60℃加热1小时灭活病毒方法对常规临床检测指标的影响研究

目的基于60℃加热1 h可有效灭活埃博拉病毒的观点,探讨用该条件加热处理对常规临床检测指标的影响。方法对40例受检者用枸橼酸钠抗凝管和EDTA-K2抗凝管分别采集静脉血,标本分为2份,检测未进行加热灭活处理(灭活前)和进行了加热灭活处理(灭活后)样本的生物化学、免疫、凝血和血常规指标,比较热灭活前后各指标的差异。结果与灭活前相比,灭活后样本的生物化学和免疫项目中总蛋白、总胆红素、肌酐、总胆固醇、甘油三酯、钠、钾、氯、C-反应蛋白、尿素氮、葡萄糖、HBs Ab水平差异无统计学意义(P均0.05);白蛋白、球蛋白、前白蛋白、丙氨酸氨基转移酶、天冬氨酸氨基转移酶和乳酸脱氢酶有较显著降低,但下降幅度不到80%且灭活前、后数据存在相关性(r0.80,P0.01),用回归方程校正后检测数据仍具有参考价值;碱性磷酸酶、γ-谷氨酰基转移酶、胆碱酯酶和肌酸激酶的测定值下降非常明显(P均0.01),且灭活前后2组数据无显著相关性。凝血检测项目中,D-二聚体在2组间的差异无统计学意义(P0.05);其余项目均受到热灭活严重影响。血常规项目中,血红蛋白、白细胞和血小板计数在2组间的差异无统计学意义(P0.05),但样本须要事先稀释。结论 60℃加热1 h灭活处理对部分生物化学、免疫和血常规项目无明显影响,部分项目检测值明显降低,但其中有6项指标经校正后仍具参考价值。     

The relationship between coagulation and the extend of surgery and postoperative infection in surgical infants below 6 months of age

AIM: First to assess coagulation changes after surgery in children below 6 months of age. Second to detect differences attributable to the extent of surgery and postoperative infection. MATERIALS AND METHODS: Blood counts, haemoglobin concentration (Hb), haematocrit (Ht), prothrombine time (PT), activated partial thromboplastine time (aPTT) and thrombelastography (TEG) were studied pre- and 2+/-1/2 d postoperatively. Patients were divided in 3 groups. I: minor surgery without access to the abdomen or thorax (n=51); II: abdominal or thoracic interventions (n=24); III: abdominal surgery with postoperative sepsis (n=11). RESULTS: Preoperative values of Hb, Ht and INR were related to the age of the infant. Postoperatively clot strength and formation rate increased in gr. I (p<0.05). In gr. II, clot formation was initiated earlier (p<0.05) even though PT decreased (p<0.05). In group III, patients postoperatively developed a tendency for hypocoagulability in all TEG-parameters, but not in plasmatic coagulation. Postoperative TEG measurements were significantly inferior in gr. III when compared to gr. I and II. CONCLUSION: Our findings suggest activation of whole blood coagulation in the uncomplicated postoperative period despite of a decrease in plasmatic coagulation. In sepsis, only thrombelastography, but not plasmatic coagulation was affected.     

Centrifugal and roller pumps — are there differences in coagulation and fibrinolysis during and after cardiopulmonary bypass?

Summary A number of hemostatic parameters reflecting the activation of coagulation and fibrinolysis were investigated in a prospective study of 24 patients undergoing cardiopulmonary bypass (CPB) during heart surgery. The patients were randomized to a group in which either a roller (group 1) or a centrifugal pump (group 2) was used. Blood samples were taken preoperatively, at the onset of and every 20min during CPB, after the administration of protamine, and 4, 20, 44, and 68 h postoperatively. The groups did not differ significantly in hematocrit, fibrinogen, factor XIII, and antithrombin III. Significant differences in favor of group 2 during and after CPB were found in prothrombin fragment F1+2, plasmin-antiplasmin complex (PAP), thrombin-antithrombin complex (TAT), and D-dimer (F1+2P < 0.01 after 80-min CPB, PAPP < 0.005 after 40-min CPB, TAT and D-dimerP < 0.05 after 100-min CPB, D-dimer and PAPP < 0.05 after protamine administration, TAT and F1+2 4h after CPB).These findings indicate the activation of fibrinolysis preceding thrombin generation during cardiopulmonary bypass. In addition, we conclude that centrifugal blood pumping is beneficial in avoiding excessive activation of both coagulation and fibrinolysis.     

