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.
Yong, K; Khwaja, A
Leucocytes express adhesion promoting receptors which mediate cell-cell and cell-matrix interactions. These adhesive interactions are crucial to the regulation of haemopoiesis and thymocyte maturation, the direction and control of leucocyte traffic and migration through tissues, and in the development of immune and non-immune inflammatory responses. Several families of adhesion receptors have been identified (Table). The leucocyte integrin family comprises 3 alpha beta heterodimeric membrane glycoproteins which share a common beta subunit, designated CD18. The alpha subunits of each of the 3 members, lymphocyte function associated antigen-1 (LFA-1), macrophage antigen-1 (Mac-1) and p150,95 are designated CD11a, b and c respectively. These adhesion molecules play a critical part in the immune and inflammatory responses of leucocytes. The leucocyte integrin family is, in turn, part of the integrin superfamily, members of which are evolutionally, structurally and functionally related. Another Integrin subfamily found on leucocytes is the VLA group, so-called because the 'very late activation antigens' VLA-1 and VLA-2 were originally found to appear late in T-cell activation. Members of this family function mainly as extracellular matrix adhesion receptors and are found both on haemopoietic and non-haemopoietic cells. They play a part in diverse cellular functions including tissue organisation, lymphocyte recirculation and T-cell immune responses. A third integrin subfamily, the cytoadhesins, are receptors on platelets and endothelial cells which bind extracellular matrix proteins. A second family of adhesion receptors is the immunoglobulin superfamily, members of which include CD2, LFA-3 and ICAM-1, which participate in T-cell adhesive interactions, and the antigen-specific receptors of T and B cells, CD4, CD8 and the MHC Class I and II molecules. A recently recognised family of adhesion receptors is the selectins, characterised by a common lectin domain. Leucocyte
Pichler, Herbert; Fritsch, Gerhard; König, Margit; Daxberger, Helga; Glogova, Evgenia; Pötschger, Ulrike; Breuer, Sabine; Lawitschka, Anita; Güclü, Ece D; Karlhuber, Susanne; Holter, Wolfgang; Haas, Oskar A; Lion, Thomas; Matthes-Martin, Susanne
The impact of persistent mixed chimerism (MC) after haematopoietic stem cell transplantation (HSCT) remains unclarified. We investigated the incidence of MC in peripheral blood beyond day +50 after HSCT and its impact on rejection, chronic graft-versus-host disease (c-GvHD) and relapse in 161 children receiving allogeneic HSCT for haematological malignancies. The 1-year incidence of late MC was 26%. Spontaneous conversion to complete donor chimerism (CC) occurred in 43% of patients as compared to 62% after donor lymphocyte infusions. No graft rejection occurred. The 1-year incidence of c-GvHD was 20 ± 7% for MC, and 18 ± 4% for CC patients (P = 0·734). The 3-year cumulative incidence of relapse (CIR) according to chimerism status at days +50 and +100 was 22 ± 4% for CC patients vs. 22 ± 8% for MC patients (day +50; P = 0·935) and 21 ± 4% vs. 20 ± 7% (day +100; P = 0·907). Three-year CIRs in patients with persistent MC and patients with CC/limited MC were comparable (8 ± 7% vs. 19 ± 4%; P = 0·960). HSCT for acute leukaemia or myelodysplastic syndrome as secondary malignancies (hazard ratio (HR) 4·7; P = 0·008), for AML (HR 3·0; P = 0·02) and from mismatched donors (HR 3·1; P = 0·03) were independent factors associated with relapse. Our data suggest that late MC neither protects from c-GvHD nor does it reliably predict impending disease relapse. PMID:26996395
Cvetkovic, A.; Mohedano, R.; Dross, O.; Hernandez, M.; Benítez, P.; Miñano, J. C.; Vilaplana, J.; Chaves, J.
In SSL general illumination, there is a clear trend to high flux packages with higher efficiency and higher CRI addressed with the use of multiple color chips and phosphors. However, such light sources require the optics provide color mixing, both in the near-field and far-field. This design problem is specially challenging for collimated luminaries, in which diffusers (which dramatically reduce the brightness) cannot be applied without enlarging the exit aperture too much. In this work we present first injection molded prototypes of a novel primary shell-shaped optics that have microlenses on both sides to provide Köhler integration. This shell is design so when it is placed on top of an inhomogeneous multichip Lambertian LED, creates a highly homogeneous virtual source (i.e, spatially and angularly mixed), also Lambertian, which is located in the same position with only small increment of the size (about 10-20%, so the average brightness is similar to the brightness of the source). This shell-mixer device is very versatile and permits now to use a lens or a reflector secondary optics to collimate the light as desired, without color separation effects. Experimental measurements have shown optical efficiency of the shell of 95%, and highly homogeneous angular intensity distribution of collimated beams, in good agreement with the ray-tracing simulations.
Fotedar, A; Mustafa, A S; Narang, B S; Talwar, G P
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
Spinelli, F; Sara, R; Milella, M; Ruffini, L; Sterzi, R; Causarano, I R; Sberna, M
The diagnosis of brain abscess is often difficult, as the clinical symptoms are not specific. Computed tomography (CT) and magnetic resonance imaging (MRI) are highly sensitive, but different cerebral lesions, especially neoplasms, can have the same ring-like contrast enhancement. Brain abscess is a severe illness requiring rapid diagnosis to choose the most appropriate therapy. Technetium-99m hexamethylpropylene amine oxime (HMPAO)-labelled leucocyte scintigraphy is commonly used to detect an inflammatory process. The aim of this study was to present the results obtained with leucocyte scintigraphy in 65 patients with intracranial mass lesions and clinical findings compatible to or suggestive of brain abscess. The final diagnosis, based on surgery, clinical findings and stereotatic puncture, was brain abscess in 17 patients, primary brain neoplasm in 22, brain metastasis in 16, lymphoma in 2, cysticercosis in 2, hematoma in 2 and cerebral infarction in 4. 99mTc-HMPAO leucocyte scintigraphy was positive in all abscess cases. The scan was negative in the rest of the patients examined, with the exception of one lesion, which was finally diagnosed as a tumour (1 false-positive). All patients who did not have false-negative scans were treated with steroids. The sensitivity, specificity and diagnostic accuracy of leucocyte scintigraphy was 100%, 97.8% and 98.4%, respectively. In conclusion, in our experience, leucocyte scintigraphy is a valuable aid in the differential diagnosis between abscess and neoplasm. PMID:10654146
Schumacher, A; Liebers, U; John, M; Gerl, V; Meyer, M; Witt, C; Wolff, G
Chronic obstructive pulmonary disease (COPD) is characterized by a dysregulated recruitment of circulating leucocytes into the lung which is associated with the onset and progress of the disease. P-selectin glycoprotein ligand-1 (PSGL-1) is expressed on leucocytes and plays an essential role in primary leucocyte-endothelial cell adhesive contacts. The present study investigated if PSGL-1 is up-regulated on leucocytes of COPD patients. Peripheral blood samples were collected from COPD patients as well as controls (smoking, nonsmoking volunteers) and subjected to analysis of PSGL-1 expression on leucocytes, i.e. neutrophils, eosinophils, monocytes and lymphocytes by flow cytometry. No significant difference was observed between healthy nonsmoking and healthy smoking control subjects. In contrast, PSGL-1 expression was found to be significantly increased on the surface of all four leucocyte populations in COPD patients compared to both control groups. The finding that PSGL-1 surface expression is up-regulated on leucocytes of COPD patients as compared to leucocytes of controls suggests PSGL-1 as a potential target for anti-inflammatory treatment. PMID:16232226
Smith, M. G. M.; Eddleston, A. L. W. F.; Dominguez, J. A.; Evans, D. B.; Bewick, M.; Williams, Roger
The leucocyte migration test, an in-vitro measure of cellular immunity, has been used to follow the changes in cell-mediated hypersensitivity to kidney and histocompatibility antigens in three patients after renal transplantation. Inhibition of leucocyte migration, indicating strong sensitization to the antigens used, occurred in each patient, starting five to seven days after transplantation. Satisfactory renal function had not been established in any of the patients at this time. In one case inhibition of leucocyte migration persisted almost continuously until the 24th day and was associated with poor renal function proved histologically to be due to rejection. Treatment with increased dosage of prednisone was associated with a rapid reversion to normal of the migration index and improvement in renal function. Later, inhibition of migration occurred again, and shortly afterwards the graft ceased to function. In the other two cases the migration index became normal without alteration in immunosuppressive therapy and a satisfactory diuresis followed. It is suggested that this simple test should prove useful in the specific diagnosis of rejection and in control of immunosuppressive therapy. ImagesFig. 3Fig. 4 PMID:4899455
Keen, P; McCarthy, D A; Passfield, L; Shaker, H A; Wade, A J
Venous blood samples were taken from eight competitors in mid-evening after a racing day, and in the early morning before the next day's race, three times during the course of the Milk Race, 1992. These were used to gather information about the changes in circulating leucocyte levels in response to the exceptionally high sustained daily workload required during a major multi-stage race. The primary objective was to provide knowledge of 'normal' values against which future clinical judgements of abnormality might be made in these unusual circumstances. During the race, estimated energy output was about 25 MJ (6000 kCal)/day. The mean total circulating leucocyte numbers (per litre of blood), and those of individual leucocyte classes (neutrophil, lymphocyte, monocyte, eosinophil and basophil) were all inside the normal range both in the morning and in the evening. Evening counts were, however, 30-50% higher than morning counts, for all classes except eosinophils. We conclude that individual clinical decisions about leucocyte levels can best be made using normal (sedentary man) values if a morning sample is taken. PMID:7788222
Scheiermann, Christoph; Frenette, Paul S.; Hidalgo, Andrés
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
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…
Cabassa, Leopoldo J.; Gomes, Arminda P.; Meyreles, Quisqueya; Capitelli, Lucia; Younge, Richard; Dragatsi, Dianna; Alvarez, Juana; Nicasio, Andel; Druss, Benjamin; Lewis-Fernández, Roberto
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
Carroll, Andrew; Mansoori, N; Li, Shundai; Lei, Lei; Vernhettes, Samantha; Visser, Richard G. F.; Somerville, Chris R; Gu, Ying; Trindade, Luisa M.
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.
Bissett, W. Paul; Meyers, Mark B.; Walsh, John J.; Mueller-Karger, Frank E.
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.
Ratinen, Ilkka Johannes
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.
Gross, K.; Alcorn, S.; Murray, A.; Morrison, R.; Nowak, B.
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.
Paul, Arun; Parmar, Harshad; Chacko, Rabin
Germ cell Tumours (GCT) are neoplasm derived from germ cells. GCT usually occurs inside the gonads. Extragonadal GCT’s are rare. Most common GCT associated with head and neck region are the teratomas. Of the few teratomas found in the head and neck, malignant transformation of a teratomatous element is very uncommon, and primary bone involvement within the head and neck is even rare. We present a case of primary malignant mixed germ cell Tumour involving the mandible, the present case presented malignant transformation of the epithelial component showing foci of squamous cell carcinoma within the GCT. PMID:26266228
Raghoebar, M.; Huisman, J.A.M.; van den Berg, W.B.; van Ginneken, C.A.M.
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.
Zielinski, Andrzej; Kronogård, Maria; Lenhoff, Håkan; Halling, Anders
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
Dein, F.J.; Wilson, A.; Fischer, D.; Langenberg, P.
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.
Background Policy efforts focus on a reorientation of health care systems towards primary prevention. To guide such efforts, we analyzed the role of primary prevention in general practice and general practitioners’ (GPs) attitudes toward primary prevention. Methods Mixed-method study including a cross-sectional survey of all community-based GPs and focus groups in a sample of GPs who collaborated with the Institute of General Practice in Berlin, Germany in 2011. Of 1168 GPs 474 returned the mail survey. Fifteen GPs participated in focus group discussions. Survey and interview guidelines were developed and tested to assess and discuss beliefs, attitudes, and practices regarding primary prevention. Results Most respondents considered primary prevention within their realm of responsibility (70%). Primary prevention, especially physical activity, healthy eating, and smoking cessation, was part of the GPs’ health care recommendations if they thought it was indicated. Still a quarter of survey respondents discussed reduction of alcohol consumption with their patients infrequently even when they thought it was indicated. Similarly 18% claimed that they discuss smoking cessation only sometimes. The focus groups revealed that GPs were concerned about the detrimental effects an uninvited health behavior suggestion could have on patients and were hesitant to take on the role of “health policing”. GPs saw primary prevention as the responsibility of multiple actors in a network of societal and municipal institutions. Conclusions The mixed-method study showed that primary prevention approaches such as lifestyle counseling is not well established in primary care. GPs used a selective approach to offer preventive advice based upon indication. GPs had a strong sense that a universal prevention approach carried the potential to destroy a good patient-physician relationship. Other approaches to public health may be warranted such as a multisectoral approach to population
Hjelmborg, Jacob B; Dalgård, Christine; Möller, Soren; Steenstrup, Troels; Kimura, Masayuki; Christensen, Kaare; Kyvik, Kirsten O; Aviv, Abraham
Background Leucocyte telomere length (LTL) is a complex trait associated with ageing and longevity. LTL dynamics are defined by LTL and its age-dependent attrition. Strong, but indirect evidence suggests that LTL at birth and its attrition during childhood largely explains interindividual LTL variation among adults. A number of studies have estimated the heritability of LTL, but none has assessed the heritability of age-dependent LTL attrition. Methods We examined the heritability of LTL dynamics based on a longitudinal evaluation (an average follow-up of 12 years) in 355 monozygotic and 297 dizygotic same-sex twins (aged 19–64 years at baseline). Results Heritability of LTL at baseline was estimated at 64% (95% CI 39% to 83%) with 22% (95% CI 6% to 49%) of shared environmental effects. Heritability of age-dependent LTL attrition rate was estimated at 28% (95% CI 16% to 44%). Individually unique environmental factors, estimated at 72% (95% CI 56% to 84%) affected LTL attrition rate with no indication of shared environmental effects. Conclusions This is the first study that estimated heritability of LTL and also its age-dependent attrition. As LTL attrition is much slower in adults than in children and given that having a long or a short LTL is largely determined before adulthood, our findings suggest that heritability and early life environment are the main determinants of LTL throughout the human life course. Thus, insights into factors that influence LTL at birth and its dynamics during childhood are crucial for understanding the role of telomere genetics in human ageing and longevity. PMID:25770094
Scammon, Debra L; Tomoaia-Cotisel, Andrada; Day, Rachel L; Day, Julie; Kim, Jaewhan; Waitzman, Norman J; Farrell, Timothy W; Magill, Michael K
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
Mathy-Hartert, M; Bourgeois, E; Grülke, S; Deby-Dupont, G; Caudron, I; Deby, C; Lamy, M; Serteyn, D
Increases of plasma concentrations of neutrophil myeloperoxidase (MPO) can be used as markers of polymorphonuclear leucocytes (PMN) activation in pathological situations (sepsis, acute lung injury, acute inflammation). To develop an assay for measurement of plasma MPO in horses during the above-mentioned infectious and inflammatory conditions, MPO was purified from equine PMN isolated from blood anticoagulated with citrate. PMN were extracted in a saline milieu (0.2 M Na acetate, 1 M NaCl, pH 4.7) to eliminate most of cellular proteins. Pellets were then extracted in the same buffer containing cationic detergent (1% cetyltrimethyl ammonium bromide). The supernatant was further purified by ion exchange chromatography (Hiload S Sepharose HP column 0.5 x 26 cm, equilibrated with 25 mM Na acetate, 0.2 M NaCl, pH 4.7) with a NaCl gradient (until 1 M). Most of the peroxidase activity of MPO (spectrophotometrically measured by the oxidation of orthodianisidine by hydrogen peroxide) was eluted at 0.65 M NaCl. MPO was further purified by gel filtration chromatography (Sephacryl S 200 column 2.6 x 42 cm with 25 mM Na acetate, 0.2 M NaCl, pH 4.7). MPO (specific activity: 74.3 U/mg) was obtained with a yield of 30% from the detergent extraction supernatant. Electrophoresis (non-reducing conditions) showed 3 bands identified, by comparison with human MPO, (i) the mature tetrameric enzyme (150 kDa) with 2 light and 2 heavy subunits, (ii) the precursor form (88 kDa) and (iii) a form of the heavy subunit without the prosthetic heme group (40 kDa). The mature enzyme and its precursor were glycosylated and possessed peroxidase activity. Equine MPO showed strong similarities with human and bovine MPO, with an absorption peak at 430 nm (Soret peak) characteristic of ferrimyeloperoxidase. Enzymatic activity was pH dependent (optimal value at pH 5.5). Images Figure 1. PMID:9553712
Ozen, Asli; Balasubramanian, Hari
At the heart of the practice of primary care is the concept of a physician panel. A panel refers to the set of patients for whose long term, holistic care the physician is responsible. A physician's appointment burden is determined by the size and composition of the panel. Size refers to the number of patients in the panel while composition refers to the case-mix, or the type of patients (older versus younger, healthy versus chronic patients), in the panel. In this paper, we quantify the impact of the size and case-mix on the ability of a multi-provider practice to provide adequate access to its empanelled patients. We use overflow frequency, or the probability that the demand exceeds the capacity, as a measure of access. We formulate problem of minimizing the maximum overflow for a multi-physician practice as a non-linear integer programming problem and establish structural insights that enable us to create simple yet near optimal heuristic strategies to change panels. This optimization framework helps a practice: (1) quantify the imbalances across physicians due to the variation in case mix and panel size, and the resulting effect on access; and (2) determine how panels can be altered in the least disruptive way to improve access. We illustrate our methodology using four test practices created using patient level data from the primary care practice at Mayo Clinic, Rochester, Minnesota. An important advantage of our approach is that it can be implemented in an Excel Spreadsheet and used for aggregate level planning and panel management decisions. PMID:23076360
Nanda, Rishi; Koul, Monika; Srivastava, Salil; Upadhyay, Vinod; Dwivedi, Rahul
Problem considered The objective was to evaluate and compare the clinical and radiographic efficacy of two different combinations of antibacterial drugs when used in non-instrumental endodontic treatment of necrosed primary teeth. Materials and method Forty teeth were randomly divided into two groups, viz. groups A and B having 20 teeth each. In Group A, 3 Mix (ciprofloxacin, metronidazole, and minocycline) and in Group B, Other Mix (ciprofloxacin, ornidazole, and minocycline) mixed with propylene glycol were used. Medication cavities of Group-A and B were filled with 3 Mix and Other Mix respectively followed by restoration of teeth. Clinical evaluation was done at 3 months whereas, clinical and radiographic evaluation was done at 6 and 12 months, respectively. Results Both the groups showed 100% clinical success whereas, radiographic success rate was 81% with 3 Mix and 92% with Other Mix but, was not statistically significant. Conclusions Both the antibacterial pastes can be used effectively in non instrumental endodontic treatment of necrosed primary teeth and require more clinical trials to prove its efficacy. PMID:25737928
Thomas, Kristin; Bendtsen, Preben; Krevers, Barbro
Background Primary care is increasingly being encouraged to integrate healthy lifestyle promotion in routine care. However, implementation has been suboptimal. Coordinated care could facilitate lifestyle promotion practice but more empirical knowledge is needed about the implementation process of coordinated care initiatives. This study aimed to evaluate the implementation of a coordinated healthy lifestyle promotion initiative in a primary care setting. Methods A mixed method, convergent, parallel design was used. Three primary care centres took part in a two-year research project. Data collection methods included individual interviews, document data and questionnaires. The General Theory of Implementation was used as a framework in the analysis to integrate the data sources. Results Multi-disciplinary teams were implemented in the centres although the role of the teams as a resource for coordinated lifestyle promotion was not fully embedded at the centres. Embedding of the teams was challenged by differences among the staff, patients and team members on resources, commitment, social norms and roles. Conclusions The study highlights the importance of identifying and engaging key stakeholders early in an implementation process. The findings showed how the development phase influenced the implementation and embedding processes, which add aspects to the General Theory of Implementation. PMID:26312058
Kuebler, W M; Kuhnle, G E; Groh, J; Goetz, A E
1. Sequestration of leucocytes in the lung is the net result of leucocyte rolling and sticking in pulmonary arterioles and venules and their retention in alveolar capillaries. 2. In order to investigate whether adhesion molecules of the selectin family contribute to these phenomena the effects of fucoidin (an inhibitor of L- and P-selectin) on microhaemodynamics and leucocyte kinetic were studied in pulmonary arterioles, capillaries and venules by means of intravital fluorescence microscopy in a rabbit model. 3. Fucoidin reduced leucocyte rolling in pulmonary arterioles and venules by 75 and 83%, respectively, without affecting leucocyte sticking. In alveolar capillaries, fucoidin reduced leucocyte retention and accelerated leucocyte passage, thus reducing the alveolar transit time of leucocytes by 62%. 4. It is concluded that rolling of leucocytes in pulmonary microvessels is mediated by selectins, whereas sticking relies on selectin-independent mechanisms. 5. Leucocyte retention in alveolar capillaries is not due solely to mechanical hindrance of leucocyte passage through narrow vessel segments, as previously hypothesized, but also depends on interaction of leucocytes with the capillary endothelium. PMID:9192309
Nicholson, David P.; Wilson, Samuel T.; Doney, Scott C.; Karl, David M.
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.
Kucera, Jan P.; Prudat, Yann; Marcu, Irene C.; Azzarito, Michela; Ullrich, Nina D.
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
Zander, Nicole E; Piehler, Thuvan; Banton, Rohan; Benjamin, Richard
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. PMID:27317559
Richards, Michael R; Polsky, Daniel
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. PMID:26443665
Kitani, Hiroshi; Yoshioka, Miyako; Takenouchi, Takato; Sato, Mitsuru; Yamanaka, Noriko
We recently developed a novel procedure to obtain liver-macrophages in sufficient number and purity using a mixed primary culture of rat and bovine hepatocytes. In this study, we aim to apply this method to the neonatal swine liver. Swine parenchymal hepatocytes were isolated by a two-step collagenase perfusion method and cultured in T75 culture flasks. Similar to the rat and bovine cells, the swine hepatocytes retained an epithelial cell morphology for only a few days and progressively changed into fibroblastic cells. After 5-13 days of culture, macrophage-like cells actively proliferated on the mixed fibroblastic cell sheet. Gentle shaking of the culture flask followed by the transfer and brief incubation of the culture supernatant resulted in a quick and selective adhesion of macrophage-like cells to a plastic dish surface. After rinsing dishes with saline, the attached macrophage-like cells were collected at a yield of 10(6) cells per T75 culture flask at 2-3 day intervals for more than 3 weeks. The isolated cells displayed a typical macrophage morphology and were strongly positive for macrophage markers, such as CD172a, Iba-1 and KT022, but negative for cytokeratin, desmin and α-smooth muscle actin, indicating a highly purified macrophage population. The isolated cells exhibited phagocytosis of polystyrene microbeads and a release of inflammatory cytokines upon lipopolysaccharide stimulation. This shaking and attachment method is applicable to the swine liver and provides a sufficient number of macrophages without any need of complex laboratory equipments. PMID:24707456
Iliffe, Steve; Robinson, Louise; Bamford, Claire; Waugh, Amy; Fox, Chris; Livingston, Gill; Manthorpe, Jill; Brown, Pat; Stephens, Barbara; Brittain, Katie; Chew-Graham, Carolyn; Katona, Cornelius
Background Case management may be a feasible solution to the problem of service fragmentation for people with dementia. Aim To adapt a US model of primary care-based case management for people with dementia and test it in general practice. Design and setting Mixed-methodology case studies of case management implementation in four general practices: one rural, one inner-city, and two urban practices. Practice nurses undertook the case manager role in the rural and inner-city practices, and were allocated one session per week for case management by their practices. A seconded social worker worked full time for the two urban practices. Method Participants were community-dwelling patients with dementia who were living at home with a family carer, and who were not receiving specialist care coordination. Case study methods included analysis of case loads and interviews with patients, carers, local NHS and other stakeholders, and case managers. Measures included numbers of patients with dementia and their carers, number of unmet needs identified, and actions taken to meet needs. Case manager records were compared with findings from interviews with patients and carers, and with other stakeholders. Results The number of eligible patients was smaller than expected. No practice achieved its recruitment target. Researchers identified more unmet needs than case managers. The practice nurse case managers reported lack of time and found research documentation burdensome. Patients and carers were positive about case management as a first point of contact with the practice, as a ‘safety net’, and for creating a one-to-one therapeutic relationship. Conclusion Further investigation is required before case management for people with dementia and their carers can be implemented in primary care. PMID:25348998
Hill, R S; Kennedy, M; Mackinder, C
Human peripheral blood leucocytes (neutrophil-rich) were collected either with heparin or acid citrate dextrose, frozen with dimethyl sulphoxide at a controlled rate, stored in liquid nitrogen at--196 degrees C and reconstituted with a solution containing dextran. After reconstitution, 20.2% of cells (in absolute numbers 1 in 5 fresh cells) showed a strongly positive nitroblue tetrazolium (NBT) reaction. The quantitative NBT test confirmed the synthesis of formazan/10(6) reconstituted neutrophilsa s15% of the fresh capacity. A slow titration reconstitution method for cells did not improve the functional capacity of thawed leucocytes as judged by the NBT test. When comparing anticoagulants, heparin increased the post-reconstitution cell yields after freezing and increased the absolute number of reconstituted cells capable of developing a positive NBT reaction. PMID:643322
Rathkolb, B; Pohlenz, J F; Wohlsein, P
A modified fixative of formalin dichromate was combined with a cold embedding procedure for the preservation of bovine leucocyte surface antigens. Fourteen monoclonal antibodies recognizing seven bovine leucocyte surface antigens (BoCD1w2, BoCD4, BoCD8, BoWC1, BoWC3, BoWC4 and BoIgM) were applied as primary antisera in a sensitive avidin-biotin-peroxidase complex detection method. The staining results were compared with those obtained in cryostat and routinely formalin-fixed sections of corresponding tissue samples. Using the modified formalin dichromate fixative and the cold embedding procedure, all the leucocyte surface antigens tested were detectable immunohistologically in paraffin sections with a generally more distinct staining than in traditionally processed tissues. Morphological structures were better preserved than in cryostat sections but, to some extent, were poorer when compared with routinely formalinfixed tissues. However, this method suggests that there are only mild masking effects and provides an alternative to the use of unfixed material, particularly for morphological-immunohistochemical investigations. PMID:9248856
Traldi, Aline; Valdrighi, Heloísa Cristina; de Souza, Luciane Zanin; Vedovello, Silvia Amélia Scudeler
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
D'Agata, I D; Paradis, K; Chad, Z; Bonny, Y; Seidman, E
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
Chen, Shih-Heng; Oyarzabal, Esteban A.; Hong, Jau-Shyong
Microglia, neurons, and macroglia (astrocytes and oligodendrocytes) are the major cell types in the central nervous system. In the past decades, primary microglia-enriched cultures have been widely used to study the biological functions of microglia in vitro. In order to study the interactions between microglia and other brain cells, neuron–glia, neuron–microglia, and mixed glia cultures were developed. The aim of this chapter is to provide basic and adaptable protocols for the preparation of these microglia-containing primary cultures from rodent. Meanwhile, we also want to provide a collection of tips from our collective experiences doing primary brain cell cultures. PMID:23813383
Davies, K. A.; Toothill, V. J.; Savill, J.; Hotchin, N.; Peters, A. M.; Pearson, J. D.; Haslett, C.; Burke, M.; Law, S. K. A.; Mercer, N. F. G.; Walport, M. J.; Webster, A. D. B.
