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PubMed1996

Seroprevalence of Helicobacter pylori infection in patients with connective tissue diseases.

To assess the possibility that Helicobacter pylori might be an etiologic agent, titers of anti-H. pylori IgG in sera of patients with connective tissue diseases [rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), polymyositis or dermatomyositis (PM/DM), progressive systemic sclerosis (PSS), mixed connective tissue disease (MCTD) and Sjögren's syndrome (SjS)] were compared with those of non-patient (healthy) volunteers and of patients with chronic pulmonary diseases (CPD) by ELISA using an extract of sonicated H. pylori as the antigen. Among patients with connective tissue diseases, those with SLE and RA had anti-H. pylori titers as low as healthy volunteers. Patients with SjS had much higher average titers than patients with CPD (P < 0.05). We previously reported that levels of myeloid calcium-binding protein (MRP8 and MRP14) were elevated in the serum of patients with connective tissue diseases. No correlation was found between serum levels of anti-H. pylori IgG and of MRP, a novel marker of inflammation. Furthermore, sera with high IgG titers were selected, and their reactivity with the H. pylori antigen were analyzed by Western blotting. H. pylori antigens with a variety of molecular masses were immunostained with sera from patients and from healthy volunteers, but a 16-kDa antigen was only immunostained by reaction with the sera of patients with MCTD and SjS, although the number of test samples was small.

Adult↗
PubMed1989

Measuring the heaviness of smoking: using self-reported time to the first cigarette of the day and number of cigarettes smoked per day.

Two simple self-report measures have been used to assess the heaviness of smoking, 'number of cigarettes per day' (CPD) and 'time to the first cigarette of the day' (TTF). Little attention, however, has been given to the precise method of scoring this information. Using biochemical indicators of heaviness of smoking (alveolar carbon monoxide and cotinine), we explore the optimum data transformations for regression analysis and categorical analysis. We suggest a four category scoring scheme for both time to the first cigarette of the day (less than or equal to 5, 6-30, 31-60 and 61+ min) and average daily consumption of cigarettes (1-10, 11-20, 21-30, 31+ cigarettes) as the most powerful and practical categorical scoring of these variables. Due to possible ceiling effects on biochemical measures, we suggest using logarithmic transformations of CPD or TTF for regression or correlation analyses.

Adolescent↗
PubMed1983

Effects of glutaraldehyde and critical point drying on the shape and size of erythrocytes in isotonic and hypotonic media.

Scanning electron microscopy (SEM) was applied to studies on osmotically swollen erythrocytes fixed by a method using increasing concentrations of glutaraldehyde (GA), OsO4 as a post fixative, and critical point drying (CPD). This fixation method prevented the osmotic effect of GA itself and preserved the cellular equilibrium shape intact in various media, i.e. discocytes in an isotonic medium and ellipsoids or spheres and ghosts in a hypotonic medium. The discocytes were reduced less in diameter and area than the ellipsoids and spheres during CPD, but the degree of volume shrinkage was of the same magnitude (approx. 73%) regardless of the cell shape. The shrinkage of the discocytes was advanced until the dry volume (approx. 29 microm3) was reached, while the shrinkage of the osmotically swollen cells ceased earlier. A miniaturization process coupled with solvent evaporation is suggested as an explanation for this. The osmotic swelling of erythrocytes was studied quantitatively with the aid of SEM micrographs. The increase in volume was about 60%, which is in agreement with the values obtained by light microscopy for unfixed and GA-fixed cells in the present work and with those reported in the literature.

Aldehydes↗
PubMed1987

Preparation of the mammalian organ of Corti for scanning electron microscopy.

The effect of various standard tissue preparation procedures for scanning microscopy on the morphology of the mammalian organ of Corti has been examined. Comparison was made of (i) different fixatives: glutaraldehyde (GA) alone, osmium (Os) alone, GA and Os together and GA followed by Os; (ii) freeze-drying (FD) versus critical point-drying (CPD); (iii) ligand binding of osmium with thiocarbohydrazide (TOTO procedure) versus sputter coating to provide a conductive coat. Results were assessed in terms of known structural parameters in fresh tissue. It is shown: (i) that double fixation, either GA, Os together or GA followed by Os, consistently produced less tissue distortion than either fixative alone, (ii) that FD produced less shrinkage of the tissue than CPD, and (iii) that the TOTO procedure appeared to rigidify the tissue and enhanced preservation of the morphology.

Animals↗
PubMed2004

Setting a standard for performance assessment of doctor-patient communication in general practice.