Impact of evacuated collection tube fill volume and mixing on routine coagulation testing using 2.5-ml (pediatric) tubes

BACKGROUND: Anecdotal observations by pharmacists monitoring anticoagulated patients indicate that blood samples collected in 2.7-mL (pediatric) evacuated tubes frequently produced falsely elevated international normalized ratio (INR) results. OBJECTIVE: To evaluate the impact of various preanalytical variables (fill volume, sample mixing, and elapsed time between sample collection and mixing) on INR test results using pediatric collection tubes in healthy volunteers and patients receiving warfarin anticoagulation therapy. Fifteen patients receiving warfarin and the 15 healthy volunteers participated in each study arm. METHODS: Multiple blood samples for coagulation testing were obtained from study subjects in full-draw pediatric collection tubes made of siliconized glass. The impact of sample mixing was evaluated by randomly varying the number of times each tube (five tubes total) was inverted following sample collection between one and five. The impact of timely sample mixing was evaluated by randomly varying the elapsed time between sample collection and mixing between 0 min and 4 min in each of five samples. The impact of incomplete collection tube filling was evaluated by randomly varying the volume of six tubes between 50% and 100%. Duplicate coagulation assays were performed on each sample by a centralized hematology laboratory, and the average result was reported. RESULTS: Statistical analysis revealed that neither sample mixing nor the elapsed time between sample collection and mixing had a statistically significant effect on INR test results. For patients receiving warfarin, tube fill volume had a statistically significant effect on the reported INR results (p < 0.001). The mean (+/- SD) INR derived from sample tubes filled 100% was 3.2 +/- 1.2, compared to 9.9 +/- 4.2 for tubes filled only 50% full (p < 0.01). Statistically significant INR elevations became apparent for sample tube fill volumes of < 90%. CONCLUSION: Pediatric blood collection tubes should be filled at least 90% full to ensure accurate INR test results. Anticoagulation therapy providers should routinely inquire about the type of collection tube used (adult vs pediatric) and the adequacy of sample collection volume before deriving therapeutic plans in asymptomatic excessively anticoagulated patients.     

Donor Safety and Plasma Quality in Automated Plasmapheresis

The level of blood cells and differential counts as well as of selected clotting and complement system components and breakdown products were measured in donor plasma of 42 polycarbonate filter (group I) and 7 nylon filter (group II) plasmapheresis procedures. Three different sampling time points were considered: (1) 1 min prior to connecting the donor to the machine (sample A); (2) 1 min after donation by a repeat venipuncture (sample B), and (3) in collected plasma (sample C). The better biocompatibility of the newly introduced nylon filters became evident on the basis of blood cell counts with significant drops of total white blood cell counts, monocytes, lymphocytes and platelets in sample B of group I, but not of group II. Similarly, complement studies revealed significant decrease of CH 50, C4 and C3 in samples B and C of group I, but only in samples C of group II. Coagulation studies showed significant increases of fibrinopeptide A and beta-thromboglobulin in samples B and C of group I; in group II beta-thromboglobulin was significantly increased in sample C compared to sample A. Plasminogen levels were decreased in samples B and C of group I but not of group II. Nonactivated partial thromboplastin time remained normal in group I. Factor VIII:C determinations in group II revealed a recovery of 86% in sample C.     