We describe a male patient with leucocyte adhesion molecule deficiency (LAD) of moderate phenotype. Although diagnosis was made only 2 years before his death, the patient survived until 19 years of age. This enabled us to perform a number of novel investigations, both in vivo and in vitro, relating to his leucocyte biology. Monocytes cultured in vitro matured into morphologically normal, phagocytically capable macrophages, which were able to recognize aged `apoptotic' neutrophils. By injection of radiolabelled autologous neutrophils we demonstrated a prolonged neutrophil half-life, but normal margination, de-margination on exercise, and splenic pooling. Neutrophile adherence in vitro to vascular endothelium was normal. Histological examination of the patient's lungs at postmortem showed intravascular aggregation of polymorphonuclear leucocytes but a paucity of cells in the interstitium and alveolar spaces. These findings indicate that the peripheral blood leucocytosis commonly observed in these patients may be due to prolonged intravascular neutrophil survival, and suggest that CD11/18 molecules have an important role in facilitating neutrophil emigration from blood vessels at sites of inflammation. ImagesFig. 1Fig. 5Fig. 7 PMID:1673876
Mudd, S; Mudd, E B
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
Andrews, Elizabeth; Pearson, David; Kelly, Charlotte; Stroud, Laura; Rivas Perez, Martin
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
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
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
Mincheff, M.S.; Meryman, H.T. )
Treatment of stimulator cells with paraformaldehyde for 60 sec or ultraviolet-B (UV-B) irradiation eliminates their ability to elicit T cell proliferation in a primary mixed leukocyte reaction. However, a T cell response equal to 20-40% of control value could be elicited by paraformaldehyde fixed or UV-B irradiated cells providing the latter are incubated at 37 degrees C for 18 hr prior to treatment. The incubation also induces a one-log increase in the density of fluorescence when the cells are stained with monoclonal antibodies against class II molecules DR and DP as well as the intercellular adhesion molecule -1. We interpret this as an increase in the membrane expression of these structures following incubation. Chloroquine and cerulenin, known to inhibit protein degradation and antigen processing and presentation do not influence the upregulation in membrane expression of these class II and adhesion molecules, but do prevent incubation from overriding the effect of paraformaldehyde treatment. Colchicine, which reduces the traffic through tubular lysosomes, also has no effect on the upregulation but enhances allopresentation. We propose that incubation of stimulator cells in the presence of chloroquine and cerulenin results in the membrane expression of class II molecules without associated peptides. The inability of stimulator cells expressing such nude MHC molecules to elicit T cell proliferation after chemical modification could be due to easier crosslinking of the allodeterminants by paraformaldehyde when the binding site is empty but could also mean that nude MHC molecules are not per se immunogenic and become so only after acquisition of a peptide. It is also possible that chloroquine, NH4Cl, and cerulenin block the expression of signals other than the class II and cell adhesion molecules that are essential for induction of T cell proliferation.
Carroll, Andrew; Mansoori, Nasim; Li, Shundai; Lei, Lei; Vernhettes, Samantha; Visser, Richard G.F.; Somerville, Chris; Gu, Ying; Trindade, Luisa M.
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. PMID:22926318
Braguglia, C M; Bagnuolo, G; Gianico, A; Mininni, G; Pastore, C; Mascolo, G
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. PMID:26520096
Sale, Joanna E. M.; Brazil, Kevin
The practice of mixed-methods research has increased considerably over the last 10 years. While these studies have been criticized for violating quantitative and qualitative paradigmatic assumptions, the methodological quality of mixed-method studies has not been addressed. The purpose of this paper is to identify criteria to critically appraise the quality of mixed-method studies in the health literature. Criteria for critically appraising quantitative and qualitative studies were generated from a review of the literature. These criteria were organized according to a cross-paradigm framework. We recommend that these criteria be applied to a sample of mixed-method studies which are judged to be exemplary. With the consultation of critical appraisal experts and experienced qualitative, quantitative, and mixed-method researchers, further efforts are required to revise and prioritize the criteria according to importance. PMID:26526412
Huf, Christina; Raggl, Andrea
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…
Negral, L; Marañón, E; Castrillón, L; Fernández-Nava, Y
Primary, secondary and mixed sludge were treated by inverted phase fermentation. This treatment results in solid-liquid separation of sludge after endogenous enzymatic hydrolysis (anaerobic conditions: 42°C, 48 hours). The soluble chemical oxygen demand (sCOD) was increased in the solid phase up to 1,800%, 21,300% and 260% in primary, secondary and mixed sludge, respectively. The corresponding increase in sCOD in the liquid phase accordingly reached values of up to 440%, 5,100% and 140%. Phase separation led to an enrichment of volatile solids in the solid phase (89-358% primary sludge, 80-102% secondary sludge and 29-133% mixed sludge). The NH4+-N values increased notably after the endogenous enzymatic hydrolysis itself. To investigate the short-term evolution following the treatment, the variation in sCOD, NH4+-N and solids was also monitored after keeping the hydrolysate at 37°C under anaerobic conditions for 24 hours. This stage showed no generalized pattern in terms of sCOD. PMID:26465310
Hannig, Madeleine; Figulla, Hans-Reiner; Sauer, Heinrich; Wartenberg, Maria
Embryonic stem (ES) cells spontaneously differentiate capillary-like structures as well as leucocytes such as monocytes/macrophages, neutrophils, natural killer (NK) cells and cytototoxic T lymphocytes. The interplay between vasculogenesis and leucocyte differentiation as well as the population of tumour tissues with ES cell-derived leucocytes and endothelial cells is, however, not sufficiently specified. In the present study, gene expression of the cell surface markers CD68 and CD14 (expressed on monocytes and macrophages), Mac-1 (CD11b) (expressed on granulocytes, monocytes and NK cells) and CD16 (expressed on neutrophils) was investigated in murine CGR8 ES cells in relation to the endothelial cell markers CD31 and vascular endothelial (VE)-cadherin. Expression of leucocyte markers increased from day 7-8 of cell culture on. Furthermore, addition of macrophage colony-stimulating factor to the cell culture medium resulted in a threefold increase in the number of CD68(+) monocytes/macrophages. Treatment of embryoid bodies with lipopolysaccharide (LPS) up-regulated CD14 thus suggesting functionality of the CD14 LPS receptor. Differentiation of vascular structures positive for CD31 and VE-cadherin preceded leucocyte differentiation by 2 days (i.e. from day 5-6 on) suggesting that vasculogenesis may be a determinant of leucocyte differentiation. Consequently the Flk-1 antagonist SU5416 which inhibits vasculogenesis of ES cells significantly blunted leucocyte differentiation. Confrontation culture of embryoid bodies with multicellular breast tumour spheroids initiated significant increase of leucocyte cell numbers and invasion of leucocytes into the tumour tissue. In summary our data demonstrate that during ES cell differentiation vasculogenesis precedes leucocyte differentiation, and point towards the direction that leucocyte cell invasion into tumour tissue may initiate the pro-inflammatory microenvironment necessary for tumour vascularization. PMID:20414974
Arndt, H; Palitzsch, K D; Anderson, D C; Rusche, J; Grisham, M B; Granger, D N
Leucocyte-endothelial cell adhesion is modulated by a variety of adhesion glycoprotein expressed on the surface of leucocytes and endothelial cells. Although in vitro studies show that these adhesion molecules mediate the decrease in leucocyte rolling velocity and the increase in leucocyte adherence and emigration associated with inflammation, there are few in vivo data to support this hypothesis. The aim of this study was to assess the role of leucocyte (CD11b/CD18) and endothelial cell (P- and E-selectin) adhesion molecules in mediating the leucocyte-endothelial cell adhesion elicited in rat mesenteric venules during a model of longlasting intestinal inflammation. Indomethacin was injected 48 and 24 hours before the experiment. The mesenteric microcirculation was observed by intravital microscopy in animals treated with monoclonal antibodies (MAb) directed against either P-selectin, E-selectin, or CD11b/CD18. Leucocyte rolling velocity, and the number of adherent and emigrated leucocytes as well as vessel diameter and erythrocyte velocity were monitored in roughly 30 micron diameter postcapillary venules. Indomethacin treatment resulted in mucosal ulceration and granulocyte infiltration, and a corresponding inflammatory response in the mesentery, which was characterised by an increase in the number of adherent (eightfold) and emigrated (sixfold) leucocytes and a reduction (80%) in leucocyte rolling velocity. The indomethacin induced leucocyte-endothelial cell adhesion in mesenteric venules was significantly reduced by treatment with MAbs against either CD11b/CD18 or E-selectin, but not by the P-selectin MAb. These results suggest that both leukocyte (CD11b/CD18) and endothelial cell (E-selectin) adhesion molecules contribute to the granulocyte accumulation in a chronic model of intestinal inflammation. PMID:7590433
Abdelhamid, Asmaa; Howe, Amanda; Stokes, Tim; Qureshi, Nadeem; Steel, Nick
Background Clinical practice guidelines are widely used in primary care, yet are not always based on applicable research. Aim To explore primary care practitioners’ views on the applicability to primary care patients of evidence underpinning National Institute for Health and Care Excellence (NICE) guideline recommendations. Design and setting Delphi survey and focus groups in primary care, England, UK. Method Delphi survey of the perceived applicability of 14 guideline recommendations rated before and after a description of their evidence base, followed by two focus groups. Results GPs significantly reduced scores for their perceived likelihood of pursuing recommendations after finding these were based on studies with low applicability to primary care, but maintained their scores for recommendations based on highly applicable research. GPs reported they were more likely to use guidelines where evidence was applicable to primary care, and less likely if the evidence base came from a secondary care population. Practitioners in the focus groups accepted that guideline developers would use the most relevant evidence available, but wanted clearer signposting of those recommendations particularly relevant for primary care patients. Their main need was for brief, clear, and accessible guidelines. Conclusion Guidelines should specify the extent to which the research evidence underpinning each recommendation is applicable to primary care. The relevance of guideline recommendations to primary care populations could be more explicitly considered at all three stages of guideline development: scoping and evidence synthesis, recommendation development, and publication. The relevant evidence base needs to be presented clearly and concisely, and in an easy to identify way. PMID:25348996
Boeni, Fabienne; Hersberger, Kurt E.; Arnet, Isabelle
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
Avila, J L; Convit, J
1. A lysosomal fraction was separated by density-gradient centrifugation from a highly purified human polymorphonuclear leucocyte suspension. 2. Some 23 different lysosomal enzymes were assayed for activity in the presence of various concentrations of glycosaminoglycans. 3. The 21 acid hydrolases assayed were strongly inhibited to different degrees by low (0-12 mmol/l) concentrations of glycosaminoglycans in a pH-dependent manner. Thus inhibitions were stronger below pH4.5, with activity returning to control values at about pH5.0. 4. On a molar basis, the inhibitory activity for the several glycosaminoglycans studied was: heparin greater than chondroitin sulphate greater than hyaluronic acid. 5. Once the glycosaminoglycan-acid hydrolase complex was formed, it was partially dissociated by slight elevations in the pH of the incubation medium, by increasing the ionic strength of the incubation medium, or by adding several cationic proteins (e.g. histone, protamine). 6. As leucocytic lysosomes contain large amounts of chondroitin sulphate, and have a strongly acid intragranular pH, we suggest that glycosaminoglycans may modify lysosomal function through the formation of complexes with lysosomal enzymes, by inhibiting the digestive activity of the acid hydrolases when the intralysosomal pH is below their pI. PMID:2162
Toes, G J; van den Dungen, J J; Haan, J; Hermens, R A; van Oeveren, W
Platelets and leucocytes are important participants in the response of the body to small diameter vascular grafts implanted into the arterial circulation. A sensitive and quick method for measuring platelet and leucocyte deposition contributes to material evaluation. With a newly developed fluorescence labeling method we examined the deposition of platelets and leucocytes onto vascular grafts in vitro. Human platelets and leucocytes were isolated and labeled with the fluorescence label Europium trichloride (EuCl3). After reconstitution of the labeled cells in plasma their functionality appeared intact and competitive with unlabeled cells. Eu-labeled platelets or leucocytes were then incubated with expanded polytetrafluoroethylene (ePTFE), Dacron and polyurethane (PU) vascular grafts in autologous plasma. Beta-thromboglobin and thromboxane release from platelets and beta-glucuronidase release from leucocytes during the incubation experiments were measured. Platelets and leucocytes deposited significantly less onto ePTFE compared to Dacron and polyurethane (P < 0.01). Our results are in accordance with results of in vivo studies using radio-active labeling to study platelet and leucocyte deposition. However, a new finding was that this reduced cell deposition may in part be due to possible toxic effects of ePTFE, shown by increased haemolysis and beta-thromboglobin release. PMID:10514073
McCollum, Rosalind; Chen, Lieping; ChenXiang, Tang; Liu, Xiaoyun; Starfield, Barbara; Jinhuan, Zheng; Tolhurst, Rachel
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. PMID:23576191
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
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. PMID:26548627
Tammen, I; Klippert, H; Kuczka, A; Treviranus, A; Pohlenz, J; Stöber, M; Simon, D; Harlizius, B
A modified DNA test, based on the polymerase chain reaction, was developed for the monogenic recessive disease bovine leucocyte adhesion deficiency (BLAD). The test was improved by the selection of new primers which facilitated the interpretation of the results. An easily scorable banding pattern makes the test useful in cattle breeding schemes and for clinical diagnosis. A total of 2381 samples was analysed over a period of three years. The carrier rate among young bulls at artificial insemination (AI) stations decreased from 11.6 per cent in 1993 to 9.9 per cent in the first five months of 1995. Continuous screening of young bulls before entering AI is still recommended unless both parents are proven to be genetically free of BLAD. The carrier rate among clinically suspect animals was not increased, and carriers are therefore not expected to be immunodeficient. Despite all efforts to eradicate the disease, calves with BLAD were still observed in 1995. PMID:8735510
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
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
King, Alissa R.
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…
Vowden, Kim James
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…
Ubha, Neerose; Cahill, Sharon
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…
Rachinsky, I; Boguslavsky, L; Goldstein, D; Golan, H; Pak, I; Katz, M; Lantsberg, S
Pelvic inflammatory disease (PID) is one of the major health problems of women of child-bearing age. Among the most serious complications of PID is the formation of a tubo-ovarian abscess (TOA). Early diagnosis of this condition may prevent serious surgical complications such as peritonitis and sepsis, which may be fatal. The purpose of this study was to investigate the efficacy of technetium-99m hexamethylpropylene amine oxime (HMPAO) leucocyte scintigraphy in the diagnosis of TOA. Twenty women with high clinical suspicion of TOA underwent 99mTc-HMPAO leucocyte scintigraphy. The labelling of leucocytes with 99mTc-HMPAO was performed according to a standard protocol. Scans were obtained at 1, 3 and 24 h following the injection of the labelled leucocytes. In eight cases the early and/or late scan was positive, in 11 cases it was negative, and in one case of ovarian cyst torsion, confirmed by laparoscopy, it showed slight uptake in the capsule of the cyst (false-positive). The sensitivity of 99mTc-HMPAO leucocyte scintigraphy was 100%, specificity 91.6%, positive predictive value 89%, negative predictive value 100% and overall accuracy 95%. It is concluded that leucocyte scintigraphy is a non-invasive, safe, physiological and accurate procedure for the diagnosis of TOA. The 24-h scan is crucial, since in some cases the abscess was not clearly visualized on the early scan. Leucocyte scintigraphy may reduce the need for CT, diagnostic laparoscopy and unnecessary invasive surgical procedures. PMID:11189939
He, Wenping; Frueh, Johannes; Wu, Zhenwei; He, Qiang
Modern drug delivery systems rely on either antibody-based single-surface recognition or on surface-hydrophobicity-based approaches. For a tumor showing various surface mutations, both approaches fail. This publication hereby presents Janus capsules based on polyelectrolyte multilayer microcapsules exhibiting human leucocyte (THP-1 cell line) cell membranes for discriminating HUVEC cells from three different cancer cell lines. Despite destroying the cellular integrity of leucocyte cells, the modified Janus capsules are able to adhere to cancer cells. Leucocyte cell-membrane-coated Janus capsules are phagocytosed with the cellular membrane part pointing to the cells. PMID:26824329
Modin, Oskar; Saheb Alam, Soroush; Persson, Frank; Wilén, Britt-Marie
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
Modin, Oskar; Saheb Alam, Soroush; Persson, Frank; Wilén, Britt-Marie
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
Vaden, Timothy D.; Song, Chen; Zaveri, Rahul A.; Imre, D.; Zelenyuk, Alla
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.
Sun, B.H.; Oh, S.; Rothe, P.H.
During an overcooling transient in a pressurized water reactor, cold water from the high pressure injection (HPI) mixes with the hot primary coolant in the cold leg. The transit time is a gauge for the assessment of the time and the velocity of the mixed flow that passes through the cold leg to the downcomer. Existing data from mixing tests at the Electric Power Research Institute (EPRI)/CREARE and EPRI/SAI facilities are analyzed. By means of models for HPI jet entrainment as well as the propagation of a gravity current, dimensionless correlations have been developed for the transit time and cold water front velocity at stagnant loop flow conditions. Based on this transit time correlation for stagnant loop flow and the limiting condition for large loop flow, a general correlation has been developed to account for the loop flow effect on transit time. These correlations unify a wide range of data obtained from five geometrically different test sections with two fluids (pure water and saline solution). In addition to the geometric factors, the governing dimensionless parameters for the transit time are the HPI jet Froude number, the Froude number for the cold-leg channel, and the ratio of loop flow to HPI flow.
Graham, Tanya; Rose, Diana; Murray, Joanna; Ashworth, Mark; Tylee, André
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
Dong, Su; Song, Xue-Song; Chen, Guang; Liu, Jia
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. PMID:26589365
Meysenburg, Rebecca; Albrecht, Julie A; Litchfield, Ruth; Ritter-Gooder, Paula K
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. PMID:24211815
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
Çakir, Tansel; Ilhan, Tolgay Tuyan; Karabagli, Pinar; Çelik, Çetin
Malignant Mixed Mullerian Tumour of the Ovary (OMMMT), also referred to as carcinosarcoma is a very rare tumour accounting for less than 1% of all ovarian cancers. Due to the rarity of OMMMT, little is known about the disease course and outcome of women with these tumours. It is important to evaluate because of its aggressive behaviour with extremely unfavourable prognosis. These tumours are composed of both malignant epithelial and mesenchymal elements. Current data in the literature is still limited to small case series and case reports, therefore, its optimal treatment is somewhat controversial. In the current report, we introduce a case of OMMMT which was successfully treated with Platinum-based combination chemotherapy after optimal cytoreductive surgery. The clinical manifestations, pathologic characteristics, diagnosis and management of these tumours are reviewed here. Although the most effective treatment is currently unknown, optimal cytoreductive surgery and platinum-based chemotherapy appears to improve the outcomes. Despite the aggressive nature of this tumour and its poor response to the treatment, management works best when cancer is found early. The stage of the disease is the most important prognostic factor. Therefore, the crucial question is how to diagnose the cancer at earlier stages rather than seeking the optimal treatment. PMID:27134951
Boros, András Mihály; Széplaki, Gábor; Perge, Péter; Jenei, Zsigmond; Bagyura, Zsolt; Zima, Endre; Molnár, Levente; Apor, Astrid; Becker, Dávid; Gellér, László; Prohászka, Zoltán; Merkely, Béla
Aims The low lymphocyte counts and high neutrophil leucocyte fractions have been associated with poor prognosis in chronic heart failure. We hypothesized that the baseline ratio of the neutrophil leucocytes to the lymphocytes (NL ratio) would predict the outcome of chronic heart failure patients undergoing cardiac resynchronization therapy (CRT). Methods and results The qualitative blood counts and the serum levels of N-terminal of the prohormone brain natriuretic peptide (NT-proBNP) of 122 chronic heart failure patients and 122 healthy controls were analysed prospectively in this observational study. The 2-year mortality was considered as primary endpoint and the 6-month reverse remodelling (≥15% decrease in the end-systolic volume) as secondary endpoint. Multivariable regression analyses were applied and net reclassification improvement (NRI) and integrated discrimination improvement (IDI) were calculated. The NL ratio was elevated in chronic heart failure patients when compared with the healthy controls [2.93 (2.12–4.05) vs. 2.21 (1.64–2.81), P < 0.0001]. The baseline NL ratio exceeding 2.95 predicted the lack of the 6-month reverse remodelling [n = 63, odds ratio = 0.38 (0.17–0.85), P = 0.01; NRI = 0.49 (0.14–0.83), P = 0.005; IDI = 0.04 (0.00–0.07), P = 0.02] and the 2-year mortality [n = 29, hazard ratio = 2.44 (1.04–5.71), P = 0.03; NRI = 0.63 (0.24–1.01), P = 0.001; IDI = 0.04 (0.00–0.08), P = 0.02] independently of the NT-proBNP levels or other factors. Conclusion The NL ratio is elevated in chronic heart failure and predicts outcome after CRT. According to the reclassification analysis, 4% of the patients would have been better categorized in the prediction models by combining the NT-proBNP with the NL ratio. Thus, a single blood count measurement could facilitate the optimal patient selection for the CRT. PMID:25972301
Evans, Andrew M; Wood, Fiona C; Carter, Ben
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
Datta, Shivani; Horwood, Jeremy; Hickman, Matthew; Sharp, Debbie
Background Hepatitis C is often asymptomatic, presenting with liver failure and cancer decades after infection. People who inject drugs (PWID) and immigrant populations from countries with a moderate-to-high prevalence of hepatitis C virus (HCV) are the main risk groups. Deaths and hospital admissions due to HCV cirrhosis tripled between 1998 and 2010, but the majority of people with chronic HCV are unaware of it. Aim To identify patients at risk of developing hepatitis C using routine GP data, to determine the proportion not tested, and to explore GPs’ views regarding testing. Design and setting Mixed-methods service evaluation (density-based selection of PWID) in six NHS practices in Bristol. Method Patients at risk of HCV were identified. The Health Protection Agency laboratory (now part of Public Health England) provided test results. Semi-structured interviews with 17 GPs were audiorecorded and thematic analyses conducted on anonymised transcripts. Results Of 3765 patients identified as being at risk of developing hepatitis C, 3051 (81%) had no test result, including 53% of PWID and 93% of the ‘ethnicity’ group. All GPs said they usually test PWID. Most GPs test for HIV and hepatitis B in immigrants more often than they test for HCV. Barriers to testing included not questioning patients about risk factors, competing priorities, the chaotic lifestyle of PWID, difficulty extracting information from computerised records, and forgetting to address HCV. Conclusion Computer prompts and GP education on whom to test are warranted. Ensuring that country of origin and drug use is included on the new-patient questionnaire might also aid case-finding for HCV. PMID:24567619
Azize, Pary M; Endacott, Ruth; Cattani, Allegra; Humphreys, Ann
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. PMID:23991687
Zhou, Chun-Hua; Hao, Zhan-Qing; He, Hong-Shi; Zhou, Dan-Hui
PnET-II (photosynthesis and evapotranspiration) model is an ecosystem process model, which requires extensive input parameters, including vegetation parameters, soil parameters and climate parameters, to simulate net primary productivity (NPP). This study estimated the total and wood (stem and branch) NPPs of Korean pine and broadleaf species in Fenglin Natural Reserve, and examined the responses of the NPPs to the variations of the input parameters in PnET-II model. The simulation results indicated that among the vegetation parameters in PnET-II model, the variation of canopy parameters, had greater effects on the simulated NPPs of Korean pine and broadleaf species, and the response of Korean pine's total NPP to vegetation parameters was larger than that of broadleaf species'. The variation of soil water holding capacity (WHC) had smaller effects on the NPPs of Korean pine and broadleaf species, and the response of Korean pine's NPP to the WHC was somewhat smaller than that of broadleaf trees'. In climate scenarios, the variation of air temperature had the greatest effects on the simulated NPP of Korean pine and broadleaf trees, followed by precipitation and radiation. Different climate scenarios had different effects on the predicted results. The total and wood NPPs of Korean pine and broadleaf trees had different responses to the input parameters. PMID:18655573
Arinola, O G; Salimonu, L S
Schistosome antigens (soluble egg antigen and adult worm antigen) and non-schistosome antigens (Bacille Calmette Guerin vaccine antigen and measles virus vaccine antigen) were used to assay for leucocyte migration inhibitory factors (LMIF) in Nigerian children with and without Schistosoma haematobium infection. The severity of S. haematobium infection was graded into light infection (1-49 eggs/ 10 ml urine) or heavy infection (more than or equal to 50 eggs/10 ml urine). The mean percentage migration indices were significantly reduced in heavily infected urinary schistosomiasis (USS) subjects compared with the controls or lightly infected subjects when non-antigens were used to stimulate LMIF production. When Schistosome antigens were used to stimulate LMIF production, there were no significant decreases in the % migration indices in heavily infected USS subjects compared with the light USS subjects. The mean percentage migration index was significantly increased when schistosome antigen was used to stimulate LMIF production in treated USS subjects compared with untreated USS subjects but the increase was not significant when non-schistosome antigen was used. The conclusion that could be drawn from this study is that LMIF assay using schistosome antigen(s) has epidemiological value in schistosomiasis. PMID:10504865
Wakeyama, H; Takeshige, K; Takayanagi, R; Minakami, S
A phagocytic vesicle fraction with high NADPH-dependent superoxide-forming activity was obtained in large quantity from pig blood polymorphonuclear leucocytes, phagocytosing oil droplets in the presence of cyanide. The activity of the homogenate of the phagocytosing cells was 40 times that of the resting cells, and 70% of the activity in the homogenate was recovered in the phagocytic vesicle fraction. Essentially all of the superoxide-forming activity was extracted by repeated extraction with a mixture containing deoxycholate and Tween 20. The extract had a superoxide-forming activity of 1 mumol/min per mg of protein with NADPH, and one-fifth of this with NADH, Km values being similar to those of the vesicle fraction (40 microM for NADPH and 400 microM for NADH). A stoichiometric relationship of 1:2 for NADPH oxidation and superoxide formation was obtained, in agreement with the reaction NADPH +2O2 leads to NADP+ + 2O2 -. + H+. The activity of the extract was enhanced 2-fold by the addition of FAD, suggesting that the flavin is a component of the enzyme system. The Km value for FAD was 0.077 microM. The activities in both vesicle fraction and extract were labile even on refrigeration, but could be kept for several months at -70 degrees C. PMID:6293459
Henricks, P A; van der Tol, M E; Verhoef, J
The process of aggregation of human polymorphonuclear leucocytes (PMN) during the uptake of bacteria was studied. Radiolabelled S. aureus were opsonized in different sera, washed, resuspended in buffer and added to the PMN. Uptake of the bacteria and aggregation of the PMN were measured simultaneously. Maximal aggregation occurred within 6 min, when 5 X 10(6) PMN had phagocytosed 2.5 X 10(8) S. aureus. Also the effects of serum concentrations and different sera for opsonization of the bacteria on PMN aggregation were studied. Despite normal uptake, aggregation of PMN was low when bacteria were opsonized in complement-deficient sera. Furthermore when PMN were treated with pronase to inactivate complement receptors on the cell surface of the PMN, and bacteria preopsonized in immune serum were added, no change in uptake occurred, although the degree of aggregation halved compared to control PMN. So, interaction between the bacteria and the complement receptor of the PMN cell membrane is needed for triggering the process of aggregation. By using dansylcadaverin and diphenylamine to modulate lysosomal enzyme release, azide or PMN from a chronic granulomatous disease patient to study the effect of the formation of oxygen species, and theophylline, DB-cAMP or 8 Br-cAMP to increase cAMP levels, it was concluded that aggregation of PMN during phagocytosis was not dependent on oxygen metabolism, degranulation or cAMP levels of PMN. PMID:6086503
Haase, Andrea; Rott, Stephanie; Mantion, Alexandre; Graf, Philipp; Plendl, Johanna; Thünemann, Andreas F.; Meier, Wolfgang P.; Taubert, Andreas; Luch, Andreas; Reiser, Georg
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
Khanassov, Vladimir; Vedel, Isabelle; Pluye, Pierre
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
Leyenaar, JoAnna K.; Bergert, Lora; Mallory, Leah A.; Engel, Richard; Rassbach, Caroline; Shen, Mark; Woehrlen, Tess; Cooperberg, David; Coghlin, Daniel
Objective 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. Methods We administered a questionnaire to PCPs referring to sixteen 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. Results Responses were received from 201 PCPs, representing a response rate of 63%. While there were no differences between referral hospital type and PCP-reported receipt of discharge communication (RR 1.61,95%CI 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 (RR 1.78,95%CI 1.26–2.51). Analysis of free text responses yielded four major themes: (i) structured discharge communication; (ii) direct personal communication; (iii) reliability and timeliness of communication; and (iv) communication for effective post-discharge care. Conclusions 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
McIlfatrick, S; Keeney, S; McKenna, H; McCarley, N; McIlwee, G
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. PMID:24004198
Chignalia, Andreia Z; Oliveira, Maria Aparecida; Debbas, Victor; Dull, Randal O; Laurindo, Francisco R M; Touyz, Rhian M; Carvalho, Maria Helena C; Fortes, Zuleica B; Tostes, Rita C
The mechanisms whereby testosterone increases cardiovascular risk are not clarified. However, oxidative stress and inflammation seem to be determinants. Herein, we sought to determine whether exogenous testosterone, at physiological levels, induces leucocyte migration, a central feature in immune and inflammatory responses and the mediating mechanisms. We hypothesized that testosterone induces leucocyte migration via NADPH oxidase (NADPHox)-driven reactive oxygen species (ROS) and cyclooxygenase (COX)-dependent mechanisms. Sixteen-week-old Wistar rats received an intraperitoneal injection (5 ml) of either testosterone (10(-7) mol/l) or saline. Rats were pre-treated with 5 ml of sodium salicylate (SS, non-selective COX inhibitor, 1.25 × 10(-3) mol/l, 1 h prior to testosterone or saline), flutamide (androgen receptor antagonist, 10(-5) mol/l), apocynin (NADPHox inhibitor, 3 × 10(-4) mol/l), N-[2-Cyclohexyloxy-4-nitrophenyl]methanesulfonamide (NS398, COX2 inhibitor, 10(-4) mol/l) or saline, 4 h before testosterone or saline administration. Leucocyte migration was assessed 24 h after testosterone administration by intravital microscopy of the mesenteric bed. Serum levels of testosterone were measured by radioimmunoassay. NADPHox activity was assessed in membrane fractions of the mesenteric bed by dihydroethidium (DHE) fluorescence and in isolated vascular smooth muscle cells (VSMC) by HPLC. NADPHox subunits and VCAM (vascular cell adhesion molecule) expression were determined by immunoblotting. Testosterone administration did not change serum levels of endogenous testosterone, but increased venular leucocyte migration to the adventia, NADPHox activity and expression (P < 0.05). These effects were blocked by flutamide. SS inhibited testosterone-induced leucocyte migration (P<0.05). Apocynin and NS398 abolished testosterone-induced leucocyte migration and NADPHox activity (P<0.05). Testosterone induces leucocyte migration via NADPHox- and COX2-dependent mechanisms and
Mangino, Massimo; Christiansen, Lene; Stone, Rivka; Hunt, Steven C; Horvath, Kent; Eisenberg, Dan T A; Kimura, Masayuki; Petersen, Inge; Kark, Jeremy D; Herbig, Utz; Reiner, Alex P; Benetos, Athanase; Codd, Veryan; Nyholt, Dale R; Sinnreich, Ronit; Christensen, Kaare; Nassar, Hisham; Hwang, Shih-Jen; Levy, Daniel; Bataille, Veronique; Fitzpatrick, Annette L; Chen, Wei; Berenson, Gerald S; Samani, Nilesh J; Martin, Nicholas G; Tishkoff, Sarah; Schork, Nicholas J; Kyvik, Kirsten Ohm; Dalgård, Christine; Spector, Timothy D; Aviv, Abraham
Background Leucocyte telomere length (LTL), which is fashioned by multiple genes, has been linked to a host of human diseases, including sporadic melanoma. A number of genes associated with LTL have already been identified through genome-wide association studies. The main aim of this study was to establish whether DCAF4 (DDB1 and CUL4-associated factor 4) is associated with LTL. In addition, using ingenuity pathway analysis (IPA), we examined whether LTL-associated genes in the general population might partially explain the inherently longer LTL in patients with sporadic melanoma, the risk for which is increased with ultraviolet radiation (UVR). Results Genome-wide association (GWA) meta-analysis and de novo genotyping of 20 022 individuals revealed a novel association (p=6.4×10−10) between LTL and rs2535913, which lies within DCAF4. Notably, eQTL analysis showed that rs2535913 is associated with decline in DCAF4 expressions in both lymphoblastoid cells and sun-exposed skin (p=4.1×10−3 and 2×10−3, respectively). Moreover, IPA revealed that LTL-associated genes, derived from GWA meta-analysis (N=9190), are over-represented among genes engaged in melanoma pathways. Meeting increasingly stringent p value thresholds (p<0.05, <0.01, <0.005, <0.001) in the LTL-GWA meta-analysis, these genes were jointly over-represented for melanoma at p values ranging from 1.97×10−169 to 3.42×10−24. Conclusions We uncovered a new locus associated with LTL in the general population. We also provided preliminary findings that suggest a link of LTL through genetic mechanisms with UVR and melanoma in the general population. PMID:25624462
García Gil, Merche; Alonso, Fernando; Alvarez Chiva, Vicente; Sánchez Crespo, Mariano; Mato, José M.