CONTEXT: Continuing professional development (CPD) of general practitioners. OBJECTIVE: Criterion-referenced standards for assessing performance in the real practice of general practitioners (GPs) should be available to identify learning needs or poor performers for CPD. The applicability of common standard setting procedures in authentic assessment has not been investigated. METHODS: To set a standard for assessment of GP-patient communication with video observation of daily practice, we investigated 2 well known examples of 2 different standard setting approaches. An Angoff procedure was applied to 8 written cases. A borderline regression method was applied to videotaped consultations of 88 GPs. The procedures and outcomes were evaluated by the applicability of the procedure, the reliability of the standards and the credibility as perceived by the stakeholders, namely, the GPs. RESULTS: Both methods are applicable and reliable; the obtained standards are credible according to the GPs. CONCLUSIONS: Both modified methods can be used to set a standard for assessment in daily practice. The context in which the standard will be used - i.e. the specific purpose of the standard, the moment the standard must be available or if specific feedback must be given - is important because methods differ in practical aspects.

Clinical Competence↗
PubMed2005

The Rad4 homologue YDR314C is essential for strand-specific repair of RNA polymerase I-transcribed rDNA in Saccharomyces cerevisiae.

Summary The Saccharomyces cerevisiae protein Rad4 is involved in damage recognition in nucleotide excision repair (NER). In RNA polymerase II-transcribed regions Rad4 is essential for both NER subpathways global genome repair (GGR) and transcription coupled repair (TCR). In ribosomal DNA (rDNA), however, the RNA polymerase I-transcribed strand can be repaired in the absence of Rad4. In Saccharomyces cerevisiae the YDR314C protein shows homology to Rad4. The possible involvement of YDR314C in NER was studied by analysing strand-specific cyclobutane pyrimidine dimer (CPD) removal in both RNA pol I- and RNA pol II-transcribed genes. Here we show that the Rad4-independent repair of rDNA is dependent on YDR314C. Moreover, in Rad4 proficient cells preferential repair of the transcribed strand of RNA pol I-transcribed genes was lost after deletion of YDR314C, demonstrating that Rad4 cannot replace YDR314C. CPD removal from the RNA pol II-transcribed RPB2 gene was unaffected in ydr314c mutants. We conclude that the two homologous proteins Rad4 and YDR314C are both involved in NER and probably have a similar function, but operate at different loci in the genome and are unable to replace each other.

Amino Acid Sequence↗
PubMed1980

Preservation of human erythrocytes in the liquid state: biological results with a new medium.

Red blood cells (RBC) were collected with citrate-phosphate-dextrose (CPD) in a blood-pack optimal additive system. After concentration to 90% hematocrit they were diluted with saline-adenine-glucose medium (SAG-RBC), and stored for 35 days. In this work the RBC were stored in the presence of leukocytes. The SAG medium allows RBC conservation during 35 days at +4 degrees C. The adenosine triphosphate (ATP) level of RBC is compatible with their survival. During the first 2 weeks, hemolysis of SAG-RBC was not greater than in CPD blood. Nevertheless, hemolysis reached 1.49% on day 35, and there was a marked increase in RBC osmotic fragility. Scanning electron-microscopic studies of 35-day RBC showed that the majority of them became echinocytes. After incubation in fresh frozen plasma, the RBC recovered satisfactory osmotic resistance and normal disc shape. The post-transfusion viability was normal with greater than 70% recovery after 48 h. The in vivo restoration of 2,3-diphosphoglycerate (2,3-DPG) was rapid in the transfused SAG-RBC, 50% of the initial 2,3-DPG level being restored in 1 h. The in vivo studies proved that the functional quality of these RBC was compatible with their use in transfusion. The most important problem concerns the supernatant hemoglobin level of the SAG-RBC to be used for massive transfusion.

Adenine↗
PubMed1980

Preparation of improved cryoprecipitated factor VIII concentrate. A controlled study of three variables affecting the yield.

The effect on cryoprecipitate factor VIII recovery of three imortant variables has been investigated. The design of the study was such as to eliminate, for the purpose of internal comparisons, spurious effects such as those due to the variation in factor VIII levels in donor plasma. The choice of anticoagulant was shown to be of particular significance, plasma collected into CPD anticoagulant giving higher cryoprecipitate factor VIII yield thoughout the study. The effect on cryoprecipitate factor VIII recovery of delay before separation and freezing of the plasma was more complex. Very fresh plasma (separated and frozen within 2 h of collection) gave the highest recovery of factor VIII, but no difference was detected between cryoprecipitate factor VIII recovery of plasma separated and frozen at 4 h and at 18 h after blood collection. Controlled rapid thawing of the plasma was also shown to be advantageous, at least for the preparation of cryoprecipitates from plasma anticoagulated with CPD anticoagulant.