Intracardiac echo-guided radiofrequency catheter ablation of atrial fibrillation in patients with atrial septal defect or patent foramen ovale repair: a feasibility, safety, and efficacy study

Introduction: Patients with atrial septal defect (ASD) are at higher risk for atrial fibrillation (AF) even after repair. Transseptal access in these patients is perceived to be difficult. We describe the feasibility, safety, and efficacy of pulmonary vein antral isolation (PVAI) in these patients. Method: We prospectively compared post‐ASD/patent foramen ovale (PFO) repair patients (group I, n = 45) with age‐gender‐AF type matched controls (group II, n = 45). All the patients underwent PVAI through a double transseptal puncture with a roving circular mapping catheter technique guided by intracardiac echocardiography (ICE). The short‐term (3 months) and long‐term (12 month) failure rates were assessed. Results: In group I, 23 (51%) had percutaneous closure devices and 22 (49%) had a surgical closure. There was no significant difference between group I and II in the baseline characteristics. Intracardiac echo‐guided double transseptal access was obtained in 98% of patients in group I and in 100% of patients in group II. PVAI was performed in all patients, with right atrial flutter ablation in 7 patients in group I and in 4 patients in group II. Over a mean follow‐up of 15 ± 4 months, group I had higher short‐term (18% vs 13%, P = 0.77) and long‐term recurrence (24% vs 18%, P = 0.6) than group II. There was no significant difference in the perioperative complications between the two groups. Echocardiography at 3 months showed interatrial communication in 2 patients in group I and 1 patient in group II, which resolved at 12 months. Conclusion: Percutaneous AF ablation using double transseptal access is feasible, safe, and efficacious in patients with ASD and PFO repairs.     

Heparan sulfate levels in mucopolysaccharidoses and mucolipidoses

Summary Glycosaminoglycans are accumulated in both mucopolysaccharidoses (MPS) and mucolipidoses (ML). MPS I, II, III and VII and ML II and ML III patients cannot properly degrade heparan sulphate (HS). In spite of the importance of HS storage in the metabolic pathway in these diseases, blood and urine HS levels have not been determined systematically using a simple and economical method. Using a new ELISA method using anti-HS antibodies, HS concentrations in blood and urine were determined in MPS and ML II and ML III patients. HS concentrations were determined in 156 plasma samples from MPS I (n = 23), MPS II (n = 26), MPS III (n = 24), MPS IV (n = 62), MPS VI (n = 5), MPS VII (n = 5), ML II (n = 8) and ML III (n = 3), and 205 urine samples from MPS I (n = 33), MPS II (n = 33), MPS III (n = 30), MPS IV (n = 82), MPS VI (n = 7), MPS VII (n = 9), ML II (n = 8) and ML III (n = 3). The ELISA method used monoclonal antibodies against HS. MPS I, II, III and VII and ML II and III patients had significant elevation in plasma HS, compared to the age-matched controls (p < 0.0001). Eighty-three out of 89 (93.3%) of individual values in the above MPS types and ML were above the mean +2SD of the controls. In urine samples, 75% of individual values in patients with those types were above the mean +2SD of the controls. In contrast to the previous understanding of the HS metabolic pathway, plasma HS levels in all five MPS VI and 15% of MPS IV patients were elevated above the mean +2SD of the controls. These findings suggest that HS concentration determined by ELISA, especially in plasma, could be a helpful marker for detection of the most severe MPS I, II, III, VI and VII and ML II, distinguishing them from normal populations.     

Piperacillin/tazobactam vs ceftazidime in the treatment of neutropenic fever in patients with acute leukemia or following autologous peripheral blood stem cell transplantation: a prospective randomized trial

Piperacillin/tazobactam was compared with ceftazidime for the empirical treatment of febrile neutropenia in patients with acute leukemia or following autologous peripheral blood stem cell transplantation. Owing to inclusion criteria, it was possible for the same patient to be randomized several times. A total of 219 individual patients were admitted to a prospective randomized clinical study: 24 patients were included twice. Patients (23.5%) remained afebrile. Patients who developed febrile neutropenia were randomized to receive intravenous ceftazidime (n = 74 patients, group I) or piperacillin/tazobactam (n = 87 patients, group II). Response to first-line antibiotic treatment was seen in 55% (group I) and 53% (group II). After the addition of vancomycin, a further 19% (group I) and 24% (group II) of the patients became afebrile. Causes of fever were: microbiologically documented infection in 36 and 34 patients of group I and II; Clostridium difficile in eight and 12 patients of group I and II, and fever of unknown origin in 30 and 41 patients of group I and II. One patient died in each group. Single-agent therapy with piperacillin/tazobactam is as effective as ceftazidime in the treatment of neutropenic fever and is well tolerated. Direct and indirect costs of both treatment regimes are equivalent.     

