Encouraging experience with intracardiac transplantation of unselected autologous bone marrow cells concomitant with coronary artery bypass surgery after myocardial infarction.

BACKGROUND: Chronic heart failure after myocardial infarction is still a serious problem without a fundamental therapy. Experimental transplantation of bone marrow cells (BMC) into infarcted myocardium resulted in regeneration and functional improvement. OBJECTIVE: Clinical investigation of safety and efficacy of intracardiac transplantation of unselected autologous BMC. METHOD: 22 patients scheduled for elective and isolated coronary artery bypass grafting (CABG) with a reduced LVEF due to myocardial infarction were included. Intraoperatively, sternal bone marrow blood was aspirated, and a sterile buffy coat was prepared and applicated. 19 age, LVEF and coronary disease matched patients served as controls. Heart function, geometry, and scar proportion were assessed by echocardiography and Gadolinium-MRI at the time of the operation and 6 months thereafter. RESULTS: Transplanted patients received a mean number of 360 × 106 BMC. We did not notice any significant differences in early or late complications in the transplant group as compared to controls. At six months follow up only the transplanted patients showed a significant improvement of NYHA classes from 2.7 to 1.5 and of LVEF from 36 to 43 %, (p < 0.05). Furthermore, only CABG concomitant with BMC-TX led to a significant reduction of left ventricular end diastolic diameter (LVEDD) from 59 to 54 mm and of scar proportion of the infarcted segments from 2.53 to 2.42, (p < 0.05). CONCLUSION: Intracardiac transplantation of unselected, autologous BMC is safe and feasible. In adjunct with coronary revascularization it leads to an improvement of ventricular geometry and function. Moreover, it reduces myocardial scar proportion and heart failure symptoms.

Effect of CYSTUS052 and green tea on subjective symptoms in patients with infection of the upper respiratory tract.

Examples of medicinal herbs that have been perpetuated along several generations based simply on a folk tradition are Cistus and green tea. The principal active constituents of the genus Cistus and green tea are polyphenolic compounds. Polyphenols exhibit a wide range of antibacterial, antifungal and antiinflammatory effects. The present work aimed to investigate the clinical effect of a Cistus extract (CYSTUS052) in comparison with green tea on 300 patients with infections of the upper respiratory tract. Due to the lack of clinical study data on their efficacy in patients, this is a report of the findings of our study on the clinical efficacy of CYSTUS052 in patients with the upper respiratory tract infections (URTIs). This study observed a total of 300 patients (277 completers) treated with CYSTUS052 given in lozenges compared with treatment with an extract of green tea. The patients scored the subjective severity of target symptoms using a predefined scale. The score of subjective symptoms decreased over the course of treatment with CYSTUS052, whereas treatment with green tea resulted in a less significant decrease of symptoms. CYSTUS052 therefore proved to be an effective adjuvant for the treatment of respiratory infections.

Cistus incanus (CYSTUS052) for treating patients with infection of the upper respiratory tract. A prospective, randomised, placebo-controlled clinical study.

In this prospective, randomized, placebo-controlled clinical study we aimed to investigate the clinical effect of a Cistus extract (CYSTUS052) in 160 patients with infections of the upper respiratory tract. The extract contains a high percentage of highly polymeric polyphenols. In cell culture and in a mouse model it exerts antiviral and antimicrobial activities. Principal active constituents of the genus Cistus are polyphenolic compounds. Plant-derived polyphenols have been shown to be strong antioxidants with potential health benefits. Various reports have appeared on the antiviral and antibacterial potential, including several reports describing the antiviral activity of polyphenols against influenza virus. Clinical studies on the effectiveness of Cistus incanus are scarce. Only one controlled application observation study demonstrated the effectiveness of a Cistus extract. The present randomised, placebo-controlled clinical study was designed to compare the symptom scores in patients with common cold treated either with CYSTUS052 or with placebo. A score of subjective symptoms decreased significantly over the course of treatment with Cistus, whereas treatment with placebo resulted in a less distinct decrease of symptoms. Among the inflammatory markers investigated, the C-reactive protein was mostly affected by Cistus and decreased significantly in the treatment group.

Depletion of autoreactive immunologic memory followed by autologous hematopoietic stem cell transplantation in patients with refractory SLE induces long-term remission through de novo generation of a juvenile and tolerant immune system.