Anticoagulants↗
PubMed1983

An integrated liquid-frozen blood banking system.

Frozen red cells have been field-tested at the Mobile Fleet Hospital deployed at 29 Palms, Calif., and at Bridgeport, Calif., and at the PACOM Blood Program Office, Okinawa, Japan, to evaluate the feasibility of using these products to support the US Navy and Marines in combat areas. We studied both nonrejuvenated and rejuvenated cryopreserved red cell concentrates. Red cells stored at 4 degrees C in CPD or CPDA-1 for 3-6 days (nonrejuvenated), and red cells stored at 4 degrees C in CPD for 22-26 days or in CPDA-1 for 36-38 days before biochemical modification (rejuvenated), were frozen with 40% w/v glycerol at -80 degrees C. A multiple-bag collection system with a 600 or 800 ml primary bag was used for blood collection, and the primary bag was used for processing from preparation of the red cell concentrate through prewash dilution. The thawed red cells were washed with 1.5 liters of a sodium chloride-glucose-phosphate solution. As part of this feasibility study, frozen red cells were shipped by air in dry ice (-79 degrees C), and previously frozen washed red cells were shipped in wet ice (4 degrees C). In vitro red cell recovery values were at least 90%. After postwash storage at 4 degrees C for as long as 3 days, the red cells had 24-hour posttransfusion survivals of at least 70%, normal or slightly impaired oxygen transport function, and minimal hemolysis.

Blood Banks↗
PubMed1986

Influence of the primary anticoagulant on the recovery of factor VIII in cryoprecipitate.

The influence of anticoagulant on overall factor VIII-yield was measured by drawing blood from one donor simultaneously in three bags containing ACD, CPD and heparin, respectively. After parallel processing factor VIII:C and factor VIII:CAg were measured. It is concluded that, under the circumstances used in this experiment, CPD gives the highest yield of factor VIII.

Adenine↗
PubMed1987

Survival of red cells stored for 21 and 35 days in a non-di-(2-ethylhexyl)phthalate plastic container.

Whole blood and red cells were stored using citrate-phosphate-dextrose (CPD) and citrate-phosphate-dextrose-adenine (CPDA-1) anticoagulants in polyvinylchloride bags made flexible with di-(2-ethylhexyl)phthalate (DEHP) or tri-(2-ethylhexyl)trimellitate (TOTM) plasticizers. After storage the posttransfusion viability of these cells was tested in autologous donors. Cells stored in TOTM-plasticized film had a survival rate less than 75% when stored for 35 days, while other systems had a survival greater than this. When compared with the red cells stored in CPD-DEHP-plasticized film, the viability of whole blood and red cells stored in CPDA-TOTM showed a statistically significant decrease (p = less than 0.01). Therefore, red cell storage in TOTM-plasticized PVC with current anticoagulant should be limited to 21 days.

Benzoates↗
PubMed1989

Centrifugation of very freshly donated blood may yield platelets unstable to storage in the new generation of containers.

The stability of platelets stored in the second generation of container has been examined further. Platelet concentrates prepared in CPD anticoagulant were found to deteriorate fairly rapidly during storage if prepared from very freshly donated blood, whereas a more than 4-hour delay between donation and platelet isolation yielded platelet concentrates much more stable to storage. Delays of more than 2 h (relative to the time of donation) in the isolation of platelets led to decreases in the aggregation response to low or moderate doses of collagen. Platelets from blood collected into CPD adenine showed decreased aggregation responses relative to equivalent cells from blood collected into adenine-free anticoagulant. It seems that a decreased aggregation response is associated with improved storage properties.

Adenine↗
PubMed1992

Influence of the cryoprotective agents glycerol and hydroxyethyl starch on red blood cell ATP and 2,3-diphosphoglyceric acid levels.