International Normalized Ratio (INR), coagulation factor activities and calibrated automated thrombin generation – influence of 24 h storage at ambient temperature

International Normalized Ratio (INR) measurements are used to monitor oral anticoagulation therapy with coumarins. Single coagulation factor activities and calibrated automated thrombin (CAT) generation are considered as more advanced methods for evaluating overall haemostatic capacity. The aims were to assess the variability of INR, coagulation factor activities, and CAT, during 24 h of storage of blood samples at ambient temperature. A total of 24 patients on stable coumarin treatment were followed prospectively for 6 weeks. INR was analyzed at 0, 6 and 24 h after blood sampling and 1-stage clotting activity of coagulation factors II, VII, IX, and X as well as CAT generation was recorded after 0 and 24 h respectively. Statistical analyses included Bland–Altman plot, 95% limits of agreement, and a variability test using a mixed effect model. The level of INR remained statistically unchanged from 0 to 6 and 24 h of storage. Coagulation factor activities and CAT revealed no significant difference induced by 24 h of storage, although the limits of agreement were wide. Patients’ individual INR, coagulation factor activities, and CAT generation were not significantly influenced by 24 h storage of blood samples, but for the CAT generation analyses a trend toward time dependency was detected.     

Storage Temperature of the Unbuffered Rapid Urease Test

Objectives: The unbuffered rapid urease test (RUT) is an accurate, rapid, and inexpensive method for detecting Helicobacter pylori. However, it is generally recommended that the reagent he prepared daily. This prospective study was undertaken to evaluate the shelf life of our unbuffered RUT when stored at 4 and –20°C. Methods : Ninety-five patients were studied. Three sets of antral (X2) and body (X1) biopsy samples were taken from each patient. The samples were subjected to histological examination, with the RUTs stored at 4 and –20°C. The RUT tubes were examined at 1 and 15 min. Results : Fifty-six patients (59%) were infected with H. pylori as defined by histological examination. The reagent was classified according to storage time (group I, ≤5 days; group II, >5 days). The mean (SD) storage time of group I (n = 59) and group II (n = 36) was 3.2 (1.4) and 9.9 (5.0) days, respectively. At 15 min, the sensitivity of our RUT stored at 4°C was significantly higher in group I than in group II (92 vs 47%). On the other hand, the sensitivity of our RUT stored at –20°C remained consistently high in both groups (15 min: group I, 92%; group II, 100%). Our RUTs stored at 4 and –20°C were highly specific in both groups. Conclusions : Our RUT remains highly sensitive and specific when it is stored at 4°C for up to 5 days. When the RUT is expected to be stored for a longer period of time, the bottles should he frozen at –20°C.     

Arterial hypertension, left ventricular hypertrophy and arrhythmias]

