membrane adsorb leucocidin but the adsorption requires the presence of several molecular species. (c) Polymerization of leucocidin induced by tenfold smaller amounts of membrane than are required to bring about the first two interactions. The polymerization is reversed by adjustment of the ionic strength. It is due to the presence of the lipid components of the membrane. Different lipids are equally effective in inducing the polymerization. 3. Each component of leucocidin will polymerize in the absence of membranes and lose biological activity at low ionic strength. This is reversed by electrolyte and it does not proceed to the same extent as in the presence of membranes. 4. The nature of the interaction of leucocidin with cells, membranes and lipids and the spontaneous polymerization indicate that each component of leucocidin can adopt different isomeric forms. 5. The relationship of the interaction with the membrane to the cytotoxic effect of leucocidin is discussed. ImagesPlate 1. PMID:5944247

  2. Inhibition of human leucocyte elastase by ursolic acid. Evidence for a binding site for pentacyclic triterpenes.

    PubMed Central

    Ying, Q L; Rinehart, A R; Simon, S R; Cheronis, J C

    1991-01-01

    Several pentacyclic triterpenoid metabolites of plant origin are inhibitors of hydrolysis of both synthetic peptide substrates and elastin by human leucocyte elastase (HLE). Ursolic acid, the most potent of these compounds, has an inhibition constant of 4-6 microM for hydrolysis of peptide substrates in phosphate-buffered saline. With tripeptide and tetrapeptide substrates, the inhibition is purely competitive, whereas with a shorter dipeptide substrate the inhibition is non-competitive, suggesting that ursolic acid interacts with subsite S3 of the extended substrate-binding domain in HLE, but not with subsites S1 and S2. The carboxy group at position 28 in the pentacyclic-ring system of the triterpenes contributes to binding to HLE, since replacement of this group with a hydroxy group, as in uvaol, the alcohol analogue of ursolic acid, reduces the potency of inhibition. The inhibitory potency of ursolic acid is also reduced by addition of 1 M-NaCl, further supporting a postulated electrostatic interaction between the negative charge on the triterpene and a positively charged residue on the enzyme, which we assign to the side chain of Arg-217, located in the vicinity of subsites S4 and S5 in HLE. These observations are consistent with a binding site for ursolic acid which extends from S3 towards S4 and S5 on the enzyme. Other triterpenes, including oleanolic acid, erythrodiol, hederagenin and 18 beta-glycyrrhetic acid, can also interact with this binding site. On the basis of these results we conclude that the extended substrate-binding domain of HLE can accommodate a variety of hydrophobic ligands, including not only such molecules as fatty acids [Ashe & Zimmerman (1977) Biochem. Biophys. Res. Commun. 75, 194-199; Cook & Ternai (1988) Biol. Chem. Hoppe-Seyler 369, 629-637], but also polycyclic molecules such as the pentacyclic triterpenoids. PMID:1859379

  3. Association of chronic fatigue syndrome with human leucocyte antigen class II alleles

    PubMed Central

    Smith, J; Fritz, E L; Kerr, J R; Cleare, A J; Wessely, S; Mattey, D L

    2005-01-01

    Background: A genetic component to the development of chronic fatigue syndrome (CFS) has been proposed, and a possible association between human leucocyte antigen (HLA) class II antigens and chronic fatigue immune dysfunction has been shown in some, but not all, studies. Aims: To investigate the role of HLA class II antigens in CFS. Methods: Forty nine patients with CFS were genotyped for the HLA-DRB1, HLA-DQA1, and HLA-DQB1 alleles and the frequency of these alleles was compared with a control group comprising 102 normal individuals from the UK. All patients and controls were from the same region of England and, apart from two patients, were white. Results: Analysis by 2 × 2 contingency tables revealed an increased frequency of HLA-DQA1*01 alleles in patients with CFS (51.0% v 35%; odds ratio (OR), 1.93; p  =  0.008). HLA-DQB1*06 was also increased in the patients with CFS (30.2% v 20.0%; OR, 1.73, p  =  0.052). Only the association between HLA-DQA1*01 and CFS was significant in logistic regression models containing HLA-DQA1*01 and HLA-DRQB1*06, and this was independent of HLA-DRB1 alleles. There was a decreased expression of HLA-DRB1*11 in CFS, although this association disappeared after correction for multiple comparisons. Conclusions: CFS may be associated with HLA-DQA1*01, although a role for other genes in linkage disequilibrium cannot be ruled out. PMID:16049290

  4. Total Leucocyte Count, C-reactive Protein and Neutrophil Count: Diagnostic Aid in Acute Appendicitis

    PubMed Central

    Shafi, Sheikh Muzamil; Afsheen, Misbha; Reshi, Farooq A.

    2009-01-01

    Background/Aim: Acute appendicitis is one of the most common acute intraabdominal affections seen in surgical departments, which can be treated easily if an accurate diagnosis is made in time. Otherwise, delay in diagnosis and treatment can lead to diffuse peritonitis. Materials and Methods: A study was conducted on 110 patients who were operated for acute appendicitis to determine the role and predictive value of the total leucocyte count (TLC), C-reactive protein (CRP) and percentage of neutrophil count in the diagnosis of acute appendicitis. Preoperative TLC, CRP and percentage of neutrophil count were determined and were compared with the results of the histopathology of the removed appendix. Results: Of all the patients studied, 92 had histopathologically positive appendicitis. The TLC was found to be significantly high in 90 patients who proved to have acute appendicitis, whereas CRP was high in only 88 patients and neutrophil percentage was raised in 91; four had a normal CRP level. Thus, TLC had a sensitivity, specificity and positive predictive value of 97.82%, 55.55% and 91.8%, respectively. CRP had a sensitivity, specificity and positive predictive value of 95.6%, 77.77% and 95.6% respectively. Percentage of neutrophil count had a sensitivity, specificity and positive predictive value of 98.9%, 38.88% and 89.21%, respectively. When used in combination, there was a marked improvement in the specificity and the positive predictive value to 88.04% and 98.7%, respectively. Conclusion: The inflammatory markers, i.e., TLC, CRP and neutrophil count can be helpful in the diagnosis when measured together as this increases their specificity and positive predictive value. PMID:19568576

  5. Human leucocyte antigen (HLA) class I and II typing in Belgian multiple sclerosis patients.

    PubMed

    Lysandropoulos, Andreas P; Racapé, Judith; Holovska, Vanda; Toungouz, Michel

    2017-03-01

    This is one of the first studies to compare the frequencies of different human leucocyte antigen (HLA) class I and II alleles and haplotype HLA-DRB1*15-DQB1*06 in a cohort of 119 patients with multiple sclerosis (MS) and a cohort of 124 healthy controls in Belgium. An association with MS was found for the HLA-DRB1*15 (odds ratio [OR] 2.60 [95% confidence interval (CI) 1.51-4.50]) and HLA-DQB1*06 (OR 1.97 [95% CI 1.18-3.29]) alleles, and for haplotype DRB1*15-DQB1*06 (OR 2.63 [95% CI 1.52-4.56]). The HLA-B*07 allele also tended to be more frequent in MS patients (OR 1.46 [95% CI 0.80-2.65]) and more frequent among MS patients with than in those without the HLA-DRB1*15 allele (26/54 [48.1%] versus 6/65 [9.2%]; p value <0.0001). Other alleles were underrepresented in MS patients, such as the HLA-DRB1*07 (OR 0.39 [95% CI 0.21-0.73]) and HLA-A*02 (OR 0.56 [95% CI 0.34-0.94]), showing a protective role against the disease. The HLA-B*44 (OR 0.58 [95% CI 0.31-1.09]) and HLA-DRB1*04 (OR 0.75 [95% CI 0.42-1.34]) alleles tended to be less frequent in MS patients. Altogether, the significant results observed in this population are in line with those from other countries and confirm that propensity to MS can be due to a complex presence of various HLA class I and class II alleles.

  6. Leucocytes are a major source of circulating nicotinamide phosphoribosyltransferase (NAMPT)/pre-B cell colony (PBEF)/visfatin linking obesity and inflammation in humans.

    PubMed

    Friebe, D; Neef, M; Kratzsch, J; Erbs, S; Dittrich, K; Garten, A; Petzold-Quinque, S; Blüher, S; Reinehr, T; Stumvoll, M; Blüher, M; Kiess, W; Körner, A

    2011-05-01

    Nicotinamide phosphoribosyltransferase (NAMPT) is a multifunctional protein potentially involved in obesity and glucose metabolism. We systematically studied the association between circulating NAMPT, obesity, interventions and glucose metabolism and investigated potential underlying inflammatory mechanisms. Fasting morning NAMPT serum levels were measured in cohorts of lean vs obese children, cohorts of intervention by lifestyle, exercise and bariatric surgery, and during an OGTT. In addition, mRNA expression, protein production and enzymatic activity of NAMPT were assessed from isolated leucocytes and subpopulations. Circulating NAMPT was significantly elevated in obese compared with lean children and declined after obesity interventions concomitantly with the decline in BMI, high-sensitivity C-reactive protein (hsCrP) and leucocyte counts. Circulating NAMPT significantly correlated with glucose metabolism and cardiovascular variables in univariate analyses, but only the association with glucose response during an OGTT was independent from BMI. We therefore assessed the NAMPT dynamic following an oral glucose load and found a significant decline of NAMPT levels to 77.0 ± 0.1% as a function of time, and insulin-to-glucose ratio during an OGTT in obese insulin-resistant adolescents. Circulating NAMPT was, however, most strongly associated with leucocyte counts (r = 0.46, p < 0.001). The leucocyte count itself determined significantly and independently from BMI insulin resistance in multiple regression analyses. We systematically evaluated NAMPT expression among several tissues and found that NAMPT was predominantly expressed in leucocytes. In subsequent analyses of leucocyte subpopulations, we identified higher NAMPT protein concentrations in lysates of granulocytes and monocytes compared with lymphocytes, whereas granulocytes secreted highest amounts of NAMPT protein into cell culture supernatant fractions. We confirmed nicotinamide mononucleotide

  7. Primary care characteristics and their association with health screening in a low-socioeconomic status public rental-flat population in Singapore- a mixed methods study.

    PubMed

    Wee, Liang En; Cher, Wen Qi; Sin, David; Li, Zong Chen; Koh, Gerald Choon-Huat

    2016-02-06

    In Singapore, subsidized primary care is provided by centralized polyclinics; since 2000, policies have allowed lower-income Singaporeans to utilize subsidies at private general-practitioner (GP) clinics. We sought to determine whether proximity to primary care, subsidised primary care, or having regular primary care associated with health screening participation in a low socioeconomic-status public rental-flat community in Singapore. From 2009-2014, residents in five public rental-flat enclaves (N = 936) and neighboring owner-occupied precincts (N = 1060) were assessed for participation in cardiovascular and cancer screening. We then evaluated whether proximity to primary care, subsidised primary care, or having regular primary care associated with improved adherence to health screening. We also investigated attitudes to health screening using qualitative methodology. In the rental flat population, for cardiovascular screening, regular primary care was independently associated with regular diabetes screening (adjusted odds ratio, aOR = 1.59, CI = 1.12-2.26, p = 0.009) and hyperlipidemia screening (aOR = 1.82, CI = 1.10-3.04, p = 0.023). In the owner-occupied flats, regular primary care was independently associated with regular hypertension screening (aOR = 9.34 (1.82-47.85, p = 0.007), while subsidized primary care was associated with regular diabetes screening (aOR = 2.94, CI = 1.04-8.31, p = 0.042). For cancer screening, in the rental flat population, proximity to primary care was associated with less participation in regular colorectal cancer screening (aOR = 0.42, CI = 0.17-0.99, p = 0.049) and breast cancer screening (aOR = 0.29, CI = 0.10-0.84, p = 0.023). In the owner-occupied flat population, for gynecological cancer screening, usage of subsidized primary care and proximity to primary care was associated with higher rates of breast cancer and cervical cancer screening; however

  8. The effects of vitamin A supplementation with measles vaccine on leucocyte counts and in vitro cytokine production.

    PubMed

    Jensen, Kristoffer Jarlov; Fisker, Ane Bærent; Andersen, Andreas; Sartono, Erliyani; Yazdanbakhsh, Maria; Aaby, Peter; Erikstrup, Christian; Benn, Christine Stabell

    2016-02-28

    As WHO recommends vitamin A supplementation (VAS) at vaccination contacts after age 6 months, many children receive VAS together with measles vaccine (MV). We aimed to investigate the immunological effect of VAS given with MV. Within a randomised placebo-controlled trial investigating the effect on overall mortality of providing VAS with vaccines in Guinea-Bissau, we conducted an immunological sub-study of VAS v. placebo with MV, analysing leucocyte counts, whole blood in vitro cytokine production, vitamin A status and concentration of C-reactive protein (CRP). VAS compared with placebo was associated with an increased frequency of CRP ≥ 5 mg/l (28 v. 12%; P=0·005). Six weeks after supplementation, VAS had significant sex-differential effects on leucocyte, lymphocyte, monocyte and basophil cell counts, decreasing them in males but increasing them in females. Mainly in females, the effect of VAS on cytokine responses differed by previous VAS: in previous VAS recipients, VAS increased the pro-inflammatory and T helper cell type 1 (Th1) cytokine responses, whereas VAS decreased these responses in previously unsupplemented children. In previous VAS recipients, VAS was associated with increased IFN-γ responses to phytohaemagglutinin in females (geometric mean ratio (GMR): 3·97; 95% CI 1·44, 10·90) but not in males (GMR 0·44; 95% CI 0·14, 1·42); the opposite was observed in previously unsupplemented children. Our results corroborate that VAS provided with MV has immunological effects, which may depend on sex and previous VAS. VAS may increase the number of leucocytes, but also repress both the innate and lymphocyte-derived cytokine responses in females, whereas this repression may be opposite if the females have previously received VAS.