Because hydroxyethyl starch (HAES) is used for volume replacement therapy and as a cryoprotectant for frozen red blood cells (RBCs), this compound, in contrast to glycerol, does not require labor-intensive removal from thawed cells prior to transfusion. We here report the effect of both glycerol and HAES on the RBC organic phosphates ATP and 2,3-diphosphoglyceric acid (2,3-DPG). The CPD-A1-stabilized RBCs of 20 healthy donors (3 females, 17 males) were separately frozen in either 40% glycerol or 6% HAES, of molecular weight 200,000. ATP and 2,3-DPG concentrations were determined in CPD-A1 RBCs before addition of cryoprotectant and in cryopreserved thawed RBCs after 24 h storage at -80 degrees C (glycerol) and -196 degrees C (HAES). It appears that HAES, but not glycerol, significantly reduces ATP concentrations whereas both lead to a reduction of 2,3-DPG concentrations; this reduction was more pronounced with glycerol than with HAES. Experiments with the blood of 6 donors demonstrated that HAES affects autohemolysis by 16%, in contrast to glycerol, after which cryoprotectant autohemolysis was affected by 3.1% only. RBC recoveries were comparable using glycerol or HAES as cryoprotectants. A distinct pattern of reduction of 2,3-DPG levels by glycerol and less by HAES, and of ATP levels by HAES but not by glycerol, emerges. Our findings may be of importance if HAES is to be introduced as a convenient cryoprotectant.

2,3-Diphosphoglycerate↗
PubMed1992

Red cell and platelet concentrates from blood collected into half-strength citrate anticoagulant: improved maintenance of red cell 2,3-diphosphoglycerate in half-citrate red cells.

This study confirms previous work suggesting equivalent in vitro properties in blood components prepared from donations collected into half-citrate preservative (HCPD) compared to components derived from donations collected into standard citrate-phosphate-dextrose (CPD) preservatives. In addition, red cell products harvested from HCPD donations showed significantly improved maintenance of pH over storage, and this was reflected in improved maintenance of intracellular 2,3-diphosphoglycerate (2,3-DPG). This effect was observed in whole blood and in red cells suspended in a phosphate-containing additive solution (Tuta AAS). Collection into HCPD also improved 2,3-DPG maintenance in red cell concentrates processed following an 18-hour hold at 22 degrees C. These improvements were less pronounced in red cells suspended in a non-phosphate-containing medium (Fenwal Adsol) in which a higher pH was maintained even in units collected in CPD. Platelets harvested from HCPD blood and suspended in plasma showed equivalent quality to platelets from standard donations. Some deterioration of platelet properties was observed when HCPD platelets were stored in a non-citrate synthetic medium. Together with data indicating improved coagulation factor stability, these results suggest that collection into HCPD improves stored blood quality and may also allow logistical benefits in blood component preparation.

2,3-Diphosphoglycerate↗
PubMed1993

Rheological evaluation of X-ray irradiated blood.

CPD blood, in glass containers and polyvinyl chloride (PVC) bags, was X-irradiated with 50 Gy (n = 4), and then stored for up to 4 weeks. Rheological, hematological and biochemical changes of the red cells during storage were examined. Changes in blood viscosity and various properties of red cells (deformability, shape, hematological indices and concentrations of 2,3-diphosphoglycerate and nucleotides) during storage at 4 degrees C were not altered by the irradiation. The rheological, biochemical and morphological properties of red cells were as well preserved in PVC bags as in glass containers. No abnormal and/or cross-linked proteins in red-cell membranes were induced by the irradiation. Irradiation of PVC bags did not affect the above properties of red cells through any changes in the bag constituents. In conclusion, X-ray irradiation with 50 Gy to CPD blood in PVC bags does not affect the rheological functions of red cells.

Blood↗
PubMed1996

Evaluation of platelet function using the in vitro bleeding time and corrected count increment of transfused platelets. Comparison between platelet concentrates derived from pooled buffy coates and apheresis.