Arterial hypertension is the most common cause of chronic pressure overload of the left ventricle. Electrocardiographic and echocardiographic signs of left ventricular hypertrophy in hypertensive patients are associated with an increased cardiovascular mortality and incidence of sudden death habitually due to ventricular arrhythmias. The significance of a normal increase in systolic blood pressure during exercise in persons without evident resting hypertension is uncertain. M-mode and 2D echocardiography, 24-hour continuous ambulatory electrocardiographic (Holter), exercise testing and 24-hour ambulatory blood pressure monitoring (ABPM) were performed on 22 normotensive patients (group I); 25 normotensives with exaggerated blood pressure response to exercise (greater than 220 mmHg) (group II) and 33 hypertensive patients (group III). None was taking cardioactive drugs. Left ventricular hypertrophy (LVH) was found on one patient of group I (4.5%), 13 of group II (52%) and 20 of group III (61%). Left ventricular mass index (LVMI) was linearly correlated with maximum exercise blood pressure (group I: r2 = 0.518, p less than 0.0002; group II: r2 = 0.098, NS; group III: r2 = 0.407, p less than 0.0001) with 24-hour systolic pressure overload (ABPM) (group I: r2 = 0.848, p less than 0.0001; group II: r2 = 0.705, p less than 0.0001; group III: r2 = 0.839, p less than 0.0001) and 24-hour diastolic pressure overload (ABPM) (group I: r2 = 0.612, p less than 0.0001; group II: r2 = 0.815, p less than 0.0001; group III: r2 = 0.807, p less than 0.0001) within each group but not between different groups. The hypertensive subjects (group III) had a higher average heart rate (p less than 0.0001) more supraventricular premature (p less than 0.0001) and ventricular premature (p less than 0.0001) beats than the normotensive (group I) and normotensive patients with abnormal increases in systolic blood pressure response to exercise (group II) (p less than 0.0001) (NS) and (p less than 0.0002), respectively. LVMI was linearly correlated with ventricular premature beats (group I: r2 = 0.072, NS; group II: r2 = 0.823, p less than 0.0001; group III: r2 = 0.691, p less than 0.0001). Frequent and complex ventricular arrhythmias were more common in patients with LVH normotensives or hypertensives than without LVI (p less than 0.0001) and the age increases their severity. We conclude that normotensives with hypertensive response to exercise have similar incidence of LVI; if those patients develop sustained hypertension, LVI was previous to arterial hypertension. There are two types of hypertrophy: secondary hypertrophy is linked to the high afterload and vasoconstriction typical in hypertension.(ABSTRACT TRUNCATED AT 400 WORDS)     

Obstructive jaundice promotes bacterial translocation in humans.

BACKGROUND/AIMS: Significant bacterial translocation was demonstrated following experimental biliary obstruction, however very little is known about the importance and the prevalence of gut-origin sepsis in obstructive jaundice patients. Therefore, the aim of this study was to investigate the concept of gut-origin sepsis in obstructive jaundiced patients and its clinical importance. METHODOLOGY: Twenty-one patients requiring laparotomy for obstructive jaundice (group I) and thirty patients operated on electively mainly for chronic cholecystitis (group II) were studied. Peritoneal swab, mesenteric lymph node, portal venous blood, liver wedge biopsy and bile were sampled for culture immediately after opening the peritoneum. Additionally, peripheral blood samples were taken pre- and post-operatively from all patients. Post-operatively, patients were monitored for infectious complications. RESULTS: The mean serum bilirubin concentration, gamma glutamyl transferase and alkaline phosphatase levels in jaundiced patients before therapeutic intervention were significantly higher than in control patients. Five patients demonstrated bacterial translocation in group I (24%), whereas only one did so in group II (3.5%, p < 0.05). Septic complications were detected in three patients, but only in two with bacterial translocation in group I. There was one patient with bacterial translocation who had septic complication in group II. CONCLUSIONS: The present study demonstrated that obstructive jaundice significantly promotes bacterial translocation in humans, however, its clinical importance has yet to be defined.     

Prevalence of daytime hypercapnia or hypoxia in patients with OSAS and normal lung function

The purpose of this study was to determine factors increasing daytime P aCO₂orP aO₂in obstructive sleep apnoea syndrome patients (OSAS) with normal pulmonary function tests. Anthropometric, pulmonary function tests, arterial blood gases and sleep polygraphic data were analysed retrospectively in 218 OSAS patients (apnoea–hypopnoea index >15 h⁻¹; 18 females, 55±11 years): 125 patients had abnormal pulmonary function tests, i.e. one or more flow or volume under 80% or above 120% of predictive value (group I) and 93 had normal pulmonary function tests (group II). Hypercapnia was defined as P aCO₂≥6·0 kPa and hypoxia asP aO₂<9·3 kPa. Patients with abnormal pulmonary function tests were more hypoxic and hypercapnic, more obese, and had a higher apnoea–hypopnoea index (P<0·05). Seventeen patients of group I and four of group II were hypercapnic (13·6% and 4·3%, respectively ). Thirty-one patients in group I (24·8%) had a P aO₂<9·3 kPa and six (6·5%) in group II. Stepwise multiple regression analysis showed that in group II, only two factors were correlated with P aCO₂: mean apnoea duration and FRC (respectively: c=0·228, P<0·001;c=0·006,P =0·0108); and only two with P aO₂: mean apnoea duration: (c=−0·218, P=0·029) and BMI (c=−3·72,P <0·0001). Daytime hypercapnia is present in 4·3% and daytime hypoxia in 6·5% of patients with occlusive sleep apnoea syndrome and normal pulmonary function tests. These alterations in blood gases in OSAS with normal pulmonary function tests should be considered as OSAS severity criteria.     