  9. Detection of DNA strand breaks by comet assay in sputum leucocytes of bitumen-exposed workers: a pilot study.

    PubMed

    Marczynski, B; Raulf-Heimsoth, M; Pesch, B; Kendzia, B; Käfferlein, H U; Vosshans, B; Borowitzki, G; Lee, E-H; Bramer, R; Brüning, T

    2010-09-01

    DNA strand breaks were determined in leucocytes of induced sputum (IS) and compared with DNA strand breaks in blood lymphocytes from 42 bitumen-exposed workers pre and post shift. Comet assay results were expressed in arbitrary units based on visual scoring (sputum leucocytes) and Olive tail moment (OTM, blood lymphocytes). DNA damage in IS leucocytes was overall high but did not change during shift. Level of DNA strand breaks in IS samples correlated with total cell count and neutrophil content (Spearman rank correlation coefficient r(s) = 0.47, p = 0.001, r(s)= 0.48, p = 0.001, respectively) and with IL-8 concentration before and after shift (r(s) = 0.31, P = 0.048, and r(s) = 0.43, P = 0.005). DNA damage in IS was not associated with DNA strand breaks in blood lymphocytes (r(s) = -0.04, p = 0.802 before shift, r(s) = 0.27, p = 0.088 after shift). A higher level of DNA strand breaks was measured in blood lymphocytes before shift (median OTM 1.7 before and 1.3 after shift, p = 0.023). A strong correlation was found between the number of neutrophils and IL-8 concentration in IS before and after shift (r(s) = 0.77 and r(s)= 0.75, p < 0.001). This study showed an association between genotoxic and inflammatory effects in the lower airways and compared simultaneously DNA strand breaks in IS and blood of bitumen-exposed workers.

  10. Determinants of Primary School Non-Enrollment and Absenteeism: Results from a Retrospective, Convergent Mixed Methods, Cohort Study in Rural Western Kenya.

    PubMed

    King, Nia; Dewey, Cate; Borish, David

    2015-01-01

    Education is a key element in the socioeconomic development required to improve quality of life in Kenya. Despite the introduction of free primary education, primary school enrollment and attendance levels remain low. Drawing on qualitative and quantitative data, this study explores the determinants of non-enrollment and absenteeism in rural western Kenya and potential mitigation strategies to address these issues. The study was conducted in Bwaliro village in rural western Kenya. A random sample of 64 students was obtained by blocking the village primary school's student population according to grade level, gender, and orphan status. Qualitative and quantitative data were collected through interviews with parents, guardians, and key informants, and focus group discussions with students. Quantitative data were compared using chi-square tests, Student's T-test, and Poisson regressions. Qualitative data were analyzed using thematic content analysis. Malaria, menstruation, and lack of money were among the most notable determinants of primary school dropout and absenteeism, and these factors disproportionately impacted orphans and female students. Potential mitigation strategies suggested by the community included provision of malaria treatment or prevention, reduction in education costs, expansion of the established school-feeding program, and provision of sanitary pads. Despite free primary education, numerous factors continue to prevent children in rural western Kenya from attending primary school. The findings suggest that interventions should primarily target orphaned and female students. Prior to implementation, suggested mitigation strategies should be assessed for cost-effectiveness.

  11. Determinants of Primary School Non-Enrollment and Absenteeism: Results from a Retrospective, Convergent Mixed Methods, Cohort Study in Rural Western Kenya

    PubMed Central

    King, Nia; Dewey, Cate; Borish, David

    2015-01-01

    Background Education is a key element in the socioeconomic development required to improve quality of life in Kenya. Despite the introduction of free primary education, primary school enrollment and attendance levels remain low. Drawing on qualitative and quantitative data, this study explores the determinants of non-enrollment and absenteeism in rural western Kenya and potential mitigation strategies to address these issues. Methods The study was conducted in Bwaliro village in rural western Kenya. A random sample of 64 students was obtained by blocking the village primary school’s student population according to grade level, gender, and orphan status. Qualitative and quantitative data were collected through interviews with parents, guardians, and key informants, and focus group discussions with students. Quantitative data were compared using chi-square tests, Student’s T-test, and Poisson regressions. Qualitative data were analyzed using thematic content analysis. Results Malaria, menstruation, and lack of money were among the most notable determinants of primary school dropout and absenteeism, and these factors disproportionately impacted orphans and female students. Potential mitigation strategies suggested by the community included provision of malaria treatment or prevention, reduction in education costs, expansion of the established school-feeding program, and provision of sanitary pads. Conclusion Despite free primary education, numerous factors continue to prevent children in rural western Kenya from attending primary school. The findings suggest that interventions should primarily target orphaned and female students. Prior to implementation, suggested mitigation strategies should be assessed for cost-effectiveness. PMID:26371885

  12. Influence of inseminate components on porcine leucocyte migration in vitro and in vivo after pre- and post-ovulatory insemination.

    PubMed

    Taylor, U; Schuberth, H J; Rath, D; Michelmann, H W; Sauter-Louis, C; Zerbe, H

    2009-04-01

    A post-breeding migration of leucocytes (PMN) into the uterus is considered to be an important reason for sperm losses. Minimizing such effects may be necessary for successful insemination with low sperm numbers, as required with sex-sorted spermatozoa. We examined the magnitude of PMN influx 3 h after pre- or post-ovulatory insemination with various combinations of seminal plasma (SP), semen extender Androhep (AH; Minitüb, Tiefenbach, Germany) and sperm preparations (S). Pre-ovulatory inseminations with preparations containing 98% AH caused a massive influx of PMN, independent of whether spermatozoa were present (628 +/- 189 x 10(6) leucocytes/uterine horn) or not (580 +/- 153 x 10(6)). Post-ovulatory, 98% AH caused a comparable immigration only in the absence of sperm cells (AH: 569 +/- 198 x 10(6), AH+S: 162 +/- 102 x 10(6)). The presence of SP significantly dampened the numbers of recruited uterine leucocytes. The reaction to all inseminates containing 98% SP both with and without spermatozoa, used before ovulation (SP: 14 +/- 6 x 10(6), SP+S: 73 +/- 27 x 10(6)) and after ovulation (SP: 60 +/- 32 x 10(6), SP+S: 51 +/- 33 x 10(6)) did not differ significantly from controls using phosphate buffered saline (PBS) (pre-ovulatory: 1 +/- 1 x 10(6), post-ovulatory: 11 +/- 9 x 10(6)). Quantitative in vitro transmigration assays with blood-derived PMN proved that AH-induced leucocyte migration into the uterus to be not as a result of direct chemotaxis, because, on account of the chelator citrate, AH significantly inhibited the transmigration towards recombinant human Interleukin-8 (rhCXCL8) (AH: 14 +/- 5% migration rate vs controls: 37 +/- 6%, p < 0.05). Supernatants of spermatozoa incubated in PBS for 1, 12 or 24 h showed neither chemoattractive nor chemotaxis-inhibiting properties. SP at > or =0.1% [v/v] significantly inhibited the in vitro transmigration of PMN. With respect to in vivo migration of neutrophils, the striking difference in the results between semen

  13. Clinical correlates of human leucocyte antigen (HLA)-G in systemic sclerosis1

    PubMed Central

    Favoino, E; Favia, I E; Vettori, S; Vicenti, C; Prete, M; Valentini, G; Perosa, F

    2015-01-01

    Human leucocyte antigen (HLA)-G has a tolerogenic function and could play a role in the pathogenesis of immune-mediated diseases, including systemic sclerosis (SSc). The aim of this study was to evaluate HLA-G serum expression (sHLA-G) and the HLA-G gene 14 base pairs (bp) insertion/deletion (del−/del+) polymorphism in patients with Ssc, to search for possible associations with clinical and laboratory variables. sHLA-G was measured by enzyme-linked immunosorbent assay (ELISA) in sera from 77 patients with SSc and 32 healthy donors (HD); the 14 bp del−/del+ polymorphism was evaluated by polymerase chain reaction (PCR) amplification of peripheral blood mononuclear cells (PBMC) genomic DNA. Receiver operating characteristics (ROC) analysis identified the HLA-G cut-off that best discriminated dichotomized clinical and serological variables, that was subsequently employed to subdivide SSc patients into HLA-G high (HLA-G+) and low (HLA-G−) profile groups. sHLA-G were not statistically different between SSc patients and HD, nor between distinct SSc autoantibody subsets. Subdividing SSc patients by HLA-G positivity or negativity yielded significant differences for the modified Rodnan skin score (mRss) (P = 0·032), ‘general’ (P = 0·031) and ‘kidney’ (P = 0·028) Medsger severity scores (MSS) and disease activity index, and especially Δ heart/lung (P = 0·005). A worse ‘general’ MSS (P = 0·002) and Δ heart/lung (P = 0·011) were more frequent in the low sHLA-G group. These two variables and mRss were associated with sHLA-G levels at logistic regression analysis. Treatment had no influence on sHLA-G. Moreover, a higher frequency of scleredema was detected in the del+/del+ than the del-/del+ group (P = 0.04). These data suggest modulatory effects of sHLA-G on SSc. Prospective studies are needed to investigate a role in predicting the disease course. PMID:25847615

  14. JC Polyomavirus Infection Is Strongly Controlled by Human Leucocyte Antigen Class II Variants

    PubMed Central

    Sundqvist, Emilie; Buck, Dorothea; Warnke, Clemens; Albrecht, Eva; Gieger, Christian; Khademi, Mohsen; Lima Bomfim, Izaura; Fogdell-Hahn, Anna; Link, Jenny; Alfredsson, Lars; Søndergaard, Helle Bach; Hillert, Jan; Oturai, Annette B.; Hemme, Bernhard

    2014-01-01

    JC polyomavirus (JCV) carriers with a compromised immune system, such as in HIV, or subjects on immune-modulating therapies, such as anti VLA-4 therapy may develop progressive multifocal leukoencephalopathy (PML) which is a lytic infection of oligodendrocytes in the brain. Serum antibodies to JCV mark infection occur only in 50–60% of infected individuals, and high JCV-antibody titers seem to increase the risk of developing PML. We here investigated the role of human leukocyte antigen (HLA), instrumental in immune defense in JCV antibody response. Anti-JCV antibody status, as a surrogate for JCV infection, were compared to HLA class I and II alleles in 1621 Scandinavian persons with MS and 1064 population-based Swedish controls and associations were replicated in 718 German persons with MS. HLA-alleles were determined by SNP imputation, sequence specific (SSP) kits and a reverse PCR sequence-specific oligonucleotide (PCR-SSO) method. An initial GWAS screen displayed a strong HLA class II region signal. The HLA-DRB1*15 haplotype was strongly negatively associated to JCV sero-status in Scandinavian MS cases (OR = 0.42, p = 7×10−15) and controls (OR = 0.53, p = 2×10−5). In contrast, the DQB1*06:03 haplotype was positively associated with JCV sero-status, in Scandinavian MS cases (OR = 1.63, p = 0.006), and controls (OR = 2.69, p = 1×10−5). The German dataset confirmed these findings (OR = 0.54, p = 1×10−4 and OR = 1.58, p = 0.03 respectively for these haplotypes). HLA class II restricted immune responses, and hence CD4+ T cell immunity is pivotal for JCV infection control. Alleles within the HLA-DR1*15 haplotype are associated with a protective effect on JCV infection. Alleles within the DQB1*06:03 haplotype show an opposite association. These associations between JC virus antibody response and human leucocyte antigens supports the notion that CD4+ T cells are crucial in the immune defence to JCV and lays

  15. Inherited human group IVA cytosolic phospholipase A2 deficiency abolishes platelet, endothelial, and leucocyte eicosanoid generation

    PubMed Central

    Kirkby, Nicholas S.; Reed, Daniel M.; Edin, Matthew L.; Rauzi, Francesca; Mataragka, Stefania; Vojnovic, Ivana; Bishop-Bailey, David; Milne, Ginger L.; Longhurst, Hilary; Zeldin, Darryl C.; Mitchell, Jane A.; Warner, Timothy D.