We have previously observed that the phagocytosis of zymosan particles coated with complement by human polymorphonuclear leucocytes is accompanied by a time- and dose-dependent inhibition of phosphatidylcholine synthesis by transmethylation [García Gil, Alonso, Sánchez Crespo & Mato (1981) Biochem. Biophys. Res. Commun. 101, 740–748]. The present studies show that phosphatidylcholine synthesis by a cholinephosphotransferase reaction is enhanced, up to 3-fold, during phagocytosis by polymorphonuclear cells. This effect was tested by both measuring the incorporation of radioactivity into phosphatidylcholine in cells labelled with [Me-14C]choline, and by assaying the activity of CDP-choline:diacylglycerol cholinephosphotransferase. The time course of CDP-choline:diacylglycerol cholinephosphotransferase activation by zymosan mirrors the inhibition of phospholipid methyltransferase activity previously reported. The extent of incorporation of radioactivity into phosphatidylcholine induced by various doses of zymosan correlates with the physiological response of the cells to this stimulus. This effect was specific for phosphatidylcholine, and phosphatidyl-ethanolamine turnover was not affected by zymosan. The purpose of this enhanced phosphatidylcholine synthesis is not to provide phospholipid molecules rich in arachidonic acid. The present studies show that about 80% of the arachidonic acid generated in response to zymosan derives from phosphatidylinositol. A transient accumulation of arachidonoyldiacylglycerol has also been observed, which indicates that a phospholipase C is responsible, at least in part, for the generation of arachidonic acid. Finally, isobutylmethylxanthine and quinacrine, inhibitors of phosphatidylinositol turnover, inhibit both arachidonic acid generation and phagocytosis, indicating a function for this pathway during this process. PMID:6181780
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
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
Vaden, Timothy D; Song, Chen; Zaveri, Rahul A; Imre, Dan; Zelenyuk, Alla
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 alpha-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 alpha-pinene products on size-selected DOP particles and by condensation of DOP on size-selected alpha-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 approximately 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
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
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
Kitani, Hiroshi; Takenouchi, Takato; Sato, Mitsuru; Yoshioka, Miyako; Yamanaka, Noriko
We report a simple and efficient method to obtain macrophage-like cells from the mixed primary cultures of adult rat liver cells. A parenchymal hepatocyte enriched fraction was prepared from adult rat livers and seeded into culture flasks. After 7 to 10 days of culture, when most hepatocytes were degenerated or transformed into fibroblastic cells, macrophage-like cells vigorously proliferated on the cell sheet. By shaking the flasks, macrophage-like cells were readily detached. Subsequent transfer and incubation in plastic dishes resulted in quick and selective adhesion of macrophage-like cells, while other contaminating cells remained suspended in the medium. After rinsing with saline, attached macrophage-like cells were harvested with 95 to 99% purity, as evaluated by flow cytometry or immunocytochemistry. These cells showed typical macrophage morphology and were strongly positive for markers of rat macrophages, such as ED-1, ED-3, and OX-41, but negative for cytokeratins and alpha-smooth muscle actin. They possessed functional properties of typical macrophages, including active phagocytosis of latex beads, proliferative response to recombinant GM-CSF, secretion of inflammatory and anti-inflammatory cytokines upon stimulation with LPS, and formation of multinucleated giant cells. As more than 10(6) cells can be recovered repeatedly from a T75 culture flask at two to three day intervals for more than two weeks, our procedure might implicate a novel alternative to obtain Kupffer cells in sufficient number and purity without complex equipment and skills. PMID:20600081
Li, Haipeng; Liu, Jia; Yao, Jianhua; Zhong, Jianfeng; Guo, Lei; Sun, Tiansheng
Fracture, a common type injury in trauma patients, often results in the development of the systemic inflammatory response syndrome (SIRS). Though the mechanism of the fracture-initiated SIRS still remains not well characterized, it is well documented that the polymorphonuclear leucocytes (PMN) play an important role in the inflammatory process. We hypothesize that fractures recruit PMN to the local tissue, which is followed by an increase in the number of peripheral PMN and initiation of SIRS. In the current study, we established a closed femoral fracture rat model. We evaluated the levels of MPO, IL-1β and CINC-1 in fractured tissue homogenate, and we measured the levels of IL-6 and IL-10, the biomarkers for systemic inflammatory response, in the rat sera. In clinical part of the study, we collected blood from patients with isolated closed femoral fractures and evaluated PMN-related chemoattractants (IL-8, IL-1β and G-CSF) and the number of peripheral PMN. We further evaluated the level of mitochondrial DNA in the local haematoma of fracture and the circulating plasma of the patients with fracture. In the animal model of closed femoral fracture, we found a significant recruitment of PMN to the local tissue after fracture, which correlates with the elevated MPO level. We also showed that the concentration of IL-1β and CINC-1 in local tissue is significantly increased and might be responsible for the PMN recruitment. Recruitment of PMN to the local tissue was accompanied with a significant increase in the systemic levels of IL-6 and IL-10 in serum. In the patients with closed femoral fracture, we observed an increase in the number of peripheral PMN and PMN-related chemoattractants, including IL-8, IL-1β and G-CSF. The level of mitochondrial DNA in the local haematoma of fracture and the circulating plasma of patients were significantly higher compared to the healthy volunteers. Our data suggest that fracture released mitochondrial DNA into the local haematoma of
Courtier, Jesse; Robbins, Elizabeth; Soares, Bruno; Horvai, Andrew; Mackenzie, John D.
We report a case of primary malignant mixed tumor (MMT) of bone in an 18-year-old boy with X-ray, CT, MR, scintigraphic, FDG PET, and pathologic correlation. Primary MMT of bone is a highly aggressive tumor and presents both a diagnostic and clinical treatment challenge. This tumor is extremely rare and to the best of our knowledge, this is the first report of the diagnostic imaging findings for primary MMT arising from bone in a patient of this age group. PMID:26909264
Honess, D. J.; Kitamoto, Y.; Rampling, M. R.; Bleehen, N. M.
Transient plugging of microcapillaries by leucocytes is a possible reason for the occurrence of acute hypoxia in tumours. We compared the abilities of nicotinamide at 1000 micrograms ml-1 and 150 micrograms ml-1 and pentoxifylline at 300 micrograms ml-1 to increase the filterability of normal and artificially activated human leucocytes through 8 microns pores, as a model for the capillary bed. Using a St George's filtrometer, filterability of treated leucocyte suspensions was compared with control for three to six sequential 60 microliters samples, normalising control values to unity. Pentoxifylline at 300 micrograms ml-1 halved the ratio of treated to control value to 0.47 +/- 0.13 (2 s.e.), P = 0.001 (i.e. an increase in filterability), and nicotinamide at 1000 micrograms ml-1 reduced it to 0.69 +/- 0.22, P = 0.04, but the clinically achievable 150 micrograms ml-1 was ineffective (0.82 +/- 0.25, P = 0.24). Filterability of artificially activated leucocytes was reduced (3.9 +/- 1.20) but was restored to control values of unity by 1000 micrograms ml-1 nicotinamide and 300 micrograms ml-1 pentoxifylline and partially restored by 150 micrograms ml-1 nicotinamide (1.2 mM), which was isoeffective with 100 micrograms ml-1 pentoxifylline (0.37 mM). Pentoxifylline is therefore more effective on a molar basis and was shown to affect both polymorphonuclear leucocytes and lymphocytes, while nicotinamide only affects lymphocytes. The data are consistent with the hypothesis that both agents modify acute hypoxia by increasing leucocyte filterability. PMID:8763888
Numata, T.; Yamamoto, S.; Yamura, T.
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.
Hu, Jia; Mansur, Rodrigo; McIntyre, Roger S.
Bipolar disorder, while commonly encountered in the primary care setting, is often misdiagnosed or undiagnosed. In the DSM-IV-TR, patients could be diagnosed as being in a mixed state only if they had concurrent manic and depressive symptoms; while this occurs in some patients, many more experience subsyndromal mixed symptoms that would disqualify a “mixed state” diagnosis. The recently released DSM-5 attempts to capture this large proportion of patients with subsyndromal mixed symptoms with the inclusion of the “mixed specifier.” The presence of such subsyndromal mixed symptoms has significant implications for both diagnosis and treatment. For those presenting with major depressive disorder with subsyndromal manic symptoms, clinicians must be vigilant for the development of full-blown bipolar disease. In treating this group, selective serotonin reuptake inhibitors and serotonin-norepinephrine reuptake inhibitors remain first-line therapy, but augmentation with other therapies is often required. If a diagnosis of bipolar disorder is confirmed and the patient is experiencing a depressive phase, traditional antidepressants should be avoided. For those presenting with mania and mixed depressive symptoms, treatment with a combination of atypical antipsychotics and mood stabilizers is best. PMID:25133063
Ford, H. C.; Carter, J. M.
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
Rees, Philippa; Edwards, Adrian; Powell, Colin; Evans, Huw Prosser; Carter, Ben; Hibbert, Peter; Makeham, Meredith; Sheikh, Aziz; Donaldson, Liam; Carson-Stevens, Andrew
Background Children are scheduled to receive 18–20 immunizations before their 18th birthday in England and Wales; this approximates to 13 million vaccines administered per annum. Each immunization represents a potential opportunity for immunization-related error and effective immunization is imperative to maintain the public health benefit from immunization. Using data from a national reporting system, this study aimed to characterize pediatric immunization-related safety incident reports from primary care in England and Wales between 2002 and 2013. Methods A cross-sectional mixed methods study was undertaken. This comprised reading the free-text of incident reports and applying codes to describe incident type, potential contributory factors, harm severity, and incident outcomes. A subsequent thematic analysis was undertaken to interpret the most commonly occurring codes, such as those describing the incident, events leading up to it and reported contributory factors, within the contexts they were described. Results We identified 1745 reports and most (n = 1077, 61.7%) described harm outcomes including three deaths, 67 reports of moderate harm and 1007 reports of low harm. Failure of timely vaccination was the potential cause of three child deaths from meningitis and pneumonia, and described in a further 113 reports. Vaccine administration incidents included the wrong number of doses (n = 476, 27.3%), wrong timing (n = 294, 16.8%), and wrong vaccine (n = 249, 14.3%). Documentation failures were frequently implicated. Socially and medically vulnerable children were commonly described. Conclusion This is the largest examination of reported contributory factors for immunization-related patient safety incidents in children. Our findings suggest investments in IT infrastructure to support data linkage and identification of risk predictors, development of consultation models that promote the role of parents in mitigating safety incidents, and improvement
Avila, J L; Convit, J
1. The activities of 30 different lysosomal enzymes were determined in vitro in the presence of the sulphated glycosaminoglycans, heparin and chondroitin sulphate, all the enzymes being measured on a density-gradient-purified lysosomal fraction. 2. Each enzyme was studied as a function of the pH of the incubation medium. In general the presence of sulphated glycosaminoglycans induced a strong pH-dependent inhibition of lysosomal enzymes at pH values lower than 5.0, with full activity at higher pH values. However, in the particular case of lysozyme and phospholipase A2 the heparin-induced inhibition was maintained in the pH range 4.0-7.0. 3. For certain enzymes, such as acid beta-glycerophosphatase, alpha-galactosidase, acid lipase, lysozyme and phospholipase A2, the pH-dependent behaviour obtained in the presence of heparin was quite different to that obtained with chondroitin sulphate, suggesting the existence of physicochemical characteristic factors playing a role in the intermolecular interaction for each of the sulphated glycosaminoglycans studied. 4. Except in the particular case of peroxidase activity, in all other lysosomal enzymes measured the glycosaminoglycan-enzyme complex formation was a temperature-and time-independent phenomenon. 5. The effects of the ionic strength and pH on this intermolecular interaction reinforce the concept of an electrostatic reversible interaction between anionic groups of the glycosaminoglycans and cationic groups on the enzyme molecule. 6. As leucocytic primary lysosomes have a very acid intragranular pH and large amounts of chondroitin sulphate, we propose that this glycosaminoglycan might act as molecular regulator of leucocytic activity, by inhibiting lysosomal enzymes when the intragranular pH is below the pI of lysosomal enzymes. This fact, plus the intravacuolar pH changes described during the phagocytic process, might explain the unresponsiveness of lysosomal enzymes against each other existing in primary lysosomes as
Kemoli, Arthur M
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
Kremsky, Isaac; Morgan, Todd E.; Hou, Xiaogang; Li, Lei; Finch, Caleb E.
Astrocytic GFAP expression increases during normal aging in many brain regions and in primary astrocyte cultures derived from aging rodent brains. As shown below, we unexpectedly found that the age-related increase of GFAP expression was suppressed in mixed glia (astrocytes + microglia). However, the age-related increase of GFAP was observed when E18 neurons were co-cultured with mixed glia. Thus, the presence of microglia can suppress the age-related increase of GFAP, in primary cultures of astrocytes. To more broadly characterize how aging and co-culture with neurons alters glial gene expression, we profiled gene expression in mixed glia from young (3 mo) and old (24 mo) male rat cerebral cortex by Affymetrix microarray (Rat230 2.0). The majority of age changes were independent of the presence of neurons. Overall, the expression of 2-fold more genes increased with age than decreased with age. The minority of age changes that were either suppressed or revealed by the presence of neurons may be useful to analyze glial-neuron interaction during aging. Some in vitro changes are shared with those of aging rat hippocampus in studies from the Landfield group (Rowe et al., 2007; Kadish et al., 2009). PMID:22226781
Ling, E A
Intravenous injections of colloidal carbon were used to label circulating mononuclear leucocytes. In the neonatal rats (3-5 days old), either 24 or 48 hours later, carbon-labelled macrophages were seen in the brain tissue. In the areas examined, notably the corpus callosum and the cerebral cortex, labelled macrophages were distributed randomly. They were either perivascular, perineuronal or lay between the nerve fibres. The labelled cells were mostly spindle-shaped with an eccentric nucleus and the cytoplasm at one pole of the cell was engorged with dark carbon particles. Abundant labelled cells were also seen over the brain surface in the layers of meninges. There was no evidence of leucocytic infiltration into the brain tissue of mature animals. It is concluded from the present work that a proportion (but not all) of the macrophages in the neonatal rat brain are derived from the blood stream. Images Figs. 1-8 PMID:632205
Keshavarzian, A.; Saverymuttu, S.H.; Chadwick, V.S.
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.
Rodríguez, A; Esteban, M A; Meseguer, J
In this paper the possible involvement of the mannose-receptor on the non-specific recognition and phagocytosis of heat killed yeast cells (Saccharomyces cerevisiae) by gilthead seabream (Sparus aurata L.) head-kidney leucocytes was established by studying the ability of different sugars to inhibit the uptake of the yeast cells by leucocytes. Leucocytes were preincubated for 30min with different concentrations of sugar (alpha-mannan, d-mannose, d-fucose, l-fucose, d-glucose, d-glucosamine and n-acetyl-glucosamine, all of them described as specific ligands of the vertebrate mannose-receptor) and afterwards incubated with FITC-labelled yeast cells for phagocytosis assays. The phagocytic ability (percentage of cells with one or more ingested yeast cells within the total cell population) and capacity (number of ingested yeast cells per cell) of leucocytes was analysed by flow cytometry. The results demonstrate the potential existence of a specific receptor-sugar or receptor-yeast cell binding process, which was saturable, specific and dose-dependent. More specifically, when leucocytes were preincubated with appropriate doses of d-mannose, d- or l-fucose, d-glucose or n-acetyl-glucosamine the phagocytosis of yeast cells by head-kidney leucocytes was partially blocked. Seabream leucocytes were also preincubated with chloroquine, a lysosomotropic drug which downregulates (in a nonspecific manner) the expression of mannose-receptors in mammals, before phagocytosis assays were performed. The results demonstrated that the phagocytosis of yeast was completely blocked by this substance. The overall results seem to corroborate the presence of the mannose-receptor in seabream phagocytes, which is involved in the non-specific binding and phagocytosis of yeast cells by head-kidney leucocytes. PMID:12711272
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…
Feinstein, G; Janoff, A
Granules of human peripheral blood leucocytes contain four well-characterized elastase isozymes and one or two slow-moving elastaselike esterases (SE) which have not been as well characterized. SE are capable of hydrolyzing typical elastase synthetic sybstrates such as N-acetyl-dl-alanine-alpha-naphthyl ester (Ac-DL-Ala-1-ONap) and N-t-butyloxycarbonyl-L-alanine-p-nitrophenyl ester (Boc-Ala-ONp), but unlike the highly basic elastase isozymes, SE barely migrate into 13% acrylamide gels during cationic electrophoresis at pH 4.3. Hydrolysis of Ac-DL-Ala-1-ONap by SE requires the presence of Triton in the gel, and hydrolysis of Boc-Ala-ONp by the same enzyme(s) is also enhanced in the presence of the detergent. Triton is not required for these activities, in the case of the elastase isozymes. Diisopropylfluorophosphate (Dip-F) inactivates both SE and the elastase isozymes, whereas Ac-(Ala)2-Pro-AlaCH2Cl (a powerful inactivator of the leucocyte elastase isozymes at 10-4 M concentration) does not inactivate SE at the same concentration. Immunochemical studies revealed antigenic cross-reaction between the rapidly migrating leucocyte elastase isozymes and SE. Two preparations of leucocyte granules from nonleukemic bone marrow cells showed no activity of the rapidly migrating elastase isozymes, but did contain SE activity. SE may be a precursor or zymogen form of the elastase isozymes, present in immature cells and partly retained through later stages of development. PMID:1265076
Shetty, Shishir; Weston, Christopher J; Adams, David H; Lalor, Patricia F
Leucocyte infiltration into human liver tissue is a common process in all adult inflammatory liver diseases. Chronic infiltration can drive the development of fibrosis and progression to cirrhosis. Understanding the molecular mechanisms that mediate leucocyte recruitment to the liver could identify important therapeutic targets for liver disease. The key interaction during leucocyte recruitment is that of inflammatory cells with endothelium under conditions of shear stress. Recruitment to the liver occurs within the low shear channels of the hepatic sinusoids which are lined by hepatic sinusoidal endothelial cells (HSEC). The conditions within the hepatic sinusoids can be recapitulated by perfusing leucocytes through channels lined by human HSEC monolayers at specific flow rates. In these conditions leucocytes undergo a brief tethering step followed by activation and firm adhesion, followed by a crawling step and subsequent transmigration across the endothelial layer. Using phase contrast microscopy, each step of this 'adhesion cascade' can be visualized and recorded followed by offline analysis. Endothelial cells or leucocytes can be pretreated with inhibitors to determine the role of specific molecules during this process. PMID:24686418
Shetty, Shishir; Weston, Christopher J.; Adams, David H.; Lalor, Patricia F.
Leucocyte infiltration into human liver tissue is a common process in all adult inflammatory liver diseases. Chronic infiltration can drive the development of fibrosis and progression to cirrhosis. Understanding the molecular mechanisms that mediate leucocyte recruitment to the liver could identify important therapeutic targets for liver disease. The key interaction during leucocyte recruitment is that of inflammatory cells with endothelium under conditions of shear stress. Recruitment to the liver occurs within the low shear channels of the hepatic sinusoids which are lined by hepatic sinusoidal endothelial cells (HSEC). The conditions within the hepatic sinusoids can be recapitulated by perfusing leucocytes through channels lined by human HSEC monolayers at specific flow rates. In these conditions leucocytes undergo a brief tethering step followed by activation and firm adhesion, followed by a crawling step and subsequent transmigration across the endothelial layer. Using phase contrast microscopy, each step of this 'adhesion cascade' can be visualized and recorded followed by offline analysis. Endothelial cells or leucocytes can be pretreated with inhibitors to determine the role of specific molecules during this process. PMID:24686418
Peter, F W; Li-Peuser, H; Vogt, P M; Muehlberger, T; Homann, H H; Steinau, H U
An intact microcirculation is essential for normal healing to occur. Wound repair may be impaired by various endogenous and exogenous factors, such as reduced microvascular perfusion, infection and debris. In the nonhealing wound, radical surgical debridement is critical. To supplement healing, various ointments are used in clinical practice. Little is known about their effects on tissue perfusion. We have therefore selected two substances widely used, the antiseptic Betadine and the enzyme combination Elase and investigated their impact on the microcirculation and on leucocyte activity, using the cremaster muscle as a model. We found that functional capillary density and arteriolar diameters were significantly reduced by Betadine, whereas leucocyte activity was not affected. In the Elase group, capillary flow and arteriolar diameters were significantly increased, and again leucocyte activity was not changed. The mechanism by which Betadine reduces microvascular flow is believed to be the same as in reperfusion injury. The positive effect of Elase on the microcirculation might be attributed to plasmin, which has been shown to dilate blood vessels. PMID:11952671
Mallory, Mark L.; Little, Catherine M.; Boyd, Ellen S.; Ballard, Jennifer; Elliott, Kyle H.; Gilchrist, H. Grant; Hipfner, J. Mark; Petersen, Aevar; Shutler, Dave
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
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.
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.
Peers, S. H.; Smillie, F.; Elderfield, A. J.; Flower, R. J.