Effects of beta-radiation using a holmium-166 coated balloon on neointimal hyperplasia in a porcine coronary stent restenosis model.

Brachytherapy is a promising method of preventing and treating coronary stent restenosis. The present study was designed to observe the therapeutic effects of a radioactive balloon loaded with Holmium-166 ((166)Ho) in a porcine coronary stent restenosis model. A radioisotope of (166)Ho was coated onto the balloon surface using polyurethane (20 Gy at 0.5 mm depth). Stent overdilation injuries were induced in 2 coronary arteries in each pig (n=8). Four weeks after the injury, control balloon dilation was performed in one coronary artery (Group I) and radiation therapy using the (166)Ho coated balloon in the other coronary artery (Group II) in each pig. Follow-up coronary angiography and histopathologic assessment were performed at 4 weeks after the radiation therapy or the control balloon dilations. With regard to complete blood cell counts, liver function tests, lipid profiles and coagulation tests, there were no differences between the baseline and after radiation. On quantitative coronary angiographic analysis, reference and target artery diameter showed no differences between the 2 groups before, or 4 and 8 weeks after stenting. On histopathologic analysis of groups I and II, the injury score was 1.34+/-0.09 and 1.32+/-0.10, the area of internal elastic lamina was 4.99+/-0.17 mm(2) and 4.82+/-0.20 mm(2), and the luminal area was 3.20+/-0.10 mm(2) and 3.45+/-0.14 mm(2), respectively (p=NS). The neointimal area was 1.78+/-0.11 mm(2) in group I and 1.36+/-0.12 mm(2) in group II (p=0.017), and the histopathologic area of stenosis was 35.1+/-1.6% in group I and 27.6+/-1.9% in group II (p=0.005). In conclusion, beta-radiation of the stented porcine coronary artery using a radioactive (166)Ho coated balloon inhibited stent restenosis without any side effects.     

Hypercoagulability after immunotherapy with corynebacterium parvum in man

Summary The effect of an immunotherapy with Corynebacterium parvum on the blood coagulation system was investigated in a randomized trial of 18 patients with metastatic breast cancer. All patients received cytostatic therapy. Additionally, C. parvum was given intravenously on day 15 of the cytostatic cycle (group I) or on day 1 (group II) or not at all (group III). Fibrinopeptide A increased within 2 h after intravenous administration of C. parvum in groups I and II and normalized after 24 h (p<0.05). Platelet counts decreased continuously in all treatment groups (p<0.05). Prothrombin time, fibrinogen concentration, factor VIII:C and factor VIIIR:Ag were not affected. The fibrinolytic activity showed a slight but not statistically significant increase after intravenous administration of C. parvum. The data suggest that plasma hypercoagulability is induced or enhanced in man even after small intravenous doses of C. parvum.Supported by grants from the Deutsche ForschungsgemeinschaftDedicated to Prof. Dr. Dr. h. c. mult. G. Schettler on his 65^th birthday     