    2016-01-01

    Eicosanoids are important vascular regulators, but the phospholipase A2 (PLA2) isoforms supporting their production within the cardiovascular system are not fully understood. To address this, we have studied platelets, endothelial cells, and leukocytes from 2 siblings with a homozygous loss-of-function mutation in group IVA cytosolic phospholipase A2 (cPLA2α). Chromatography/mass spectrometry was used to determine levels of a broad range of eicosanoids produced by isolated vascular cells, and in plasma and urine. Eicosanoid release data were paired with studies of cellular function. Absence of cPLA2α almost abolished eicosanoid synthesis in platelets (e.g., thromboxane A2, control 20.5 ± 1.4 ng/ml vs. patient 0.1 ng/ml) and leukocytes [e.g., prostaglandin E2 (PGE2), control 21.9 ± 7.4 ng/ml vs. patient 1.9 ng/ml], and this was associated with impaired platelet activation and enhanced inflammatory responses. cPLA2α-deficient endothelial cells showed reduced, but not absent, formation of prostaglandin I2 (prostacyclin; control 956 ± 422 pg/ml vs. patient 196 pg/ml) and were primed for inflammation. In the urine, prostaglandin metabolites were selectively influenced by cPLA2α deficiency. For example, prostacyclin metabolites were strongly reduced (18.4% of control) in patients lacking cPLA2α, whereas PGE2 metabolites (77.8% of control) were similar to healthy volunteer levels. These studies constitute a definitive account, demonstrating the fundamental role of cPLA2α to eicosanoid formation and cellular responses within the human circulation.—Kirkby, N. S., Reed, D. M., Edin, M. L., Rauzi, F., Mataragka, S., Vojnovic, I., Bishop-Bailey, D., Milne, G. L., Longhurst, H., Zeldin, D. C., Mitchell, J. A., Warner, T. D. Inherited human group IVA cytosolic phospholipase A2 deficiency abolishes platelet, endothelial, and leucocyte eicosanoid generation. PMID:26183771

  16. Primary haemostasis: newer insights.

    PubMed

    Berndt, M C; Metharom, P; Andrews, R K

    2014-05-01

    At the same time as biophysical and omics approaches are drilling deeper into the molecular details of platelets and other blood cells, as well as their receptors and mechanisms of regulation, there is also an increasing awareness of the functional overlap between human vascular systems. Together, these studies are redefining the intricate networks linking haemostasis and thrombosis with inflammation, infectious disease, cancer/metastasis and other vascular pathophysiology. The focus of this state-of-the-art review is some of the newer advances relevant to primary haemostasis. Of particular interest, platelet-specific primary adhesion-signalling receptors and associated activation pathways control platelet function in flowing blood and provide molecular links to other systems. Platelet glycoprotein (GP)Ibα of the GPIb-IX-V complex and GPVI not only initiate platelet aggregation and thrombus formation by primary interactions with von Willebrand factor and collagen, respectively, but are also involved in coagulation, leucocyte engagement, bacterial or viral interactions, and are relevant as potential risk markers in a range of human diseases. Understanding these systems in unprecedented detail promises significant advances in evaluation of individual risk, in new diagnostic or therapeutic possibilities and in monitoring the response to drugs or other treatment. © 2014 John Wiley & Sons Ltd.

  17. Classification of platelet concentrates: from pure platelet-rich plasma (P-PRP) to leucocyte- and platelet-rich fibrin (L-PRF).

    PubMed

    Dohan Ehrenfest, David M; Rasmusson, Lars; Albrektsson, Tomas

    2009-03-01

    The topical use of platelet concentrates is recent and its efficiency remains controversial. Several techniques for platelet concentrates are available; however, their applications have been confusing because each method leads to a different product with different biology and potential uses. Here, we present classification of the different platelet concentrates into four categories, depending on their leucocyte and fibrin content: pure platelet-rich plasma (P-PRP), such as cell separator PRP, Vivostat PRF or Anitua's PRGF; leucocyte- and platelet-rich plasma (L-PRP), such as Curasan, Regen, Plateltex, SmartPReP, PCCS, Magellan or GPS PRP; pure plaletet-rich fibrin (P-PRF), such as Fibrinet; and leucocyte- and platelet-rich fibrin (L-PRF), such as Choukroun's PRF. This classification should help to elucidate successes and failures that have occurred so far, as well as providing an objective approach for the further development of these techniques.

  18. Biodistribution of canine leucocytes labelled with technetium-99m stannous fluoride colloid in whole blood and their ability to localise to sites of induced inflammation.

    PubMed

    Abushhiwa, Mohamed H; Salehi, Nouria S; Lichtenstein, Meir; Lording, Peter M; Finnin, Peter J; Whitton, Robert C; Charles, Jennifer A; Parry, Bruce W

    2010-08-01

    This study assessed the biodistribution of autologous leucocytes radiolabelled with technetium-99m stannous fluoride colloid (99mTcSnC) for detection of foci of induced inflammation in dogs. Venous blood was collected from seven healthy dogs and incubated with 99mTcSnC for 1h at room temperature. Radiolabelled samples were injected intravenously (IV) and the dogs were scanned using a gamma camera. Another seven healthy dogs were injected intradermally with tumour necrosis factor alpha and then IV with 99mTcSnC radiolabelled autologous blood 3h later before being scanned. The radiolabelled leucocytes localised to sites of inflammation by 30 min post-injection. IV injection of autologous leucocytes radiolabelled with 99mTcSnC appears to be a sensitive method for localisation of induced foci of inflammation in dogs.

  19. In vitro effects of metals on isolated head-kidney and blood leucocytes of the teleost fish Sparus aurata L. and Dicentrarchus labrax L.

    PubMed

    Morcillo, Patricia; Meseguer, José; Esteban, M Ángeles; Cuesta, Alberto

    2016-07-01

    The in vitro use of fish leucocytes to test the toxicity of aquatic pollutants, and particularly the immutoxicological effects, could be a valuable alternative to fish bioassays but has received little attention. In this study, head-kidney and peripheral blood leucocytes (HKLs and PBLs, respectively) from gilthead seabream (Sparus aurata L.) and European sea bass (Dicentrarchus labrax L.) specimens were exposed to Cd, MeHg (methylmercury), Pb or As for 24 h being evaluated the resulting cytotoxicity. Exposure to metals produced a dose-dependent reduction in the viability, and MeHg showed the highest toxicity followed by Cd, As and Pb. Interestingly, leucocytes from European sea bass are more resistant to metal exposure than those from gilthead seabream. Similarly, HKLs are always more sensitive than those isolated from blood from the same fish species. Moreover, fish leucocytes incubated with metals exhibited alterations in gene expression profiles that were more pronounced in the HKLs in general, being Pb the metal provoking less effects. Concretely, genes related to cellular protection (metallothionein), stress (heat shock protein 70) and oxidative stress (superoxide dismutase, catalase and glutathione reductase) were, in general, down-regulated in seabream HKLs but up-regulated in seabream PBLs and sea bass HKLs and PBLs. In addition, this profile leads to the increase of expression in genes related to apoptosis (Bcl2 associated X protein and caspase 3). Finally, transcription of genes involved in immunity (interleukin-1β and immunoglobulin M) was down-regulated, mainly in seabream leucocytes. This study points to the benefits for evaluating the toxicological mechanisms of marine pollution using fish leucocytes in vitro and insight into the mechanisms at gene level.

  20. Harmonization of light scatter and fluorescence flow cytometry profiles obtained after staining peripheral blood leucocytes for cell surface-only versus intracellular antigens with the Fix & Perm reagent.

    PubMed

    da Costa, Elaine Sobral; Peres, Rodrigo Tosta; Almeida, Julia; Lécrevisse, Quentin; Arroyo, María Elena; Teodósio, Cristina; Pedreira, Carlos Eduardo; van Dongen, Jacques J M; Orfao, Alberto

    2010-01-01

    Staining for intracellular markers with the Fix & Perm reagent is associated with variations in the scatter properties of leucocytes, limiting automated analysis of flow cytometry (FCM) data. Here, we investigated those variables significantly contributing to changes in the light scatter, autofluorescence, and bcl2 staining characteristics of peripheral blood (PB) leucocytes, after fixation with Fix & Perm. Our major aim was to evaluate a new mathematical approach for automated harmonization of FCM data from datafiles corresponding to aliquots of a sample treated with cell-surface-only versus Fix & Perm intracellular staining techniques. Overall, neither the anticoagulant used nor sample storage for <24 h showed significant impact on the light scatter and fluorescence properties of PB leucocytes; similarly, the duration of the fixation period (once >15 min were used) had a minimum impact on the FCM properties of PB leucocytes. Conversely, changes in cell/protein concentrations and the fixative/sample (vol/vol) ratio had a clear impact on the light scatter features of some populations of leucocytes. Accordingly, lower cell/protein concentrations were associated with lower scatter values, particularly for the neutrophils. Such changes could be partially corrected through the use of higher fixative to sample volume ratios. Despite the variable changes detected between aliquots of the same sample treated with cell surface-only versus intracellular staining procedures, the new mathematical approach here proposed and evaluated for automated harmonization of common parameters in both datafiles, could correct the FCM profiles of leucocytes derived from cells undergoing conventional fixation/permeabilization procedures, and made them indistinguishable from those corresponding to aliquots of the same sample treated with cell-surface-only staining techniques.

  1. The Effect of Process Oriented Writing Activities on the Achievement and Attitude of the Preservice Primary School Teachers: An Example of Mixed Method Study

    ERIC Educational Resources Information Center

    Özenç, Emine Gül

    2016-01-01

    The purpose of this study is to find out whether process oriented writing exercises/activities have any effect on the achievement and attitude of preservice teachers as well as to set forth the opinions of primary preservice teachers on process oriented writing approach. In the research one classroom was designated as experimental group (N = 35)…

  2. Effect of glucose intake on human leucocyte /sup 86/Rb influx and (/sup 3/H)-ouabain binding

    SciTech Connect

    Turaihi, K.; Baron, D.N.; Dandona, P.

    1988-02-01

    /sup 86/Rb influx and (/sup 3/H) ouabain binding by human leucocytes were measured in eight normal nonobese fasting subjects before and after a challenge with 75 g glucose orally. The mean ouabain-sensitive /sup 86/Rb influx increased significantly from 194 to 283 mmol/kg protein/h (P less than .01), and (/sup 3/H)-ouabain binding increased from 236 to 403 fmol/mg protein. The mean plasma potassium concentration fell from 4.2 to 3.9 mmol/L (P less than .05). Following intravenous glucose infusion, the median /sup 86/Rb transport increased from 186 to 267 mmol/kg protein/h, while median plasma potassium concentration fell from 4.3 to 3.9 mmol/L. Therefore, glucose intake acutely increases Na-K ATPase units, stimulates potassium (Rb) transport, and causes a concomitant fall in plasma potassium concentrations. Nutritional intake is probably an important determinant of Na-K ATPase units and activity in the human leucocyte.

  3. Chemokines, chemokine receptors and adhesion molecules on different human endothelia: discriminating the tissue-specific functions that affect leucocyte migration

    PubMed Central

    HILLYER, P; MORDELET, E; FLYNN, G; MALE, D

    2003-01-01

    The selective accumulation of different leucocyte populations during inflammation is regulated by adhesion molecules and chemokines expressed by vascular endothelium. This study examined how chemokine production and the expression of adhesion molecules and chemokine receptors vary between endothelia from different vascular beds. Human saphenous vein endothelium was compared with lung and dermal microvascular endothelia and with umbilical vein endothelium and a bone-marrow endothelial cell line. All endothelia produced CCL2 and CXCL8 constitutively, whereas CXCL10 and CCL5 were only secreted after tumour necrosis factor (TNF)-α or interferon (IFN)-γ stimulation. In combination with TNF-α, IFN-γ suppressed CXCL8 but enhanced CCL5 and CXCL10, whereas transforming growth factor (TGF)-β reduced secretion of all chemokines. Basal chemokine secretion was higher from umbilical vein than other endothelial cells. Chemokine receptors, CXCR1, CXCR3 and CCR3, were present on all endothelia but highest on saphenous vein. CCR4, CCR5, CCR6, CXCR2, CXCR4 and CXCR5 were also detected at variable levels on different endothelia. The variation between endothelia in chemokine secretion was much greater than the variations in adhesion molecules, both on resting cells and following cytokine stimulation. These results indicate that it is the tissue-specific variations in endothelial chemokine secretion rather than variations in adhesion molecules that can explain the different patterns of inflammation and leucocyte traffic seen in non-lymphoid tissues. PMID:14632748

  4. A Melanoma Lymph Node Metastasis with a Donor-Patient Hybrid Genome following Bone Marrow Transplantation: A Second Case of Leucocyte-Tumor Cell Hybridization in Cancer Metastasis

    PubMed Central

    Duvall, Eric; Grasmick, Zachary; Haedicke, Kay

    2017-01-01

    Background Metastatic disease is the principal cause of mortality in cancer, yet the underlying mechanisms are not fully understood. Macrophage-cancer cell fusion as a cause of metastasis was proposed more than a century ago by German pathologist Prof. Otto Aichel. Since then this theory has been confirmed in numerous animal studies and recently in a patient with metastatic melanoma. Methods Here we analyzed tumor DNA from a 51-year-old man who, 8 years following an allogeneic BMT from his brother for treatment of chronic myelogenous leukemia (CML), developed a nodular malignant melanoma on the upper back with spread to an axillary sentinal lymph node. We used laser microdissection to isolate FFPE tumor cells free of leucocytes. They were genotyped using forensic short tandem repeat (STR) length-polymorphisms to distinguish donor and patient genomes. Tumor and pre-transplant blood lymphocyte DNAs were analyzed for donor and patient alleles at 15 autosomal STR loci and the sex chromosomes. Results DNA analysis of the primary melanoma and the nodal metastasis exhibit alleles at each STR locus that are consistent with both the patient and donor. The doses vary between these samples indicative of the relative amounts of genomic DNA derived from the patient and donor. Conclusion The evidence supports fusion and hybridization between donor and patient cells as the initiator of metastasis in this patient. That this phenomenon has now been seen in a second case suggests that fusion is likely to play a significant role for melanoma and other solid tumor metastasis, perhaps leading to new avenues of treatment for this most problematic disease. PMID:28146572

