Haemostatic function and biomarkers of endothelial damage before and after platelet transfusion in patients with acute myeloid leukaemia.

OBJECTIVES: The beneficial effect of platelet transfusion on haemostasis is well established, but there is emerging evidence that platelet transfusion induces an inflammatory response in vascular endothelial cells. BACKGROUND: We investigated haemostatic function and endothelial biomarkers before and after platelet transfusion in patients with acute myeloid leukaemia. MATERIALS AND METHODS: Blood was sampled before, 1 and 24 h after platelet transfusion. Primary and secondary haemostasis was evaluated by whole blood aggregometry (Multiplate) and thromboelastography (TEG). Endothelial biomarkers (sICAM-1, syndecan-1, sThrombomodulin, sVE-Cadherin) and platelet activation biomarkers (sCD40L, TGF-beta) were investigated along with haematology/biochemistry analyses. RESULTS: Twenty-two patients were included. Despite continued low platelet counts, platelet transfusion normalised the median values of most TEG parameters and slightly increased platelet aggregation (all P < 0·05). Endothelial biomarkers were not significantly affected by transfusion. The 1 h sCD40L level correlated positively with Syndecan-1 and soluble thrombomodulin delta values, biomarkers of endothelial damage (both P = 0·005). CONCLUSION: Platelet transfusion improved haemostasis, whereas post-transfusion increases in sCD40L were associated with endothelial damage, indicating that transfused platelets and platelet-derived pro-inflammatory mediators may have opposite effects on the endothelium.

Haemostatic function and biomarkers of endothelial damage before and after RBC transfusion in patients with haematologic disease.

BACKGROUND AND OBJECTIVES: Transfusion of red blood cells (RBC) is beneficial for the patient but can also be harmful, as randomized trials have demonstrated increased infection rates, bleeding and mortality. The study aim was to investigate the response of the vascular system (the haemostatic function and the endothelium) to RBC transfusion. MATERIALS AND METHODS: Blood was sampled from patients with various transfusion-dependent haematologic diseases before 1 and 24 h after RBC transfusion. Primary and secondary haemostasis was evaluated by whole-blood impedance aggregometry (Multiplate) and by thromboelastography (TEG). Samples were analysed by ELISA for biomarkers reflecting endothelial activation and damage (sICAM-1, syndecan-1, sThrombomodulin (sTM), sVE-Cadherin), platelet activation (sCD40L) and inflammation (hsCRP). RESULTS: A total of 58 patients were enrolled in the study. Median age was 71 years. Compared to before transfusion, patients had slightly reduced coagulability 1 h after RBC transfusion, assessed by TEG. However, transfusion of older RBC products (>14 days) was associated with increased coagulability (all P < 0·05). The level of syndecan-1 increased slightly 24 h after transfusion (median 12·4 (IQR 9-23) vs. 13·2 (9-25) ng/ml, P < 0·01), indicating increased glycocalyx degradation. CONCLUSION: Overall, RBC transfusion was associated with reduced coagulability and endothelial glycocalyx degradation. Transfusion of older RBCs was however associated with increased coagulability. The changes observed were small to moderate and the clinical relevance of these findings should be investigated in larger studies.

Temperature effect on high salinity depuration of Vibrio vulnificus and V. parahaemolyticus from the Eastern oyster (Crassostrea virginica).