Clinical trials have indicated that immunoablation followed by autologous hematopoietic stem cell transplantation (ASCT) has the potential to induce clinical remission in patients with refractory systemic lupus erythematosus (SLE), but the mechanisms have remained unclear. We now report the results of a single-center prospective study of long-term immune reconstitution after ASCT in 7 patients with SLE. The clinical remissions observed in these patients are accompanied by the depletion of autoreactive immunologic memory, reflected by the disappearance of pathogenic anti-double-stranded DNA (dsDNA) antibodies and protective antibodies in serum and a fundamental resetting of the adaptive immune system. The latter comprises recurrence of CD31(+)CD45RA(+)CD4(+) T cells (recent thymic emigrants) with a doubling in absolute numbers compared with age-matched healthy controls at the 3-year follow-up (P = .016), the regeneration of thymic-derived FoxP3(+) regulatory T cells, and normalization of peripheral T-cell receptor (TCR) repertoire usage. Likewise, responders exhibited normalization of the previously disturbed B-cell homeostasis with numeric recovery of the naive B-cell compartment within 1 year after ASCT. These data are the first to demonstrate that both depletion of the autoreactive immunologic memory and a profound resetting of the adaptive immune system are required to reestablish self-tolerance in SLE.

Direct assessment of thymic reactivation after autologous stem cell transplantation.

Methods to quantify Th cell reconstitution after immunosuppressive therapies such as hematopoietic stem cell transplantation are becoming a key issue since persistent Th cell deficiencies may result in severe complications and adverse events. We employed here cytometric monitoring of CD31+ thymus-naive Th cells for the direct assessment of human thymic function in 10 patients undergoing autologous stem cell transplantation for severe autoimmune diseases. High frequencies of posttransplant recurring naive Th cells coexpressed CD31 and stable long-term reconstitution with elevated absolute counts of CD31+ thymus-naive Th cells that were enriched with T cell receptor excision circles was demonstrated. Cytometric monitoring of CD31+ thymus-naive Th cells enables to directly evaluate human thymic function ex vivo.

Cryopreservation of platelets at the end of their conventional shelf life leads to severely impaired in vitro function.

Storage time for platelet concentrates (PCs) is limited to five days due to 'aging' of the platelets and an increasing risk of bacterial proliferation. Storage time can be prolonged by cryopreservation. We investigated in vitro function of six consecutive PCs at the end of their conventional shelf life followed by cryopreservation for 24 hours. Spontaneous, adenosine diphosphate (ADP)-induced and collagen-induced activation before and after cryopreservation were determined by flow cytometry. Additionally, ADP- and collagen-induced aggregation was measured. After cryopreservation two-thirds of the platelets were spontaneously activated, twice as many as before the procedure (p < 0.001). ADP-induced activation was significantly reduced (p = 0.014). Collagen-induced activation was unchanged. Aggregation stimulated by ADP and collagen was significantly reduced (p = 0.005 and p = 0.009, respectively). Our results show severely impaired in vitro function of platelets after storage at 22 degrees C for five days followed by cryopreservation. Cryopreservation of PCs after a storage time of five days cannot be recommended.

Hemoglobin screening in prospective blood donors: comparison of different blood samples and different quantitative methods.

Portable photometers are increasingly used for hemoglobin screening in blood centers. In a total of 500 unselected prospective blood donors, venous hemoglobin concentrations were measured using a hematology analyzer, and fingerstick hemoglobin concentrations were measured on both a HemoCue B and a Biotest hemoglobin photometer. In 100 of the donors, earstick hemoglobin levels were also measured on the HemoCue system. The mean hemoglobin levels in the fingerstick samples were slightly higher than those in the venous samples. The deviation exceeded the limit of +/-10g l(-1) in 9% of samples tested by HemoCue and in 20% of those investigated by Biotest. The use of earstick samples led to considerable overestimation of the actual hemoglobin concentration.

Rapid identification of iron deficiency in blood donors with red cell indexes provided by Advia 120.