  5. Altered gene expression in T-cell receptor signalling in peripheral blood leucocytes in acute coronary syndrome predicts secondary coronary events

    PubMed Central

    Takashima, Shin-ichiro; Usui, Soichiro; Kurokawa, Keisuke; Kitano, Teppei; Kato, Takeshi; Murai, Hisayoshi; Furusho, Hiroshi; Oda, Hiroyuki; Maruyama, Michiro; Nagata, Yoshiki; Usuda, Kazuo; Kubota, Koji; Takeshita, Yumie; Sakai, Yoshio; Honda, Masao; Kaneko, Shuichi; Takamura, Masayuki

    2016-01-01

    Objective Comprehensive profiling of gene expression in peripheral blood leucocytes (PBLs) in patients with acute coronary syndrome (ACS) as a prognosticator is needed. We explored the specific profile of gene expression in PBLs in ACS for long-term risk stratification. Methods 30 patients with ACS who underwent primary percutaneous coronary intervention (PCI) and 15 age-matched adults who participated in medical check-ups were enrolled from three centres. Peripheral blood samples were collected to extract RNA for microarray analyses. Results During the 5-year follow-up, 36% of this cohort developed the expected non-fatal coronary events (NFEs) of target lesion revascularisation (TLR) and PCI for a de novo lesion. Class comparison analysis (p<0.005) demonstrated that 83 genes among 7785 prefiltered genes (41 upregulated vs 42 downregulated genes) were extracted to classify the patients according to the occurrence of NFE. Pathway analysis based on gene ontology revealed that the NFEs were associated with altered gene expression regarding the T-cell receptor signalling pathway in ACS. Univariate t test showed that the expression level of death-associated protein kinase1 (DAPK1), known to regulate inflammation, was the most significantly negatively regulated gene in the event group (0.61-fold, p<0.0005). Kaplan-Meier curve analysis and multivariate analysis adjusted for baseline characteristics or clinical biomarkers demonstrated that lower DAPK1 expression in PBL emerged as an independent risk factor for the NFEs (HR: 8.73; CI 1.05 to 72.8, p=0.045). Conclusions Altered gene expression in T-cell receptor signalling in PBL in ACS could be a prognosticator for secondary coronary events. Trial registration number UMIN000001932; Results. PMID:27403330

  6. Suppression of polymorphonuclear leucocyte chemotaxis by Pseudomonas aeruginosa elastase in vitro: a study of the mechanisms and the correlation with ring abscess in pseudomonal keratitis.

    PubMed Central

    Ijiri, Y.; Matsumoto, K.; Kamata, R.; Nishino, N.; Okamura, R.; Kambara, T.; Yamamoto, T.

    1994-01-01

    Bacteria, or the culture supernatants of an elastase non-producing strain of Pseudomonas aeruginosa, elicited a chemotactic response from polymorphonuclear leucocytes (PMN) in vitro. The chemoattractive capacity was diminished under the presence of Boc-Phe-Leu-Phe-Leu-Phe, a receptor antagonist of N-formyl-Met-Leu-Phe (fMLP) which is a bacterial chemotactic peptide to PMN. This indicated that the chemoattractant derived from Pseudomonas aeruginosa was a fMLP-like molecule(s). In contrast, culture supernatants of an elastase producing strain of Pseudomonas aeruginosa produced negligible chemotactic response from PMN. Indeed, an inhibitory effect of the culture supernatants or of purified Pseudomonas aeruginosa elastase (PAE) on PMN chemotaxis was observed when fMLP was used as a chemoattractant. Another fMLP-induced function of PMN, respiratory burst activation, was also diminished by pretreatment of PMN with PAE. PAE hydrolysed fMLP at the Met-Leu bond and diminished the chemoattractant capacity. In addition, a receptor analysis with fML-3H-P demonstrated a decrease in numbers of fMLP receptors on PMN without changing the dissociation constant values after the treatment of the cells with PAE. In the primary structure of the fMLP receptor previously reported, a preferential amino acid sequence for cleavage by PAE was identified in what was believed to be an extracellular portion of the receptor molecule. These results suggested that PAE could diminish PMN infiltration in response to Pseudomonas aeruginosa in vivo by cleavage of the fMLP-like pseudomonal chemotactic ligand and the receptors on PMN. Images Figure 4 PMID:7734333

  7. Generation of β cell-specific human cytotoxic T cells by lentiviral transduction and their survival in immunodeficient human leucocyte antigen-transgenic mice

    PubMed Central

    Babad, J; Mukherjee, G; Follenzi, A; Ali, R; Roep, B O; Shultz, L D; Santamaria, P; Yang, O O; Goldstein, H; Greiner, D L; DiLorenzo, T P

    2015-01-01

    Several β cell antigens recognized by T cells in the non-obese diabetic (NOD) mouse model of type 1 diabetes (T1D) are also T cell targets in the human disease. While numerous antigen-specific therapies prevent diabetes in NOD mice, successful translation of rodent findings to patients has been difficult. A human leucocyte antigen (HLA)-transgenic mouse model incorporating human β cell-specific T cells might provide a better platform for evaluating antigen-specific therapies. The ability to study such T cells is limited by their low frequency in peripheral blood and the difficulty in obtaining islet-infiltrating T cells from patients. We have worked to overcome this limitation by using lentiviral transduction to ‘reprogram’ primary human CD8 T cells to express three T cell receptors (TCRs) specific for a peptide derived from the β cell antigen islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP265–273) and recognized in the context of the human class I major histocompatibility complex (MHC) molecule HLA-A2. The TCRs bound peptide/MHC multimers with a range of avidities, but all bound with at least 10-fold lower avidity than the anti-viral TCR used for comparison. One exhibited antigenic recognition promiscuity. The β cell-specific human CD8 T cells generated by lentiviral transduction with one of the TCRs released interferon (IFN)-γ in response to antigen and exhibited cytotoxic activity against peptide-pulsed target cells. The cells engrafted in HLA-A2-transgenic NOD-scid IL2rγnull mice and could be detected in the blood, spleen and pancreas up to 5 weeks post-transfer, suggesting the utility of this approach for the evaluation of T cell-modulatory therapies for T1D and other T cell-mediated autoimmune diseases. PMID:25302633

  8. Utility of 111In-labelled leucocyte scintigraphy in patients with fever of unknown origin in an era of changing disease spectrum and investigational techniques.

    PubMed

    Seshadri, Nagabhushan; Solanki, Chandra K; Balan, Kottekkattu

    2008-03-01

    (111)In-labelled leucocyte, imaging is often used to investigate patients with fever of unknown origin (FUO). Its diagnostic performance, however, has been variable and a broad range of sensitivities and specificities have been reported. The purpose of this investigation was to evaluate the usefulness of (111)In-labelled leucocytes scintigraphy in the detection of a cause of FUO in the light of a changing spectrum of diseases causing it and advances in investigational techniques. Sixty-one patients with a clinical diagnosis of FUO underwent whole-body (111)In-troponolate-labelled leucocyte scintigraphy in our department over a 2-year period between February 2004 and February 2006. Of these, 54 patients were retrospectively reviewed to identify a cause of FUO. Other parameters such as C-reactive protein (CRP), leucocyte count and radiological findings were also evaluated. Leucocyte scintigraphy was found to be true positive in 12 patients, true negative in 24 patients, false positive in 10 patients and false negative in eight patients. The overall sensitivity of scintigraphy was 60%, specificity 71%, positive predictive value 55%, and negative predictive value 75%. There was no difference in the scintigraphic sensitivity between patients with spontaneous FUO and those with post-operative FUO although the latter showed a higher specificity and PPV. CRP and leucocyte count did not differ significantly between patients with true positive and true negative scintigrams. Overall, 83% of patients with abnormal radiological examinations had positive findings on scintigraphy and 87% of patients with negative findings on radiology had normal scintigraphy. Despite changes in disease spectrum and advances in investigational techniques, our results suggest that (111)In-leucocyte scintigraphy is still a useful technique in establishing the cause of FUO. A higher PPV of this test in post-operative situations makes it especially applicable in this category of patients. Equally, the

  9. The trans-stilbene oxide-active glutathione transferase in human mononuclear leucocytes is identical with the hepatic glutathione transferase mu.

    PubMed Central

    Seidegård, J; Guthenberg, C; Pero, R W; Mannervik, B

    1987-01-01

    A glutathione transferase from human mononuclear leucocytes with high activity towards trans-stilbene oxide (GT-tSBO) was purified. GT-tSBO is expressed in only about 50% of the individuals studied. As judged from activity measurements, immunological studies and the fact that only those individuals who express glutathione transferase mu have high activity towards trans-stilbene oxide, it is concluded that the hepatic transferase mu is identical with the glutathione transferase (GT-tSBO) in mononuclear leucocytes. PMID:3689332

  10. The prevalence of various dental characteristics in the primary and mixed dentition in patients born with non-syndromic unilateral cleft lip with or without cleft palate.

    PubMed

    Pegelow, Marie; Alqadi, Nadia; Karsten, Agneta Linder-Aronson

    2012-10-01

    This retrospective study was carried out on consecutively collected dental casts, x-rays, and photos of 129 Swedish children who had been born with non-syndromic unilateral (U) cleft lip (CL), cleft lip and alveolus (CLA), or cleft lip and palate (CLP). The following dental characteristics were investigated in the primary and permanent dentitions: 1. the presence, eruption, position, and shape of the lateral incisor; 2. the prevalence of rotation and enamel hypoplasia of the permanent central incisor; 3. the occurrence of hypodontia outside the cleft region; and 4. the transition from the primary to the succeeding permanent lateral incisor in the cleft region. Patients with clefts involving the palate (UCLP) exhibited a high frequency of hypodontia. In patients with clefts, which did not include the palate, malformed lateral incisors were a common finding. In the primary and permanent dentition, the lateral incisor had erupted distal to the cleft in 31.8 and 24.8 per cent of the UCLA and UCLP patients, respectively. No significant pattern was seen regarding the transition from the primary to the succeeding permanent lateral incisor (P = 0.15). The central incisor was rotated in 55 per cent of the patients and this positional deviation was particularly frequent in cases where the lateral incisor was missing in the premaxilla (P < 0.05). Hypodontia outside the cleft region was recorded in 15.5 per cent of the patients. Patients with UCLP had more often crossbite than patients with a UCL or a UCLA phenotype (P < 0.001).

  11. Using information to deliver safer care: a mixed-methods study exploring general practitioners' information needs in North West London primary care.

    PubMed

    Mastellos, Nikolaos; Car, Josip; Majeed, Azeem; Aylin, Paul

    2014-12-08

    The National Health Service in England has given increasing priority to improving inter-professional communication, enabling better management of patients with chronic conditions and reducing medical errors through effective use of information. Despite considerable efforts to reduce patient harm through better information usage, medical errors continue to occur, posing a serious threat to patient safety. This study explores the range, quality and sophistication of existing information systems in primary care with the aim to capture what information practitioners need to provide a safe service and identify barriers to its effective use in care pathways. Data were collected through semi-structured interviews with general practitioners from surgeries in North West London and a survey evaluating their experience with information systems in care pathways. Important information is still missing, specifically discharge summaries detailing medication changes and changes in the diagnosis and management of patients, blood results ordered by hospital specialists and findings from clinical investigations. Participants identified numerous barriers, including the communication gap between primary and secondary care, the variable quality and consistency of clinical correspondence and the inadequate technological integration. Despite attempts to improve integration and information flow in care pathways, existing systems provide practitioners with only partial access to information, hindering their ability to take informed decisions. This study offers a framework for understanding what tools should be in place to enable effective use of information in primary care.

  12. A FIRE-ACE/SHEBA Case Study of Mixed-Phase Arctic Boundary Layer Clouds: Entrainment Rate Limitations on Rapid Primary Ice Nucleation Processes

    NASA Technical Reports Server (NTRS)

    Fridlin, Ann; vanDiedenhoven, Bastiaan; Ackerman, Andrew S.; Avramov, Alexander; Mrowiec, Agnieszka; Morrison, Hugh; Zuidema, Paquita; Shupe, Matthew D.

    2012-01-01

    Observations of long-lived mixed-phase Arctic boundary layer clouds on 7 May 1998 during the First International Satellite Cloud Climatology Project (ISCCP) Regional Experiment (FIRE)Arctic Cloud Experiment (ACE)Surface Heat Budget of the Arctic Ocean (SHEBA) campaign provide a unique opportunity to test understanding of cloud ice formation. Under the microphysically simple conditions observed (apparently negligible ice aggregation, sublimation, and multiplication), the only expected source of new ice crystals is activation of heterogeneous ice nuclei (IN) and the only sink is sedimentation. Large-eddy simulations with size-resolved microphysics are initialized with IN number concentration N(sub IN) measured above cloud top, but details of IN activation behavior are unknown. If activated rapidly (in deposition, condensation, or immersion modes), as commonly assumed, IN are depleted from the well-mixed boundary layer within minutes. Quasi-equilibrium ice number concentration N(sub i) is then limited to a small fraction of overlying N(sub IN) that is determined by the cloud-top entrainment rate w(sub e) divided by the number-weighted ice fall speed at the surface v(sub f). Because w(sub c)< 1 cm/s and v(sub f)> 10 cm/s, N(sub i)/N(sub IN)<< 1. Such conditions may be common for this cloud type, which has implications for modeling IN diagnostically, interpreting measurements, and quantifying sensitivity to increasing N(sub IN) (when w(sub e)/v(sub f)< 1, entrainment rate limitations serve to buffer cloud system response). To reproduce observed ice crystal size distributions and cloud radar reflectivities with rapidly consumed IN in this case, the measured above-cloud N(sub IN) must be multiplied by approximately 30. However, results are sensitive to assumed ice crystal properties not constrained by measurements. In addition, simulations do not reproduce the pronounced mesoscale heterogeneity in radar reflectivity that is observed.

