Identification of a novel centrosome/microtubule-associated coiled-coil protein involved in cell-cycle progression and spindle organization

Here we describe the identification of a novel vertebrate-specific centrosome/spindle pole-associated protein (CSPP) involved in cell-cycle regulation. The protein is predicted to have a tripartite domain structure, where the N- and C-terminal domains are linked through a coiled-coil mid-domain. Experimental analysis of the identified domains revealed that spindle association is dependent on the N-terminal and the coiled-coil mid domain. The expression of CSPP at the mRNA level was detected in all tested cell lines and in testis tissue. Ectopic expression of CSPP in HEK293T cells blocked cell-cycle progression in early G1 phase and in mitosis in a dose-dependent manner. Interestingly, mitosis-arrested cells contained aberrant spindles and showed impairment of chromosome congression. Inhibition of CSPP gene expression by small interfering RNAs induced cell-cycle arrest/delay in S phase. This phenotype was characterized by elevated levels of cyclin A, decreased levels of cyclin E and hyperphosphorylation of the S-phase checkpoint kinase Chk1. The activation of Chk1 may indicate a replication stress response due to an inappropriate G1/S-phase transition. Taken together, we demonstrate that CSPP is associated with centrosomes and microtubules and may play a role in the regulation of G(1)/S-phase progression and spindle assembly.     

Effective training for patients with intermittent claudication

OBJECTIVE: Current guidelines for treatment of intermittent claudication (IC) do not include a specific recommendation for the intensity of exercise therapy. Thus, the purpose of this study was to determine the relative effectiveness of high versus low intensity exercise for patients with IC, and further to study the effect of such training on blood flow to the legs during exercise. DESIGN: The effect of eight weeks of supervised endurance training was examined in 16 patients with IC. The patients were randomly assigned to training at intensities corresponding to either 60% or 80% of their peak oxygen consumption (VO2peak), respectively. RESULTS: VO2peak and time to exhaustion increased significantly (9% and 16%, respectively) more in the high intensity group (p<0.05). Blood flow to the legs did not change after training in any of the groups. CONCLUSION: High intensity training gave larger improvements in VO2peak and time to exhaustion than low intensity training. As blood flow did not change after the exercise program, it is likely that the observed different increase of VO2peak was due to changed mitochondrial oxidative capacity and/or skeletal muscle diffusive capacity.     

Heparin-coated circuits (Duraflo II) with reduced versus full anticoagulation during coronary artery bypass surgery

BACKGROUND: Introduction of completely heparin-coated cardiopulmonary bypass (CPB) circuits combined with reduced systemic anticoagulation has been shown to reduce postoperative bleeding and requirements for allogeneic transfusions after cardiac surgery. However, some uncertainty exists whether this effect is due to the reduced amount of heparin or to the heparinized surface itself. Therefore, a retrospective study was undertaken, comparing two different anticoagulation protocols applied to coronary artery bypass patients treated with identical heparin-coated CPB equipment. METHOD: Over a 12 month period all coronary artery bypass patients operated with extracorporeal circulation were subjected to a Duraflo II heparin-coated circuit (Baxter Healthcare Corp, Bentley Laboratories Division, Irvine, Calif) and full heparin dose (activated clotting time [ACT] > 480 seconds; Group F, n = 651). Over the next 24 months, all coronary patients who were treated with an identical circuit combined with reduced systemic heparinization (ACT > 250 seconds) were included in Group R (n = 675). Except for the different anticoagulation protocols, all treatment regimens before, during, and after the operation remained unchanged throughout the study period. RESULTS: There were no statistically significant differences in any major demographic or operative parameters. In Group R, the postoperative bleeding was mean 665 +/- 257 ml versus 757 +/- 367 ml in Group F (p < 0.0001), and the perioperative decrease in hemoglobin concentration was significantly lower in Group R (22 +/- 1.2 gm/L versus 25 +/- 1.3 gm/L, p < 0.0001). The time for postoperative ventilatory support was shorter in Group R (1.7 +/- 1.3 hours versus 1.9 +/- 1.1 hours in Group F, p = 0.0006), and the incidence of new episodes of atrial fibrillation after the operation was lower (26.4% in Group R versus 32.8% in Group F, p = 0.01). There were no significant differences in the incidences of perioperative myocardial infarction, stroke, transient neurological disturbances, physical rehabilitation, or mortality. No technical or coagulation problems were recorded in either group. CONCLUSION: The use of Duraflo II coated circuits for CPB combined with reduced anticoagulation decrease postoperative bleeding and hemoglobin loss compared with full heparin dose treatment. In addition, the intubation time was shorter and the incidence of postoperative atrial fibrillation was lower in the patients treated with low heparin doses.     