Erectile and endothelial dysfunction in Type II diabetes: a possible link

Aims/hypothesis: The aim of this study was to evaluate the relation between erectile dysfunction and endothelial functions, coagulation activation, peripheral and autonomic neuropathy in men with Type II (non-insulin-dependent) diabetes mellitus. Methods: We studied 30 Type II diabetic patients with symptomatic erectile dysfunction and 30 potent diabetic patients matched for age and disease. Endothelial functions were assessed with the l-arginine test, plasma thrombomodulin and cell adhesion molecules circulating concentrations. Haemostasis was evaluated with markers of thrombin activation and fibrinolysis. Quantitative sensory testing (vibratory, warming, and heat-pain thresholds), cardiovascular reflex tests and 24-h blood pressure monitoring were used to assess peripheral or autonomic neuropathy. Results: Mean erectile score and HbA_{1 c} were 10.5 ± 5.8 and 8.3 ± 1.6 % in patients with erectile dysfunction, and 24.0 ± 0.7 and 6.8 ± 1.4 % in those without erectile dysfunction, respectively (p < 0.001); there was a significant relation between HbA_{1 c} and erectile function score in patients with erectile dysfunction (r = –0.45, p = 0.02). The decrease in blood pressure and platelet aggregation in response to l-arginine was lower (p < 0.05–0.02) in patients with erectile dysfunction, whereas soluble thrombomodulin, P-selectin and intercellular cell ahhesion molecule-1 concentrations were higher (p < 0.05–0.02). Indices of coagulation activation (F1 + 2 and d-dimers) and reduced fibrinolysis (PAI-1) were also found to be higher in erectile dysfunction patients. Heat-pain and warm perception thresholds, as well as cardiovascular reflex tests, were most commonly abnormal in patients with erectile dysfunction (p < 0.05). In multivariate analysis, HbA_{1 c}, MBP response to l-arginine, P-selectin, indices of coagulation, and quantitative sensory testing were independent predictors of erectile function score. Conclusion/interpretation: Erectile dysfunction in diabetic men correlates with endothelial dysfunction. A reduced nitric oxide activity might provide a unifying explanation. [Diabetologia (2001) 44: 1155–1160] Received: 18 April 2001 and in revised form: 21 April 2001     

Maintaining fetal normoglycemia prevents the increase in myometrial activity and uterine 13,14-dihydro-15-keto-prostaglandin F2 alpha production during food withdrawal in late pregnancy in the ewe

Food deprivation during pregnancy leads to an increase in maternal and fetal prostaglandin (PG) production and increased uterine contractility. We investigated the effect of maintaining fetal normoglycemia during food withdrawal-induced maternal hypoglycemia on uterine 13,14-dihydro-15-keto-prostaglandin F2 alpha (PGFM) production and myometrial activity in late pregnant sheep. Pregnant sheep were surgically instrumented with fetal and maternal catheters and electromyogram leads under halothane anesthesia. Maternal and fetal blood plasma samples were obtained once a day at 0900 h, 24 h before (baseline sample) and after 48 h of food withdrawal. Food, but not water, was withdrawn from ewes in group I (n = 5). During food withdrawal in group II (n = 5), glucose was infused into a fetal vein to maintain fetal normoglycemia. All data were normalized to the concentration in the baseline sample in each animal as 100%. After 48 h of food withdrawal, maternal whole blood glucose fell by 42.2 +/- 4.4% (mean +/- SEM: group I) and 31.4 +/- 6.2% (group II). These values were not significantly different. Fetal blood glucose fell by 40.4 +/- 5.7% (group I). In group II, fetal blood glucose was maintained in the normal range (99.6 +/- 1.6% of baseline). Maternal uterine electromyogram activity, uterine venous estrone sulfate, and uterine veno-arterial difference in PGFM rose significantly during food withdrawal in group I ewes, but not in group II ewes. Maternal and fetal arterial plasma ACTH and cortisol did not change in group II animals. We conclude that maintenance of fetal normoglycemia during 48 h of food withdrawal in sheep prevents the increase in myometrial activity, maternal plasma estrogens, and uterine PGFM production during food withdrawal in late pregnancy.     