  13. Human leucocyte antigen class I‐redirected anti‐tumour CD4+ T cells require a higher T cell receptor binding affinity for optimal activity than CD8+ T cells

    PubMed Central

    Tan, M. P.; Dolton, G. M.; Gerry, A. B.; Brewer, J. E.; Bennett, A. D.; Pumphrey, N. J.; Jakobsen, B. K.

    2016-01-01

    Summary CD4+ T helper cells are a valuable component of the immune response towards cancer. Unfortunately, natural tumour‐specific CD4+ T cells occur in low frequency, express relatively low‐affinity T cell receptors (TCRs) and show poor reactivity towards cognate antigen. In addition, the lack of human leucocyte antigen (HLA) class II expression on most cancers dictates that these cells are often unable to respond to tumour cells directly. These deficiencies can be overcome by transducing primary CD4+ T cells with tumour‐specific HLA class I‐restricted TCRs prior to adoptive transfer. The lack of help from the co‐receptor CD8 glycoprotein in CD4+ cells might result in these cells requiring a different optimal TCR binding affinity. Here we compared primary CD4+ and CD8+ T cells expressing wild‐type and a range of affinity‐enhanced TCRs specific for the HLA A*0201‐restricted NY‐ESO‐1‐ and gp100 tumour antigens. Our major findings are: (i) redirected primary CD4+ T cells expressing TCRs of sufficiently high affinity exhibit a wide range of effector functions, including cytotoxicity, in response to cognate peptide; and (ii) optimal TCR binding affinity is higher in CD4+ T cells than CD8+ T cells. These results indicate that the CD4+ T cell component of current adoptive therapies using TCRs optimized for CD8+ T cells is below par and that there is room for substantial improvement. PMID:27324616

  14. Formation and loss of O6-methyldeoxyguanosine in human leucocyte DNA following sequential DTIC and fotemustine chemotherapy.

    PubMed

    Lee, S M; Margison, G P; Thatcher, N; O'Connor, P J; Cooper, D P

    1994-05-01

    There is increasing evidence to indicate that O6-methyldeoxyguanosine (O6-MedG) formation in DNA is a critical cytotoxic event following exposure to certain anti-tumour alkylating agents and that the DNA repair protein O6-alkylguanine-DNA alkyltransferase (ATase) can confer resistance to these agents. We recently demonstrated a wide inter-individual variation in the depletion and subsequent regeneration of ATase in human peripheral blood lymphocytes following sequential DTIC (400 mg m-2) and fotemustine (100 mg m-2) treatment, with the nadir ATase activity occurring approximately 4 h after DTIC administration. We have now measured the formation and loss of O6-methyldeoxyguanosine (O6-MedG) in the DNA of peripheral leucocytes of eight patients receiving this treatment regimen. O6-MedG could be detected within 1 h and maximal levels occurred approximately 3-5 h after DTIC administration. Following the first treatment cycle, considerable inter-individual variation was observed in the peak O6-MedG levels, with values ranging from 0.71 to 14.3 mumol of O6-MedG per mol of dG (6.41 +/- 5.53, mean +/- s.d.). Inter- and intra-individual variation in the extent of O6-MedG formation was also seen in patients receiving additional treatment cycles. This may be a consequence of inter-patient differences in the capacity for metabolism of DTIC to release a methylating intermediate and could be one of the determinants of clinical response. Both the pretreatment ATase levels and the extent of ATase depletion were inversely correlated with the amount of O6-MedG formed in leucocyte DNA when expressed either as peak levels (r = -0.59 and -0.75 respectively) or as the area under the concentration-time curve (r = -0.72 and -0.73 respectively). One complete and one partial clinical response were seen, and these occurred in the two patients with the highest O6-MedG levels in the peripheral leucocyte DNA, although the true significance of this observation has yet to be established.

  15. Formation and loss of O6-methyldeoxyguanosine in human leucocyte DNA following sequential DTIC and fotemustine chemotherapy.

    PubMed Central

    Lee, S. M.; Margison, G. P.; Thatcher, N.; O'Connor, P. J.; Cooper, D. P.

    1994-01-01

    There is increasing evidence to indicate that O6-methyldeoxyguanosine (O6-MedG) formation in DNA is a critical cytotoxic event following exposure to certain anti-tumour alkylating agents and that the DNA repair protein O6-alkylguanine-DNA alkyltransferase (ATase) can confer resistance to these agents. We recently demonstrated a wide inter-individual variation in the depletion and subsequent regeneration of ATase in human peripheral blood lymphocytes following sequential DTIC (400 mg m-2) and fotemustine (100 mg m-2) treatment, with the nadir ATase activity occurring approximately 4 h after DTIC administration. We have now measured the formation and loss of O6-methyldeoxyguanosine (O6-MedG) in the DNA of peripheral leucocytes of eight patients receiving this treatment regimen. O6-MedG could be detected within 1 h and maximal levels occurred approximately 3-5 h after DTIC administration. Following the first treatment cycle, considerable inter-individual variation was observed in the peak O6-MedG levels, with values ranging from 0.71 to 14.3 mumol of O6-MedG per mol of dG (6.41 +/- 5.53, mean +/- s.d.). Inter- and intra-individual variation in the extent of O6-MedG formation was also seen in patients receiving additional treatment cycles. This may be a consequence of inter-patient differences in the capacity for metabolism of DTIC to release a methylating intermediate and could be one of the determinants of clinical response. Both the pretreatment ATase levels and the extent of ATase depletion were inversely correlated with the amount of O6-MedG formed in leucocyte DNA when expressed either as peak levels (r = -0.59 and -0.75 respectively) or as the area under the concentration-time curve (r = -0.72 and -0.73 respectively). One complete and one partial clinical response were seen, and these occurred in the two patients with the highest O6-MedG levels in the peripheral leucocyte DNA, although the true significance of this observation has yet to be established. PMID

  16. Modulating phenotype and cytokine production of leucocytic retinal infiltrate in experimental autoimmune uveoretinitis following intranasal tolerance induction with retinal antigens

    PubMed Central

    Laliotou, B.; Dick, A.

    1999-01-01

    BACKGROUND/AIM—Nasal administration of retinal antigens induces systemic tolerance which results in suppression of experimental autoimmune uveoretinitis (EAU) when subsequently exposed to antigen. The aim was to establish if tolerance induction alters retinal infiltrating leucocyte phenotype and cytokine profile in tolerised animals when there is significantly reduced tissue destruction despite immunisation with retinal antigen.
METHODS—Female Lewis rats were tolerised by intranasal administration with retinal extract (RE) before immunisation with RE to induce EAU. Control animals were administered phosphate buffered saline (PBS) intranasally. Post immunisation, daily clinical responses were recorded and at the height of disease, retinas were removed and either infiltrating leucocytes isolated for flow cytometric phenotype assessment and intracellular cytokine production, or chorioretina processed for immunohistochemistry. Fellow eyes were assessed for cytokine mRNA by semiquantitative RT-PCR.
RESULTS—Flow cytometric analysis showed that before clinical onset of EAU there is no evidence of macrophage infiltration and no significant difference in circulating T cell populations within the retina. By day 14 a reduced retinal infiltrate in tolerised animals was observed and in particular a reduction in numbers of "activated" (with respect to CD4 and MHC class II expression) macrophages. Immunohistochemistry confirmed these findings and additionally minimal rod outer segment destruction was observed histologically. Cytokine analysis revealed that both IL-10 mRNA and intracellular IL-10 production was increased in tolerised eyes 7 days post immunisation. Although by day 14 post immunisation, IL-10 production was equivalent in both groups, a reduced percentage of IFN-γ+ macrophages and IFN-γ+ CD4+ T cells with increased percentage of IL-4+ CD4+ T cells were observed in tolerised animals.
CONCLUSIONS—Leucocytic infiltrate is not only reduced in number

  17. The protective effects of Achillea L. species native in Turkey against H(2)O(2)-induced oxidative damage in human erythrocytes and leucocytes.

    PubMed

    Konyalioglu, Sibel; Karamenderes, Canan

    2005-11-14

    The aim of the present study was to investigate the protective effects of infusions prepared from 15 Achillea (Asteraceae) species against H(2)O(2)-induced oxidative damage in human erythrocytes and leucocytes used in traditional Turkish medicine. CAT, SOD and GPx activities, effects of LPO and GSH levels of the infusions on erythrocytes and leucocytes were assessed. The results indicated that all infusions of Achillea species were effective on antioxidant enzyme systems of erythrocytes and leucocytes when compared with H(2)O(2) group. Achillea falcata was the most effective one on CAT, GPx and SOD enzyme systems of erythrocytes. Among plant infusions, Achillea crithmifolia and Achillea nobilis subsp. neilrechii showed the highest activities on CAT, while Achillea millefolium subsp. pannonica on SOD, Achillea teretifolia on GPx and Achillea nobilis subsp. sipylea on LPO enzyme systems of leucocytes. The present results demonstrate that infusions of Achillea species are a potential source of natural antioxidants for treatment and prevention of diseases in which LPO takes place.

  18. Improving Ebola infection prevention and control in primary healthcare facilities in Sierra Leone: a single-group pretest post-test, mixed-methods study.

    PubMed

    Ratnayake, Ruwan; Ho, Lara S; Ansumana, Rashid; Brown, Hannah; Borchert, Matthias; Miller, Laura; Kratz, Thomas; McMahon, Shannon A; Sahr, Foday

    2016-01-01

    Accomplishing infection prevention and control (IPC) in health facilities in Sub-Saharan Africa is challenging. Owing to poor IPC, healthcare workers (HCWs) were frequently infected during Sierra Leone's Ebola epidemic. In late 2014, IPC was rapidly and nationally scaled up. We carried out workshops in sampled facilities to further improve adherence to IPC. We investigated HCW experiences and observed practice gaps, before and after the workshops. We conducted an uncontrolled, before and after, mixed-methods study in eight health facilities in Bo and Kenema Districts during December 2014 and January 2015. Quantitative methods administered to HCWs at baseline and follow-up included a survey on attitudes and self-efficacy towards IPC, and structured observations of behaviours. The intervention involved a workshop for HCWs to develop improvement plans for their facility. We analysed the changes between rounds in survey responses and behaviours. We used interviews to explore attitudes and self-efficacy throughout the study period. HCWs described IPC as 'life-saving' and personal protective equipment (PPE) as uncomfortable for providers and frightening for patients. At baseline, self-efficacy was high (median=4/strongly agree). Responses reflecting unfavourable attitudes were low for glove use (median=1/strongly disagree, IQR, 1-2) and PPE use with ill family members (median=1, IQR, 1-2), and mixed for PPE use with ill HCWs (median=2/disagree, IQR, 1-4). Observations demonstrated consistent glove reuse and poor HCW handwashing. The maintenance of distance (RR 1.09, 95% CI 1.02 to 1.16) and patient handwashing (RR 1.19, 95% CI 1.3 to 1.25) improved to >90%. We found favourable attitudes towards IPC and gaps in practice. Risk perceptions of HCWs and tendencies to ration PPE where chronic supply chain issues normally lead to PPE stock-outs may affect practice. As Sierra Leone's Ebola Recovery Strategy aims to make all facilities IPC compliant, socio

  19. Quality of life, sense of coherence and experiences with three different treatments in patients with psychological distress in primary care: a mixed-methods study.

    PubMed

    Arvidsdotter, Tina; Marklund, Bertil; Taft, Charles; Kylén, Sven

    2015-04-26

    Psychological distress is associated with impaired health-related quality of life (HRQL) and poor sense of coherence (SOC). In a previous study, we found that therapeutic acupuncture (TA) and an integrative treatment that combined TA with person-centred approach in a salutogenic dialogue (IT) alleviated anxiety and depression significantly more than conventional treatment (CT) in primary care patients. Here, we report on secondary analyses regarding the HRQL and SOC from that previous pragmatic randomised controlled trial (RCT). Quantitative and qualitative design. One hundred twenty patients were referred for psychological distress. Quantitative analyses were performed at baseline and after 8 weeks of treatment using the SF-36 mental component summary (MCS), physical component summary (PCS) and the Sense of Coherence-13 (SOC) questionnaires. Qualitative manifest content analyses were based on open-ended questions-"Have you experienced any changes since the start of the treatment? Will you describe these changes?" No baseline differences were found. At 8 weeks, both the IT and TA groups had statistically better scores and greater improvement from baseline on the MCS and SOC than the CT group. The effect sizes were large. No significant differences were found between the IT and TA groups or in relation to the PCS. SOC was highly correlated with the MCS but not with the PCS. Dropout rates were low. The experiences of the intervention resulted in four categories: Being heading back; Status quo; Feeling confirmed; and Feeling abandoned, with 13 related subcategories. IT and TA seem to improve sense of coherence and mental health status in primary care patients with psychological distress, whereas CT appears to be less beneficial. IT and TA appear to be well-accepted and may serve as useful adjunct treatment modalities to standard primary care. Our results are consistent with much of the previous research in highlighting a strong relationship between SOC and mental

  20. Deleterious effect of Brij 35 on alkyl 2-pyrones and other hydrophobic inhibitors of human sputum and leucocyte elastase.