Quality of intraoperative autologous blood withdrawal used for retransfusion after cardiopulmonary bypass

BACKGROUND: Intraoperative autologous blood withdrawal protects the pooled blood from the deleterious effects of cardiopulmonary bypass. Following reinfusion after cardiopulmonary bypass, the fresh autologous blood contributes to less coagulation abnormalities and reduces postoperative bleeding and the need for allogeneic blood products. However, few data have been available concerning the quality and potential activation of fresh blood stored at room temperature in the operating room. METHODS: Forty coronary artery bypass grafting patients undergoing a consistent intraoperative and postoperative autotransfusion protocol had a median of 1,000 mL of autologous blood withdrawn before cardiopulmonary bypass. After heparinization the blood was drained from the venous catheter via venous cannula into standard blood bags and stored in the operating room until termination of cardiopulmonary bypass. Samples for hemostatic and inflammatory markers were taken from the pooled blood immediately before it was returned to the patient. RESULTS: There was some activation of platelets in the stored autologous blood, as measured by an increase of beta-thromboglobulin. Indications of thrombin formation, as assessed by plasma levels of thrombin-antithrombin complex and prothrombin fragment 1.2 were not seen, and there was no fibrinolytic activity. The red blood cells remained intact, indicated by the absence of plasma free hemoglobin. As for the inflammatory response, the levels of the terminal complement complex remained stable, and the cytokines tumor necrosis factor-alpha and interleukin 6 levels were not increased during storage. The complement activation products increased minimally, but remained within normal ranges. CONCLUSIONS: Except for slight activation of platelets, there was no indication of coagulation, hemolysis, fibrinolysis, or immunologic activity in the autologous blood after approximately 1 hour of operating room storage. The autologous blood was preserved in a condition of high quality, and retransfusion after cardiopulmonary bypass represents an uncomplicated and almost costless procedure for blood conservation.     

Heparin-coated circuits and reduced systemic anticoagulation applied to 2500 consecutive first-time coronary artery bypass grafting procedures

BACKGROUND: In contrast to the widespread popularity of off-pump techniques for coronary artery bypass grafting, our institution has chosen a different strategy, emphasizing improvements in the technology for extracorporeal circulation, as well as simplifying surgical and clinical management. The clinical short-term results of this approach were analyzed. METHODS: The on-pump strategy includes routine use of heparin-coated circuits combined with low systemic heparinization (activated coagulation time of more than 250 seconds), intention of total revascularization within limited ischemic times and pump times, minimal use of blood transfusions, early extubation, and rapid postoperative recovery. The data from the first 2,500 consecutive first-time coronary artery bypass grafting patients (January 1998 to February 2002) treated with this protocol were retrospectively analyzed. RESULTS: There were 487 female (median age 68 years) and 2013 male (median age 64 years) patients. A median of four (one to nine) (mean 4.5 +/- 1.2) distal anastomoses were created, and the median aortic cross-clamp time and pump time were 34 and 54 minutes, respectively. At least one internal mammary artery was used in 99.7% of the patients. Blood or bank blood products were given to 118 patients (4.7%). Median extubation time was 1.5 hours. The stroke rate was 0.8%, transient neurologic deficits occurred in 0.6% of the patients, and the incidence of perioperative myocardial infarction was 1.1%. By the fifth day, 91% of the patients were ready for discharge. Seven patients (0.28%) died during their hospital stay. CONCLUSIONS: Coronary artery bypass grafting with heparin-coated cardiopulmonary bypass circuits and reduced systemic anticoagulation resulted in excellent clinical results, with minimal blood transfusions and rapid postoperative mobilization. The high number of grafted coronary arteries indicates complete revascularization in most patients, which is known to be a significant predictor of long-term event-free survival.     

D‐dimer levels and stroke progression in patients with acute ischemic stroke and atrial fibrillation

Krarup L‐H, Sandset EC, Sandset PM, Berge E. D‐dimer levels and stroke progression in patients with acute ischemic stroke and atrial fibrillation.
Acta Neurol Scand: 2011: 124: 40–44.
© 2010 John Wiley & Sons A/S. Background – Patients with acute ischemic stroke and atrial fibrillation are at increased risk of stroke progression and recurrence. We sought to assess whether D‐dimer and other markers of hemostatic activation could predict these adverse events in such patients. Method – Blood samples were obtained from patients included in the Heparin in Acute Embolic Stroke Trial. Stroke progression was defined as a ≥3‐point worsening on the Scandinavian Stroke Scale during the first 48 h after randomization. Blood samples were analyzed for D‐dimer, prothrombin fragment 1 + 2, soluble fibrin monomer, and C‐reactive protein. Results – A total of 382 patients were included in the analyses. Levels of D‐dimer and other markers of hemostatic activation were not significantly higher in patients with stroke progression than in other patients (D‐dimer median values: 1025 ng/ml vs 970 ng/ml, P = 0.73). The same was true for recurrent stroke (D‐dimer: 720 ng/ml vs 973 ng/ml, P = 0.96), and the combined endpoint of stroke progression, recurrent stroke, and death (D‐dimer: 991 ng/ml vs 970 ng/ml, P = 0.91). Multivariable analyses did not alter the results. Conclusion – D‐dimer and other markers of hemostatic activation were not associated with stroke progression, recurrent stroke, or death in patients with acute ischemic stroke and atrial fibrillation.     

Why several truths can be true

In this paper, we offer a perspective on complementarity, acknowledging that it is not possible for human perception and cognition to grasp reality with unambiguous concepts or theories. Therefore, multiple concepts and perspectives are valid when they are not exaggerated beyond reasonable limits and do not claim exclusive validity. We recommend a humble stance enabling respectful dialogue between different perspectives in medical science and practice.

• KEY POINTS

• No single perspective in clinical or scientific medicine can exhaustively explain medical phenomena.

• Scientific attitude is characterised by a willingness to look for objections against what we prefer as truths.

• Complementarity or unifying contradictions are concepts that allow for humility and pluralism in clinical and scientific medicine.