The use of Vacutainer tubes for collection and storage of blood for coagulation testing

Summary. A study has been performed to evaluate the suitability of Vacutainer tubes in blood specimen collection for coagulation tests and to compare them with the conventional syringe technique employed in UK hospitals. Blood was collected from healthy volunteers, an ante-natal group and patients on long-term oral anticoagulants. Samples were stored at two different temperatures; 4|MoC and ambient room temperature (RT). Prothrombin times, factor VII assays and APTT were performed at baseline and after 2 h and 4 h storage. There was significant activation of the extrinsic system in the blood samples collected by Vacutainer when stored at 4|MoC which became more significant on prolonged storage. The effect was less pronounced when the Vacutainer tubes were stored at RT. In contrast, the blood collected by the syringe method did not show these changes with the exception of the ante-natal specimens where a lesser degree of activation than in the Vacutainer tubes was observed after 4 h at 4|MoC. The activation of the Vacutainer samples at 4|MoC is considered undesirable and could be of clinical significance in oral anticoagulant dosage.     

Non-interferon-based therapy: an option for amelioration of necro-inflammation in hepatitis C patients who cannot afford interferon therapy.

Objectives: Interferon (IFN) therapy is not affordable by the majority of Egyptian patients. Our aim was to tailor an effective and inexpensive regimen that ameliorates hepatic necro‐inflammatory activity among chronic hepatitis C (CHC) patients. Methods: One hundred and seventy naïve CHC patients with elevated alanine aminotransferase (ALT) (>1.5‐fold) and detectable hepatitis C virus (HCV)‐RNA by polymerase chain reaction, who cannot afford IFN‐based therapy were randomly allocated either to non‐interferon‐based therapy (N‐IFN‐BT) (group I) or silymarin therapy (group II). Group I comprised 87 patients (biopsy proved chronic hepatitis in 62 patients) who were administered a daily combination of ribavirin (600–800 mg) plus amantadine (200 mg) and ursodeoxycholic acid (UDCA) (500 mg) for 24 weeks. Group II comprised 83 patients who were administered Silymarin 450 mg/day for 24 weeks. Results: Statistical evaluation was conducted on 82 patients from group I and 72 from group II because of the withdrawal of five and 11 patients from Groups I and II, respectively. Age, sex, social status and biochemical parameters were comparable in both groups. Normalization of ALT at the end of treatment was achieved in 58.5% and 15.3% (P<0.001), whereas end of treatment virologic response (ETVR) was achieved in 2.4% and 0% of Groups I and II, respectively. Twenty‐four weeks after cessation of therapy, sustained biochemical response (SBR) was achieved in 28% and 2.8% (P<0.001), while sustained virologic response (SVR) was maintained in 2.4% and 0% of the patients in Groups I and II, respectively. In Group I, histopathological examination revealed a decreased activity index by an average score of 1.5 points among 38/62 of the rebiopsied patients. Conclusion: Twenty‐four weeks N‐IFN‐BT achieved a fourfold‐higher ETBR and a tenfold‐higher SBR compared with silymarin therapy, which reflects an improvement of necroinflammatory activity as proven by repeat histopathology.     

Stability of coagulation proteins in frozen plasma.

This study reports on the frozen stability of all commonly measured coagulation proteins in normal citrated plasma: activated partial thromboplastin time, prothrombin time (%), thrombin time and fibrinogen (Clauss); clotting assays for factors II, V, VII, VIII, IX, X, XI and XII; functional assays for protein C (clotting), protein S (clotting), antithrombin (chromogenic) and plasminogen (chromogenic); and immunological assays for von Willebrand factor and D-dimer. All these factors listed are stable for up to 3 months if frozen at -24 degrees C or lower. At -74 degrees C, all these factors (allowing for 10% variation) were stable for at least 18 months, most were stable for 24 months. The number of proteins showing > 5% variation over baseline after 6 months storage indicates that some decay does occur even at -74 degrees C. There was no clear advantage in snap freezing at -74 degrees C and then storing at -24 degrees C over both freezing and storing at -24 degrees C; therefore, the freezing process itself is not responsible for the loss of stability. The best stability, especially at -24 degrees C, was obtained when small samples (1 ml) were stored in screw-cap tubes with a minimum dead space. The decrease in stability of the coagulation proteins directly correlates with the effect of temperature and time.