1. We have studied the occurrence, distribution and disposition of lipocortins (annexins) 1, 2 and 5 in mixed peritoneal leucocytes obtained from rats in which glucocorticoid levels were altered by adrenalectomy, administration of the glucocorticoid antagonist, RU486, or by injection of dexamethasone or hydrocortisone, as well as from rats in which the peritoneal cells were elicited by inflammatory stimuli. 2. In cells obtained from untreated rats with an intact adrenal cortex, lipocortins 1, 2 and 5 were readily detectable: the majority of each of the proteins was apparently located intracellularly with much smaller amounts in the membrane. Lipocortin 1 and to a lesser extent lipocortin 5 were also seen in a Ca(2+)-dependent association with the external plasma membrane. Following administration of RU486 (2 x 20 mg kg-1) the amounts of lipocortin 1 and 2 in cells were greatly reduced. Conversely, injection of hydrocortisone (1 mg kg-1) or dexamethasone (0.08 mg kg-1) caused an increase in the amount of lipocortin 1 and 2 in peritoneal cells within 30 min. Lipocortin 5 was unchanged by any manipulation of glucocorticoid levels. 3. Lipocortins 1 and 2 were elevated in both intracellular and membrane-associated fractions of macrophages elicited by intraperitoneal injection in inflammogens. This phenomenon also occurred in adrenalectomized animals. 4. Our data indicate that glucocorticoids control the synthesis of some members of the lipocortin family in rat mixed peritoneal cells but also suggest the existence of a separate system for controlling the generation of this protein. The significance of these observations is considered in relation to the mechanism of glucocorticoid hormone action on eicosanoid production. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 PMID:8428216
Vannoy, Steven D; Robins, Lynne S
Objective To characterise suicide-risk discussions in depressed primary-care patients. Design Secondary analysis of recordings and self reports by physicians and patients. Descriptive statistics of depression and suicide-related discussion, with qualitative extraction of disclosure, enquiry and physician response. Setting 12 primary-care clinics between July 2003 and March 2005. Participants 48 primary-care physicians and 1776 adult patients. Measures Presence of depression or suicide-related discussions during the encounter; patient and physician demographics; depression symptom severity and suicide ideation as measured by the Patient Health Questionnaire (PHQ9); physician's decision-making style as measured by the Medical Outcomes Study Participatory Decision-Making Scale; support for autonomy as measured by the Health Care Climate Questionnaire; trust in their physician as measured by the Primary Care Assessment Survey; physician response to suicide-related enquiry or disclosure. Results Of the 1776 encounters, 128 involved patients scoring >14 on the PHQ9. These patients were seen by 43 of the 48 physicians. Suicide ideation was endorsed by 59% (n=75). Depression was discussed in 52% of the encounters (n=66). Suicide-related discussion occurred in only 11% (n=13) of encounters. 92% (n=12) of the suicide discussions occurred with patients scoring <2 on PHQ9 item 9. Suicide was discussed in only one encounter with a male. Variation in elicitation and response styles demonstrated preferred and discouraged interviewing strategies. Conclusions Suicide ideation is present in a significant proportion of depressed primary care patients but rarely discussed. Men, who carry the highest risk for suicide, are unlikely to disclose their ideation or be asked about it. Patient-centred communication and positive healthcare climate do not appear to increase the likelihood of suicide related discussion. Physicians should be encouraged to ask about suicide ideation in their
Bowie, Paul; Forrest, Eleanor; Price, Julie; Verstappen, Wim; Cunningham, David; Halley, Lyn; Grant, Suzanne; Kelly, Moya; Mckay, John
ABSTRACT Background: The systems-based management of laboratory test ordering and results handling is a known source of error in primary care settings worldwide. The consequences are wide-ranging for patients (e.g. avoidable harm or poor care experience), general practitioners (e.g. delayed clinical decision making and potential medico-legal implications) and the primary care organization (e.g. increased allocation of resources to problem-solve and dealing with complaints). Guidance is required to assist care teams to minimize associated risks and improve patient safety. Objective: To identify, develop and build expert consensus on ‘good practice’ guidance statements to inform the implementation of safe systems for ordering laboratory tests and managing results in European primary care settings. Methods: Mixed methods studies were undertaken in the UK and Ireland, and the findings were triangulated to develop ‘good practice’ statements. Expert consensus was then sought on the findings at the wider European level via a Delphi group meeting during 2013. Results: We based consensus on 10 safety domains and developed 77 related ‘good practice’ statements (≥ 80% agreement levels) judged to be essential to creating safety and minimizing risks in laboratory test ordering and subsequent results handling systems in international primary care. Conclusion: Guidance was developed for improving patient safety in this important area of primary care practice. We need to consider how this guidance can be made accessible to frontline care teams, utilized by clinical educators and improvement advisers, implemented by decision makers and evaluated to determine acceptability, feasibility and impacts on patient safety. PMID:26339831
Radford, Sholto R; Crane, Rebecca Susan; Eames, Catrin; Gold, Eluned; Owens, Gareth Wyn
Background Mindfulness-based cognitive therapy (MBCT) is an intervention developed for the prevention of recurrent depression which is now being applied to widening numbers of clinical populations. Despite evidence for its effectiveness in preventing relapse in depression, less is known about its efficacy within routine clinical practice for groups of patients with more varied mental health problems, despite this being a potentially promising context for its application. Aims This pilot study aimed to investigate whether MBCT would be feasible and effective when delivered in a primary care context for patients who are vulnerable to recurrent depression and anxiety. Results Attrition from the programme was low and both attendance and engagement with home practices (during and after the intervention) were comparable with or higher than those observed in the existing literature. Improvements in self-reported depression, anxiety, rumination, self-compassion and well-being were evident over the 8-week programme and at 6-month post intervention follow-up. Conclusions Despite limitations in terms of sample size and the absence of a control group, the results demonstrate that the promising research results of MBCT for depression are transferable from a research to a practice setting, and demonstrate that it may be an effective and feasible intervention when delivered in a primary care setting for a range of mental health problems. PMID:23997825
Godycki-Cwirko, Maciek; Esmail, Aneez; Dovey, Susan; Wensing, Michel; Parker, Dianne; Kowalczyk, Anna; Błaszczyk, Honorata; Kosiek, Katarzyna
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
Marquart, H V; Rasmussen, J M; Leslie, R G
Previous studies from this laboratory have shown that normal peripheral blood B cells are capable of activating complement via the alternative pathway (AP), that the activation is associated with complement receptor type 2 (CR2) expression, and that erythrocytes at normal blood levels partially inhibit the activation. The purpose of the present study was to investigate whether factor I (FI) deficiency, which leads to continued formation of the AP convertase (C3bBb) resulting in the consumption of factor B and C3 and large scale generation of C3b fragments, affects the phenotype and/or function of the patients' B cells. Using flow cytometry, peripheral blood leucocytes (PBL) from two FI-deficient patients were investigated for expression of complement receptors and complement regulatory proteins, in vivo-deposited C3 fragments and in vitro complement-activating ability. CR1 levels on B cells were significantly lower in FI-deficient patients than in normal individuals, whereas CR2 levels were found to be reduced, although not to a significant extent. CR1 levels on monocytes and polymorphonuclear leucocytes (PMN) were found to be normal or slightly raised. All leucocyte subpopulations were found to be covered in vivo with C3b fragments. AP activation on B cells from FI-deficient patients in homologous serum was significantly reduced compared with that for normal individuals, whereas no in vitro activation was seen in autologous serum. In addition, the in vivo-bound C3b fragments were degraded to C3d,g when the patients' PBL were incubated in homologous serum containing EDTA. Finally, the patients, erythrocytes failed to exert any inhibition on AP activation in homologous serum. PMID:9301541
Gerardi, A S
Bovine leucocyte adhesion deficiency (BLAD) is a genetic disease of cattle which affects the hematopoietic system. In the last decade BLAD has become a disease of economic importance in the dairy industry. This review describes the chronological developments and thinking that led to the elucidation of BLAD as a disease distinct from previous models in canine and human populations. All species affected show signs of chronic and recurrent infections. Necrotic and/or gangrenous infections of soft tissues are prevalent, in addition to secondary infections with bacteria or fungi. Low birthweight and unthriftiness are key signs in all species affected by leucocyte adhesion deficiency (LAD). Dermatomycoses and impaired pus formation are also common findings. The physiological basis for BLAD is a deficiency in the chemotactic and phagocytic properties of leucocytes and particularly neutrophils. The inhibition of diapedesis in the inflammatory response prevents normal immune reactions to invading pathogens. Chronic infections are a consequence of the faulty immune mechanisms. The biochemical aetiology of BLAD involves cell surface glycoprotein molecules known as integrins. These are responsible for the cell-cell interactions necessary for neutrophils to adhere to vascular endothelium in a normal individual. Experiments with monoclonal antibodies to block LFA-1, Mac-1, and p150,95 (three integrins vital for cell-cell interactions) mimic BLAD symptomatology and have led to the discovery of the reciprocal intercellular adhesion molecule (ICAM). Through pedigree analysis and biochemical detection with restrictive endonucleases, BLAD has been isolated genetically to a single gene locus. The economic significance and prophylaxis of the disease are briefly discussed. In addition, the beneficial aspects of the study of BLAD are considered. There are advantages in producing a BLAD-like state for preventing transplant rejection, ischaemia-reperfusion injury and other problems arising
PONGCHAROEN, S; BULMER, J N; SEARLE, R F
Complete hydatidiform moles are totally paternally derived and represent complete allografts that might be expected to provoke maternal immune rejection. Our previous and other studies have shown expression of Fas by increased numbers of activated decidual CD4+ T cells in both complete and partial molar pregnancy as well as increased FasL+ expression by molar trophoblasts compared with trophoblasts in normal pregnancies. As the Fas/FasL system represents a major apoptotic pathway that can play a role in immune privilege, the aim of this study was to investigate whether apoptosis of decidual immune cells, particularly T cells, could be responsible for maternal immune tolerance in molar pregnancy. Using terminal deoxynucleotidyl transferase (TdT)-mediated nick end-labelling (TUNEL), a significant increase in TUNEL+ cells was demonstrated in decidua associated with partial (P = 0·0052) and complete (P = 0·0096) hydatidiform mole compared with normal early pregnancy. Co-labelling immunoperoxidase studies showed that the TUNEL+ cells in both normal and molar pregnancies were not activated CD45RO+ immune cells, CD3+ T cells, CD56+ uterine natural killer (NK) cells or CD14+ CD68+ macrophages. Double immunohistochemical labelling with antiactive caspase-3 and leucocyte markers confirmed the lack of leucocyte apoptosis. Double immunostaining with anticytokeratin to detect trophoblast and M30 CytoDeath, which detects a neoepitope of cytokeratin 18 revealed after caspase-mediated cleavage, revealed apoptotic extravillous trophoblast cells within decidual tissue. We conclude that there is no evidence that apoptosis of decidual leucocytes plays a role in maintaining maternal tolerance in either normal or molar pregnancy. PMID:15498045
Lecoeur, Hervé; Giraud, Emilie; Prévost, Marie-Christine; Milon, Geneviève; Lang, Thierry
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
Background The accuracy of the World Health Organization method of estimating malaria parasite density from thick blood smears by assuming a white blood cell (WBC) count of 8,000/μL has been questioned in several studies. Since epidemiological investigations, anti-malarial efficacy trials and routine laboratory reporting in Papua New Guinea (PNG) have all relied on this approach, its validity was assessed as part of a trial of artemisinin-based combination therapy, which included blood smear microscopy and automated measurement of leucocyte densities on Days 0, 3 and 7. Results 168 children with uncomplicated malaria (median (inter-quartile range) age 44 (39–47) months) were enrolled, 80.3% with Plasmodium falciparum monoinfection, 14.9% with Plasmodium vivax monoinfection, and 4.8% with mixed P. falciparum/P. vivax infection. All responded to allocated therapy and none had a malaria-positive slide on Day 3. Consistent with a median baseline WBC density of 7.3 (6.5-7.8) × 109/L, there was no significant difference in baseline parasite density between the two methods regardless of Plasmodium species. Bland Altman plots showed that, for both species, the mean difference between paired parasite densities calculated from assumed and measured WBC densities was close to zero. At parasite densities <10,000/μL by measured WBC, almost all between-method differences were within the 95% limits of agreement. Above this range, there was increasing scatter but no systematic bias. Conclusions Diagnostic thresholds and parasite clearance assessment in most PNG children with uncomplicated malaria are relatively robust, but accurate estimates of a higher parasitaemia, as a prognostic index, requires formal WBC measurement. PMID:24739250
Xie, Xin; Nie, Xiaorong; Yu, Bingbin; Zhang, Xu
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.
The aim of this study was to describe a young man with probably the largest pheochromocytoma associated with MEN 2A, described till date. The patient, a non-vegetarian, fifth of eight siblings, married, having five children, presented with episodes of difficult-to-control hypertension requiring over five antihypertensives. He was referred to us with an abdominal CT scan that revealed a 16 cm left-sided adrenal mass. Biochemical testing confirmed a catecholamine secreting pathology. Histopathology confirmed the mass as a pheochromocytoma weighing 1.8 kg. Further evaluation suggested a parathormone-dependent hypercalcemia and a left-sided thyroid mass. Histopathology confirmed parathyroid hyperplasia and medullary carcinoma of the thyroid mixed with papillary carcinoma of thyroid. Putting all the findings together showed that the patient was suffering from multiple endocrine neoplasia 2. Multiple endocrine neoplasia 2A is a rare syndrome. The case is unique in the way it presented, with all the three tumors at the same time. The management was bold and addressed all the three lesions in the same hospital admission. We are also reporting the largest described case of pheochromocytoma from India. PMID:23961501
Puschel, Klaus; Thompson, Beti
Summary Breast cancer has the highest incidence of all cancers among women in Chile. In 2005, a national health program progressively introduced free mammography screening for women aged 50 and older; however, three years later the rates of compliance with mammographic screening was only 12% in Santiago, the capital city of Chile. This implementation article combines the findings of two previous studies that applied qualitative and quantitative methods to improve mammography screening in an area of Santiago. Socio-cultural and accessibility factors were identified as barriers and facilitators during the qualitative phase of the study and then applied to the design of a quantitative randomized clinical trial. After six months of intervention, 6% of women in the standard care group, 51.8% in the low intensity intervention group, and 70.1% in the high intensity intervention group had undergone a screening mammogram. This review discusses how the utilization of mixed methods research can contribute to the improvement of the implementation of health policies in local communities. PMID:21334897
Identification of monoclonal antibodies with specificity to alpha- or beta-chains of beta 2-integrins using peripheral blood leucocytes of normal and bovine leucocyte adhesion deficient (BLAD) cattle.
Rutten, V P; Hoek, A; Müller, K E
The reactivity of monoclonal antibodies entered in the panels of the Third Workshop on Ruminant Leucocyte Antigens with lymphocytes and monocytes of normal and beta 2-integrin deficient (Bovine Leucocyte Adhesion Deficiency: BLAD) animals was determined by flow cytometry to investigate potential specificities to alpha- or beta-chains of beta 2-integrins. From the 13 monoclonal antibodies that were entered as having specificity for CD11/CD18 antigens, ten were confirmed correct, but three had reactivity with cells from BLAD animals. We conclude that our approach provides an easy way to reliably identify the majority of beta 2-integrin specific monoclonal antibodies. PMID:8896223
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
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
Mairal, L; de Lima, P A; Martin-Comin, J; Baliellas, C; Xiol, X; Roca, M; Ricart, Y; Ramos, M
Technetium-99m hexamethylpropylene amine oxime (HMPAO) labelled leucocytes and indium-111 polyclonal immunoglobulin (IgG) were simultaneously injected into a group of 27 patients routinely referred for the investigation of inflammatory bowel disease (IBD). Ten-minute anterior abdomen and tail on detector views were obtained at 30 min, 4 h and 24 h p.i. of both tracers. The diagnosis of IBD was obtained in all cases by endoscopy with biopsy and/or surgery. Images were blindly evaluated by two experienced observers who only knew of the clinical suspicion of IBD. IBD was confirmed in 20 patients (12 with Crohn's disease and eight with ulcerative colitis). Sensitivity, specificity and accuracy were 100%, 85% and 96% respectively for labelled leucocytes and 70%, 85% and 74% for IgG. Both IgG and leucocyte scans were normal in six out of seven patients in whom a diagnosis of IBD was excluded; the remaining patient, with ischaemic colitis, was falsely positive with both agents. As far as disease extension is concerned, the IgG study localized 27 diseased segments, whereas 49 were seen with the leucocyte study. Eighty-four segments were normal and 25 showed tracer uptake with both agents. Twenty-four were positive only with the leucocyte study and two were positive only with the IgG study. Agreement between the agents was 80.7%. These results confirm that 111In-human polyclonal scintigraphy is less sensitive than 99mTc-HMPAO scintigraphy both for the diagnosis of IBD and in the evaluation of disease extension. Nevertheless, if leucocyte labelling is not available, labelled IgG can be used only for diagnostic purposes. PMID:7498228
Berliner, S; Fried, M; Caspi, D; Weinberger, A; Yaron, M; Pinkhas, J; Aronson, M
Previous work has shown that leucocyte adhesiveness/aggregation (LAA), as measured by the leukergy test, correlates well with disease severity in rheumatic patients. As LAA is probably a manifestation of the acute phase reaction various components of the acute phase reaction were measured in order to identify the best marker of disease activity. In addition to LAA, the following variables were measured in 79 patients with various rheumatic diseases and in 10 controls: white blood cell and platelet counts, erythrocyte sedimentation rate, haptoglobin, fibrinogen, C reactive protein, albumin, globulin, caeruloplasmin, alpha 1, alpha 2, beta, and gamma globulin, and haemoglobin concentrations. Patients were graded according to the state of their disease as mild, moderate, or severe. The extent of leucocyte adhesiveness/aggregation in peripheral blood proved to be the best laboratory variable for the grading of disease activity. Correct grading was obtained in 63% of the patients by means of the LAA, compared with 48% with C reactive protein, 41% with caeruloplasmin, 40% with haptoglobin, and 32% with haemoglobin. It is suggested that LAA of the peripheral blood during inflammation may be used as a reliable marker of disease severity. PMID:3260093
Turaihi, K.; Baron, D.N.; Dandona, P.
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.
Belen, Burcu; Polat, Meltem
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
De Feo, G; Galasso, M; Landi, R; Donnarumma, A; De Gisi, S
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). PMID:24191462
Ashorn, R G; Räsänen, L; Marnela, K M; Krohn, K J
The effect of human transfer factor (TF) or its components L-serine and/or glycine in tuberculin (PPD), or leucoagglutinin (LA) induced leucocyte migration inhibitory factor (LIF) secretion was studied. Augmentation of LIF secretion was seen with low concentration ( = 0.078 g/l) of TF when lymphocytes were cultured in minimum essential medium for suspension cultures (MEM-S), a culture medium lacking L-serine and glycine. High concentrations (0.3125-5.0 g/l dry weight) of TF were inhibitory in MEM-S. In RPMI 1640, a culture medium containing L-serine and glycine, TF was either inhibitory or had no effect. The combination of L-serine and glycine, at concentrations equivalent or lower than the optimum of TF, had an augmenting effect on LIF secretion identical to that of TF, but no inhibition at higher concentrations was seen. The results indicate that human TF contains components which have suppressive or augmenting effects on LIF secretion in vitro. The augmenting effect may be mainly due to L-serine and glycine and thus not related to TF's activity in vivo. PMID:385187
Gauthier, David T.; Cartwright, Deborah D.; Densmore, Christine L.; Blazer, Vicki; Ottinger, Christopher A.
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.
Haller, D; Bode, C; Hammes, W; Pfeifer, A; Schiffrin, E; Blum, S
BACKGROUND AND AIM—Intestinal epithelial cells (IEC) are thought to participate in the mucosal defence against bacteria and in the regulation of mucosal tissue homeostasis. Reactivity of IEC to bacterial signals may depend on interactions with immunocompetent cells. To address the question of whether non-pathogenic bacteria modify the immune response of the intestinal epithelium, we co-cultivated enterocyte-like CaCO-2 cells with human blood leucocytes in separate compartments of transwell cultures. METHODS—CaCO-2/PBMC co-cultures were stimulated with non-pathogenic bacteria and enteropathogenic Escherichia coli. Expression of tumour necrosis factor alpha (TNF-α), interleukin (IL)-1β, IL-8, monocyte chemoattracting protein 1 (MCP-1), and IL-10 was studied by enzyme linked immunosorbent assays (cytokine secretion) and by semiquantitative reverse transcription-polymerase chain reaction. RESULTS—Challenge of CaCO-2 cells with non-pathogenic E coli and Lactobacillus sakei induced expression of IL-8, MCP-1, IL-1β, and TNF-α mRNA in the presence of underlying leucocytes. Leucocyte sensitised CaCO-2 cells produced TNF-α and IL-1β whereas IL-10 was exclusively secreted by human peripheral blood mononuclear cells. CaCO-2 cells alone remained hyporesponsive to the bacterial challenge. Lactobacillus johnsonii, an intestinal isolate, showed reduced potential to induce proinflammatory cytokines but increased transforming growth factor beta mRNA in leucocyte sensitised CaCO-2 cells. TNF-α was identified as one of the early mediators involved in cellular cross talk. In the presence of leucocytes, discriminative activation of CaCO-2 cells was observed between enteropathogenic E coli and non-pathogenic bacteria. CONCLUSION—The differential recognition of non-pathogenic bacteria by CaCO-2 cells required the presence of underlying leucocytes. These results strengthen the hypothesis that bacterial signalling at the mucosal surface is dependent on a network of
Ribaldi, E.; Mezzasoma, A. M.; Francescangeli, E.; Prosdocimi, M.; Nenci, G. G.; Goracci, G.; Gresele, P.
1. A phospholipase A2 (PLA2) represents the key enzyme in the remodelling pathway of platelet-activating factor (PAF) synthesis in human polymorphonuclear (PMN) leucocytes. 2. PLA2 activation is also the rate-limiting step for the release of the arachidonic acid utilized for the synthesis of leukotrienes in stimulated leucocytes; however, it is unknown whether the PLA2s involved in the two biosynthetic pathways are identical. 3. Cloricromene (8-monochloro-3-beta-diethylaminoethyl-4-methyl-7-ethoxy- carbonylmethoxy coumarin) is an antithrombotic coumarin derivative which inhibits platelet and leucocyte function and suppresses arachidonic acid liberation by interfering with PLA2 activation. 4. The aim of the present study was to assess whether chloricromene inhibits PAF synthesis by stimulated human polymorphonuclear leucocytes (PMNs). 5. Cloricromene (50-500 microM) inhibited in a concentration-dependent manner the release of PAF, as measured by h.p.l.c. bioassay, from A23187-stimulated PMNs. Significant inhibition (45%) of PAF-release was obtained with 50 microM cloricromene and the IC50 was 85 microM. Mepacrine (500 microM), a non-specific PLA2 inhibitor, strikingly reduced PAF release. 6. The incorporation of [3H]-acetate into [3H]-PAF induced by serum-treated zymosan in human PMNs was also inhibited concentration-dependently by cloricromene, with an IC50 of 105 microM. Mepacrine also suppressed [3H]-acetate incorporation into [3H]-PAF. 7. Cloricromene did not affect the activities of the enzymes involved in PAF-synthesis acetyltransferase or phosphocholine transferase. 8. Our data demonstrate that cloricromene, an inhibitor of PLA2-activation in human leucocytes, reduces the synthesis of PAF by stimulated PMNs. This finding has a twofold implication: the PLA2s (or the mechanisms that regulate their activation) involved in PAF synthesis and arachidonate release in human leucocytes are either identical or else indistinguishable by their sensitivity to cloricromene
Zhao, Lian; Gaudry, Leonie; Dunkley, Scott; Brighton, Tim; Guo, Zhi Xin; Ye, Zheng Liang; Luo, Run Zhi; Chesterman, Colin N
Platelet and leucocyte activity are important in the acute development of thrombosis and in the pathogenesis of ischaemic vascular disease. Dan Shen Di Wan (DS, Cardiotonic Pill or Dantonic(R) Pill) is one of the most commonly used Chinese herbal formulations for treating patients with atherosclerotic disease in China and several Asian countries. We studied the effect of DS on platelet and leucocyte function and compared the effects with conventional antiplatelet agents, cangrelor (ADP P2Y(12) receptor antagonist) and aspirin (acetyl salicylic acid, ASA). Measurements were made by platelet aggregation (%) and activation (CD62P %), platelet-monocyte conjugate formation (P/M, CD42a median fluorescence, mf), platelet-neutrophil conjugate formation (P/N, mf), and leucocyte activation (CD11b median fluorescence on monocytes and neutrophils, mf) in response to 3.3 micromol/L adenosine diphosphate (ADP), 1.0 micromol/L platelet activating factor (PAF), 5.0 micromol/L adrenaline and 0.5 microg/mL collagen. We also evaluated the effect of its main component, water soluble extract of salvia miltiorrhiza (SME) on intracellular calcium mobilization in platelets triggered by 10 micromol/L ADP, 10 micromol/L PAF, 2 microg/mL collagen and 15 micromol/L thrombin receptor activating peptide (TRAP). Overall DS showed inhibition of platelet aggregation, platelet activation, platelet-leucocyte conjugate formation and leucocyte activation in response to all the agonists apart from adrenaline (all p < 0.01). DS showed inhibition of platelet aggregation and leucocyte activation equivalent to cangrelor 100 nmol/L and ASA 100 micromol/L. SME dose-dependently inhibited intracellular calcium mobilization in platelets following stimulation with all the platelet agonists with maximum effective at 0.36 mg/mL (all p < 0.01). When used at 0.18 mg/mL its inhibitory effect was equivalent to cangrelor and ASA. We conclude that DS is a potential inhibitor of both platelet and leucocyte activation
Kownatzki, E; Kapp, A; Uhrich, S
Human mononuclear leucocytes isolated from the peripheral blood by centrifugation on Ficoll-Hypaque cushions and adherent on plastic petri dishes, produced a chemotactic factor that attracted human neutrophilic granulocytes to the same extent as did optimal concentrations of the complement split product C5a and the leukotriene B4. The active component eluted from a Sephadex G-50 gel filtration column as a single peak with an apparent molecular weight of 10,000. The chemotactic activity was resistant to reductive cleavage of disulfide bonds and heating at 100 degrees C for 30 min but was lost when reduction and heating were combined. Digestion with a proteolytic enzyme eliminated the attractive potential. The data suggest that this is a novel chemotactic peptide. It is conceivable that it has been seen previously and was mistaken for a lymphokine or interleukin 1. PMID:3731527
Nagahata, H; Matsuki, S; Higuchi, H; Inanami, O; Kuwabara, M; Kobayashi, K
Bone marrow transplantation (BMT) was performed in a 9-month-old heifer with bovine leucocyte adhesion deficiency (BLAD). Clinical and haematological findings, selected neutrophil function and CD18 expression of neutrophils in a B2 integrin-deficient heifer were examined. Twelve months after BMT, a small fluorescent region in the CD18-positive area of a flow cytometric profile was demonstrated and estimated to represent 0.3-0.5% of the CD18-positive neutrophils as measured by flow cytometric analysis following immunomagnetic separation. The animal's clinical condition appeared to improve and stabilize over our observation period of 28 months following BMT. Newly expressed CD18 seemed to play an important role in ameliorating the clinical signs of BLAD in this heifer. PMID:9691847
Sedgwick, A D; Morris, T; Russell, B A; Lees, P
Percoll gradients have been used to separate relatively pure populations of viable equine polymorphonuclear (PMN) and mononuclear (MN) cells. In preliminary studies, a continuous density gradient of 70% Percoll solution was used to separate two distinct leucocyte-rich bands. After measurement of the density of each band on the continuous gradient, discontinuous Percoll gradients, using 60% and 75% Percoll solutions, were used to provide a rapid means of separating PMN and MN cells. The yield of viable cells per ml of blood was 3.0 X 10(6) and 3.2 X 10(6) for MN and PMN cells, respectively. Corresponding values for recovery were 45% and 72%. The purity was 94% for PMNs and 99% for MNs. PMID:3798735
Lawrence, R H; Sorrelli, T C
Evidence that leukotriene B4 (LTB4) is a significant inflammatory mediator in chronic pseudomonal respiratory disease was sought in adolescents and young adults with cystic fibrosis. Specific chemotaxis of peripheral blood polymorphonuclear leucocytes (PMN) was used as an indirect measure of remote in vivo exposure to LTB4. PMN from 17 patients showed a significant decrease in chemotaxis to 10(-7)-10(-9) M LTB4, but normal responses to 10(-8) M n-formyl-methionyl-leucyl-phenylalanine and 4 mg/ml casein, when compared with 17 healthy age- and sex-matched controls. This result is consistent with chronic production of LTB4, and specific deactivation of circulating PMN receptors for LTB4 in patients with cystic fibrosis. Pharmacologic inhibition of LTB4 production in vivo may help elucidate its role in the pathogenesis of lung damage in cystic fibrosis. PMID:1322257
Csiba, A; Whitwell, H L; Moore, M
Monoclonal antibodies (McAbs) directed against the framework determinants of Class I and Class II products of the major histocompatibility complex (MHC) and against leucocyte differentiation antigens were used in an indirect immunoperoxidase technique to study their expression in normal, benign (adenomatous polyps) and malignant disease of the colon. Class I products (detected by the McAb 2A1) were strongly expressed on all cell types in normal and benign tissues but some carcinomas exhibited a heterogenous pattern of epithelial cell staining and 4/15 were completely negative. Class II products (detected by TDR31.1) were strongly expressed on cells (mainly B lymphocytes) within the lamina propria. In carcinomas TDR31.1 staining was mainly interstitial, but in 2/15, DR + epithelial cells were also detected. In normal and benign tissues, leucocytes (reactive with 2D1) found predominantly in the lamina propria, comprised T cells mainly of the helper/inducer (OKT4) subset, DR + cells in approx. equivalent proportion and a few OKM1+ cells mostly of macrophage morphology. Occasional intraepithelial lymphocytes were of cytotoxic/suppressor (OKT8) phenotype. In malignant neoplasms, there was wide inter and intra-tumour variation in the proportion of leucocytes which were heterogeneous with respect to cell type and confined mainly to the stroma. T cells were consistently predominant, but B cells and macrophages were also present. Two neoplasms showed unequivocal evidence of a shift (relative to peripheral blood) in favour of the OKT8+ subset, but in the majority of tumours OKT4+; and OKT8+ cells were present in roughly similar proportions. Natural killer cells (monitored with Leu7, HNK1) were virtually undetectable in both normal and malignant tissues. There were no apparent correlations between the extent and type of leucocyte infiltration, tumour differentiation or expression of MHC products. Some implications for the extrapolation of in vitro data on leucocyte function
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
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
Marti, J. H.; Thomson, D. M.