Vibrio vulnificus (Vv) and Vibrio parahaemolyticus (Vp) are opportunistic human pathogens naturally associated with the Eastern oyster Crassostrea virginica. The abundances of both pathogens in oysters are positively correlated with temperature, thus ingestion of raw oysters during the warm summer months is a risk factor for contracting illness from these bacteria. Current post-harvest processing (PHP) methods for elimination of these pathogens are expensive and kill the oyster, changing their organoleptic properties and making them less appealing to some consumers. High salinity has proven effective in reducing Vv numbers in the wild and our research aims at developing an indoor recirculating system to reduce pathogenic Vibrios while maintaining the taste and texture of live oysters. The goal of this study was to determine the influence of temperature on the efficacy of high salinity depuration. Vv was enumerated as most probable number (MPN) per gram of oyster tissue using the FDA-approved modified cellobiose polymyxin colistin (mCPC) protocol and with an alternative Vibrio specific media CHROMagar™ Vibrio (CaV). CaV was also used to quantify Vp. Oysters were held at 35 psu for 10 days at three temperatures: low (20°C), mid (22.5°C) and high (25°C). There was no difference in MPN/g of Vv between media; however more Vv isolates were obtained from mCPC than CaV. There was no significant effect of temperature on reduction of Vv or Vp throughout depuration but there was a tendency for low temperatures to be less effective than the higher ones. High salinity resulted in a significant decrease in Vv by day 3 and again by day 10, and a decrease in Vp by day 3. Oyster condition indices were maintained throughout depuration and mortality was low (4% across three trials). Overall these results support the use of mCPC for Vv enumeration and demonstrate the promise of high salinity depuration for PHP of the Eastern oyster. The trend for lower temperatures to be less effective is surprising and indicates a potential interaction between salinity and temperature that should be further investigated.

Characterization of the gut microbiota of three commercially valuable warmwater fish species.

AIMS: Due to the strong influence of the gut microbiota on fish health, dominant bacterial species in the gut are strong candidates for probiotics. This study aimed to characterize the gut microbiota of channel catfish Ictalurus punctatus, largemouth bass Micropterus salmoides and bluegill Lepomis macrochirus to provide a baseline for future probiotic studies. METHODS AND RESULTS: The gut microbiota of five pooled individuals from each fish species was identified using 16S rRNA pyrosequencing. Microbiota differed significantly between fish species in terms of bacterial species evenness. However, all gut communities analysed were dominated by the phylum Fusobacteria, specifically the species Cetobacterium somerae. Relatively high abundances of the human pathogens Plesiomonas shigelloides and Fusobacterium mortiferum, as well as members of the genus Aeromonas, suggest these species are normal inhabitants of the gut. CONCLUSIONS: The overwhelming dominance of the genus Cetobacterium in all species warrants further investigation into its role in the fish gut microbiota. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides the first characterization of the gut microbiota of three economically significant fishes and establishes a baseline for future probiotic trials.

Specialized psychosocial treatment plus treatment as usual (TAU) versus TAU for patients with cannabis use disorder and psychosis: the CapOpus randomized trial.

BACKGROUND: Cannabis abuse in psychotic patients is associated with rehospitalizations, reduced adherence and increased symptom severity. Previous psychosocial interventions have been ineffective in cannabis use, possibly because of low sample sizes and short interventions. We investigated whether adding CapOpus to treatment as usual (TAU) reduces cannabis use in patients with cannabis use disorder and psychosis. Method A total of 103 patients with psychosis and cannabis use disorder were centrally randomized to 6 months of CapOpus plus TAU (n = 52) or TAU (n = 51). CapOpus consisted mainly of motivational interviewing and cognitive behaviour therapy (CBT). TAU was targeted primarily at the psychotic disorder. The primary outcome was self-reported days with cannabis use in the preceding month. RESULTS: Pre-randomization cannabis use frequency was 14.9 [95% confidence interval (CI) 12.7-17.1] days/month. Post-treatment, the ratio of days/month with cannabis use in CapOpus versus TAU was 0.76 (95% CI 0.38-1.50) (p = 0.42), and 0.80 (95% CI 0.21-3.10) (p = 0.75) at the 4-month follow-up. From 46.4 (95% CI 36.4-56.3) monthly joints pre-randomization, consumption fell to 27.3 (95% CI 12.6-41.9) joints in CapOpus and 48.2 (95% CI 31.8-64.6) in TAU (p = 0.06). Follow-up amounts were 28.4 (95% CI 13.5-43.2) and 41.6 (95% CI 25.2-58.0) joints (p = 0.23). Several subgroup analyses suggested benefits of CapOpus. CONCLUSIONS: CapOpus did not reduce the frequency, but possibly the amount, of cannabis use. This is similar to the findings of previous trials in this population. Implementation of CapOpus-type interventions is thus not warranted at present but subgroup analyses call for further trials.