    PubMed

    Cook, L; Ternai, B

    1988-10-01

    Brij 35 significantly reduced the inhibitory activity of hydrophobic alkyl 2-pyrones, oleic acid and alkyl peptides towards human sputum and leucocyte elastase, whereas 4-methoxy-6-(2'-hydroxy-2'-(carbobutyloxy)-vinyl)-2-pyrone, alpha-1-proteinase inhibitor and a sulfated chitosan were unaffected. The effect of Brij 35 on elastase appeared to be irreversible, since dialysis against Brij-free buffer was not accompanied by a return to inhibitory activity by the first group of inhibitors. However, passage through an ionic-exchange column was effective in removing the detergent from the enzyme. Brij 35 is also an activator of the elastases: kcat for Boc-Ala-4-nitrophenyl ester and methylsuccinyl-Ala-Ala-Pro-Val-4-nitroanilide increased by 20% and 40%, respectively in the presence of 0.015% Brij 35. Binding of the substrates to the enzyme is unaffected, since Km is unchanged.

  1. An analysis of myeloma plasma cell phenotype using antibodies defined at the IIIrd International Workshop on Human Leucocyte Differentiation Antigens.

    PubMed Central

    Jackson, N; Ling, N R; Ball, J; Bromidge, E; Nathan, P D; Franklin, I M

    1988-01-01

    Fresh bone marrow from 43 cases of myeloma and three cases of plasma cell leukaemia has been phenotyped both by indirect immune-rosetting and, on fixed cytospin preparations, by indirect immunofluorescence. Both clustered and unclustered B cell associated antibodies from the IIIrd International Workshop on Human Leucocyte Differentiation Antigens were used. The results confirm the lack of many pan-B antigens on the surface of myeloma plasma cells, i.e. CD19-23, 37, 39, w40. Strong surface reactivity is seen with CD38 antibodies and with one CD24 antibody (HB8). Weak reactions are sometimes obtained with CD9, 10 and 45R. On cytospin preparations CD37, 39 and w40 are sometimes weakly positive, and anti-rough endoplasmic reticulum antibodies are always strongly positive. Specific and surface-reacting antiplasma cell antibodies are still lacking. PMID:3048803

  2. Cytometric analysis of surface molecules of leucocytes and phagocytic activity of granulocytes and monocytes/macrophages in cows with pyometra.

    PubMed

    Brodzki, P; Kostro, K; Brodzki, A; Niemczuk, K; Lisiecka, U

    2014-10-01

    Pyometra is a serious problem in dairy cow herds, causing large economic losses due to infertility. The development of pyometra depends mainly on the immunological status of the cow. The aim of the study was a comparative evaluation of selected indicators involving non-specific and specific immunity in cows with pyometra and in cows without inflammation of the uterus. The study was performed in 20 cows, which were divided into two groups: pyometra group and healthy group, each comprising 10 cows, based on the results of cytological and ultrasonographic tests. A flow cytometric analysis was performed for the surface molecules CD4, CD8, CD14, CD21, CD25 and CD4(+) CD25(+) on leucocytes, and the phagocytic activity was determined from granulocytes and monocytes/macrophages in the peripheral blood and uterine washings, respectively. It was demonstrated that the percentage of phagocytic granulocytes and monocytes/macrophages in both the peripheral blood and uterine washings was significantly lower in cows with pyometra compared with the healthy group (p < 0.001). Significantly (p ≤ 0.001) lower percentage of CD4(+) , CD14(+) , CD25(+) and CD4(+) CD25(+) phenotype leucocytes was also observed in the peripheral blood of cows from the pyometra group, along with a significantly higher (p < 0.001) percentage of CD8(+) and CD21(+) lymphocytes as compared to the healthy group. The results of work indicate that disfunction of cell immunity coexisting with pyometra may be caused by a bacterial infection and the presence of blocking agents (IL-10), released by the increasing number of CD8(+) lymphocytes what leads to the advanced inflammation of uterus.

  3. Differential patterns of endothelial and leucocyte activation in ‘typhus-like’ illnesses in Laos and Thailand

    PubMed Central

    Paris, D H; Jenjaroen, K; Blacksell, S D; Phetsouvanh, R; Wuthiekanun, V; Newton, P N; Day, N P J; Turner, G D H

    2008-01-01

    Scrub typhus is responsible for a large proportion of undifferentiated fevers in south-east Asia. The cellular tropism and pathophysiology of the causative agent, Orientia tsutsugamushi, remain poorly understood. We measured endothelial and leucocyte activation by soluble cell adhesion molecule enzyme-linked immunosorbent assays in 242 Lao and Thai patients with scrub or murine typhus, leptospirosis, dengue, typhoid and uncomplicated falciparum malaria on admission to hospital. Soluble E-selectin (sE-selectin) levels were lowest in dengue, sL-selectin highest in scrub typhus with a high sE-selectin to sL-selectin ratio in leptospirosis patients. In scrub typhus patients elevated sL-selectin levels correlated with the duration of skin rash (P = 0·03) and the presence of eschar (P = 0·03), elevated white blood cell (WBC) count (P = 0·007), elevated lymphocyte (P = 0·007) and neutrophil counts (P = 0·015) and elevated levels of sE-selectin correlated with the duration of illness before admission (P = 0·03), the presence of lymphadenopathy (P = 0·033) and eschar (P = 0·03), elevated WBC (P = 0·005) and neutrophil counts (P = 0·0003). In comparison, soluble selectin levels in murine typhus patients correlated only with elevated WBC counts (P = 0·03 for sE-selectin and sL-selectin). Soluble intercellular adhesion molecule-1 and soluble vascular adhesion molecule-1 levels were not associated significantly with any clinical parameters in scrub or murine typhus patients. The data presented suggest mononuclear cell activation in scrub typhus. As adhesion molecules direct leucocyte migration and induce inflammatory and immune responses, this may represent O. tsutsugamushi tropism during early dissemination, or local immune activation within the eschar. PMID:18505434

  4. Bothrops asper snake venom and its metalloproteinase BaP-1 activate the complement system. Role in leucocyte recruitment.

    PubMed Central

    Farsky, S H; Gonçalves, L R; Gutiérrez, J M; Correa, A P; Rucavado, A; Gasque, P; Tambourgi, D V

    2000-01-01

    The venom of the snake Bothrops asper, the most important poisonous snake in Central America, evokes an inflammatory response, the mechanisms of which are not well characterized. The objectives of this study were to investigate whether B. asper venom and its purified toxins--phospholipases and metalloproteinase--activate the complement system and the contribution of the effect on leucocyte recruitment. In vitro chemotaxis assays were performed using Boyden's chamber model to investigate the ability of serum incubated with venom and its purified toxins to induce neutrophil migration. The complement consumption by the venom was evaluated using an in vitro haemolytic assay. The importance of complement activation by the venom on neutrophil migration was investigated in vivo by injecting the venom into the peritoneal cavity of C5-deficient mice. Data obtained demonstrated that serum incubated with crude venom and its purified metalloproteinase BaP-1 are able to induce rat neutrophil chemotaxis, probably mediated by agent(s) derived from the complement system. This hypothesis was corroborated by the capacity of the venom to activate this system in vitro. The involvement of C5a in neutrophil chemotaxis induced by venom-activated serum was demonstrated by abolishing migration when neutrophils were pre-incubated with antirat C5a receptor antibody. The relevance of the complement system in in vivo leucocyte mobilization was further demonstrated by the drastic decrease of this response in C5-deficient mice. Pre-incubation of serum with the soluble human recombinant complement receptor type 1 (sCR 1) did not prevent the response induced by the venom, but abolished the migration evoked by metalloproteinase-activated serum. These data show the role of the complement system in bothropic envenomation and the participation of metalloproteinase in the effect. Also, they suggest that the venom may contain other component(s) which can cause direct activation of C5a. PMID:11200361

  5. Effect of vegetable consumption on the association between peripheral leucocyte telomere length and hypertension: a case–control study

    PubMed Central

    Lian, Fuzhi; Wang, Jinquan; Huang, Xianhong; Wu, Yinyin; Cao, Yifei; Tan, Xiaohua; Xu, Xianrong; Hong, Yu; Yang, Lei; Gao, Xiang

    2015-01-01

    Objectives Peripheral leucocyte telomere length has been suggested to be inversely associated with hypertension risk. Both telomere length and hypertension risk can be modified by certain dietary factors, such as fruit and vegetables. This study was to examine the potential effect of these dietary factors on the association between telomere length and hypertension risk. Study design A community-based case–control study. Participants 271 hypertensive patients and 455 normotensive controls aged 40–70 years and living in Yinzhou, Zhejiang Province, China. Outcome measures Peripheral leucocyte relative telomere length (RTL) was measured using quantitative real-time PCR. Dietary intake was assessed by a brief semiquantitative food frequency questionnaire. The association between RTL and hypertension risk was analysed using logistic regression, and the modulatory effect of dietary intake on RTL-related hypertension risk was analysed using likelihood ratio tests. Results Among controls, longer age-adjusted RTL was associated with higher vegetable intake (p=0.01). Individuals with longer age-adjusted RTL (based on median value) were 30% less likely to have hypertension (OR=0.70, 95% CI 0.52 to 0.96; p=0.03). The observed RTL-hypertension relationship appeared to be modified by vegetable intake—longer RTL was significantly associated with lower hypertension risk only in those with greater vegetable consumption (OR=0.28, 95% CI 0.14 to 0.57; p<0.001), but not in those with lower vegetable intake (P-interaction=0.008). Conclusions Certain dietary factors might modify telomere-related hypertension risk. PMID:26560064

  6. User Manuals for a Primary Care Electronic Medical Record System: A Mixed Methods Study of User- and Vendor-Generated Documents

    PubMed Central

    Dow, Rustam; Barnsley, Jan; Tu, Karen; Domb, Sharon; Jadad, Alejandro R.; Lemieux-Charles, Louise

    2015-01-01

    Research problem Tutorials and user manuals are important forms of impersonal support for using software applications including electronic medical records (EMRs). Differences between user- and vendor documentation may indicate support needs, which are not sufficiently addressed by the official documentation, and reveal new elements that may inform the design of tutorials and user manuals. Research question What are the differences between user-generated tutorials and manuals for an EMR and the official user manual from the software vendor? Literature review Effective design of tutorials and user manuals requires careful packaging of information, balance between declarative and procedural texts, an action and task-oriented approach, support for error recognition and recovery, and effective use of visual elements. No previous research compared these elements between formal and informal documents. Methodology We conducted an mixed methods study. Seven tutorials and two manuals for an EMR were collected from three family health teams and compared with the official user manual from the software vendor. Documents were qualitatively analyzed using a framework analysis approach in relation to the principles of technical documentation described above. Subsets of the data were quantitatively analyzed using cross-tabulation to compare the types of error information and visual cues in screen captures between user- and vendor-generated manuals. Results and discussion The user-developed tutorials and manuals differed from the vendor-developed manual in that they contained mostly procedural and not declarative information; were customized to the specific workflow, user roles, and patient characteristics; contained more error information related to work processes than to software usage; and used explicit visual cues on screen captures to help users identify window elements. These findings imply that to support EMR implementation, tutorials and manuals need to be customized and

  7. Mixed cryoglobulinemia

    PubMed Central

    Ferri, Clodoveo

    2008-01-01

    Mixed cryoglobulinemia (MC), type II and type III, refers to the presence of circulating cryoprecipitable immune complexes in the serum and manifests clinically by a classical triad of purpura, weakness and arthralgias. It is considered to be a rare disorder, but its true prevalence remains unknown. The disease is more common in Southern Europe than in Northern Europe or Northern America. The prevalence of 'essential' MC is reported as approximately 1:100,000 (with a female-to-male ratio 3:1), but this term is now used to refer to a minority of MC patients only. MC is characterized by variable organ involvement including skin lesions (orthostatic purpura, ulcers), chronic hepatitis, membranoproliferative glomerulonephritis, peripheral neuropathy, diffuse vasculitis, and, less frequently, interstitial lung involvement and endocrine disorders. Some patients may develop lymphatic and hepatic malignancies, usually as a late complication. MC may be associated with numerous infectious or immunological diseases. When isolated, MC may represent a distinct disease, the so-called 'essential' MC. The etiopathogenesis of MC is not completely understood. Hepatitis C virus (HCV) infection is suggested to play a causative role, with the contribution of genetic and/or environmental factors. Moreover, MC may be associated with other infectious agents or immunological disorders, such as human immunodeficiency virus (HIV) infection or primary Sjögren's syndrome. Diagnosis is based on clinical and laboratory findings. Circulating mixed cryoglobulins, low C4 levels and orthostatic skin purpura are the hallmarks of the disease. Leukocytoclastic vasculitis involving medium- and, more often, small-sized blood vessels is the typical pathological finding, easily detectable by means of skin biopsy of recent vasculitic lesions. Differential diagnoses include a wide range of systemic, infectious and neoplastic disorders, mainly autoimmune hepatitis, Sjögren's syndrome, polyarthritis, and B