Tumour antigen-induced inhibition of leucocyte adherence was modified for use in glass test tubes (Tube LAI assay) for the study of cell-mediated anti-tumour immunity to human malignant melanoma. Peripheral blood leucocytes (PBL) of 20 out of 25 patients (80%) with active malignant melanoma responded to an extract of malignant melanoma with LAI, whereas only 4-5% of 475 control subjects showed a response. The malignant melanoma patients reacted to both allogeneic and autologous extracts of malignant melanoma which indicates a common cross-reacting antigen. Malignant melanoma patients did not respond to unrelated tumour extracts. The LAI was mediated by PBL (monocytes) "armed" with cytophilic anti-tumour antibody specific for the sensitizing tumour antigen. The anti-tumour response of the malignant melanoma patients was dependent on the stage of the cancer, and 11 out of 13 Stage I patients had a positive NAI, whereas patients with disseminated cancer had decreased response. The diminished LAI in patients with large tumour burdens appeared to be the result of release of tumour antigen systemically. Also, surgery and chemotherapy depressed LAI. Although LAI was depressed after surgical excision of the cutaneous melanoma, most patients showed LAI 1-3 months later. Tumour-free melanoma patients monitored for one year by the Tube LAI assay showed a decline in their anti-tumour immunity 5-6 months after surgery. The NAI was low or negative after the 8th post-surgical month in tumour-free patients. Patients with residual malignant melanoma showed persistent or recurrent LAI after the 8th post-surgical month. LAI reactivity monitored after "curative" surgery for malignant melanoma may assist in determining whether the patient is tumour-free or has a recurrence. PMID:962991
Su, Yang; Lei, Xi; Wu, Lingyun; Liu, Lixin
Methylglyoxal (MG) is a reactive dicarbonyl metabolite formed during glucose, protein and fatty acid metabolism. In hyperglycaemic conditions, increased MG level has been linked to the development of diabetes and its vascular complications at the macrovascular and microvascular levels where inflammation plays a role. To study the mechanism of MG-induced inflammation in vivo, we applied MG locally to healthy mice and used intravital microscopy to investigate the role of endothelial cell adhesion molecules in MG-induced leucocyte recruitment in cremasteric microvasculature. Administration of MG (25 and 50 mg/kg) to the tissue dose-dependently induced leucocyte recruitment at 4·0–5·5 hr, with 84–92% recruited cells being neutrophils. Such MG treatment up-regulated the expression of endothelial cell adhesion molecules P-selectin, E-selectin, intercellular adhesion molecule-1, but not vascular cell adhesion molecule-1. Activation of the nuclear factor-κB signalling pathway contributed to MG-induced up-regulation of these adhesion molecules and leucocyte recruitment. The role of the up-regulated endothelial cell adhesion molecules in MG-induced leucocyte recruitment was determined by applying specific functional blocking antibodies to MG-treated animals and observing changes in leucocyte recruitment parameters. Our data demonstrate that the up-regulation of P-selectin, E-selectin and intercellular adhesion molecule-1 contributes to the increased leucocyte rolling flux, reduced leucocyte rolling velocity, and increased leucocyte adhesion, respectively. Our results reveal the role of endothelial cell adhesion molecules in MG-induced leucocyte recruitment in microvasculature, an inflammatory condition related to diabetic vascular complications. PMID:22681228
Leblanc, Kori; Valentinis, Alissia; Kavanagh, Doug; Zahr, Nina; Ivers, Noah M
Background Atrial fibrillation (AF) is a common and preventable cause of stroke. Barriers to reducing stroke risk through appropriate prescribing have been identified at the system, provider, and patient levels. To ensure a multifaceted initiative to address these barriers is effective, it is essential to incorporate user-centered design to ensure all intervention components are optimized for users. Objective To test the usability of an electronic medical record (EMR) toolkit for AF in primary care with the goal of further refining the intervention to meet the needs of primary care clinicians. Methods An EMR-based toolkit for AF was created and optimized through usability testing and iterative redesign incorporating a human factors approach. A mixed-methods pilot study consisting of observations, semi-structured interviews, and surveys was conducted to examine usability and perceived impact on patient care and workflow. Results A total of 14 clinicians (13 family physicians and 1 nurse practitioner) participated in the study. Nine iterations of the toolkit were created in response to feedback from clinicians and the research team; interface-related changes were made, additional AF-related resources were added, and functionality issues were fixed to make the toolkit more effective. After improvements were made, clinicians expressed that the toolkit improved accessibility to AF-related information and resources, served as a reminder for guideline-concordant AF management, and was easy to use. Most clinicians intended to continue using the toolkit for patient care. With respect to impact on care, clinicians believed the toolkit increased the thoroughness of their assessments for patients with AF and improved the quality of AF-related care received by their patients. Conclusions The positive feedback surrounding the EMR toolkit for AF and its perceived impact on patient care can be attributed to the adoption of a user-centered design that merged clinically important
De Vecchi, E; Villa, F; Bortolin, M; Toscano, M; Tacchini, L; Romanò, C L; Drago, L
Analysis of joint fluid is of paramount importance for the diagnosis of prosthetic joint infections. Different markers of inflammation and/or infection in joint fluid have been proposed for diagnosis of these infections. In this study we evaluated the performance of leucocyte esterase, C-reactive protein (CRP) and glucose assays in synovial fluids from 129 patients with septic (n = 27) or aseptic (n = 102) prosthetic joint failure. Samples were collected in serum tubes and centrifuged to limit the presence of corpuscle interfering with the assays. Determinations of leucocyte esterase and glucose were carried out by means of enzymatic colorimetric reactions performed on strips for urine analysis. Tests were considered positive when graded + or ++ whereas traces or absence of colour were considered negative. CRP was measured using an automated turbidimetric method and considered suggestive for infections when >10 mg/L. Leucocyte esterase was positive in 25/27 infected patients and negative in 99/102 not infected patients (sensitivity 92.6%, specificity 97.0%). CRP was higher than the threshold in 22/27 infected patients and in 6/102 not infected patients (sensitivity: 81.5%; specificity: 94.1%) whereas glucose showed the lowest sensitivity (77.8%) and specificity (81.4%), being negative in 21/27 and 19/102 infected and not infected patients, respectively. CRP led to a correct diagnosis in 19 of 22 patients with discordant esterase and glucose results. In conclusion, evaluation of leucocyte esterase, glucose and CRP may represent a useful tool for rapid diagnosis of prosthetic joint infections. PMID:27040804
Gagné, François; Fortier, Marlène; Fournier, Michel; Smyth, Shirley-Anne
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. PMID:24218853
Muñoz-Atienza, Estefanía; Araújo, Carlos; Lluch, Nuria; Hernández, Pablo E; Herranz, Carmen; Cintas, Luis M; Magadán, Susana
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. PMID:25707601
Sánchez-Barrena, María José; Vallis, Yvonne; Clatworthy, Menna R.; Doherty, Gary J.; Veprintsev, Dmitry B.; Evans, Philip R.; McMahon, Harvey T.
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
A type of microscopical chamber is described which enables monolayers of cells to be examined at 37°C by phase-contrast microscopy at high magnification and which can be perfused semi-automatically. Such chambers have been used to observe morphological changes in the basophil leucocytes of atopic subjects when challenged with an extract of Timothy grass pollen. The appearance of basophil leucocytes in monolayers prepared from both washed and defibrinated blood cell suspensions has been studied. Basophils taken from non-atopic subjects or atopic subjects who were not hypersensitive to grass pollen showed no reaction to Timothy grass pollen extract. By contrast, basophils taken from pollen sensitive atopic subjects reacted to Timothy grass pollen extract with an acute change in motility and many degranulated. The morphological changes observed are described and illustrated in detail. No significant changes were seen in other types of cell. Some immunological and cellular mechanisms which may underlie this degranulation of human basophil leucocytes are discussed. ImagesFIG. 2FIG. 3FIG. 4 PMID:4924942
Brogan, T. D.
The phagocytic activity of human polymorphonuclear leucocyte preparations, which were free from plasma, has been estimated by direct determination under phase contrast of the number of living cells containing test particles. Spores of Aspergillus fumigatus were phagocytosed in the absence of added serum but phagocytosis of paraffin wax particles occurred only in the presence of serum containing the heat-labile and C′4 components of complement. In view of the unreactive nature of the paraffin hydrocarbons, it was considered unlikely that natural antibody played any part in the phenomenon. Although no phagocytosis of wax particles occurred in the absence of serum, almost 100 per cent of cells were phagocytic in preparations containing adequate concentrations of serum. It was therefore possible to determine the serum concentration necessary for 50 per cent of the polymorphs to phagocytose wax particles. By this means it was demonstrated that the addition of the carbohydrate components of sputum had a small but significant inhibitory effect on phagocytosis and that dextran had no such effect. The sputum mucoprotein depressed the complement titre of serum and this might have accounted for the reduction in the ability of a serum to promote phagocytosis when this complex was added. The sputum mucopolysaccharide had no such effect on the complement titre of serum and must have exerted its inhibitory action in some other way. ImagesFIG. 2 PMID:14239838
Pyle, Angela; Taylor, Robert W; Durham, Steve E; Deschauer, Marcus; Schaefer, Andrew M; Samuels, David C; Chinnery, Patrick F
Background The 3243A→G MTTL1 mutation is the most common heteroplasmic mitochondrial DNA (mtDNA) mutation associated with disease. Previous studies have shown that the percentage of mutated mtDNA decreases in blood as patients get older, but the mechanisms behind this remain unclear. Objectives and method To understand the dynamics of the process and the underlying mechanisms, an accurate fluorescent assay was established for 3243A→G heteroplasmy and the amount of mtDNA in blood with real‐time polymerase chain reaction was determined. The amount of mutated and wild‐type mtDNA was measured at two time points in 11 subjects. Results The percentage of mutated mtDNA decreases exponentially during life, and peripheral blood leucocytes in patients harbouring 3243A→G are profoundly depleted of mtDNA. Conclusions A similar decrease in mtDNA has been seen in other mitochondrial disorders, and in 3243A→G cell lines in culture, indicating that depletion of mtDNA may be a common secondary phenomenon in several mitochondrial diseases. Depletion of mtDNA is not always due to mutation of a nuclear gene involved in mtDNA maintenance. PMID:16950816
Windsor, A C; Hobbs, C B; Treby, D A; Gupta, C B
Twenty-seven elderly men aged 69-94 years had tyrosine tolerance tests measured before and after receiving ascorbic acid 1 g daily for four days. There was a significant rise in the fasting and half-hourly mean plasma tyrosine levels after ascorbic acid administration in those subjects with an initial leucocyte ascorbic-acid level (LAA) below 15 micrograms/10(8) w.c.c. The peak of the tyrosine tolerance curve was also advanced following administration of ascorbic acid in those subjects with an initial LAA below 15 micrograms/10(8) w.c.c. A further ten elderly men aged 73-89 years had simultaneous measurements of the circadian rhythms of LAA, plasma cortisol and plasma tyrosine levels before and after receiving ascorbic acid 1 g daily for four days. All ten subjects showed a peak LAA level at 5 p.m. when saturated with vitamin C but there was no significant correlation between plasma cortisol changes and plasma tyrosine changes. Possible explanations for the results are discussed. PMID:803133
Thude, H; Schorner, U; Helfricht, C; Loth, M; Maak, B; Barz, D
Neonatal alloimmune thrombocytopenia (NAIT) occurs when maternal alloantibodies to antigens presented on foetal platelets cause their immune destruction. Whether human leucocyte antigen (HLA) antibodies can cause NAIT is controversial. Here, a patient was described who suffered from a NAIT caused by an HLA-B27 antibody. Sera from the mother and the newborn were tested for human platelet antigen antibodies and HLA antibodies by monoclonal antibody-specific immobilization of platelet antigens (MAIPA) assay, solid phase-linked immunosorbent assay (ELISA), lymphocytotoxicity assay (LCT) and flow cytometric analysis. No antibodies against cluster designation (CD)109 and platelet glycoproteins of the father were found in patient's and mother's serum. However, HLA ELISA was used to identify HLA antibody in both sera. The antibody was specified as HLA-B27 antibody. Typing results showed that the father descended HLA-B27 antigen on patient and his brother. The mother was HLA-B27 negative. It is most conceivable that the previous pregnancy of the mother induced the production of anti-HLA-B27 antibody, which crossed the placenta and subsequently caused an NAIT in the case presented. PMID:16623921
Dupont, Aurélie; De Pauw-Gillet, Marie-Claire; Schnitzler, Joseph; Siebert, Ursula; Das, Krishna
Methylmercury (MeHg) is highly immunotoxic and can alter the health status of the harbour seal, Phoca vitulina, from the North Sea. To investigate the mechanism of MeHg-induced toxicity in harbour seal lymphocytes, Concanavalin A (ConA)-stimulated peripheral blood leucocytes were exposed in vitro to sublethal concentrations of MeHgCl (0.2, 1, and 2 µM) for 72 h and then analysed for their viability and ultrastructure. After 72 h of incubation, cells were counted with a propidium iodide staining technique, a metabolic MTS assay was performed, and cells exposed to 1 µM of MeHgCl were observed by transmission electron microscopy (TEM). Alive cell numbers decreased with increased MeHgCl concentrations. In presence of ConA and 1 µM of MeHgCl, TEM images revealed a higher frequency of apoptotic cells. Exposed cells displayed condensation of the chromatin at the nuclear membrane and mitochondrial damages. The results suggest that in vitro MeHgCl-induced apoptosis in harbour seal lymphocytes through a mitochondrial pathway. PMID:26264045
Whitney, J L; Krockenberger, M B; Day, M J; Beatty, J A; Dhand, N K; Barrs, V R
Leucocyte populations in the sinonasal mucosa of cats with and without upper respiratory tract aspergillosis were compared using immunohistochemistry and computer-aided morphometry. Inflammation was identified in the nasal mucosa of all affected cats, comprising predominantly of lymphoplasmacytic infiltration of the lamina propria associated with epithelial proliferation and degeneration. There was intense and diffuse expression of class II antigens of the major histocompatibility complex, associated with sites of hyphal invasion with hyperplasia and ulceration of the epithelium adjacent to fungal elements. Significantly more CD79b(+) cells, total lymphocytes, immunoglobulin (Ig)-expressing cells and MAC387(+) cells infiltrated the epithelium and more IgG(+) cells and total Ig-expressing cells infiltrated the lamina propria in affected cats compared with controls. Importantly, the inflammatory profile in affected cats was not consistent with the T helper (Th)1 and Th17 cell-mediated response that confers protective acquired immunity against invasive aspergillosis in dogs and people and in murine models of the infection. This finding may help to explain the development of invasive aspergillosis in systemically immunocompetent cats. PMID:27576043
Guo, R; Lim, C K; Peters, T J
Rapid, sensitive and specific high-performance liquid chromatographic assays are described for protoporphyrinogen oxidase and ferrochelatase in human leucocytes. The enzyme reaction products were separated and quantitated by reversed-phase high-performance liquid chromatography with fluorescence detection. The optimal pH for the protoporphyrinogen oxidase assay was 8.6 and the Michaelis constant for protoporphyrinogen IX was 9.78 +/- 0.96 microM (mean +/- S.D.). The mean (+/- S.D.) activity of protoporphyrinogen oxidase in fourteen apparently healthy subjects was 0.146 +/- 0.023 nmol protoporphyrin IX per min per mg protein. In one patient with variegate porphyria, the activity was 0.028 nmol protoporphyrin IX per min per mg protein. The optimal pH for ferrochelatase was 7.4 and with protoporphyrin and Zn2+ as substrates, the Michaelis constants were 1.49 and 8.33 microM, respectively. The mean activity of ferrochelatase in ten control subjects was 0.24 nM Zn-protoporphyrin or 2.05 nM Zn-mesoporphyrin formed per h per mg protein. PMID:1939451
Tournoy, K G; Depraetere, S; Pauwels, R A; Leroux-Roels, G G
The innate immune system of severe combined immunodeficient (SCID) mice represents an important barrier to the successful engraftment of human cells. Different genetic and pharmacological strategies improve the graft survival. Non-obese diabetic (NOD)-SCID mice are better hosts for reconstitution with human peripheral blood leucocytes (Hu-PBL) because of their reduced natural killer cell and macrophage activity next to defective T and B cell functions. We investigated effects of TM-β1, a rat monoclonal antibody recognizing the mouse IL-2 receptor β-chain, on Hu-PBL survival and function in NOD-SCID and SCID mice. Relative to untreated littermates, TM-β1 improved Hu-PBL survival in SCID and NOD-SCID mice. Moreover, TM-β1-pretreated NOD-SCID mice displayed significantly better Hu-PBL survival and tissue distribution than TM-β1-pretreated SCID mice. Irradiation of NOD-SCID mice further enhanced the effects of TM-β1. However, these animals died within 3 weeks post-grafting due to graft-versus-host disease. Secondary immune responses were evaluated with Hu-PBL from a donor immune to hepatitis B surface antigen (HBsAg). In TM-β1-pretreated NOD-SCID mice, human HBsAg-specific memory B cells produced high titres of anti-HBsAg immunoglobulin irrespective of the administration of a secondary antigen booster dose. This contrasts with secondary immune responses in TM-β1-pretreated SCID mice where high titred antigen-specific immunoglobulins were produced when the appropriate antigen booster was given. In conclusion, reducing the function of the innate immune system in immunodeficient mice improves survival of the human graft and can result in an activation of the memory B cells without the need for recall antigen exposure. PMID:10606988
Guerrieri, Diego; Tateosian, Nancy L; Maffía, Paulo C; Reiteri, Romina M; Amiano, Nicolás O; Costa, María J; Villalonga, Ximena; Sanchez, Mercedes L; Estein, Silvia M; Garcia, Verónica E; Sallenave, Jean-Michel; Chuluyan, Héctor E
Serine leucocyte proteinase inhibitor (SLPI) is the main serine proteinase inhibitor produced by epithelial cells and has been shown to be a pleiotropic molecule with anti-inflammatory and microbicidal activities. However, the role of SLPI on the adaptive immune response is not well established. Therefore, we evaluated the effect of SLPI on lymphocyte proliferation and cytokine production. Human peripheral blood mononuclear cells (PBMC) were treated with mitogens plus SLPI and proliferation was assessed by [(3) H]thymidine uptake. The SLPI decreased the lymphocyte proliferation induced by interleukin-2 (IL-2) or OKT3 monoclonal antibodies in a dose-dependent manner. Inhibition was not observed when depleting monocytes from the PBMC and it was restored by adding monocytes and SLPI. SLPI-treated monocyte slightly decreased MHC II and increased CD18 expression, and secreted greater amounts of IL-4, IL-6 and IL-10 in the cell culture supernatants. SLPI-treated monocyte culture supernatant inhibited the CD4(+) lymphocyte proliferation but did not affect the proliferation of CD8(+) cells. Moreover, IL-2 increased T-bet expression and the presence of SLPI significantly decreased it. Finally, SLPI-treated monocyte culture supernatant dramatically decreased interferon-γ but increased IL-4, IL-6 and IL-10 in the presence of IL-2-treated T cells. Our results demonstrate that SLPI target monocytes, which in turn inhibit CD4 lymphocyte proliferation and T helper type 1 cytokine secretion. Overall, these results suggest that SLPI is an alarm protein that modulates not only the innate immune response but also the adaptive immune response. PMID:21574992
Guerrieri, Diego; Tateosian, Nancy L; Maffía, Paulo C; Reiteri, Romina M; Amiano, Nicolás O; Costa, María J; Villalonga, Ximena; Sanchez, Mercedes L; Estein, Silvia M; Garcia, Verónica E; Sallenave, Jean-Michel; Chuluyan, Héctor E
Serine leucocyte proteinase inhibitor (SLPI) is the main serine proteinase inhibitor produced by epithelial cells and has been shown to be a pleiotropic molecule with anti-inflammatory and microbicidal activities. However, the role of SLPI on the adaptive immune response is not well established. Therefore, we evaluated the effect of SLPI on lymphocyte proliferation and cytokine production. Human peripheral blood mononuclear cells (PBMC) were treated with mitogens plus SLPI and proliferation was assessed by [3H]thymidine uptake. The SLPI decreased the lymphocyte proliferation induced by interleukin-2 (IL-2) or OKT3 monoclonal antibodies in a dose-dependent manner. Inhibition was not observed when depleting monocytes from the PBMC and it was restored by adding monocytes and SLPI. SLPI-treated monocyte slightly decreased MHC II and increased CD18 expression, and secreted greater amounts of IL-4, IL-6 and IL-10 in the cell culture supernatants. SLPI-treated monocyte culture supernatant inhibited the CD4+ lymphocyte proliferation but did not affect the proliferation of CD8+ cells. Moreover, IL-2 increased T-bet expression and the presence of SLPI significantly decreased it. Finally, SLPI-treated monocyte culture supernatant dramatically decreased interferon-γ but increased IL-4, IL-6 and IL-10 in the presence of IL-2-treated T cells. Our results demonstrate that SLPI target monocytes, which in turn inhibit CD4 lymphocyte proliferation and T helper type 1 cytokine secretion. Overall, these results suggest that SLPI is an alarm protein that modulates not only the innate immune response but also the adaptive immune response. PMID:21574992
Theodorou, Georgios; Politis, Ioannis
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. PMID:27600972
Kverneland, Anders H; Streitz, Mathias; Geissler, Edward; Hutchinson, James; Vogt, Katrin; Boës, David; Niemann, Nadja; Pedersen, Anders Elm; Schlickeiser, Stephan; Sawitzki, Birgit
Flow cytometry is now accepted as an ideal technology to reveal changes in immune cell composition and function. However, it is also an error-prone and variable technology, which makes it difficult to reproduce findings across laboratories. We have recently developed a strategy to standardize whole blood flow cytometry. The performance of our protocols was challenged here by profiling samples from healthy volunteers to reveal age- and gender-dependent differences and to establish a standardized reference cohort for use in clinical trials. Whole blood samples from two different cohorts were analyzed (first cohort: n = 52, second cohort: n = 46, both 20-84 years with equal gender distribution). The second cohort was run as a validation cohort by a different operator. The "ONE Study" panels were applied to analyze expression of >30 different surface markers to enumerate proportional and absolute numbers of >50 leucocyte subsets. Indeed, analysis of the first cohort revealed significant age-dependent changes in subsets e.g. increased activated and differentiated CD4(+) and CD8(+) T cell subsets, acquisition of a memory phenotype for Tregs as well as decreased MDC2 and Marginal Zone B cells. Males and females showed different dynamics in age-dependent T cell activation and differentiation, indicating faster immunosenescence in males. Importantly, although both cohorts consisted of a small sample size, our standardized approach enabled validation of age-dependent changes with the second cohort. Thus, we have proven the utility of our strategy and generated reproducible reference ranges accounting for age- and gender-dependent differences, which are crucial for a better patient monitoring and individualized therapy. © 2016 International Society for Advancement of Cytometry. PMID:27144459
Precipitation is a primary determinant of terrestrial ecosystem structure and function and as such, precipitation-use efficiency (PUE) is a key determinant of aboveground net primary production (ANPP). We used 76 datasets to contrast ANPP and PUE estimates between northern (southeast Montana) and s...
Henriksson, R.; Nilsson, S.; Colleen, S.; WersÃ¤ll, P.; Helsing, M.; Zimmerman, R.; Engman, K.