BACKGROUND: A new generation of automated hematology analyzers allows the rapid determination of various red cell (RBC) indexes, including the percentage of hypochromic mature RBCs (HYPOm) and the hemoglobin (Hb) content of reticulocytes (CHr). These indexes have not yet been validated as measures for the detection of iron deficiency in blood donors. STUDY DESIGN AND METHODS: Iron status was evaluated in a total of 1142 unselected prospective blood donors based on measurement of serum ferritin, soluble transferrin receptor, and Hb compared to RBC indexes provided by an automated hematology analyzer (Advia 120, Bayer HealthCare) including HYPOm and CHr. RESULTS: Assuming that the most precise measure for body iron storage is related to the logarithm of the ratio of soluble transferrin receptor to ferritin, the sensitivity of ferritin for the diagnosis of iron depletion was 89 percent compared to 57 percent for HYPOm and CHr, respectively, to 69 percent for the combination of both RBC indexes, and to 26 percent for Hb concentration. CONCLUSION: The RBC indexes HYPOm und CHr are significantly better screening measures for identification of iron depletion in blood donors than Hb.

Marburg I polymorphism of factor VII-activating protease is associated with idiopathic venous thromboembolism.

The factor VII-activating protease (FSAP) variant Marburg I is known to attenuate the profibrinolytic system in vitro and was recently shown to be a significant predictor for the evolution and progression of carotid stenosis. The objective of this case-control study was to assess FSAP Marburg I's role in the occurrence of venous thromboembolism (VTE). The frequency of FSAP Marburg I was significantly increased in patients with a history of VTE (17 of 213 patients, 8.0%, P = .014) or idiopathic VTE (12 of 103 patients, 11.7%, P = .002) compared to healthy controls (5 of 213 controls, 2.3%). Logistic regression analysis confirmed FSAP Marburg I to be an independent risk factor for VTE (odds ratio, 3.5; 95% confidence interval [CI], 1.2-10.0) and idiopathic VTE (odds ratio, 6.2; 95% CI, 2.0-18.9).

The functional integrity of platelets in volume-reduced platelet concentrates.

Premature and low-birth-weight infants usually require small-volume platelet transfusions to treat thrombocytopenia. Also, infants undergoing open-heart surgery with extracorporeal circulation and with compromised cardiac function are at risk for excessive intravascular volume. The small-volume platelet substitution can be achieved by dispensing an aliquot from the unit of a standard single-donor platelet concentrate (PC). Alternatively, there is an indication for volume reduction of PCs to maximize the number of platelets transfused in the smallest possible volume. We determined the spontaneous and induced activation of platelets before and after volume reduction in 20 consecutive single-donor-apheresis PCs. After a mean storage time of 2 days, the PCs were plasma-depleted by centrifugation. Spontaneous, adenosine diphosphate (ADP)-induced, and collagen-induced activation were determined by flow cytometry. Furthermore, ADP- and collagen-induced aggregation were measured. A total of 33.8% of platelets in standard PCs were activated spontaneously. Volume reduction of PCs led to a mild but significant increase of spontaneous activation of platelets (43.2%). Additionally, volume reduction resulted in an impaired ADP-induced aggregability of platelets, whereas collagen induction was unaffected. Transfusion of volume-reduced PCs is an effective alternative to use of standard PCs in patients at frequent risk for excessive intravascular volume, because equal volumes increase the platelet count twice as effectively.

Daily doses of 20 mg of elemental iron compensate for iron loss in regular blood donors: a randomized, double-blind, placebo-controlled study.

BACKGROUND: A considerable number of regular blood donors develops an iron deficiency, and the exact amount of iron required to compensate for the iron loss from whole-blood donation in males and females is still unknown. STUDY DESIGN AND METHODS: A total of 526 regular blood donors (289 male and 237 female) were randomly assigned to treatment with either 40 mg, 20 mg, or 0 mg per day of elemental iron as ferrous gluconate for a period of 6 months, during which one unit of whole blood was collected on four occasions (males) or three occasions (females). Hemoglobin level, serum ferritin, and soluble transferrin receptor levels were measured before each donation. RESULTS: Daily doses of either 40 mg or 20 mg of elemental iron adequately compensated for iron loss in males, who gave blood at 2-month intervals, but did not result in a positive iron balance or an increase in storage iron as reflected by the logarithm of the ratio of transferrin receptor to ferritin concentration. In females, who donated at 3-month intervals, the same daily doses not only restored the iron balance but also led to an increase in storage iron. The number of gastrointestinal side effects due to iron supplementation (12%) was only slightly higher in both iron groups than in the placebo group. CONCLUSION: The results of this study indicate that 20 mg of elemental iron per day can adequately compensate for iron loss in males and females who donate whole blood up to four (females) or six times per year (males).

Iron supplementation and 2-unit red blood cell apheresis: a randomized, double-blind, placebo-controlled study.