Quality of root fillings performed with two root filling techniques. An in vitro study using micro-CT.

OBJECTIVE: The aim of this study was to compare the presence of voids in root fillings performed in oval and ribbon-shaped canals with two root filling techniques, lateral compaction technique (LCT) or hybrid technique (HT), a combination of a gutta-percha masterpoint and thermoplastic gutta-percha. Furthermore, the obturation time for the two techniques was evaluated. MATERIALS AND METHODS: Sixty-seven roots with oval and ribbon-shaped canals were prepared using Profile Ni-Ti rotary files. After preparation, the roots were randomly allocated to two groups according to root filling technique. All roots were filled with AH plus and gutta-percha. Group 1 was filled using LCT (n = 34) and group 2 was filled using HT (n = 33). The obturation time was measured in 30 cases evenly distributed between the two techniques. Voids in relation to the root canal fillings were assessed using cross-section images from Micro-computed Tomography scans. RESULTS: All root canal fillings had voids. Permutation test showed no statistically significant difference between the two root filling techniques in relation to presence of voids (p = 0.092). A statistically significant difference in obturation time between the two techniques was found (p < 0.001). CONCLUSION: The present study found no statistically significant difference in percentage of voids between two root filling techniques. A 40% reduction in obturation time was found for the HT compared to the LCT.

Reduced probability of smoking cessation in men with increasing number of job losses and partnership breakdowns.

BACKGROUND: Unemployment and partnership breakdowns are common stressful life events, but their association with smoking cessation has been investigated in only a few studies. OBJECTIVE: To investigate how history of employment and cohabitation affects the probability of smoking cessation and to study joint exposure to both. METHODS: Birth cohort study of smoking cessation of 6232 Danish men born in 1953 with a follow-up at age 51 (response rate 66.2%). History of unemployment and cohabitation was measured annually using register data. Information on smoking cessation was obtained by a questionnaire. RESULTS: The probability of smoking cessation decreased with the number of job losses (ranging from 1 OR 0.54 (95% CI 0.46 to 0.64) to 3+ OR 0.41 (95% CI 0.30 to 0.55)) and of broken partnerships (ranging from 1 OR 0.74 (95% CI 0.63 to 0.85) to 3+ OR 0.50 (95% CI 0.39 to 0.63)). Furthermore, smoking cessation was associated with the duration of the periods of unemployment (ranging from 1-5 years (OR 0.75, 95% CI 0.65 to 0.85) to 10-23 years (OR 0.29, 95% CI 0.22 to 0.38)) and with living without a partner for > 5 years (ranging from 6-9 years to 10-23 years (OR 0.80, 95% CI 0.66 to 0.97) to 10-23 years (OR 0.44, 95% CI 0.37 to 0.52)). Those who never cohabited and experienced one or more job losses had a particular low chance of smoking cessation (OR 0.19, 95% CI 0.12 to 0.30). CONCLUSION: The numbers of job losses and of broken partnerships were both inversely associated with probability of smoking cessation.

Measurement delay associated with the Guardian RT continuous glucose monitoring system.