Metastatic renal cell carcinoma (RCC) has a poor prognosis. Conventional treatment strategies, including chemotherapy and hormonal therapy, have limited value. Although encouraging results have been achieved in terms of objective response using immunological manipulations, no conclusive studies yet exist with a controlled comparative evaluation of survival. Therefore, the present study was undertaken, which compared one of the present (and presumed best) treatments, interleukin 2/interferon-alpha (IL-2/IFN-alpha) and tamoxifen, with a control arm of tamoxifen only. Tamoxifen has been shown to potentiate in vivo anti-tumour activity of IL-2, and because of its non-toxic behaviour it was included in both groups. The study was open, randomized and included seven institutions in Sweden. The patients were stratified according to the different centres involved. An interim analysis was planned when a minimum of 100 patients were evaluable. The 128 patients finally included had a histologically documented metastatic RCC, with a life expectancy of more than 3 months, a performance status WHO 0-2 and no prior chemo- or immunotherapy. Informed consent was obtained from each patient. The patients randomized to the control arm (n = 63) received only tamoxifen 40 mg p.o. daily for at least 1 year or until progression. The patients (n = 65) randomized to biotherapy received subcutaneous recombinant IL-2, leucocyte IFN-alpha in a treatment cycle of 42 days, as well as tamoxifen p.o. In the absence of undue toxicity or disease progression, these patients received one additional treatment cycle of 42 days followed by maintenance treatment, consisting of 5 days therapy every 4 weeks, for 1 year, or until proven progression. Only two patients in the tamoxifen-only group received immunotherapy when the disease progressed, but without any beneficial effect. All patients received appropriate local treatment when indicated. The interim analysis demonstrated no survival advantage for
Jensen, Kristoffer Jarlov; Fisker, Ane Bærent; Andersen, Andreas; Sartono, Erliyani; Yazdanbakhsh, Maria; Aaby, Peter; Erikstrup, Christian; Benn, Christine Stabell
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. PMID:26678511
Daish, A; Starling, G C; McKenzie, J L; Nimmo, J C; Jackson, D G; Hart, D N
A new monoclonal antibody, CMRF-35, has been generated that recognized a 224 amino acid cell surface protein which is a novel member of the immunoglobulin gene superfamily. The antibody, raised against large granular lymphocytes (LGL), stains LGL, monocytes, macrophages and granulocytes but not platelets or erythrocytes. In addition, a subset of peripheral blood T lymphocytes (26.6 +/- 13.4% CD5+ cells) and B lymphocytes (13.7 +/- 6.8% CD20+ cells) stained with CMRF-35 but tonsil T and B cells were essentially negative. Expression of the CMRF-35 antigen (Ag) on different leucocyte populations was markedly influenced by stimulation of the cells with mitogens and cytokines. Activation of peripheral blood T cells with phytohaemagglutinin (PHA), or phorbol myristate acetate (PMA) and calcium ionophore (CaI) led to a decrease in the proportion of CMRF-35+ T lymphocytes. In contrast, PHA activation of tonsil T lymphocytes resulted in an increase in CMRF-35 Ag expression (47.1 +/- 1.5% CD5 cells at 6 days). An increase in CMRF-35 Ag was also seen on phorbol ester and CaI-activated tonsil B cells. No change in CMRF-35 expression on natural killer (NK) cells occurred following activation with interleukin-2 (IL-2) but the CMRF-35 Ag was down-regulated following Fc receptor stimulation. A moderate increase in CMRF-35 expression occurred during monocyte-macrophage differentiation and the expression of the Ag on monocytes was differentially regulated by interferon-gamma (IFN-gamma). This regulation of the CMRF-35 Ag on the leucocyte surface suggests that the molecule has an important function common to diverse leucocyte types. PMID:8509141
Birnbaum, Jürgen; Klotz, Edda; Spies, Claudia D; Lorenz, Björn; Stuebs, Patrick; Hein, Ortrud Vargas; Gründling, Matthias; Pavlovic, Dragan; Usichenko, Taras; Wendt, Michael; Kox, Wolfgang J; Lehmann, Christian
Introduction Dopexamine may be a therapeutic option to improve hepatosplanchnic perfusion in sepsis. To investigate this possibility, we administered dopexamine in an experimental sepsis model in rats. Methods This prospective, randomized, controlled laboratory study was conducted in 42 Wistar rats. The animals were divided into three groups. Group 1 served as the control group (CON group). The animals in both groups 2 (LPS group) and 3 (DPX group) received an endotoxin (lipopolysaccharide from Escherichia coli – LPS) infusion (20 mg/kg for 15 minutes). DPX group additionally received dopexamine (0.5 μg/kg per minute over four hours). One half of the animals in each group underwent studies of intestinal microvascular blood flow (IMBF) using laser Doppler fluxmetry. In the other half an intravital microscopic evaluation of leucocyte-endothelial cell interaction in intestinal microcirculation was conducted. Functional capillary density (FCD) in the intestinal mucosa and in the circular as well as longitudinal muscle layer was estimated. Results One hour after endotoxin challenge, IMBF decreased significantly in LPS group to 51% compared with baseline (P < 0.05). In DPX group (endotoxin plus dopexamine) we found IMBF values significantly higher than those in LPS group (approximately at the level of controls). The impaired FCD following endotoxin challenge was improved by dopexamine in the longitudinal muscle layer (+33% in DPX group versus LPS group; P < 0.05) and in the circular muscle layer (+48% in DPX group versus LPS group; P < 0.05). In DPX group, dopexamine administration reduced the number of firmly adherent leucocytes (-31% versus LPS group; P < 0.05). Plasma levels of tumour necrosis factor-α were reduced by dopexamine infusion (LPS group: 3637 ± 553 pg/ml; DPX group: 1933 ± 201 pg/ml) one hour after endotoxin challenge. Conclusion Dopexamine administration improved IMBF and FCD (markers of intestinal microcirculation) and reduced leucocyte activation
King, Nia; Dewey, Cate; Borish, David
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
The influence of enzyme induction on acute toxicity of aniline and 4-chloroaniline to rainbow trout (Salmo gairdneri) was investigated. or these two xenobiotics, bioactivation reaction are known to occur in mammals. nduction of cytochrome P450 mixed-function oxidase was obtained ...
Favoino, E; Favia, I E; Vettori, S; Vicenti, C; Prete, M; Valentini, G; Perosa, F
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
Lepidi, H; Benoliel, A M; Mege, J L; Bongrand, P; Capo, C
Uniform concentrations of chemoattractants such as formylpeptides induced a morphological polarization of human polymorphonuclear leucocytes (PMNs) and a concentration of F-actin at the cell front. They also induced a transient increase in filamentous actin (F-actin) which preceded the cell shape change. We combined fluorescence microscopy and image analysis to study the localization of F-actin, as revealed by a specific probe (bodipyTM phallacidin) in suspended PMNs stimulated by chemoattractants. F-actin exhibited remarkable concentration in focal points after a 30 s exposure to 10(-8) M formylmethionyl-leucyl-phenylalanine (fMet-Leu-Phe), although no shape change of PMNs was detectable. A 10-min incubation with formylpeptide (10(-6) to 10(-9) M) induced the morphological polarization of PMNs and the appearance of a principal focus of F-actin in the cell head region and a secondary focus in the cell posterior end. The distribution of F-actin-associated fluorescence in 2D images of polarized PMNs might be due to an actual concentration of F-actin in privileged areas, to a local concentration of plasma membrane drawing filamentous actin or to variations in the cell volume. Then, we studied the distribution of a cytoplasmic marker, fluorescein diacetate and a membrane probe, TMA-DPH, in unstimulated rounded PMNs and in spherical and morphologically polarized PMNs stimulated by formylpeptide. The distribution of neither of these probes was correlated with F-actin distribution, especially in rounded PMNs stimulated 30 s with 10(-8) M fMet-Leu-Phe, suggesting that F-actin was concentrated in two foci located in the cell head region and in the cell posterior end. In addition, zymosan-activated serum induced the morphological polarization of PMNs and the appearance of two foci of filamentous actin, demonstrating that binding of formylpeptide to its specific receptor was not required for F-actin reorganization. We conclude that the accumulation of F-actin probably
Atherton, Anne; Born, G. V. R.
1. A new simple method is described for quantitating the adhesiveness of circulating polymorphonuclear leucocytes, or granulocytes, to the walls of blood vessels. The cheek pouch of anaesthetized hamsters or a small part of the mesentery of anaesthetized mice were prepared for continuous microscopic observation of selected venules. Those granulocytes which moved sufficiently slowly to be individually visible were counted for 1 or 2 min periods as they rolled past a selected point on one side of a vessel. The velocity distribution of these cells was determined by analysing films. Films were used also to measure mean blood flow velocity in the venules by observing embolizing platelet thrombi induced by the iontophoretic application of adenosine diphosphate. Emigration of granulocytes into the tissues was quantitated by enumerating them in standard areas of stained histological sections. 2. In control experiments with hamster cheek pouch venules, the rolling granulocyte count usually passed through a maximum shortly after the preparation was set up and then fell to a low constant value. In mouse mesentery venules the count remained at a low approximately constant value from the beginning for at least 3 hr. 3. The mean velocity of blood flow in the venules was between 900 and 200 μ/sec. All rolling granulocytes moved much more slowly; in hamster cheek pouch venules the mean velocity was about 20 μ/sec and in mouse mesentery venules about 10 μ/sec. Around these means the velocity distribution of individual cells was narrow. 4. Rolling of granulocytes was abolished by superfusing ethylenediamine tetra-acetate (EDTA, 0·1 M) suggesting that the phenomenon depends on calcium or magnesium ions. 5. Agents were applied locally to the observed venules. Human serum albumin, trypsin or histamine in high concentrations did not affect the rolling granulocyte count. 6. The rolling granulocyte count was increased during the application of Hammarsten casein or Escherichia coli
Wang, Yonghong; Yang, Xukui; Yang, Yuanyuan; Wang, Wenjun; Zhao, Meiling; Liu, Huiqiang; Li, Dongyan; Hao, Min
Objective: To identify the specific microRNA (miRNA) biomarkers of preeclampsia (PE), the miRNA profiles analysis were performed. Study Design: The blood samples were obtained from five PE patients and five normal healthy pregnant women. The small RNA profiles were analyzed to identify miRNA expression levels and find out miRNAs that may associate with PE. The quantitative reverse transcriptase–PCR (qRT-PCR) assay was used to validate differentially expressed peripheral leucocyte miRNAs in a new cohort. Result: The data analysis showed that 10 peripheral leucocyte miRNAs were significantly differently expressed in severe PE patients. Four differently expressed miRNAs were successfully validated using qRT-PCR method. Conclusion: We successfully constructed a model with high accuracy to predict PE. A combination of four peripheral leucocyte miRNAs has great potential to serve as diagnostic biomarkers of PE. PMID:26675000
Morcillo, Patricia; Meseguer, José; Esteban, M Ángeles; Cuesta, Alberto
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. PMID:27041665
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
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
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
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
Ijiri, Y.; Matsumoto, K.; Kamata, R.; Nishino, N.; Okamura, R.; Kambara, T.; Yamamoto, T.
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
Schöngen, R N; Giannetti, B M; van de Leur, E; Reinards, R; Greiling, H
1. The influence of diacetylrhein on the luminol-induced chemiluminescence of zymosan-activated polymorphonuclear leucocytes (PMNL) was investigated. At a concentration of 4 x 10(-5) mol/l diacetylrhein an inhibition of about 40% was found. 2. A model for the degradation of hyaline cartilage by frustrated phagocytosis was developed, in which human polymorphonuclear leucocytes cause a release of glycosaminoglycan peptides from hyaline cartilage slices (bovine nasal septum). We observed a 20% inhibition of this release at a concentration of 10(-4) mol/l diacetylrhein. 3. Human synovial fibroblasts synthesize the glycosaminoglycan hyaluronate. As a parameter of the rate of hyaluronate synthesis we measured the incorporation of 14C-glucosamine into hyaluronate. At a concentration of 2 x 10(-4) mol/l diacetylrhein a 4-fold increase of 14C-glucosamine incorporation in the membrane fraction of the synovial cells (tryptic fraction) and a 1.6-fold elevation of glucosamine release into the medium was measured. The synovial fibroblasts show a higher (1.5-fold) glucose consumption and lactate production in the presence of diacetylrhein (2 x 10(-4) mol/l). PMID:3415721
Turaihi, K.; Baron, D.N.; Dandona, P.
/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.
Giro, S N; Joshi, B; Peoples, S A
1. The effects of intraperitoneal injections of actinomycin D on the temporal characteristics of the accumulation of the inflammatory exudate and cells into the peritoneal and pleural cavities were studied in male Sprague Dawley rats. 2. A measurable quantity of the exudate appeared in both cavities within 24 h and reached maxima in the peritoneal and pleural cavities on the fourth and third days, respectively. Thereafter, the accumulated volume of liquid decreased progressively in the peritoneal cavity but stayed more or less at about the same level in the pleural cavity until the sixth day. 3. The pooled peritoneal and pleural exudates contained neutrophils, macrophages, mast cell and eosinophils. The leucocyte infiltration occurred in two phases, the maximum cell numbers being found on the third and fifth days. A precipitous fall in the number of leucocytes occurred on the fourth day. Neutrophils and macrophages accounted for 85-95% of the total number of leucocytes. 4. The supernatant of the inflammatory exudate after centrifugation at 3,000 g contained histamine and the soluble lysosomal enzyme proteins, acid phosphatase and beta-glucuronidase until the sixth day following the initial dose of actinomycin D. 5. It is suggested that the release of lysosomal enzymes in the exudate, subsequent to leucocyte mobilization and the release of histamine from the mast cells, are probably involved in the genesis of inflammatory conditions induced by actinomycin D. PMID:50158
Fridlin, Ann; vanDiedenhoven, Bastiaan; Ackerman, Andrew S.; Avramov, Alexander; Mrowiec, Agnieszka; Morrison, Hugh; Zuidema, Paquita; Shupe, Matthew D.
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.
Jackson, N; Ling, N R; Ball, J; Bromidge, E; Nathan, P D; Franklin, I M
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
Cason, J; Ainley, C C; Wolstencroft, R A; Thompson, R P
The adherence of polymorphonuclear leucocytes (PMN) to nylon fibre was investigated in patients with Crohn's disease, ulcerative proctocolitis, and anorexia nervosa, and compared with changes of circulating PMNs, C reactive protein concentrations, erythrocyte sedimentation rates, and clinical assessment of disease activity. PMN adherence was in excess of the maximum value detected for healthy subjects in 14 of 25 patients with Crohn's disease and two of 10 with proctocolitis, but it was within the normal range for all eight with anorexia nervosa. High adherence in Crohn's disease, however, was not associated with quantitative or qualitative changes of PMN populations, absolute concentrations of C reactive protein, erythrocyte sedimentation rates, disease severity, drug regimens, malnutrition, or zinc deficiency. High PMN adherence in Crohn's disease may therefore reflect the activation in vivo of normal PMN by humoral factors. PMID:3360954
Gladbach, Anja; Gladbach, David Joachim; Quillfeldt, Petra
The analysis of plasma biochemistry and haematology to monitor the condition of birds in the wild has been found a useful tool in ecological research. Despite biparental investment in most wild birds studied, some studies of condition indices found sex differences, and attributed these to the costs of egg formation or brooding in females or a higher contribution of males to chick rearing. We studied the natural variation of haematological and plasma biochemistry parameters (namely leucocyte, lymphocyte and heterophil counts, H/L ratio and plasma concentrations of proteins, triglycerides and carotenoids) in relation to the different measures of parental investment in males and females in the Upland goose (Chloephaga picta leucoptera), a socially monogamous species. We found no sex differences in haematological and most plasma biochemistry parameters, but a relation to different aspects of parental investment in breeding male and female Upland geese. H/L ratios were related to body condition and capture date in males while leucocyte counts, plasma protein and plasma carotenoid concentrations varied with clutch measures and hatching date in females. Higher H/L ratios of males in a low body condition and later in the year may reflect stress associated with the investment into the establishment and defence of the breeding territory. Females with higher clutch volumes had lower total leucocyte and lymphocyte numbers and higher levels of plasma protein. Earlier hatching dates were associated with lower numbers of all leucocyte types and higher values of plasma carotenoid concentrations. This indicates that differences in health state are reflected in reproductive performance in female Upland geese. We also found sexual differences in the repeatability of haematological and plasma biochemistry parameters between years and therefore suggest that their potential as a measure of individual quality differs between male and female Upland geese. Finally, numbers of leucocyte
Lindstedt, Sandra; Malmsjö, Malin; Ingemansson, Richard
Right ventricular heart rupture is a devastating complication associated with negative pressure wound therapy (NPWT) in cardiac surgery. The use of a rigid barrier disc (HeartShield™) has been suggested to offer protection against this lethal complication by preventing the heart from being drawn up by the negative pressure and damaged by the sharp sternum bone edges. Seven patients treated with conventional NPWT and seven patients treated with NPWT with a protective barrier disc (HeartShield) were compared with regard to bacterial clearance and infection parameters including C-reactive protein levels and leucocyte counts. C-reactive protein levels and leucocyte counts dropped faster and bacterial clearance occurred earlier in the HeartShield® group compared with the conventional NPWT group. Negative biopsy cultures were shown after 3·1 ± 0·4 NPWT dressing changes in the HeartShield group, and after 5·4 ± 0·6 NPWT dressing changes in the conventional NPWT group (P < 0·001). All patients were followed up with clinical check-up after 3 months. None of the patients in the HeartShield group had any signs of reinfection such as deep sternal wound infection (DSWI) or sternal fistulas, whereas in the conventional NPWT group, two patients had signs of sternal fistulas that demanded hospitalisation. HeartShield hinders the right ventricle to come into contact with the sharp sternal edges during NPWT and thereby protects from heart damage. This study shows that using HeartShield is beneficial in treating patients with DSWI. Improved wound healing by HeartShield may be a result of the efficient drainage of wound effluents from the thoracic cavity. PMID:23651118
Weckmann, M; Schultheiss, C; Hollaender, A; Bobis, I; Rupp, J; Kopp, M V
Recent data suggest a possible relationship between cystic fibrosis (CF) pharmacotherapy, Aspergillus fumigatus colonization (AC) and/or allergic bronchopulmonary aspergillosis (ABPA). The aim of this study was to determine if anti-fungal defence mechanisms are influenced by CF pharmacotherapy, i.e. if (1) neutrophils form CF and non-CF donors differ in their ability to produce chitotriosidase (CHIT-1); (2) if incubation of isolated neutrophils with azithromycin, salbutamol, prednisolone or rhDNase might influence the CHIT-1 activity; and (3) if NETosis and neutrophil killing efficiency is influenced by rhDNase. Neutrophils were isolated from the blood of CF patients (n = 19; mean age 26·8 years or healthy, non-CF donors (n = 20; 38·7 years) and stimulated with phorbol-12-myristate-13-acetate (PMA), azithromycin, salbutamol, prednisolone or rhDNase. CHIT-1 enzyme activity was measured with a fluorescent substrate. NETosis was induced by PMA and neutrophil killing efficiency was assessed by a hyphae recovery assay. Neutrophil CHIT-1 activity was comparable in the presence or absence of PMA stimulation in both CF and non-CF donors. PMA stimulation and preincubation with rhDNase increased CHIT-1 activity in culture supernatants from non-CF and CF donors. However, this increase was significant in non-CF donors but not in CF patients (P < 0·05). RhDNase reduced the number of NETs in PMA-stimulated neutrophils and decreased the killing efficiency of leucocytes in our in-vitro model. Azithromycin, salbutamol or prednisolone had no effect on CHIT-1 activity. Stimulation of isolated leucocytes with PMA and treatment with rhDNase interfered with anti-fungal defence mechanisms. However, the impact of our findings for treatment in CF patients needs to be proved in a clinical cohort. PMID:27324468
Brodzki, P; Kostro, K; Brodzki, A; Niemczuk, K; Lisiecka, U
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. PMID:25124985
Acosta, Andrés Martin; Hamedani, Farid Saei; Kajdacsy-Balla, Andre; Wiley, Elizabeth Louise
Mixed adenoneuroendocrine carcinomas (MANECs) of the biliary tract are rare tumors, and to date only a few cases arising in the gallbladder have been reported. Their histogenesis is a matter of debate, since the biliary tract normally lacks neuroendocrine cells. However, the immunohistochemical identification of nonneoplastic neuroendocrine cells in both biliary adenocarcinomas and intestinal metaplasia has been documented. Here we report a case of a 55-year-old female patient presenting with right upper quadrant pain, cholelithiasis, and a gallbladder mass identified after cholecystectomy. The histopathologic examination showed a MANEC, composed of an intestinal-type adenocarcinoma and a large cell neuroendocrine carcinoma, arising in a background of enteric metaplasia with extensive high-grade dysplasia. Moreover, we report the presence of focal pancreatic intraepithelial neoplasia-like epithelial lesions, which has not been described for these tumors yet. The histopathologic features of this case provide further support for the theory that MANECs arise following a metaplasia-dysplasia-carcinoma pathway. PMID:25838330
Williams, Christopher; Holbrook, Rachel; Lindner, Robyn; Reeve, James; Das, Anurina; Maher, Christopher
Background Many patients with back pain do not receive health care in accordance with best practice recommendations. Implementation trials to address this issue have had limited success. Despite the known effectiveness of clinical decision support systems (CDSS), none of these are available for back pain management. Objective The objective of our study was to develop a Web-based CDSS to support Australian general practitioners (GPs) to diagnose and manage back pain according to guidelines. Methods Asking a panel of international experts to review recommendations for sixteen clinical vignettes validated the tool. It was then launched nationally as part of National Pain Week and promoted to GPs via a media release and clinic based visits. Following this, a mixed methods evaluation was conducted to determine tool feasibility, acceptability, and utility. The 12 month usage data were analyzed, and in-depth, semistructured interviews with 20 GPs were conducted to identify barriers and enablers to uptake. Results The tool had acceptable face validity when reviewed by experts. Over a 12 month period there were 7125 website visits with 4503 (63.20%) unique users. Assuming most unique users are GPs, around one quarter of the country’s GPs may have used the tool at least once. Although usage was high, GP interviews highlighted the sometimes complex nature of management where the tool may not influence care. Conversely, several “touch-points”, whereby the tool may exert its influence, were identified, most notably patient engagement. Conclusions A novel CDSS tool has the potential to assist with evidence-based management of back pain. A clinical trial is required to determine its impact on practitioner and patient outcomes. PMID:24694921
Ganeshpandian, Mani; Ramakrishnan, Sethu; Palaniandavar, Mallayan; Suresh, Eringathodi; Riyasdeen, Anvarbatcha; Akbarsha, Mohammad Abdulkadher
A series of mononuclear mixed ligand copper(II) complexes of the type [Cu(L)(2,9-dmp)](ClO4)21-4, where L is a tridentate 3N ligand such as diethylenetriamine (L1) (1) or N-methyl-N'-(pyrid-2-yl-methyl)ethylenediamine (L2) (2) or di(2-picolyl)amine (L3) (3) or bis(pyrid-2-ylmethyl)-N-methylamine (L4) (4) and 2,9-dmp is 2,9-dimethyl-1,10-phenanthroline, has been isolated and characterized. The complexes 1 and 3 possess square-based pyramidal coordination geometry. Absorption spectral studies reveal that the intrinsic DNA binding affinity varies as 1>2>3>4. The higher DNA binding affinity of 1 arises from L1, which offers lower steric hindrance toward intercalation of 2,9-dmp co-ligand into DNA base pairs and is involved in hydrogen bonding interaction with DNA. Interestingly, all the complexes cleave pUC19 supercoiled DNA in the absence of an activating agent. They also exhibit oxidative (H2O2) DNA cleavage ability, which varies as 1>2>3>4, the highest cleavage efficiency of 1 being due to the largest amount of ROS it generates. The tryptophan emission-quenching experiment reveals that the stronger binding of 3 and 4 with bovine serum albumin (BSA) in the hydrophobic region, which is in line with DNA viscosity measurements. The IC50 values of 1-4 for MCF-7 breast cancer cell line are lower than that of cisplatin. Flow cytometry analysis reveals that 1 mediates the arrest of S and G2/M phases in the cell cycle progression at 24h harvesting time, which progresses into apoptosis. Hoechst 33258 staining studies indicate the higher potency of 1 to induce apoptosis. PMID:25151036
Pardo, Javier; Aisa, Gregorio; de Alava, Enrique; Sola, Jesús J; Panizo, Angel; Rodríguez-Spiteri, Natalia; García, Juan L; Torre, Wenceslao
Carcinosarcomas are malignant tumors with a mixture of carcinomatous and differentiated sarcomatous elements. We investigate the morphology, immunohistochemistry, and comparative genomic hybridization analysis of 3 mixed squamous carcinoma and osteosarcoma of the lung. All patients were male and their ages were 72, 43, and 58 years. The sizes of the neoplasms were 7, 5, and 5 cm in maximum diameter, respectively. Two patients died of the disease 9 and 14 months after surgery; and one is alive 6 months later. By light microscopy, all cases had both squamous and osteosarcomatous structures. Immunohistochemistry was positive for AE3AE1, p63, 34 E12, CAM 5.2 (2/3 cases), CK-7 (2/3 cases), epithelial membrane antigen, E-cadherin, p53, and carcinogenic embryonic antigen in carcinomatous areas, and for vimentin and CD-68 in sarcomatous component. Areas of transition positive for both cytokeratins and vimentin were seen in all cases. A total of 55 copy number changes were detected with a median of 18 abnormalities per case: 48 gains, 6 losses, and 1 high-level amplification. Chromosome alterations in osteosarcomatous areas were similar to those found in lung metastatic osteosarcoma, comparable to those found in carcinomatous areas and to lung squamous carcinomas. Coincidences between carcinomatous areas and osteosarcomatous zones were found as gains in chromosomes 1q, 3q, 5p, 8q, and 12p. These findings provide arguments that favor a common origin for both types of cells, supported by the mixture of cells, the existence of undifferentiated cells positive to both cytokeratin and vimentin markers, and the CGH overlaps of chromosomal gains between carcinomatous and sarcomatous areas. PMID:18382357
Dow, Rustam; Barnsley, Jan; Tu, Karen; Domb, Sharon; Jadad, Alejandro R.; Lemieux-Charles, Louise
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
Jiang, Hailiang; Ying, Liyan; Hao, Yueli; Zhou, Saichun; Shen, Haoyu; Tang, Ying; Bao, Yazhen
Here, we report on a procedure in which ultrasonic waves were used to mix a water-ionic liquid two-phase system to simultaneously derivatize amines with dansyl chloride and extract, and the resulting dansyl amines into an ionic liquid (IL) at room temperature within 15 min. The ILs used were room-temperature ionic liquids (RTILs), such as 1-alkyl-3-methylimidazolium hexafluorophosphate ([C(n)MIM][PF(6)], n = 4, 6, 8), which are insoluble in water, and can extract more than 88% of the total dansyl amines (Dns-amines) present in aqueous solution. The Dns-amines formed were separated by chromatography in 20 µL of RTIL in a mobile-phase gradient of acetonitrile-water flowing through an Agilent Zorbax SB-C18 column. The eluents were analyzed at 333 nm using a UV detector. The optimal analytical conditions were achieved by using the following: RTILs with a 4-carbon alkyl chain; a 7-mL water sample and 1 mL each of 5 mM dansyl chloride in acetone and 20 mM sodium tetraborate buffer for use with a fixed volume (1 mL) of IL; buffer pH of 10.4; duration of 15 min; and an acetonitrile-water ratio of 50:50 from zero time to 3 min, 80:20 from 4 to 10 min, and 50:50 from 11 to 16 min. In comparison to the single-phase hot-water (70°C) derivatization process reported earlier, the simultaneous process developed here showed a higher derivatization efficiency at the tested concentration. The linear range of analysis (0.14-7.14 mg/L) was determined, and the correlation coefficient (R) ranged from 0.9878 to 0.9980. Furthermore, the estimated percentage of recovery from amine-spiked river-water samples ranged from 76 to 108%. The method presented here is simple and fast, and does not involve the use of volatile organic extractants. PMID:21233559
Ford, John A; Jones, Andrew P; Wong, Geoff; Clark, Allan B; Porter, Tom; Shakespeare, Tom; Swart, Ann Marie; Steel, Nicholas
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
Liu, Qi; Cai, Hui-Ying; Jin, Guang-Ze
To accurately quantify forest carbon density and net primary productivity (NPP) is of great significance in estimating the role of forest ecosystems in global carbon cycle. By using the forest inventory and allometry approaches, this paper measured the carbon density and NPP of the virgin broadleaved-Korean pine (Pinus koraiensis) forest and of the broadleaved-Korean pine forest after 34 years selective-cutting (the cutting intensity was 30%, and the cutting trees were in large diameter class). The total carbon density of the virgin and selective-cutting broadleaved-Korean pine forests was (397.95 +/- 93.82) and (355.61 +/- 59.37) t C x hm(-2), respectively. In the virgin forest, the carbon density of the vegetation, debris, and soil accounted for 31.0%, 3.1%, and 65.9% of the total carbon pool, respectively; in the selective-cutting forest, the corresponding values were 31.7%, 2.9%, and 65.4%, respectively. No significant differences were observed in the total carbon density and the carbon density of each component between the two forests. The total NPP of the virgin and selective-cutting forests was (36.27 +/- 0.36) and (6.35 +/- 0.70) t C x hm(-2) x a(-1), among which, the NPP of overstory, understory, and fine roots in virgin forest and selective-cutting forest accounted for 60.3%, 2.0%, and 37.7%, and 66.1%, 2.0%, and 31.2%, respectively. No significant differences were observed in the total NPP and the contribution rate of each component between the two forests. However, the ratios of the needle and broadleaf NPPs of the virgin and selective-cutting forests were 47.24:52.76 and 20.48:79.52, respectively, with a significant difference. The results indicated that the carbon density and NPP of the broadleaved-Korean pine forest after 34 years selective-cutting recovered to the levels of the virgin broadleaved-Korean pine forest. PMID:24483061
Dryden, Alexander W S; Rana, Mrinal; Pandey, Pravin
A 49-year-old diabetic man presented with a 2-day history of a painful right eye associated with a purulent discharge. Prior to becoming symptomatic, he reported that someone spat at him, resulting in direct contact between the saliva and his affected eye. Gram stain revealed numerous leucocytes with Gram-negative diplococci, and culture yielded Neisseria meningitidis (serogroup C). There was no evidence of any systemic infection, and blood cultures were negative for any growth. He was treated for primary meningococcal conjunctivitis (PMC) with intensive topical antibiotic eyedrops as well as systemic antibiotics. One week after commencing treatment he remained systemically well and his symptoms had fully resolved. PMID:27330479
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
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 leucocytes. We conclude that HNG 1) protects CP-induced loss of male germ cells and leucocytes, 2) enhances CP-induced suppression of cancer metastases, and 3) acts as a caloric-restriction mimetic by suppressing IGF-1 levels. Our findings suggest that humanin analogues may be promising adjuvants to chemotherapy. PMID:26384090
Koivuranta-Vaara, P; Leirisalo-Repo, M; Repo, H
Polymorphonuclear leucocyte (PMN) functions (migration in vitro, chemiluminescence, O-2 production, binding of chemotactic peptide, and aggregation) were studied in HLA-B27 positive patients with previous yersinia arthritis (YA). PMNs of patients whose disease had been severe showed chemokinetic and chemiluminescence responses significantly higher than the PMNs of those with a mild disease. The results support the view that enhanced PMN function contributes to inflammatory symptoms in patients with YA. PMID:3592787
de Bruin, T; de Rooster, H; van Bree, H; Cox, E
Proliferation assays performed on peripheral blood mononuclear cells (PBMC) are commonly used in experimental and clinical immunology. A prerequisite for an in vitro assay is the ability to obtain relatively pure populations of mononuclear cells from whole blood, as contaminating polymorphonuclear cells may affect the proliferation of lymphocytes. Purification of canine leucocytes from whole blood is associated with difficulties in obtaining pure lymphocytes in high yields. The aim of this study was to optimize the lymphocyte purification from canine whole blood in terms of total cell recovery and purity, while not influencing the proliferation capacity of the isolated cells. To acquire optimal isolation of canine lymphocytes several density gradient media of different densities and osmolalities were examined. For optimal phagocyte removal, pre-treatment of whole blood with carbonyl iron/arabic gum and/or adherence to fibrinogen pre-coated polystyrene tissue flasks were examined. Lectin-induced proliferation was used as measurement of cell activity of the obtained cell fractions after the different separation procedures. Canine blood pre-treated with carbonyl iron/arabic gum followed by density gradient centrifugation with medium 'G' (density: 1.079 g/cm(3), osmolality: 256 mOsm) and adherence to pre-coated polystyrene tissue flask obtained the best PBMC cultures with a median lymphocyte purity of 88% and a median yield of recovered lymphocytes of 54%. This culture also resulted in the highest proliferation and subsequently the highest stimulation index upon lectin stimulation. PMID:16268957
Virella, G; Lopes-Virella, M F; Shuler, C; Sherwood, T; Espinoza, G A; Winocour, P; Colwell, J A
Human polymorphonuclear leucocytes (PMN) incubated with surface-bound immune complexes (IC) release a substance that induces platelet aggregation and serotonin-release. This substance was identified as platelet-activating factor (PAF) on the basis of its sensitivity to phospholipase A2 and of its purification by thin-layer chromatography in identical conditions to those used to purify zymosan-induced PAF. We used two types of substrates to absorb our IC:Sepharose particles to which we coupled human serum albumin, and which were later incubated with specific rabbit antiserum to form surface-bound immune complexes, and human erythrocytes, to which soluble IC can be passively adsorbed. Both types of surface-bound IC were found to stimulate the release of PAF by human PMN in the absence of complement. These results suggest that PMN may play a central role in the early stages of IC-induced inflammation: they recognize IC adsorbed to red cells or to any other cell able to adsorb IC, and they induce the activation of platelets and release of vasoactive amines, which leads to the increase of vascular permeability believed to be essential for extravascular IC deposition. PMID:6885111
Cury-Boaventura, M F; Gorjão, R; Martins de Lima, T; Fiamoncini, J; Godoy, A B P; Deschamphs, F C; Soriano, F G; Curi, R
Lipid emulsion (LE) containing medium/ω-6 long chain triglyceride-based emulsion (MCT/ω-6 LCT LE) has been recommended in the place of ω-6 LCT-based emulsion to prevent impairment of immune function. The impact of MCT/ω-6 LCT LE on lymphocyte and neutrophil death and expression of genes related to inflammation was investigated. Seven volunteers were recruited and infusion of MCT/ω-6 LCT LE was performed for 6 h. Four volunteers received saline and no change was found. Blood samples were collected before, immediately afterwards and 18 h after LE infusion. Lymphocytes and neutrophils were studied immediately after isolation and after 24 and 48 h in culture. The following determinations were carried out: plasma-free fatty acids, triacylglycerol and cholesterol concentrations, plasma fatty acid composition, neutral lipid accumulation in lymphocytes and neutrophils, signs of lymphocyte and neutrophil death and lymphocyte expression of genes related to inflammation. MCT/ω-6 LCT LE induced lymphocyte and neutrophil death. The mechanism for MCT/ω-6 LCT LE-dependent induction of leucocyte death may involve changes in neutral lipid content and modulation of expression of genes related to cell death, proteolysis, cell signalling, inflammatory response, oxidative stress and transcription. PMID:21682721
Jones, J; Morgan, B P
The comparative susceptibility of lymphocyte subsets, monocytes and polymorphonuclear leucocytes (PMN) to killing by murine perforin was measured using physical separation techniques, cell-surface phenotyping and scatter characteristics to isolate cell types, together with propidium iodide (PI) uptake as a measure of cell death. In the majority of individuals, PMN were more resistant to perforin than other peripheral blood cells including natural killer (NK) cells and CD8+ lymphocytes. Among the lymphocytes, CD4+ cells were the most susceptible subset, followed by CD19+, CD8+ and CD56+ lymphocytes respectively. The human promyelocytic leukaemia cell line, HL-60, and the human histiocytic lymphoma cell line, U937, were readily killed by perforin. When HL-60 were differentiated to either macrophage- or neutrophil-like end cells, and U937 differentiated to macrophage-like end cells, there was no difference between differentiated and undifferentiated cells in their relative susceptibility to perforin. The relative resistance of PMN to perforin may be important in protecting them from damage in in vivo situations where both NK cells and neutrophils are localized in the same inflammatory areas. PMID:7835917
Guilloteau, L.; Pepin, M.; Pardon, P.; Le Pape, A. )
The recruitment of polymorphonuclear leucocytes (PMNs) during the development of experimental pyogranulomas induced by Corynebacterium pseudotuberculosis was followed in nine male lambs by scintigraphic examination. Autologous blood PMNs were labelled with 99m-technetium or 111-indium and were re-injected intravenously into infected lambs. The functional properties of the labelled cells were monitored (1) in vitro by measuring their phagocytic and bactericidal activity against C. pseudotuberculosis and their chemotaxis under agarose, and (2) in vivo by following scintigraphically their capacity to accumulate in an inflammatory focus induced by intradermal injection of latex beads coated with Salmonella abortus equi lipopolysaccharide. Following inoculation of corynebacteria into the right ear of lambs, radioactive foci were observed to be localized in the right ear and in the draining lymph nodes during the 4 days following inoculation. Histopathological examination performed 32 h after inoculation confirmed the intense accumulation of PMNs at these sites. With the exception of one animal, which presented visible foci in the neck 14 days postinoculation, no radioactive foci were observed during the later phases of experimental infection, despite the presence of multiple pyogranulomas which were confirmed by bacteriological examination after necropsy of the lambs. Histopathological examination of these lesions revealed layers of fibroblasts, lymphocytes, and macrophages surrounding a necrotic centre. The results of these studies suggest that the contribution of PMNs during the chronic phase of inflammation is considerably reduced in comparison with the acute inflammatory phase of the infectious process.