BACKGROUND: The benefits of 2-unit red blood cell (RBC) apheresis are evident, but iron depletion may be a limiting factor in using this technology. Regular iron supplementation may allow a better utilization of this technique. STUDY DESIGN AND METHODS: In this study, 260 regular blood donors donated 2-unit RBCs on each of a total of seven visits at intervals of 8 to 10 weeks. The volunteers were randomly assigned to receive 100 mg of iron(II) or placebo daily. Group A received iron capsules after the first three donations, and Group B after the second three donations, respectively. Hemoglobin, serum ferritin, and serum iron were measured before each donation. RESULTS: Mean serum ferritin concentration decreased after each donation in the placebo phase of both treatment groups, but it remained largely constant during the iron phase in Group A, and even increased during the iron phase in Group B. CONCLUSION: Regular iron supplementation prevents iron depletion in the majority of donors after 2-unit RBC apheresis within an 8- to 10-week period.

Platelet activity in washed platelet concentrates.

Life-threatening anaphylaxis or febrile nonhemolytic transfusion reactions after transfusion of platelet concentrates (PCs) is a serious clinical problem caused by the sensitizing of recipients to plasma components, such as immunoglobulin A, or by cytokines. There is a possible indication for washing of PCs in these thrombocytopenic patients. However, only platelets that show activation after physiological stimulation are useful. We determined the spontaneous and induced activation of platelets before and after washing. We investigated 11 consecutive single-donor-apheresis PCs. After production and leukocyte-depletion the PCs were washed in 15%, acid-citrate-dextrose-solution. The spontaneous and the adenosine diphosphate (ADP)-induced, as well as collagen-induced activation, were determined by flow cytometry. Additionally, ADP- and collagen-induced aggregation were measured. Unwashed platelets (16.1%) were activated spontaneously. The washing of PCs led to a threefold increase of spontaneous activation of platelets (47.4%). Because of increased spontaneous activation after washing we could demonstrate cytometrically a loss of induced activation of washed platelets. Furthermore, washing resulted in an impaired ADP-induced aggregability of platelets. These results have led us to reduce the frequency of washing of PCs in our institution, where the only current indication for washing of PCs is in patients with a history of severe nonhemolytic transfusion reactions.

A practical concept for preoperative identification of patients with impaired primary hemostasis.

The findings of a large prospective study designed to identify primary and/or secondary hemostatic disorders before surgical interventions are presented. A total of 5649 unselected adult patients were enrolled to identify impaired hemostasis before surgical interventions. Each patient was asked to answer a standardized questionnaire concerning bleeding history. Activated partial thromboplastin time (aPTT), prothrombin time (PT), and platelet counts (PC) including PFA-100 (platelet function analyzer): collagen-epinephrine (C/E), and collagen-ADP (C/ADP) were routinely done in all patients. Additional tests, bleeding time (BT), and von Willebrand factor (vWF: Ag) were performed only in patients with a positive bleeding history and/or evidence of impaired hemostasis; e.g., drug ingestion. The bleeding history was negative in 5021 patients (88.8%) but positive in the remaining 628 (11.2%). Impaired hemostasis could be verified only in 256 (40.8%) of these patients. The vast majority were identified with PFA-100: C/E (n=250; 97.7%). The other six patients with impaired hemostasis were identifiable solely based on the PT (n=2), PFA-100: C/ADP (n=2), and vWF: Ag (n=2). The PFA-100: C/ADP detected 199 patients (77.7%). The only abnormality found among patients with a negative bleeding history was a prolonged aPTT due to lupus anticoagulant in nine patients (0.2%). The sensitivity of the PFA-100: collagen-epinephrine was the highest (90.8%) in comparison to the other screening tests (BT, aPTT, PT, vWF: Ag). The positive predictive value of the PFA-100: collagen-epinephrine was high (81.8%), but the negative predictive value was higher (93.4%). The use of a standardized questionnaire and, if indicated, the PFA-100: C/E and/or other specific tests not only ensure the detection of impaired hemostasis in almost every case but also a significant reduction of the cost.

A practical concept for preoperative management of patients with impaired primary hemostasis.