  8. Mixed cryoglobulinemia.

    PubMed

    Ferri, Clodoveo

    2008-09-16

    Mixed cryoglobulinemia (MC), type II and type III, refers to the presence of circulating cryoprecipitable immune complexes in the serum and manifests clinically by a classical triad of purpura, weakness and arthralgias. It is considered to be a rare disorder, but its true prevalence remains unknown. The disease is more common in Southern Europe than in Northern Europe or Northern America. The prevalence of 'essential' MC is reported as approximately 1:100,000 (with a female-to-male ratio 3:1), but this term is now used to refer to a minority of MC patients only. MC is characterized by variable organ involvement including skin lesions (orthostatic purpura, ulcers), chronic hepatitis, membranoproliferative glomerulonephritis, peripheral neuropathy, diffuse vasculitis, and, less frequently, interstitial lung involvement and endocrine disorders. Some patients may develop lymphatic and hepatic malignancies, usually as a late complication. MC may be associated with numerous infectious or immunological diseases. When isolated, MC may represent a distinct disease, the so-called 'essential' MC. The etiopathogenesis of MC is not completely understood. Hepatitis C virus (HCV) infection is suggested to play a causative role, with the contribution of genetic and/or environmental factors. Moreover, MC may be associated with other infectious agents or immunological disorders, such as human immunodeficiency virus (HIV) infection or primary Sjögren's syndrome. Diagnosis is based on clinical and laboratory findings. Circulating mixed cryoglobulins, low C4 levels and orthostatic skin purpura are the hallmarks of the disease. Leukocytoclastic vasculitis involving medium- and, more often, small-sized blood vessels is the typical pathological finding, easily detectable by means of skin biopsy of recent vasculitic lesions. Differential diagnoses include a wide range of systemic, infectious and neoplastic disorders, mainly autoimmune hepatitis, Sjögren's syndrome, polyarthritis, and B

  9. Prevention of the β-amyloid peptide-induced inflammatory process by inhibition of double-stranded RNA-dependent protein kinase in primary murine mixed co-cultures

    PubMed Central

    2011-01-01

    Background Inflammation may be involved in the pathogenesis of Alzheimer's disease (AD). There has been little success with anti-inflammatory drugs in AD, while the promise of anti-inflammatory treatment is more evident in experimental models. A new anti-inflammatory strategy requires a better understanding of molecular mechanisms. Among the plethora of signaling pathways activated by β-amyloid (Aβ) peptides, the nuclear factor-kappa B (NF-κB) pathway could be an interesting target. In virus-infected cells, double-stranded RNA-dependent protein kinase (PKR) controls the NF-κB signaling pathway. It is well-known that PKR is activated in AD. This led us to study the effect of a specific inhibitor of PKR on the Aβ42-induced inflammatory response in primary mixed murine co-cultures, allowing interactions between neurons, astrocytes and microglia. Methods Primary mixed murine co-cultures were prepared in three steps: a primary culture of astrocytes and microglia for 14 days, then a primary culture of neurons and astrocytes which were cultured with microglia purified from the first culture. Before exposure to Aβ neurotoxicity (72 h), co-cultures were treated with compound C16, a specific inhibitor of PKR. Levels of tumor necrosis factor-α (TNFα), interleukin (IL)-1β, and IL-6 were assessed by ELISA. Levels of PT451-PKR and activation of IκB, NF-κB and caspase-3 were assessed by western blotting. Apoptosis was also followed using annexin V-FITC immunostaining kit. Subcellular distribution of PT451-PKR was assessed by confocal immunofluorescence and morphological structure of cells by scanning electron microscopy. Data were analysed using one-way ANOVA followed by a Newman-Keuls' post hoc test Results In these co-cultures, PKR inhibition prevented Aβ42-induced activation of IκB and NF-κB, strongly decreased production and release of tumor necrosis factor (TNFα) and interleukin (IL)-1β, and limited apoptosis. Conclusion In spite of the complexity of the

  10. The costs of training a nurse practitioner in primary care: the importance of allowing for the cost of education and training when making decisions about changing the professional-mix.

    PubMed

    Curtis, Lesley; Netten, Ann

    2007-05-01

    What is already known on this topic * Cost containment through the most effective mix of staff achievable within available resources and organisational priorities is of increasing importance in most health systems. However, there is a dearth of information about the full economic implications of changing skill mix. * In the UK a major shift in the primary care workforce is likely in response to the rapidly developing role of nurse practitioners and policies aimed to encourage GP practices to transfer some of their responsibilities to other, less costly, professionals. * Previous research has developed an approach to incorporating the costs of qualifications, and thus the investment required to develop a skilled workforce, for a variety of health service professionals including GPs. What this study adds * This paper describes a methodology of costing nurse practitioners that incorporates the human capital cost implications of developing a skilled nurse practitioner workforce. With appropriate sources of data the method could be adapted for use internationally. * Including the full cost of qualifications results in nearly a 24 per cent increase in the unit cost of a Nurse Practitioner. * Allowing for all investment costs and adjusting for length of consultation, the cost of a GP consultation was nearly 60 per cent higher than that of a Nurse Practitioner.

  11. An electronic clinical decision support tool to assist primary care providers in cardiovascular disease risk management: development and mixed methods evaluation.

    PubMed

    Peiris, David P; Joshi, Rohina; Webster, Ruth J; Groenestein, Patrick; Usherwood, Tim P; Heeley, Emma; Turnbull, Fiona M; Lipman, Alexandra; Patel, Anushka A

    2009-12-17

    Challenges remain in translating the well-established evidence for management of cardiovascular disease (CVD) risk into clinical practice. Although electronic clinical decision support (CDS) systems are known to improve practitioner performance, their development in Australian primary health care settings is limited. Study aims were to (1) develop a valid CDS tool that assists Australian general practitioners (GPs) in global CVD risk management, and (2) preliminarily evaluate its acceptability to GPs as a point-of-care resource for both general and underserved populations. CVD risk estimation (based on Framingham algorithms) and risk-based management advice (using recommendations from six Australian guidelines) were programmed into a software package. Tool validation: Data from 137 patients attending a physician's clinic were analyzed to compare the tool's risk scores with those obtained from an independently programmed algorithm in a separate statistics package. The tool's management advice was compared with a physician's recommendations based on a manual review of the guidelines. Field test: The tool was then tested with 21 GPs from eight general practices and three Aboriginal Medical Services. Customized CDS-based recommendations were generated for 200 routinely attending patients (33% Aboriginal) using information extracted from the health record by a research assistant. GPs reviewed these recommendations during each consultation. Changes in CVD risk factor measurement and management were recorded. In-depth interviews with GPs were conducted. Validation testing: the tool's risk assessment algorithm correlated very highly with the independently programmed version in the separate statistics package (intraclass correlation coefficient 0.999). For management advice, there were only two cases of disagreement between the tool and the physician. Field test: GPs found 77% (153/200) of patient outputs easy to understand and agreed with screening and prescribing

  12. Improving access to high-quality primary care for socioeconomically disadvantaged older people in rural areas: a mixed method study protocol

    PubMed Central

    Ford, John A; Jones, Andrew P; Wong, Geoff; Clark, Allan B; Porter, Tom; Shakespeare, Tom; Swart, Ann Marie; Steel, Nicholas

    2015-01-01

    Introduction The UK has an ageing population, especially in rural areas, where deprivation is high among older people. Previous research has identified this group as at high risk of poor access to healthcare. The aim of this study is to generate a theory of how socioeconomically disadvantaged older people from rural areas access primary care, to develop an intervention based on this theory and test it in a feasibility trial. Methods and analysis On the basis of the MRC Framework for Developing and Evaluating Complex Interventions, three methods will be used to generate the theory. First, a realist review will elucidate the patient pathway based on existing literature. Second, an analysis of the English Longitudinal Study of Ageing will be completed using structural equation modelling. Third, 15 semistructured interviews will be undertaken with patients and four focus groups with health professionals. A triangulation protocol will be used to allow each of these methods to inform and be informed by each other, and to integrate data into one overall realist theory. Based on this theory, an intervention will be developed in discussion with stakeholders to ensure that the intervention is feasible and practical. The intervention will be tested within a feasibility trial, the design of which will depend on the intervention. Lessons from the feasibility trial will be used to refine the intervention and gather the information needed for a definitive trial. Ethics and dissemination Ethics approval from the regional ethics committee has been granted for the focus groups with health professionals and interviews with patients. Ethics approval will be sought for the feasibility trial after the intervention has been designed. Findings will be disseminated to the key stakeholders involved in intervention development, to researchers, clinicians and health planners through peer-reviewed journal articles and conference publications, and locally through a dissemination event. PMID

  13. Investigating the feasibility, acceptability and appropriateness of outreach case management in an urban Aboriginal and Torres Strait Islander primary health care service: a mixed methods exploratory study.

    PubMed

    Askew, Deborah A; Togni, Samantha J; Schluter, Philip J; Rogers, Lynne; Egert, Sonya; Potter, Nichola; Hayman, Noel E; Cass, Alan; Brown, Alex D H

    2016-05-13

    The disparities in health and life expectancy of Aboriginal and Torres Strait Islander peoples compared to non-Indigenous Australians are well documented. Chronic diseases are a leading contributor to these disparities. We aimed to determine the feasibility, acceptability and appropriateness of a case management approach to chronic disease care integrated within an urban Aboriginal and Torres Strait Islander primary health care service. The Home-based, Outreach case Management of chronic disease Exploratory (HOME) Study provided holistic, patient centred multidisciplinary care for Aboriginal and Torres Strait Islander people with chronic disease. A developmental evaluation approach supported the implementation and ongoing adaptations in the delivery of the model of care, and ensured its alignment with Aboriginal and Torres Strait Islander peoples' understandings of, and approaches to, health and wellbeing. In-depth, semi-structured interviews were conducted with nine patient participants (one interview also included a participant's spouse) and 15 health service staff and key themes were identified through an iterative reflective process. Quantitative data were collected directly from patient participants and from their medical records at baseline, 3 and 6 months. Patient participants' baseline characteristics were described using frequencies and percentages. Attrition and patterns of missing values over time were evaluated using binomial generalized estimating equation (GEE) models and mean differences in key clinical outcomes were determined using normal GEE models. Forty-one patients were recruited and nine withdrew over the 6 month period. There was no evidence of differential attrition. All participants (patients and health service staff) were very positive about the model of care. Patient participants became more involved in their health care, depression rates significantly decreased (p = 0.03), and significant improvements in systolic blood pressure (p

  14. Flow cytometry detection of infectious pancreatic necrosis virus (IPNV) within subpopulations of Atlantic salmon (Salmo salar L.) leucocytes after vaccination and during the time course of experimental infection.

    PubMed

    Rønneseth, Anita; Haugland, Gyri Teien; Wergeland, Heidrun I

    2013-05-01

    In the present study, intracellular infectious pancreatic necrosis virus (IPNV) in salmon leucocytes was detected by flow cytometry after experimental cohabitant challenge. IPNV vaccinated, non-vaccinated and intraperitoneally (i.p.) infected salmon (virus shedders) were analysed at different times throughout the period when mortality occurred. Fish that had survived 61 days post challenge (carriers) were also analysed. In particular, we analysed the presence of IPNV in B-cells (C7G7+cells) and in neutrophils (E3D9+ cells) in head kidney leucocytes (HKL) and in peripheral blood leucocytes (PBL). IPNV was present in HKL and PBL from all challenged fish groups at all samplings, including carriers. IPNV was also found intracellular in other leucocytes than B-cells and neutrophils. During the time course of infection there were changes in proportion of B-cells and neutrophils and in proportions of IPNV+ cells. In vaccinated fish, a delay in the changes observed in the proportion of IPNV+ cells and in the proportions of the two subpopulations was identified. The vaccinated fish were protected against disease as no fish died compared to 30.8% of non-vaccinated cohabitant fish. All i.p. infected fish, except one, survived the challenge. This is consistent with previous studies and confirmed that the routes of infection can influence mortality. The analyses in this study could not identify any factors enlightening this absence of mortality in i.p. infected fish, but both flow cytometry and qRT-PCR showed that i.p. infected fish were carriers of IPNV. The present study also found that IPNV was present in both B-cells and neutrophils as well as in other leucocytes in all carriers after cohabitant challenge. These fish had survived 9 weeks post challenge and 4 weeks after mortality has ceased. The fish harbouring virus within their leucocytes might become life long carriers and represent a risk for disease outbreaks, being virus shedders. Such fish are protected from later

  15. Normal platelet function in platelet concentrates requires non-platelet cells: a comparative in vitro evaluation of leucocyte-rich (type 1a) and leucocyte-poor (type 3b) platelet concentrates

    PubMed Central

    Parrish, William R; Roides, Breana; Hwang, Julia; Mafilios, Michael; Story, Brooks; Bhattacharyya, Samir

    2016-01-01

    Background Therapeutic success of platelet-rich plasma (PRP) may vary based on the composition and preparation method. The objective of this study was to evaluate the cellular components of platelet concentrates produced by a leucocyte-rich (LR-PRP) and a leucocyte-poor PRP systems (LP-PRP). Methods Parameters evaluated included platelet recovery, platelet concentration, red blood cell (RBC) and white blood cell (WBC) composition, platelet growth factor release and stimulation of human tendon cell proliferation in vitro. Results Platelet recoveries were 52% for LP-PRP and 89% for LR-PRP. LR-PRP demonstrated greater reproducibility with a 4.2% coefficient of variation (CV) compared with 19.4% for LP-PRP (p<0.001). LR-PRP demonstrated a greater increase in platelet concentration (7.9-fold) than LP-PRP (2.2-fold; p<0.001). LP-PRP showed 5.0-fold reductions in WBCs, while LR-PRP showed a 4.0-fold increase (p<0.001). LP-PRP reduced RBCs to a haematocrit of 0.25, while LR-PRP reduced haematocrit to 11.8. LP-PRP did not coagulate robustly on reactivation with CaCl2, and released significantly lower levels of epidermal growth factor (EGF) and transforming growth factor β1 (TGF-β1) than whole blood (p<0.03). LP-PRP also did not stimulate tendon cell proliferation greater than whole blood. In contrast, LR-PRP showed increases in each growth factor on activation with CaCl2 (p<0.01) and stimulated greater proliferation (p<0.05) compared with whole blood. Forced activation of LP-PRP with exogenous thrombin rescued the coagulation deficiency and induced greater growth factor release than comparable whole blood (p<0.03). Conclusions These data suggest that non-platelet cellular components in platelet concentrates are important for proper platelet function, including thrombin generation, growth factor release and clot retraction. PMID:27900155

  16. Atomization and Mixing Study

    NASA Technical Reports Server (NTRS)

    Ferrenberg, A.; Hunt, K.; Duesberg, J.