The functional capacity of transfused platelets was evaluated with in vitro bleeding time (IVBT) and corrected count increment (CCI) in order to compare platelet concentrates (PCs) derived from pooled buffy coats (BC-PCs) with PCs collected by apheresis (A-PCs). The suspension medium in the BC-PCs was 30% CPD plasma and 70% of an additive solution (containing sodium and potassium chloride, sodium citrate and phosphate, mannitol), and in the A-PCs the medium was 100% CPD plasma. IVBT was evaluated using a Thrombostat 4000/2. BC-PC and A-PC were transfused 57 and 41 times, respectively to 36 patients with chemotherapy-induced thrombocytopenia. PCs transfused within 2 days of donation were considered fresh, and those transfused within 3-5 days were considered stored. IVBT was determined before, as well as 10-30 min and 24 h after transfusion; CCI was determined 10-30 min and 24 h after transfusion. The median pretransfusion IVBT value was 486 s. It was measurable in 21 of 98 (21%) of the transfusions, i.e. below the cutoff limit of 486 s. Ten to 30 min after transfusion, the IVBT showed a measurable reduction in 90% of the transfusions with fresh BC-PCs, 92% of those with fresh a-PCs, 63% of those with stored BC-PCs and 79% of those with stored A-PCs. After 24 h, the corresponding values were 63% for fresh BC-PCs, 50% for fresh A-PCs, 26% for stored BC-PCs and 38% for stored A-PCs. The median value of CCI 10-30 min after transfusion was 20 for fresh BC-PCs, 17 for fresh A-PCs, 16 for stored BC-PCs and 14 for stored A-PCs. The difference in IVBT between fresh and stored BC-PCs was significant (p = 0.032), unlike that between fresh and stored A-PC. After 24 h the corresponding values were 7 for fresh BC-PCs, 4 for fresh A-PCs, 4 for stored BC-PCs and 3 for stored A-PCs. When all transfusions with fresh PCs (BC-PCs + A-PCs) were compared with all transfusions with stored PCs, a statistical difference was demonstrated in both CCI (p = 0.027) and IVBT (p = 0.043). Spearman's rank correlation coefficient (rs) was -0.41 between CCI and IVBT < 486 s 10-30 min after transfusion, and -0.55 between the posttransfusion platelet count and IVBT, indicating a relatively poor correlation between CCI and IVBT, and a slightly better correlation between platelet count and IVBT. In conclusion, BC-PCs showed a slightly higher CCI and a better response in IVBT than A-PCs. No statistical difference was demonstrated between BC-PCs and A-PCs transfused within 2 days after donation, with respect to function and recovery in vivo. BC-PCs stored for 3 days or more showed about the same CCI and IVBT as stored A-PC but significantly lower CCI and higher IVBT than fresh BC-PCs. This may indicate that the preparation and/or storage conditions were not optimal. IVBT seems to be a useful possibility to test the in vivo behavior of transfused platelets.

Adolescent↗
PubMed2005

In-vitro evaluation of the PALL Leukotrap Affinity Prion Reduction Filter as a secondary device following primary leucoreduction.

BACKGROUND AND OBJECTIVES: A filter (PRF1) designed to remove abnormal prion proteins from red-cell units has been developed. The purpose of this study was to evaluate the quality of red cells produced using this device. MATERIALS AND METHODS: Leucocyte-depleted red-cell units (CPD, CPD-A1 and CPD/SAGM) processed according to standard UK practices were filtered using PRF1. Filtered and control units were stored and sampled on day 1, day 7 and on the date of expiry and were tested using standard measures of red-cell quality. RESULTS: Filtered units were found to have significantly higher percentage haemolysis levels, lower haemoglobin levels and a smaller volume compared with controls. All results, however, were well within the permitted 0.8% haemolysis level at the end of storage and all units met the UK guidelines for haemoglobin and volume. The other test parameters measured showed no significant differences between the test and control units. CONCLUSIONS: The PRF1 filter was found to be easy to use and resulted in red-cell units that met all relevant UK and European Guidelines.

Erythrocyte Transfusion↗
PubMed2006

Carboxypeptidase E in the mouse placenta.

Carboxypeptidase E (CPE) has important functions in processing of endocrine pro-peptides, such as pro-insulin, pro-opiomelanocortin, or pro-gonadotropin-releasing hormone, as evidenced by the hyper-pro-insulinemia, obesity, and sterility of Cpe mutant mice. Down-regulation of Cpe in enlarged placentas of interspecific hybrid (interspecies hybrid placental dysplasia (IHPD)) and cloned mice suggested that reduced CPE enzyme and receptor activity could underlie abnormal placental phenotypes. In this study, we have explored the role of Cpe in murine placentation by determining its expression at various stages of gestation, and by phenotypic analysis of Cpe mutant placentas. Our results show that Cpe and Carboxypeptidase D (Cpd), another carboxypeptidase with a very similar function, are strictly co-localized in the mouse placenta from late mid-gestation to term. We also show that absence of CPE causes a sporadic but striking placental phenotype characterized by an increase in giant and glycogen cell numbers and giant cell hypertrophy. Microarray-based transcriptional profiling of Cpe mutant placentas identified only a very small number of genes with altered expression, including Dtprp, which belongs to the prolactin gene family. Concordant deregulation of Cpe and Cpd in abnormal placentas of interspecies hybrids before the onset of IHPD phenotype and recapitulation of some phenotypes of IHPD hyperplastic placentas in Cpe mutant placentas suggests that these two genes are causally involved in IHPD and may function as speciation genes in the genus Mus.

Animals↗