In a prospective study, 254 of 5649 unselected patients scheduled for surgery at our hospital were identified preoperatively as having either acquired (n=182) or inherited (n=72) impaired primary hemostasis (platelet dysfunction including von Willebrand disease). All patients were initially pretreated with desmopressin (DDAVP). Response to DDAVP or subsequent treatment(s) was defined as correction of any one of the abnormal PFA-100 platelet function tests. The non-responders were additionally treated with tranexamic acid or aprotinin; those with von Willebrand disease (vWD) received factor VIII concentrates with von Willebrand factor (vWF). Those still unresponsive to therapy received conjugated estrogens and, as a last attempt, a platelet transfusion. The administration of DDAVP led to a correction of platelet dysfunction in 229 of the 254 patients treated (90.2%). Tranexamic acid was effective in 12 of 16, aprotinin in 3 of 5, and factor VIII concentrates with vWF in all 4 patients with unresponsive to DDAVP. The remaining 6 patients were pretreated with conjugated estrogens, and 2 of these patients were additionally treated with platelet transfusion. The frequency of blood transfusion was lower, but not statistically significant (9.4% vs. 12.2%: p = 0.202) in preoperatively treated patients with impaired hemostasis than in patients without impaired hemostasis. In a retrospective group, the frequency of blood transfusion was statistically significant higher (89.3% vs. 11.3%: p < 0.001) in patients without preoperative correction of impaired hemostasis than in patients without impaired hemostasis. Preoperative correction of impaired primary hemostasis is possible in nearly all patients affected, and results in a reduction of homologous blood transfusions.

A single recombinant anti-RhD IgG prevents RhD immunization: association of RhD-positive red blood cell clearance rate with polymorphisms in the FcgammaRIIA and FcgammaIIIA genes.

A single recombinant immunoglobulin G1 (IgG1) anti-RhD antibody (MonoRho) was compared with a currently used polyclonal anti-RhD product (Rhophylac) in a phase 1 study for safety, efficacy of Rhesus D (RhD)-positive red blood cell (RBC) clearance, and prevention of RhD immunization in RhD-negative men challenged with 15 mL RhD-positive RBCs. Both the polyclonal product and recombinant anti-RhD effectively cleared RhD-positive RBCs after intravenous and intramuscular injection. The recombinant anti-RhD demonstrated a slower clearance rate compared with the polyclonal anti-RhD. There was no dose response, and there was considerable variation among subjects who received the same dose of recombinant anti-RhD. Interestingly, RhD-positive RBC clearance rates were strongly associated with Fcgamma receptor IIA (FcgammaRIIA) and FcgammaIIIA but not with FcgammaIIIB polymorphisms. Subjects homozygous for FcgammaRIIA-131H or FcgammaRIIIA-158V allotypes showed a faster clearance rate compared with both the heterozygote and the corresponding alternative homozygote allotypes. A similar but less marked trend was seen for the polyclonal anti-RhD. Despite the variation in clearance rates there was no evidence of anti-RhD alloantibodies in any of the subjects at +6 months after the RBC challenge.

Universal leukodepletion of blood components results in a significant reduction of febrile non-hemolytic but not allergic transfusion reactions.

BACKGROUND: Universal leukodepletion of blood components to prevent acute non-hemolytic transfusion reactions (NHTRs) is still a subject of debate. PATIENTS AND METHODS: Transfusion-associated NHTRs observed at our hospital in the last 6 years were retrospectively analyzed. Buffy-coat depleted red blood cells (bc-RBCs), and if indicated, leucodepleted post-storage (ld-RBCs) were initially used. In April 1997, universal leukodepletion was implemented at our hospital, and thereafter only prestorage ld-RBCs were used. All platelet concentrates transfused during this time were prestorage filtered single-donor apheresis platelets (SDAPs). RESULTS: A total of 163,090 blood products were transfused from April 1995 to April 2001 (bc-RBC: n=34,040 units; ld-RBC: n=66,967; SDAP: n=14,516; FFP: n=47,567). The number of post-transfusion febrile NHTRs occurring with each blood product was 65 (0.19%) for bc-RBCs, 8 (0.16%) for post-storage ld-RBCs, 16 (0.03%) for prestorage ld-RBCs, 16 (0.11%) for SDAPs, and 10 (0.02%) for FFP. Allergic reactions (n=116) were most frequently observed after SDAP transfusion (0.32%) and occurred at a similarly low rate after transfusion of all other blood components (0.03-0.08%). CONCLUSION: In conclusion, acute NHTRs rarely occur after the use of leukodepleted blood components. Prestorage appears to be more effective than post-storage leukodepletion in preventing febrile reactions following a blood transfusion.