Wakeyama, H; Takeshige, K; Minakami, S
NADPH-dependent 2,6-dichlorophenol-indophenol (DCIP) reductase activity in the homogenate of phagocytosing pig polymorphonuclear leucocytes was twice that of the resting cells and the activity in the phagocytic vesicles corresponded to the activity increment due to phagocytosis. The apparent Km value of the reductase activity in the vesicles for NADPH was 30 microM, which is similar to that of the NADPH-dependent superoxide (O2-) formation. Increasing the DCIP reductase activity by increasing the DCIP concentration caused a decrease in the O2- -forming activity, the NADPH oxidation rate being constant and independent of the dye concentration. p-Chloromercuribenzoate and cetyltrimethylammonium bromide at low concentrations inhibited the O2- -forming activity of the vesicles without inhibiting the DCIP reductase. Quinacrine inhibited both O2- formation and DCIP reduction. The DCIP reductase activity could be extracted with a mixture of deoxycholate and Tween-20, which extracts the O2- -forming activity. The reductase activity in the extract was enhanced 2-fold by the addition of FAD, and its apparent Km was 0.085 microM. These results indicate that the NADPH-dependent DCIP reductase activity of the phagocytic vesicles is catalysed by a flavin-containing component of the O2- -forming system. PMID:6860311
Nagamatsu, Y; Tsujioka, Y; Hashimoto, M; Giddings, J C; Yamamoto, J
Unanswered questions remain with regard to the therapeutic use of aspirin and the selective inhibition of thromboxane A2 and prostacyclin in platelets and endothelial cells. In the present study, the effects of aspirin on platelets and endothelial cells in vivo were examined using a helium-neon (He-Ne) laser-induced thrombosis model. Single intravenous injections of aspirin at concentrations of more than 0.5 mg/kg body weight mediated a dose dependent inhibition of thrombus formation in arterioles but not in venules. This antithrombotic effect was optimum after 15 min and declined after 90 min. Potent antithrombotic activity in arterioles was manifest at doses of 2.5 mg/kg to 50 mg/kg, and initial inhibition of thrombogenesis in vivo was most pronounced at high doses. Oral aspirin also inhibited thrombus formation in arterioles but not in venules, although the antithrombotic effects were delayed and prolonged. Maximum inhibition of ex vivo, collagen induced platelet aggregation by aspirin was observed approximately 180 min after intravenous injection. The results demonstrated that, although aspirin might have differential effects on platelets and endothelial cells, potent antithrombotic activity was manifest in arterioles at all concentrations. The findings suggest that the concept of the aspirin dilemma might be ignored for therapeutic purposes in many clinical circumstances. The antithrombotic effects of aspirin were unchanged in granulocyte-depleted animals, indicating that leucocyte-related mechanisms including neutrophil superoxide anion production did not modulate the potency of aspirin in this model. PMID:10197261
Dufer, J; Trentesaux, C; Desplaces, A
Esterases of human leucocytes and platelets were studied by cytochemical methods. The aim of the study was to clarify the cellular distribution and possible nature of esterases types differing in their substrate specificity and/or their inhibitor sensitivity. 3 substrates (alpha-naphthyl acetate: ANA; naphthol AS-D chloroacetate: NASDCA; and N-acetyl DL-alanine alpha-naphthyl ester: NACALA) were used and the effects of 2 inhibitors (sodium fluoride and the serine protease inhibitor phenyl methyl sulfonyl fluoride: PMSF) were evaluated. 4 enzyme types were described: Type I, present in granulocytes, was detected using NASDCA and NACALA and was resistant to fluoride but sensitive to PMSF. Other types were detected using ANA as substrate. Type II, present in monocytes, was inhibited by both fluoride and PMSF. Type III, present in platelets and plasma cells, was inhibited by fluoride but resistant to PMSF. Type IV, present in lymphocytes, was resistant to both fluoride and PMSF. The specific aims and possible areas for application of these results are discussed. PMID:6364322
Baldwin, C.E.; Stakebake, J.; Peters, M.
Radioactive mixed wastes containing a radioactive component subject to the Atomic Energy Act (AEA) and hazardous waste subject to resource Conservation and Recovery Act (RCRA) are generated, treated, and stored at the Rocky Flats Plant (RFP) and are subject to federal and state statutory and regulatory requirements. The US Environmental Protection Agency (EPA) and the Colorado Department of Health (CDH) are the two primary regulatory agencies which enforce these requirements. This paper describes the mechanism by which RFP will characterize mixed wastes within the LDR provisions of RCRA and the LDR FFCA as well as for meeting the waste acceptance criteria for disposal.
Baldwin, C.E.; Stakebake, J.; Peters, M.
Radioactive mixed wastes containing a radioactive component subject to the Atomic Energy Act (AEA) and hazardous waste subject to resource Conservation and Recovery Act (RCRA) are generated, treated, and stored at the Rocky Flats Plant (RFP) and are subject to federal and state statutory and regulatory requirements. The US Environmental Protection Agency (EPA) and the Colorado Department of Health (CDH) are the two primary regulatory agencies which enforce these requirements. This paper describes the mechanism by which RFP will characterize mixed wastes within the LDR provisions of RCRA and the LDR FFCA as well as for meeting the waste acceptance criteria for disposal.
Ferrenberg, A.; Hunt, K.; Duesberg, J.
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.
Sawai, T; Ikai, K; Uehara, M
We determined the cyclic adenosine monophosphate phosphodiesterase (cAMP-PDE) activity in peripheral blood mononuclear leucocytes from 100 patients with atopic dermatitis (AD) aged 13-57 years (mean +/- SD, 29.8 +/- 17.7 years). The correlation between cAMP-PDE activity and clinical parameters such as the severity of eczema and a personal or family predisposition to atopic respiratory diseases (ARD) (asthma or allergic rhinitis) was examined. Although the enzymic activity varied from normal to very high in the AD patients, cAMP-PDE activity was significantly (P < 0.005) elevated in AD patients (42.1 +/- 22.0 units) as compared with the normal controls (12.4 +/- 5.6) and clinical control subjects (13.4 +/- 9.5). In contrast, we found no correlation between cAMP-PDE activity and the severity of eczema when AD patients were classified into four categories (remission, mild, moderate and severe) according to the extent of their skin involvement. Furthermore, we found that systemic corticosteroid therapy in severe AD patients did not alter the cAMP-PDE activity. cAMP-PDE activity was significantly (P < 0.01) higher in those AD patients who had a personal history of ARD (47.2 +/- 11.2) than in AD patients with a family history of ARD (37.2 +/- 17.4) and those without a personal or family history ('pure' AD) (34.4 +/- 19.8). Nevertheless, the cAMP-PDE activity was significantly higher even in 'pure' AD patients than in the controls. These results suggest that an elevation of cAMP-PDE activity is closely related to a predisposition to respiratory atopy, and does not follow inflammation in AD patients. PMID:9666832
Ozakbas, S; Idiman, E; Yulug, B; Pakoz, B; Bahar, H; Gulay, Z
The aetiology of multiple sclerosis (MS) is still not fully understood. Infectious agents are believed to play a role in the development of this multifactorial disease. Cases in which this disease occurs after administration of both plasma-derived and recombinant hepatitis B vaccines have been reported. In this study, we compared a group of 11 MS patients who developed first clinical symptoms after hepatitis B vaccination (group I) with 71 MS patients who were never vaccinated against hepatitis B and were negative for hepatitis B serology (group II), and 20 healthy controls (group III). Mean age was 27.75 years (19-39) in group I, 30.16 years (18-50) in group II, and 34.4 years (18-50) in group III. Mean attack rate after 2 years was 1.5 in group I and 1.63 in group II. Mean Expanded Disability Status Scale score after 2 years was 1.31 in group I and 1.89 in group II. Human leucocyte antigen (HLA) typing and serology for hepatitis B surface antigen were performed in all groups. In groups I and II, HLA-DR2 was more frequent than in normal healthy subjects. This reflects the general role of HLA in the pathogenesis of MS but suggests that antigen presentation by different HLA is not involved in the development of MS after hepatitis B vaccination. Since there was no difference in the clinical features between vaccinated and nonvaccinated MS patients, this study supports recent reports that hepatitis B vaccination is safe in MS patients and that hepatitis B vaccination is not involved in the development of MS. PMID:16948644
Foureau, D M; Walling, T L; Maddukuri, V; Anderson, W; Culbreath, K; Kleiner, D E; Ahrens, W A; Jacobs, C; Watkins, P B; Fontana, R J; Chalasani, N; Talwalkar, J; Lee, W M; Stolz, A; Serrano, J; Bonkovsky, H L
Drug-induced liver injury (DILI) is often caused by innate and adaptive host immune responses. Characterization of inflammatory infiltrates in the liver may improve understanding of the underlying pathogenesis of DILI. This study aimed to enumerate and characterize leucocytes infiltrating liver tissue from subjects with acute DILI (n = 32) versus non-DILI causes of acute liver injury (n = 25). Immunostains for CD11b/CD4 (Kupffer and T helper cells), CD3/CD20 (T and B cells) and CD8/CD56 [T cytotoxic and natural killer (NK) cells] were evaluated in biopsies from subjects with acute DILI, either immunoallergic (IAD) or autoimmune (AID) and idiopathic autoimmune (AIH) and viral hepatitis (VH) and correlated with clinical and pathological features. All biopsies showed numerous CD8(+) T cells and macrophages. DILI cases had significantly fewer B lymphocytes than AIH and VH and significantly fewer NK cells than VH. Prominent plasma cells were unusual in IAD (three of 10 cases), but were associated strongly with AIH (eight of nine) and also observed in most with AID (six of nine). They were also found in five of 10 cases with VH. Liver biopsies from subjects with DILI were characterized by low counts of mature B cells and NK cells in portal triads in contrast to VH. NK cells were found only in cases of VH, whereas AIH and VH both showed higher counts of B cells than DILI. Plasma cells were associated most strongly with AIH and less so with AID, but were uncommon in IAD. PMID:25418487
Arons, Evgeny; Adams, Sharon; Venzon, Venzon, David J; Pastan, Ira; Kreitman, Robert J.
Frequencies of human leucocyte antigens (HLA) were determined in 287 classic hairy cell leukaemia (HCL) patients. With respect to both population (n=287) and allele (2n=574) frequency, respectively, the most common HLA class I and II antigens expressed were HLA-A*02 (49.1% and 28.6%), HLA-B*07 (21.3% and 11.1%), HLA-C*07 (46.7 and 28.2%), HLA-DQB1*03 (62.7% and 37.3%), HLA-DRB1*11 (30.0% and 16.0%) and HLA-DRB4*01 (45.3% and 29.6%). In comparing 6–14 databases of control Caucasians to 267 Caucasian HCL patients, only HLA-DRB1*11 was consistently over-represented in HCL, 31.1% of patients vs 17–19.9% of controls (p=0.0055 to <0.0001) and 16.5% of alleles vs 6.5–12.3% of control alleles (p=0.022 to <0.0001). HLA-DRB1*11 is a known risk factor for acquired thrombotic microangiopathy. Anti-CD22 recombinant immunotoxin BL22 in HCL was associated with a 12% incidence of completely reversible grade 3–4 haemolytic uraemic syndrome (HUS), mainly during the second or third retreatment cycle. Of 49 HCL patients receiving ≥2 cycles of BL22, 7 (14%) had HUS and HLA-DRB1*11 was expressed in 71% of 7 with HUS compared with only 21% of 42 without (p=0.015). These data suggest that DBR1*11 may be a marker for increased susceptibility to HCL and, among HCL patients, could be a risk factor for BL22-induced HUS. PMID:24931452
Williams, J D; Topley, N; Alobaidi, H M; Harber, M J
Unopsonized particulate zymosan and its major carbohydrate component glucan were phagocytosed under serum-free conditions by adherent polymorphonuclear leucocytes (PMN) in a dose- and time-dependent manner. Preincubation of PMN monolayers with mannan did not cause a reduction in the phagocytosis of either particle. The phagocytic response was inhibited by preincubation of the cells with trypsin at a concentration that did not inhibit the phagocytosis of sheep erythrocytes coated with IgG or of latex particles. Homology of the recognition mechanisms for glucan and zymosan was confirmed when cells cultured on fixed glucan or on fixed zymosan failed to ingest either particle to more than 40% of control phagocytosis. Similarly, zymosan and glucan activated PMN in suspension, in a dose- and time-dependent manner, to generate reactive oxygen species which were measured as luminol-dependent chemiluminescence (CL). There was, however, a four-fold greater CL response to zymosan. Preincubation of PMN with mannan resulted in a significantly decreased CL response to zymosan, while the response to glucan was unaffected. The CL response was also sensitive to a range of concentrations of trypsin. In contrast, two other complex polysaccharide particles (barley-derived beta-glucan and algae-derived laminarin) were not phagocytosed by PMN, nor did they cause the generation of CL, despite the fact that they possessed the capacity, in common with zymosan and glucan, to activate the alternative pathway of complement. The identification of a trypsin-sensitive recognition mechanism on the surface of human PMN for unopsonized zymosan and glucan represents a response not hitherto characterized. Furthermore, our data indicate that the phagocytosis of unopsonized zymosan by human PMN is dependent primarily on its glucan content, but that its capacity to activate the respiratory burst may involve mannan and the recruitment of a second cell surface recognition mechanism. PMID:3710519
... bodies , What Is Alzheimer's? NIA-Funded Memory & Aging Project Reveals Mixed Dementia Common Data from the first ... disease. For example, in the Memory and Aging Project study involving long-term cognitive assessments followed by ...
Hawk, C. W.; Landrum, D. B.; Muller, S.; Turner, M.; Parkinson, D.
The Strutjet approach to Rocket Based Combined Cycle (RBCC) propulsion depends upon fuel-rich flows from the rocket nozzles and turbine exhaust products mixing with the ingested air for successful operation in the ramjet and scramjet modes. It is desirable to delay this mixing process in the air-augmented mode of operation present during low speed flight. A model of the Strutjet device has been built and is undergoing test to investigate the mixing of the streams as a function of distance from the Strutjet exit plane during simulated low speed flight conditions. Cold flow testing of a 1/6 scale Strutjet model is underway and nearing completion. Planar Laser Induced Fluorescence (PLIF) diagnostic methods are being employed to observe the mixing of the turbine exhaust gas with the gases from both the primary rockets and the ingested air simulating low speed, air augmented operation of the RBCC. The ratio of the pressure in the turbine exhaust duct to that in the rocket nozzle wall at the point of their intersection is the independent variable in these experiments. Tests were accomplished at values of 1.0, 1.5 and 2.0 for this parameter. Qualitative results illustrate the development of the mixing zone from the exit plane of the model to a distance of about 10 rocket nozzle exit diameters downstream. These data show the mixing to be confined in the vertical plane for all cases, The lateral expansion is more pronounced at a pressure ratio of 1.0 and suggests that mixing with the ingested flow would be likely beginning at a distance of 7 nozzle exit diameters downstream of the nozzle exit plane.
Hawk, C. W.; Landrum, D. B.; Muller, S.; Turner, M.; Parkinson, D.
The Strutjet approach to Rocket Based Combined Cycle (RBCC) propulsion depends upon fuel-rich flows from the rocket nozzles and turbine exhaust products mixing with the ingested air for successful operation in the ramjet and scramjet modes. It is desirable to delay this mixing process in the air-augmented mode of operation present during low speed flight. A model of the Strutjet device has been built and is undergoing test to investigate the mixing of the streams as a function of distance from the Strutjet exit plane during simulated low speed flight conditions. Cold flow testing of a 1/6 scale Strutjet model is underway and nearing completion. Planar Laser Induced Fluorescence (PLIF) diagnostic methods are being employed to observe the mixing of the turbine exhaust gas with the gases from both the primary rockets and the ingested air simulating low speed, air augmented operation of the RBCC. The ratio of the pressure in the turbine exhaust duct to that in the rocket nozzle wall at the point of their intersection is the independent variable in these experiments. Tests were accomplished at values of 1.0, 1.5 and 2.0 for this parameter. Qualitative results illustrate the development of the mixing zone from the exit plane of the model to a distance of about 19 equivalent rocket nozzle exit diameters downstream. These data show the mixing to be confined in the vertical plane for all cases, The lateral expansion is more pronounced at a pressure ratio of 1.0 and suggests that mixing with the ingested flow would be likely beginning at a distance of 7 nozzle exit diameters downstream of the nozzle exit plane.
Minz, Ranjana W; Chhabra, Seema; Aggarwal, Ritu; Das, Ashim; Saikia, Biman; Chawla, Yogesh K
A 42 year old asymptomatic female detected as incipient Primary Biliary Cirrhosis/Autoimmune Hepatitis overlap during routine checkup. The biochemical profile showed evolution from a mildly deranged liver function test in 2004 along with increased erythrocyte sedimentation rate to a 4 times elevation of alkaline phosphatase in 2006 with mildly deranged alanine transaminase. Autoimmune markers demonstrable were Anti mitochondrial antibody M(2) and sp100. Histopathology showed dual features, dominant findings were of autoimmune heptatitis. Features consistent with Primary Biliary Cirrhosis were minimal with an occasional portal tract showing paucity of bile ducts and occasional bile duct proliferation. Human leucocyte antigen DR/DQ genotype was as follows: DRB1*03, DRB1*07, DQB1*02, DQB1*04. PMID:19829977
Minz, Ranjana W; Aggarwal, Ritu; Das, Ashim; Saikia, Biman; Chawla, Yogesh K
A 42 year old asymptomatic female detected as incipient Primary Biliary Cirrhosis/Autoimmune Hepatitis overlap during routine checkup. The biochemical profile showed evolution from a mildly deranged liver function test in 2004 along with increased erythrocyte sedimentation rate to a 4 times elevation of alkaline phosphatase in 2006 with mildly deranged alanine transaminase. Autoimmune markers demonstrable were Anti mitochondrial antibody M2 and sp100. Histopathology showed dual features, dominant findings were of autoimmune heptatitis. Features consistent with Primary Biliary Cirrhosis were minimal with an occasional portal tract showing paucity of bile ducts and occasional bile duct proliferation. Human leucocyte antigen DR/DQ genotype was as follows: DRB1*03, DRB1*07, DQB1*02, DQB1*04. PMID:19829977
Bamberger, Judith A.; Meyer, Perry A.
This paper describes the physical properties for defining the operation of a pulse jet mixing system. Pulse jet mixing operates with no moving parts located in the vessel to be mixed. Pulse tubes submerged in the vessel provide a pulsating flow due to a controlled combination of applied pressure to expel the fluid from the pulse tube nozzle followed by suction to refill the pulse tube through the same nozzle. For mixing slurries nondimensional parameters to define mixing operation include slurry properties, geometric properties and operational parameters. Primary parameters include jet Reynolds number and Froude number; alternate parameters may include particle Galileo number, particle Reynolds number, settling velocity ratio, and hindered settling velocity ratio. Rating metrics for system performance include just suspended velocity, concentration distribution as a function of elevation, and blend time.
Harmsen, C N
The author examines the extent and characteristics of mixed marriages in the Netherlands. "Nine out of ten married persons born in Turkey or Morocco have a partner who was born in the same country. The majority of married Surinamese also have a partner originating from the same country. Those who spend (a part of) their youth in Indonesia (the former Dutch East Indies), on the other hand, are mostly married to someone born in the Netherlands." (EXCERPT) PMID:12294179
Suppression of development of glomerulonephritis in NZB x NZWF1 mice by persistent infection with lactic dehydrogenase virus: relations between intercellular adhesion molecule-1 expression on endothelial cells and leucocyte accumulation in glomeruli.
Kameyama, Y.; Hayashi, T.