AIMS: Using compartment modelling, we assessed the time delay between blood glucose and sensor glucose measured by the Guardian RT continuous glucose monitoring system in young subjects with Type 1 diabetes (T1D). METHODS: Twelve children and adolescents with T1D treated by continuous subcutaneous insulin infusion (male/female 7/5; age 13.1 +/- 4.2 years; body mass index 21.9 +/- 4.3 kg/m(2); mean +/- sd) were studied over 19 h in a Clinical Research Facility. Guardian RT was calibrated every 6 h and sensor glucose measured every 5 min. Reference blood glucose was measured every 15 min using a YSI 2300 STAT Plus Analyser. A population compartment model of sensor glucose-blood glucose kinetics was adopted to estimate the time delay, the calibration scale and the calibration shift. RESULTS: The population median of the time delay was 15.8 (interquartile range 15.2, 16.5) min, which was corroborated by correlation analysis between blood glucose and 15-min delayed sensor glucose. The delay has a relatively low intersubject variability, with 95% of individuals predicted to have delays between 10.4 and 24.3 min. Population medians (interquartile range) for the scale and shift are 0.800 (0.777, 0.823) (unitless) and 1.66 (1.47, 1.84) mmol/l, respectively. CONCLUSIONS: In young subjects with T1D, the total time delay associated with the Guardian RT system was approximately 15 min. This is twice that expected on physiological grounds, suggesting a 5- to 10-min delay because of data processing. Delays above 25 min are rarely to be observed.

[Diagnostic transmission electron microscopy]. / Diagnostisk transmisjonselektronmikroskopi.

BACKGROUND: Transmission electron microscopy (TEM) is an important diagnostic tool in surgical pathology. MATERIAL AND METHODS: We give an overview, based on our own experience, of the application of TEM in diagnostic pathology. RESULTS: In spite of its potential for high-grade resolution, the actual use of TEM in diagnostic pathology is rather limited. TEM is mandatory in the examination of kidney and muscle biopsies and is often indicated when a metabolic disease is suspected. However, in many centres its importance in tumour pathology has declined because of the advances in the field of immunohistochemistry. TEM requires a great deal of resources and high levels of skills in tissue processing and diagnostic interpretation. INTERPRETATION: The TEM diagnosis must be integrated with light microscopical and, often, immunohistopathological findings, as well as with the clinical data. We therefore recommend close collaboration between the clinician and the laboratory with regard to biopsy indication, handling of samples, and the final diagnosis.

Human herpesvirus-6 immunoglobulin G antibodies in patients with multiple sclerosis.

OBJECTIVE: To further investigate a possible correlation between human herpesvirus-6 (HHV-6) infection and multiple sclerosis by analyzing the level of HHV-6 antibodies in MS patients and healthy controls. MATERIALS AND METHODS: A total of 189 serum samples from patients with multiple sclerosis (MS) in different disease stages and 190 serum samples from healthy controls matched for age and sex were analyzed for HHV-6 antibodies using a competitive ELISA. RESULTS: There was no difference between HHV-6 IgG titers in MS patients and controls. Two of the controls were seronegative for HHV-6 versus to none of the MS-patients. There was no apparent difference in HHV-6 titers from patients in different disease stages. CONCLUSION: This study cannot support the theory that HHV-6 is a contributing factor to the development of MS, although a seroprevalence study like this would not disclose whether a late primary infection (in puberty) with HHV-6 might affect the development of MS.

Glycoprotein IIb-IIIa on platelet-derived microparticles, and microparticle structures studied by electron microscopy, confocal laser microscopy and crossed radio-immunoelectrophoresis.

Shedding of microparticles from the platelet surface is usually associated with exposure of platelet procoagulant activity. Platelet-derived microparticles have been detected in blood in various disease states. In vitro, platelet stimulation with a number of different agonists results in formation of microparticles. In the present study, microparticles induced by platelet stimulation by calcium ionophore or by membrane incorporation of the terminal complement complex C5b-9 were studied using electron microscopy, confocal laser microscopy, flow cytometry and radio-immunoelectrophoresis. When studied by electron microscopy, microparticle morphology was found to be dependent upon the induction method. Platelet stimulation with the calcium ionophore resulted in smaller, more homogeneous and electron dense microparticles than those induced by insertion of the terminal complement complex. With flow cytometry and confocal laser immunofluorescence microscopy, microparticle GPIIb-IIIa was demonstrated using a FITC-conjugated antibody to GPIIIa. Surface-bound GPIIb-IIIa was demonstrated on the microparticles by immunoelectron microscopy. Crossed immunoelectrophoresis of detergent-solubilized microparticles visualized a very prominent GPIIb-IIIa immunoprecipitate arc, and binding of [(125)1]fibrinogen to microparticle GPIIb-IIIa was demonstrated by radio-immunoelectrophoresis. This suggests that the activated GPIIb-IIIa complex is preserved intact during the shedding of microparticles from the platelet surface.