    1985-01-01

    The primary objective was the obtainment of atomization and mixing performance data for a variety of typical liquid oxygen/hydrocarbon injector element designs. Such data are required to establish injector design criteria and to provide critical inputs to liquid rocket engine combustor performance and stability analysis, and computational codes and methods. Deficiencies and problems with the atomization test equipment were identified, and action initiated to resolve them. Test results of the gas/liquid mixing tests indicated that an assessment of test methods was required. A series of 71 liquid/liquid tests were performed.

  17. Leucocytes, cytokines and satellite cells: what role do they play in muscle damage and regeneration following eccentric exercise?

    PubMed

    Paulsen, Gøran; Mikkelsen, Ulla Ramer; Raastad, Truls; Peake, Jonathan M

    2012-01-01

    Exercise-induced muscle damage is an important topic in exercise physiology. However several aspects of our understanding of how muscles respond to highly stressful exercise remain unclear In the first section of this review we address the evidence that exercise can cause muscle damage and inflammation in otherwise healthy human skeletal muscles. We approach this concept by comparing changes in muscle function (i.e., the force-generating capacity) with the degree of leucocyte accumulation in muscle following exercise. In the second section, we explore the cytokine response to 'muscle-damaging exercise', primarily eccentric exercise. We review the evidence for the notion that the degree of muscle damage is related to the magnitude of the cytokine response. In the third and final section, we look at the satellite cell response to a single bout of eccentric exercise, as well as the role of the cyclooxygenase enzymes (COX1 and 2). In summary, we propose that muscle damage as evaluated by changes in muscle function is related to leucocyte accumulation in the exercised muscles. 'Extreme' exercise protocols, encompassing unaccustomed maximal eccentric exercise across a large range of motion, generally inflict severe muscle damage, inflammation and prolonged recovery (> 1 week). By contrast, exercise resembling regular athletic training (resistance exercise and downhill running) typically causes mild muscle damage (myofibrillar disruptions) and full recovery normally occurs within a few days. Large variation in individual responses to a given exercise should, however be expected. The link between cytokine and satellite cell responses and exercise-induced muscle damage is not so clear The systemic cytokine response may be linked more closely to the metabolic demands of exercise rather than muscle damage. With the exception of IL-6, the sources of systemic cytokines following exercise remain unclear The satellite cell response to severe muscle damage is related to

  18. Markers of endothelial dysfunction and leucocyte activation in Saudi and non-Saudi haplotypes of sickle cell disease.

    PubMed

    Al Najjar, Salwa; Adam, Soheir; Ahmed, Nessar; Qari, Mohamed

    2017-01-01

    Sickle cell disease (SCD) is an autosomal recessive inherited hemoglobinopathy, characterized by chronic hemolysis and recurrent vaso-occlusive crisis (VOC). This study investigates changes in leucocyte subsets and the relationship between cell adhesion molecule expression and disease manifestations in patients during steady state and acute VOC. We compared soluble E-selectin and P-selectin levels in 84 SCD patients, in steady state and during VOC to 84 healthy controls. Using immunophenotyping, we also compared lymphocyte subsets in these three groups. Further, we compared E-selectin and P-selectin levels in patients of Saudi ethnicity to non-Saudi patients, in all three groups. Lymphocyte subsets showed high percentages of total T lymphocytes, T helper and suppressor lymphocytes, B lymphocytes as well as NK cells in patients with SCD during steady state, while B lymphocytes and NK cells were significantly higher during acute VOC crisis. High levels of both soluble E-selectin (sE-selectin) and soluble P-selectin (sP-selectin) markers were demonstrated in the serum of patients with SCD during both steady state and acute VOC. Levels of selectins were significantly higher in acute VOC. The immunophenotypic expression of L-selectin, on leucocytes, was high in SCD both during steady state and during acute VOC in comparison to normal control subjects. There was no significant difference in all three study groups between Saudi and non-Saudi patients. These findings suggest that patients with SCD have increased expression of adhesion molecules: E-selectin and P-selectin, which play an important role in the pathogenesis of VOC. Despite the distinct phenotype of Saudi patients with SCD, there was no significant difference in levels of soluble E-selectin and soluble P-selectin between Saudi and non-Saudi patients in all three groups. While sickle cell disease is a well-recognized state of chronic inflammation, the role of specific adhesion molecules is steadily unraveling

  19. Discrediting microscopic pyuria and leucocyte esterase as diagnostic surrogates for infection in patients with lower urinary tract symptoms: results from a clinical and laboratory evaluation.

    PubMed

    Kupelian, Anthony S; Horsley, Harry; Khasriya, Rajvinder; Amussah, Rasheedah T; Badiani, Raj; Courtney, Angela M; Chandhyoke, Nihil S; Riaz, Usama; Savlani, Karishma; Moledina, Malik; Montes, Samantha; O'Connor, Dominic; Visavadia, Rakhee; Kelsey, Michael; Rohn, Jennifer L; Malone-Lee, James

    2013-07-01

    WHAT'S KNOWN ON THE SUBJECT? AND WHAT DOES THE STUDY ADD?: Microscopic pyuria is widely used as a surrogate marker of infection, although there is little data supporting its use in patients who present with non-acute LUTS. The effects of urinary storage, preservation, and the use of laboratory methods to enhance leucocyte detection, are also unclear. This large, prospective study highlights the poor performance of dipstick urine analysis, and direct microscopy, as surrogate markers of UTI in patients with LUTS. A series of laboratory analyses also examine the effects of urine handling and processing on test integrity, which have important implications for clinical practice. To evaluate the diagnostic performance of pyuria as a surrogate marker of urinary tract infection (UTI) in patients with chronic lower urinary tract symptoms (LUTS), and determine the impact of sample storage, cytocentrifugation, and staining techniques, on test performance. Between 2008 and 2011, we recruited 1223 patients (120 men; 1103 women; mean age 54 years) with one or more LUTS from a specialist urological outpatient service. We conducted a prospective observational study to determine the performance of microscopic pyuria ≥10 wbc/μL as a surrogate marker of UTI in patients with LUTS. All patients provided clean-catch midstream urine (MSU) samples for analysis, and routine microbiological cultures were used as our reference standard. We also scrutinised the performance of dipstick leucocyte esterase ≥ 'trace' in the detection of microscopic pyuria. The influence of sample handling and processing on test performance was examined in a series of laboratory studies. The effects of storage on leucocyte decay were determined using repeated microscopic assessments of individual urine samples, to plot temporal changes in leucocyte numbers. This study used varied storage conditions (≈20 °C and 4 °C), and boric acid preservation. Paired microscopic assessments were used to determine

  20. Levels of expression of complement regulatory proteins CD46, CD55 and CD59 on resting and activated human peripheral blood leucocytes

    PubMed Central

    Christmas, Stephen E; de la Mata Espinosa, Claudia T; Halliday, Deborah; Buxton, Cheryl A; Cummerson, Joanne A; Johnson, Peter M

    2006-01-01

    The cell surface complement regulatory (CReg) proteins CD46, CD55 and CD59 are widely expressed on human lymphoid and non-lymphoid cells. This study aimed to compare systematically levels of CReg expression by different leucocyte subsets and to determine whether levels were increased following activation in vitro. Levels of each CReg protein were similar on freshly isolated monocytes and all major lymphocyte subsets, except that CD4+ cells expressed significantly less CD46 than CD8+ cells (P < 0·05) while the reverse was observed for CD55 (P < 0·02). CD56+ cells, predominantly natural killer cells, expressed significantly lower levels of CD59 than T cells (P < 0·02). CD45RO+ cells had higher levels of surface CD46 and CD59, but lower levels of CD55, than CD45RO– cells (P < 0·02); CD25+ cells also expressed significantly less CD55 than CD25− cells (P < 0·002). Neutrophils expressed higher levels of CD59, but lower levels of CD55, than monocytes. Following activation with phytohaemagglutinin, CD46 was up-regulated on all leucocyte subsets with the exception of CD56+ cells. Both CD55 and CD59 were also markedly up-regulated on monocytes, and CD55 expression was greater on CD8+ than CD4+ cells following activation (P < 0·02). Lipopolysaccharide treatment did not significantly alter B-cell expression of CReg proteins whereas CD55 and CD59, but not CD46, were significantly up-regulated on monocytes (P < 0·02). These observations that CReg proteins are up-regulated on certain activated leucocyte subsets indicate that levels would be increased following immune responses in vivo. This could enhance both protection against local complement activation at inflammatory sites and also the immunoregulatory properties of these leucocytes. PMID:16999828

  1. In vitro effects of prostaglandin E2 on leucocytes from sticklebacks (Gasterosteus aculeatus) infected and not infected with the cestode Schistocephalus solidus.

    PubMed

    Kutyrev, Ivan A; Franke, Frederik; Büscher, Janine; Kurtz, Joachim; Scharsack, Jörn P

    2014-12-01

    Many helminth parasites have evolved strategies to evade the immune response of their hosts, which includes immunomodulation. Prostaglandin E2 (PGE2) is one of the best-described immunomodulators in mammalian helminth parasite infections. We hypothesized that also in teleost fish anti-helminthic immune responses are regulated via PGE2. We used a model system consisting of the tapeworm Schistocephalus solidus and its host, the three-spined stickleback (Gasterosteus aculeatus), to investigate in vitro effects of PGE2 on head kidney leucocytes (HKL) derived from sticklebacks that were experimentally infected with S. solidus. PGE2 was tested alone or in combination with either S. solidus antigens or bacterial lipopolysaccharides (LPS). After in vitro culture, cell viability and changes in leucocyte subpopulations (granulocytes to lymphocytes ratios) were monitored by flow cytometry and HKL were tested for their capacity to produce reactive oxygen species (ROS) with a chemiluminescence assay. In short term (2 h) HKL cultures PGE2 did not change the total numbers of live HKL, but the production of ROS decreased significantly with high (0.1 μmol L(-1)) PGE2 concentrations. In long-term (96 h) cultures high PGE2 concentrations induced a sharp decrease of leucocytes viability, while low (0.1 pmol L(-1)) and intermediate (0.1 nmol L(-1)) concentrations of PGE2 caused elevated leucocyte viability compared to controls. This coincided with reduced ROS production in cultures with high PGE2 and elevated ROS production in cultures with low PGE2. Granulocyte to lymphocyte ratios increased with high PGE2 concentrations alone and in combination with S. solidus antigens and LPS, most prominently with HKL from S. solidus infected sticklebacks. The present study supports the hypothesis that PGE2 might be an immunomodulator in tapeworm-fish parasite-host interactions.

  2. The Potent Humanin Analogue (HNG) Protects Germ Cells and Leucocytes While Enhancing Chemotherapy-Induced Suppression of Cancer Metastases in Male Mice.

    PubMed

    Lue, YanHe; Swerdloff, Ronald; Wan, Junxiang; Xiao, Jialin; French, Samuel; Atienza, Vince; Canela, Victor; Bruhn, Kevin W; Stone, Brian; Jia, Yue; Cohen, Pinchas; Wang, Christina

    2015-12-01

    Humanin is a peptide that is cytoprotective against stresses in many cell types. We investigated whether a potent humanin analogue S14G-humanin (HNG) would protect against chemotherapy-induced damage to normal cells without interfering with the chemotherapy-induced suppression of cancer cells. Young adult male mice were inoculated iv with murine melanoma cells. After 1 week, cancer-bearing mice were randomized to receive either: no treatment, daily ip injection of HNG, a single ip injection of cyclophosphamide (CP), or CP+HNG and killed at the end of 3 weeks. HNG rescued the CP-induced suppression of leucocytes and protected germ cell from CP-induced apoptosis. Lung metastases were suppressed by HNG or CP alone, and further suppressed by CP+HNG treatment. Plasma IGF-1 levels were suppressed by HNG with or without CP treatment. To investigate whether HNG maintains its protective effects on spermatogonial stem cells, sperm output, and peripheral leucocytes after repeated doses of CP, normal adult male mice received: no treatment, daily sc injection of HNG, 6 ip injections of CP at 5-day intervals, and the same regimens of CP+HNG and killed at the end of 4 weeks of treatment. Cauda epididymal sperm counts were elevated by HNG and suppressed by CP. HNG rescued the CP-induced suppression of spermatogonial stem cells, sperm count and peripheral