The development of glomerulonephritis (GN) in autoimmune NZB x NZWF1 mice was suppressed by persistent lactic dehydrogenase virus (LDV) infection. In this study the expression of intercellular adhesion molecule-1 (ICAM-1) on endothelial cells in glomeruli was examined during the development of GN. ICAM-1 expression on endothelial cells preceded the accumulation of leucocytes within glomeruli. The uninfected mice exhibited an age-related and profound increase in ICAM-1 expression associated with the development of a GN as evidenced by deposits of IgG and C3. Uninfected mice also showed increased accumulation of leucocytes, such as polymorphonuclear leucocytes (PMNs), macrophages, T and CD4+ cells, which express the lymphocyte function-associated antigen-1 (LFA-1) within glomeruli during the development of GN. These changes were strongly suppressed by LDV infection. Our findings suggest that the expression of ICAM-1 in glomerular endothelial cells may, at least in part, contribute to the development of GN. Suppressed expression of ICAM-1 in LDV-infected mice may be responsible for the suppression of GN seen in these animals. Thus there may be a pathogenetic role for ICAM-1 expression and for intraglomerular accumulation of leucocytes, especially PMNs, which express LFA-1 in the development of GN. Images Figure 1 Figure 2 Figure 7 Figure 9 Figure 11 PMID:7947231
The analysis of peptide binding to porcine major histocompatibility complex (MHC) class I molecules has not been extensively performed. Critical to understanding the adaptive immune response of swine to infection is characterization of Swine Leucocyte Antigens (SLA) class I and class II peptide bind...
A period of research with Helmut Sies in the 1980s is recalled. Our experiments aimed at an in-depth understanding of metabolic changes due to oxidative challenges under near-physiological conditions, i.e. perfused organs. A major focus were alterations of the glutathione and the NADPH/NADP(+) system by different kinds of oxidants, in particular formation of glutathione mixed disulfides with proteins. To analyze mixed disulfides, a test was adapted which is widely used until today. The observations in perfused rat livers let us believe that glutathione-6-phosphate dehydrogenase (G6PDH), i.a. might be activated by glutathionylation. Although we did not succeed to verify this hypothesis for the special case of G6PDH, the regulation of enzyme/protein activities by glutathionylation today is an accepted posttranslational mechanism in redox biology in general. Our early experimental approaches are discussed in the context of present knowledge. PMID:27095221
Romano, E L; Rossi Devivo, M L; Soyano, A; Linares, J
Studies were undertaken to investigate the antibody-dependent cell-mediated cytotoxicity (ADCC) activity of mononuclear leucocytes (MNL) from cord and healthy adult blood and that from infants with ABO haemolytic disease. The ADCC levels of MNL from both types of newborn blood were found to be higher than that of MNL from adult blood. The extent of ADCC was positively related to the degree of antibody sensitization of the red cells and to the effector cell target cell ratio. The ADCC activity was effected mainly by the adherent cell fraction and could be inhibited by cytochalasin B, hydrocortisone and also by high concentrations (more than 0.5 mg/ml) of non-specific free human IgG. Phagocytosis was also demonstrated to be an important mechanism in the destruction of IgG anti-A coated red cells by the MNL. PMID:6538121
Pallawela, Sanjeeva N S; Sonnex, Christopher; Burdett, Julia; Cooper, Dawn; Nethercott, Katrina; Thomas, Catherina M; Goon, Peter; Webb, Hayley; Carne, Christopher
Recent evidence suggests that asymptomatic nonspecific urethritis (NSU), which is not routinely tested for, is a clinically significant pathology.The aim of this pilot study was to determine if testing for urinary threads, leucocyte esterase (LE) or both in asymptomatic men is a good screening tool for NSU. Of the126 asymptomatic men, 8% met microscopic criteria for the diagnosis of NSU. The positive predictive value for NSU was 71% (95% confidence interval (CI): 29.3-95.5%) and the negative predictive value was 96% (95% CI: 92.8-99.5%). The absence of threads and negative LE makes urethritis highly unlikely, making urinary chlamydia (Chlamydia trachomatis) and gonorrhoea (Neisseria gonorrhoeae) testing sufficient. Incidental findings of further pathology occurred in 7%. PMID:24717166
Presents an art activity for fifth-grade students in which they learn about basic shapes and what happens when shapes overlap, draw seven overlapping geometric shapes, review the use of acrylic paint and mixing colors, and finally paint with primary colors. (CMK)
Antonelli, Jean; Warner, Noel; Brown, Kenneth Alun; Wright, John; Simpson, A John; Rennie, Jillian; Hulme, Gillian; Lewis, Sion Marc; Mare, Tracey Anne; Cookson, Sharon; Weir, Christopher John; Dimmick, Ian; Keenan, Jim; Rossi, Adriano Giorgio; Shankar-Hari, Manu; Walsh, Timothy S
Introduction Sepsis is an acute illness resulting from infection and the host immune response. Early identification of individuals at risk of developing life-threatening severe sepsis could enable early triage and treatment, and improve outcomes. Currently available biomarkers have poor predictive value for predicting subsequent clinical course in patients with suspected infection. Circulating leucocytes provide readily accessible tissues that reflect many aspects of the complex immune responses described in sepsis. We hypothesise that measuring cellular markers of immune responses by flow cytometry will enable early identification of infected patients at risk of adverse outcomes. We aim to characterise leucocyte surface markers (biomarkers) and their abnormalities in a population of patients presenting to the hospital emergency department with suspected sepsis, and explore their ability to predict subsequent clinical course. Methods and analysis We will conduct a prospective, multicentre, clinical, exploratory, cohort observational study. To answer our study question, 3 patient populations will be studied. First, patients with suspected sepsis from the emergency department (n=300). To assess performance characteristics of potential tests, critically ill patients with established sepsis, and age and gender matched patients without suspicion of infection requiring hospital admission (both n=100) will be recruited as comparator populations. In all 3 groups, we plan to assess circulating biomarker profiles using flow cytometry. We will select candidate biomarkers by cross-cohort comparison, and then explore their predictive value for clinical outcomes within the cohort with suspected sepsis. Ethics and dissemination The study will be carried out based on the principles in the Declaration of Helsinki and the International Conference on Harmonisation Good Clinical Practice. Ethics approval has been granted from the Scotland A Research Ethics Committee (REC) and Oxford C
Reyes-Becerril, Martha; Ascencio-Valle, Felipe; Hirono, Ikuo; Kondo, Hidehiro; Jirapongpairoj, Walissara; Esteban, Maria Angeles; Alamillo, Erika; Angulo, Carlos
The purpose of this study was to characterize the TLR21 gene from yellowtail (Seriola lalandi) and its functional activity using TLR agonist stimulation and Aeromonas antigens. The TLR21 nucleotide sequence from yellowtail was obtained using the whole-genome shotgun sequencing method and bioinformatics tools. Basal TLR21 gene expression was analyzed in several tissues. Subsequently, the gene expression of TLR21 and cytokines IL-1β and TNF-α was evaluated in TLR agonist (CpG-ODN2006, LPS, and Poly I:C) exposing head kidney leucocytes, which were then subjected to Aeromonas antigen stimulation. The yellowtail full-length cDNA sequence of SlTLR21 was 3615 bp (980 aa) showing a high degree of similarity with the counterparts of other fish species and sharing the common structural architecture of the TLR family, including LRR domains, one C-terminal LRR region, and a TIR domain. Gene expression studies revealed the constitutive expression of TLR21 mRNA in all the analyzed tissues; the highest levels were observed in spleen and head kidney where they play an important role in the fish immune system. Transcripts of TLR21 and the downstream IL-1β and TNF-α cytokine genes were most strongly up-regulated after exposure to the TLR agonists following Aeromonas antigen stimulation, suggesting they are involved in immune response. The results indicated that TLR agonists, in combination with Aeromonas antigens in head kidney leucocytes, synergistically enhance TLR21 and cytokines in yellowtail. PMID:26987525
Newby, A C
1. The role of adenosine deaminase (EC 126.96.36.199), ecto-(5'-nucleotidase) (EC 188.8.131.52) and ecto-(non-specific phosphatase) in the CN-induced catabolism of adenine nucleotides in intact rat polymorphonuclear leucocytes was investigated by inhibiting the enzymes in situ. 2. KCN (10mM for 90 min) induced a 20-30% fall in ATP concentration accompanied by an approximately equimolar increase in hypoxanthine, ADP, AMP and adenosine concentrations were unchanged, and IMP and inosine remained undetectable ( less than 0.05 nmol/10(7) cells). 3. Cells remained 98% intact, as judged by loss of the cytoplasmic enzyme lactate dehydrogenase (EC 184.108.40.206). 4. Pentostatin (30 microM), a specific inhibitor of adenosine deaminase, completely inhibited hypoxanthine production from exogenous adenosine (55 microM), but did not black CN-induced hypoxanthine production or cause adenosine accumulation in intact cells. This implied that IMP rather than adenosine was an intermediate in AMP breakdown in response to cyanide. 5. Antibodies raised against purified plasma-membrane 5'-nucleotidase inhibited the ecto-(5'-nucleotidase) by 95-98%. Non-specific phosphatases were blocked by 10 mM-sodium beta-glycerophosphate. 6. These two agents together blocked hypoxanthine production from exogenous AMP and IMP (200 microM) by more than 90%, but had no effect on production from endogenous substrates. 7. These data suggest that ectophosphatases do not participate in CN-induced catabolism of intracellular AMP in rat polymorphonuclear leucocytes. 8. A minor IMPase, not inhibited by antiserum, was detected in the soluble fraction of disrupted cells. PMID:6249264
Heyvaert, Mieke; Hannes, Karin; Maes, Bea; Onghena, Patrick
In several subdomains of the social, behavioral, health, and human sciences, research questions are increasingly answered through mixed methods studies, combining qualitative and quantitative evidence and research elements. Accordingly, the importance of including those primary mixed methods research articles in systematic reviews grows. It is…
Dryden, Alexander W.S.; Rana, Mrinal; Pandey, Pravin
Summary A 49-year-old diabetic man presented with a 2-day history of a painful right eye associated with a purulent discharge. Prior to becoming symptomatic, he reported that someone spat at him, resulting in direct contact between the saliva and his affected eye. Gram stain revealed numerous leucocytes with Gram-negative diplococci, and culture yielded Neisseria meningitidis (serogroup C). There was no evidence of any systemic infection, and blood cultures were negative for any growth. He was treated for primary meningococcal conjunctivitis (PMC) with intensive topical antibiotic eyedrops as well as systemic antibiotics. One week after commencing treatment he remained systemically well and his symptoms had fully resolved. PMID:27330479
Primary syphilis; Secondary syphilis; Late syphilis; Tertiary syphilis ... Syphilis is a sexually transmitted, infectious disease caused by the spirochete bacterium Treponema pallidum . This bacterium causes ...
Al Qassabi, Salim S; Al-Harthi, Saud M; Al-Osali, Magdi E
Gynecomastia is an enlargement of male breast resulting from a proliferation of its glandular component, and it is usually due to an altered estrogen-androgen balance. It should be differentiated from pseudogynecomastia, which is characterized by fat deposition without glandular proliferation and from breast carcinoma. Gynecomastia could be physiological in neonates and pubertal or pathological due to drug intake, chronic liver, or renal disease, hyperthyroidism, testicular or adrenal neoplasms, and hypogonadism whether primary, or secondary. Properly organized work-up is needed to reach the cause of gynecomastia. Here, we reported a case of a young Omani man with gynecomastia with the aim of creating awareness of the occurrence of Klinefelter's syndrome (KS) in patients with gynecomastia, to observe any differences in clinical presentation of KS from those reported in the literature, and highlight the needed diagnostic work-up and treatment. PMID:26318471
Shang, X Z; Issekutz, A C
Accumulation of leucocytes in inflamed lung tissue and alveolar space involves their migration through vascular endothelium and then lung connective tissue. As a model of this process, we investigated human polymorphonuclear leucocyte (PMNL) and monocyte migration through a biological barrier of human lung fibroblasts (HLF) grown on polycarbonate filters. Very few PMNL (1-2%) or monocytes (3-8%) migrated through the HLF barriers spontaneously. Migration increased to 48-53% of added PMNL and 17-24% of added monocytes, when a C5a chemotactic gradient was present. The monocyte migration induced by C5a was not inhibited by monoclonal antibodies (mAb) to CD18 (beta 2 integrins). This CD18-independent migration was partially inhibited (35%) by mAb to gamma 5 of VLA-5 and completely inhibited by the combination of mAb to gamma 4 of VLA-4 with mAb to VLA-5, in the presence of mAb to CD18. In contrast, PMNL migration across HLF induced by C5a was partially inhibited by mAb to CD18 alone, but even with the addition of mAb to VLA-4, VLA-5 beta 1 and VLA-6, the greatest degree of inhibition was only 60%. Blocking the function of CD18 was not required to observe the inhibition by mAb to VLA-4, although the inhibitory effect of mAb to VLA-5 and VLA-6 alone or in combination was only observed when CD18 mechanisms were also blocked with anti-CD18 mAb. These results demonstrate that (a) both monocytes and PMNL can use either CD11/CD18 (beta 2 integrin) or beta 1 (CD49/CD29) integrins to migrate through HLF barriers; (b) in the case of monocytes, the VLA-4 and VLA-5 integrins account for essentially all the CD11/CD18-independent migration mechanisms; and (c) in contrast to monocytes, PMNL CD18-independent migration is mediated not only by VLA-4 and VLA-5, but also by VLA-6, and up to 40% of the migration appears to be via yet to be defined PMNL surface molecules. PMID:9497495
Gao, J X; Issekutz, A C
Accumulation of leucocytes in inflammation involves their migration through vascular endothelium and then in the connective tissue. We investigated human polymorphonuclear leucocyte (PMNL) migration through a biological barrier of human dermal fibroblasts grown on microporous filters, as a model of PMNL migration in the connective tissue. PMNL did not migrate through a fibroblast monolayer unless a chemotactic factor, e.g. C5a, interleukin-8 (IL-8) or zymosan-activated plasma (ZAP; C5adesArg), was added. This migration was partially inhibited (35-70%, depending on the stimulus) by treatment of PMNL with monoclonal antibody (mAb) to CD18 (beta 2-integrins). Most of the CD18-independent migration was inhibited by mAb to beta 1-integrins (CD29). Inhibition by mAb to beta 1 was observed when the PMNL, but not the fibroblasts, were treated with mAb. The role of beta 1-integrins in PMNL transfibroblast migration was detectable only when the function of the CD11-CD18 complex was blocked, because mAb to beta 1-integrin alone had no significant effect on PMNL migration. Migration induced by C5a was more CD18-independent compared to IL-8 or C5adesArg. The CD18-independent migration was also inhibited by mAb to the beta 1-integrin subunits alpha 5 (of very late antigens-5; VLA-5) and alpha 6 (of VLA-6). Treatment of the fibroblasts (4 hr) with tumour necrosis factor-alpha (TNF-alpha) or IL-1 alpha enhanced C5a-induced PMNL transfibroblast migration and increased the proportion of migration utilizing the CD11-CD18 mechanism. However, TNF-alpha treatment had no effect on the degree of beta 1-integrin-dependent migration. These findings suggest that in response to the chemotactic factors C5a, IL-8 and C5adesArg, PMNL migration in the connective tissue is mediated by both CD11-CD18 (beta 2) and beta 1-integrins on the PMNL. The VLA-5 and VLA-6 members of beta 1-integrins are involved in this process. This is in contrast to PMNL migration across endothelium in this system, which
... in which the bone marrow produces too many platelets. Platelets are a part of the blood that aids ... Primary thrombocythemia is caused by the overproduction of platelets. If untreated, this condition gets worse over time. ...
The primary objective of this work is to quantify the mixing time when two miscible fluids are mixed by one recirculation pump and to evaluate adequacy of 2.5 hours of pump recirculation to be considered well mixed in SRS tanks, JT-71/72. The work scope described here consists of two modeling analyses. They are the steady state flow pattern analysis during pump recirculation operation of the tank liquid and transient species transport calculations based on the initial steady state flow patterns. The modeling calculations for the mixing time are performed by using the 99% homogeneity criterion for the entire domain of the tank contents.
Manzotte, Thais; Guindalini, Camila; Mazzotti, Diego R; Palombini, Luciana; de Souza, Altay L; Poyares, Dalva; Bittencourt, Lia R A; Tufik, Sergio
Human leucocyte antigen (HLA) DQB1*0602 allele, a well-known genetic risk factor for narcolepsy, has been associated with sleep parameters in healthy subjects. We aimed to assess the association of this allele with daytime sleepiness and altered sleep electroencephalogram characteristics in the general population and in patients with obstructive sleep apnoea syndrome (OSAS). Eight hundred and ninety-four individuals from the Epidemiologic Study of Sleep were genotyped for the HLA DQB1*0602 allele. Full-night polysomnography was performed, and daytime sleepiness was analysed according to the Epworth Sleepiness Scale. HLA-DQB1*0602 allele-positive and -negative subjects in the general population, as well as in patients with OSAS, exhibited similar sleep parameters and levels of daytime sleepiness. However, spectral analysis showed that allele-positive individuals with OSAS exhibited higher theta power during sleep Stage 1 (P < 0.05) in occipital derivations, and lower delta power during sleep Stages 1 and 2 (P < 0.01) compared with individuals negative for the allele, even after correction for potential confounders as age, sex, body mass index and European ancestry. No significant differences in the electroencephalogram variables were found in individuals without OSAS. The data highlight the HLA-DQB1*0602 as a potential genetic factor influencing sleep physiology in individuals diagnosed with OSAS. PMID:23136848
Turner, K J; Holt, P G; Holt, B J; Cameron, K J; Hobday, J D; Thompson, P; Phillips, M
Fractionation of human peripheral blood leucocytes (PBL) B cells by differential sedimentation on a discontinuous Percoll gradient separates B cell subpopulations which vary markedly in rates of spontaneous IgE synthesis, often revealing the presence of active IgE secreting cells which are totally suppressed within unfractionated PBL B cell preparations. The production in vitro of IgE by separated B cell populations from the same individual may respond disparately to an identical population of autologous T cells and to pokeweed mitogen. Kinetic studies revealed major differences in both the rates of release of cell-associated IgE between these B cell populations, and their rates of de novo IgE synthesis. From a methodological viewpoint, the use of this B cell fractionation technique is demonstrated to improve greatly the efficiency of detection of T cell-responsive IgE producing B cells in peripheral blood from atopics, and from a mechanistic standpoint raises the possibility that B cell heterogeneity may modulate the functional expression of IgE-regulatory T cells signals. PMID:3498575
Lunn, D P; McClure, J T; Schobert, C S; Holmes, M A
Severe combined immunodeficiency (SCID) is a fatal autosommal disease of Arabian horses that leads to failure of maturation of T- and B-lymphocyte populations, although natural killer (NK) cells are unaffected. Thymic and lymph node tissues from two foals suffering from SCID were examined in an immunohistological study using a panel of monoclonal antibodies recognising equine leucocyte differentiation antigens. In both foals, the majority of cells in lymphoid tissues had an EqCD3-EqCD4-EqCD8+ phenotype, although rare EqCD3+ cells were also detected. The EqCD3-EqCD4-EqCD8+ cells may represent an abnormal lymphocyte differentiation product resulting from the SCID defect, or alternatively may be a normal equine NK cell population. We suggest that the evidence favours the latter proposal, and that equine NK cells in normal horses therefore may be identified by an EqCD3-EqCD8+ phenotype. The implications for the nature of the equine SCID defect are discussed. Images Figure 1 PMID:7751035
Apoptosis is associated with reduced expression of complement regulatory molecules, adhesion molecules and other receptors on polymorphonuclear leucocytes: functional relevance and role in inflammation.
Jones, J; Morgan, B P
Human polymorphonuclear leucocytes (PMN) express proteins that protect them from damage by homologous complement. Protection may be particularly important when these cells migrate to inflammatory sites where complement activation is taking place. Resolution of inflammation involves removal of these PMN. The major mechanism of removal is likely to involve PMN apoptosis followed by recognition and engulfment by macrophages. However, little attention has been paid to the possible relevance of apoptosis to PMN susceptibility to immune effectors. Here we describe a reduction in cell surface expression of two complement regulatory proteins, CD59, an inhibitor of the membrane attack complex and CD55 (decay accelerating factor), an inhibitor of the C3/C5 convertase, on a subpopulation of PMN aged in culture. Loss of these proteins, both attached to the membrane by glycosyl phosphatidylinositol (GPI) anchors, correlated closely with the appearance of apoptotic morphology. We also observed a marked reduction in expression of the GPI-anchored molecule CD16 on apoptotic PMN. Reduced expression of membrane proteins was not confined to those anchored through GPI--several transmembrane molecules including CD11a CD11b and CD18 were also reduced on apoptotic PMN, whilst other were little changed (CD35, CD46). The precipitous fall in CD16 surface expression on PMN was not specific for apoptosis--in vitro incubation of PMN with lipopolysaccharide-inhibited apoptosis but caused a reduction in CD16 expression to 'apoptotic' levels. Images Figure 2 PMID:8567034
Viryasova, Galina M.; Galkina, Svetlana I.; Gaponova, Tatjana V.; Romanova, Julia M.; Sud’ina, Galina F.
In the present study we have presented data on the regulation of LT (leukotriene) and 5-oxo-ETE (5-oxo-6,8,11,14-eicosatetraenoic acid) syntheses in human neutrophils upon interaction with OZ (opsonized zymosan) or Salmonella typhimurium. Priming of neutrophils with PMA (phorbol 12-myristate 13-acetate) and LPS (lipopolysaccharide) elicits 5-oxo-ETE formation in neutrophils exposed to OZ, and the addition of AA (arachidonic acid) significantly increases 5-oxo-ETE synthesis. We found that NO (nitric oxide)-releasing compounds induce 5-oxo-ETE synthesis in neutrophils treated with OZ or S. typhimurium. Exposure of neutrophils to zymosan or bacteria in the presence of the NO donor DEA NONOate (1,1-diethyl-2-hydroxy-2-nitroso-hydrazine sodium) considerably increased the conversion of endogenously formed 5-HETE (5S-hydroxy-6,8,11,14-eicosatetraenoic acid) to 5-oxo-ETE. To our knowledge, this study is the first to demonstrate that NO is a potent regulator of 5-oxo-ETE synthesis in human polymorphonuclear leucocytes exposed to Salmonella typhimurium and zymosan. PMID:24712762
De Guise, S; Erickson, K; Blanchard, M; DiMolfetto, L; Lepper, H D; Stott, J L; Ferrick, D A
The specificity of F21.A, a monoclonal antibody raised against bottlenose dolphin leucocytes, was characterized in killer whale on the basis of immunoprecipitation of a protein of 94 kDa, as well as flow cytometric analysis. While minimally expressed on resting cells, F21.A labeled a homologue to beta-2 integrin in 89-97% of PMA-activated neutrophils, 53-66% of activated monocytes, and activated B cells but not T cells. Activation of neutrophils reached its maximum 10 min after PMA stimulation. F21.A did not label intracellular stores as did both cross-reacting anti-canine CD11b and CD18, suggesting that an activation-induced conformational change would expose a neoepitope recognized by F21.A. F21.A labeling was largely inhibited by pre-incubation with plasma, suggesting a binding site closely related to that for fibrinogen. In vitro phagocytosis and respiratory burst were almost fully inhibited upon pre-incubation with F21.A, demonstrating its functional importance. This antibody is foreseen as a possible valuable diagnostic and research tool in cetacean immunology. PMID:14741138
Camina, M. M.; Iannone, P.
Recent UK policy has emphasised both the development of socially mixed communities and the creation of balanced school intakes. In this paper, we use a case study of an area of mixed tenure in eastern England to explore policy in practice and the extent to which mechanisms of segregation impact on both the creation of socially mixed neighbourhoods…
Dias, Fabrício C.; Mendes-Junior, Celso T.; Silva, Maria C.; Tristão, Fabrine S. M.; Dellalibera-Joviliano, Renata; Soares, Edson G.; Menezes, Jean G.; Schmidt, André; Dantas, Roberto O.; Marin-Neto, José A.; Silva, João S.; Donadi, Eduardo A.
Genetic susceptibility factors, parasite strain, and an adequate modulation of the immune system seem to be crucial for disease progression after Trypanosoma cruzi infection. HLA-G and its murine functional homolog Qa2 have well-recognized immunomodulatory properties. We evaluated the HLA-G 3′ untranslated region (3′UTR) polymorphic sites (associated with mRNA stability and target for microRNA binding) and HLA-G tissue expression (heart, colon, and esophagus) in patients presenting Chagas disease, stratified according to the major clinical variants. Further, we investigated the transcriptional levels of Qa2 and other pro- and anti-inflammatory genes in affected mouse tissues during T. cruzi experimental acute and early chronic infection induced by the CL strain. Chagas disease patients exhibited differential HLA-G 3′UTR susceptibility allele/genotype/haplotype patterns, according to the major clinical variant (digestive/cardiac/mixed/indeterminate). HLA-G constitutive expression on cardiac muscle and colonic cells was decreased in Chagasic tissues; however, no difference was observed for Chagasic and non-Chagasic esophagus tissues. The transcriptional levels of Qa2 and other anti and proinflammatory (CTLA-4, PDCD1, IL-10, INF-γ, and NOS-2) genes were induced only during the acute T. cruzi infection in BALB/c and C57BL/6 mice. We present several lines of evidence indicating the role of immunomodulatory genes and molecules in human and experimental T. cruzi infection. PMID:25688175
Madkhali, Tarıq; Alhefdhi, Amal; Chen, Herbert; Elfenbein, Dawn
Primary hyperparathyroidism is a common endocrine disorder caused by overactivation of parathyroid glands resulting in excessive release of parathyroid hormone. The resultant hypercalcemia leads to a myriad of symptoms. Primary hyperparathyroidism may increase a patient’s morbidity and even mortality if left untreated. During the last few decades, disease presentation has shifted from the classic presentation of severe bone and kidney manifestations to most patients now being diagnosed on routine labs. Although surgery is the only curative therapy, many advances have been made over the past decades in the diagnosis and the surgical management of primary hyperparathyroidism. The aim of this review is to summarize the characteristics of the disease, the work up, and the treatment options. PMID:26985167
Chang, Chein-Chi; Chapman, Tom; Siverts-Wong, Elena; Wei, Li; Mei, Ying
This section covers research published during the calendar year 2015 on mixing and transport processes. The review covers mixing of anaerobic digesters, mixing of heat transfer, and environmental fate and transport. PMID:27620101
Pallan, Shelley; Khan, Aliya
Abstract Objective To discuss the presentation, diagnosis, and management of primary hyperparathyroidism (PHPT) in family medicine. Quality of evidence MEDLINE was searched from 2002 to 2009 using the terms presentation, diagnosis, and treatment of PHPT. Proceedings and guidelines from the Third International Workshop on Primary Hyperparathyroidism in May 2008 were reviewed in detail. Most studies offered level II and III evidence, although there were a number of single randomized controlled trials on PHPT (level I evidence). References from pertinent papers were also searched for relevant articles. Articles most relevant to family medicine and primary care practitioners are presented. Main message Primary hyperparathyroidism is the most common cause of hypercalcemia in outpatients. In the Western world, most patients with PHPT present with nonspecific symptoms such as fatigue, mood disturbances, and cognitive impairments. Diagnosis is established when intact parathyroid hormone levels are elevated or at the high end of the normal range in the setting of elevated total or ionized calcium levels (following exclusion of conditions that can mimic PHPT). Criteria for surgery have recently been modified. Surgery is always a suitable option in those with symptomatic PHPT and no contraindications. Those with contraindications or with asymptomatic PHPT not meeting the criteria for surgery can generally be safely monitored and considered for medical management. This might include treatment with bisphosphonates, hormone replacement therapy, raloxifene, or calcimimetic agents; however, there are currently no fracture data for any of these options. Conclusion The definitive therapy for symptomatic and asymptomatic PHPT is parathyroidectomy. In patients with asymptomatic PHPT not meeting the criteria for surgery, monitoring is safe and medical management designed to target skeletal protection or lower serum calcium is a suitable option. PMID:21321169