Medical expenditures of terminal cancer patients during the last year of life.

To measure the patterns of medical expenditures under conventional modes of cancer care for the terminally ill, we examined Blue Cross and Blue Shield Plan claims records for care during the 12 months preceding the deaths of 1,054 nonelderly patients diagnosed as having breast, colon-rectal, or lung cancer. Expenditures averaged +21,219 for the terminal year and grew exponentially as death approached, with +6,161 (29%) being spent in the final month alone. We found very few significant differences in spending or utilization by diagnosis, age, or sex of the terminal patient, but sizable differences by state of residence and between nonmetropolitan and metropolitan area residents. Our findings represent a baseline against which experience under new hospital payment strategies and alternative delivery systems might be compared.

Chido, Rodgers and C4. In vivo and in vitro coating of red blood cells, grouping and antibody detection.

C4 sucrose/low ionic strength (LIS)-coated red blood cells (RBC) are excellent for the detection of the previously 'nebulous' antibodies, anti-Chido and anti-Rodgers, as well as for serum/plasma typing of these antigens by an inhibition technique. By enzyme treatment of such cells, it is confirmed that the Ch and Rg antigens reside on the C4d part of the C4 molecule. Freshly taken RBC from normal individuals were examined with a sensitive Auto-Analyzer technique with anti-Chido, anti-C4 and anti-C3 sera. All normal RBC were shown to have C4d and C3d components on their surface. The technique was also very sensitive for the detection of the Ch antigen, which was detected on the RBC of all Chido-positive individuals, and which did not show great variation in strength by this method. The mode of in vivo C4 fixation on normal RBC seems to be different from the fixation in LIS or by RBC antibody-mediated activation.

Stimulation of human lymphocytes by cytomegalovirions and dense bodies.

Lymphocytes from healthy adult individuals were examined for their ability to incorporate thymidine in the presence of cytomegalovirus (CMV) and dense bodies. It was found that lymphocytes from donors with antibodies to CMV were stimulated to incorporate thymidine-14C both by preparation of CMV and CMV dense bodies. Lymphocytes from CMV seronegative donors did not respond. Dilution experiments and the dose-response curve indicate that the stimulation induced by the dense body preparation was not caused by the small amounts of contaminating CMV particles. These results indicate that in healthy human adults there is a correlation between CMV seropositivity and in vitro lymphocyte transformation, induced by either CMV or by dense bodies.

Identification of thorium dioxide in human liver cells by electron microscopic x-ray microanalysis.

Thirty-two years after injection of thorium dioxide (Thorotrast) for diagnostic x-ray studies in a female patient deposits were found by light microscopy in the liver macrophages (Kupffer cells). They were shown by electron microscopy to be located inside secondary lysosomes, and by autoradiography and x-ray microanalysis they were identified as thorium.

The rare blood group antigen, Wu.

The inheritance of the Wu antigen in three families is described. The observations suggest that the antigen is inherited as a Mendelian dominant character. Wu segregates independently of sex and of the ABO, MNSs, Rh, Lutheran, Duffy and Kidd blood group systems. Independence of Kell is also quite likely. Wu is a very rare antigen. The corresponding antibody is not as infrequent. Anti-Wu occurs as a 'naturally occurring' antibody, often together with antibodies against other rare antigens.