Subsecond calcium dynamics in ADP- and thrombin-stimulated platelets: a continuous-flow approach using indo-1

The regulation and kinetics (less than 5 seconds) of cytosolic calcium changes ([Ca2+]i) in stimulated blood platelets have been investigated under physiological blood flow conditions. Using a newly-developed continuous-flow approach with indo-1-loaded human platelets, adenosine diphosphate (ADP, 10 mumol/L) and thrombin (5 U/mL) were equally effective in significantly increasing [Ca2+]i by 0.5 seconds. ADP induced a transient [Ca2+]i peak of 1 to 2 mumol/L near 2 seconds, whereas thrombin caused a sustained and larger response. The first phase (less than 2 seconds) was not influenced by a lack of extracellular Ca2+, in contrast to the subsequent [Ca2+]i increase that only reached about 0.7 mumol/L for either ADP or thrombin. The shear rates used in our continuous-flow apparatus were physiological (less than 1,258 sec-1) and only slightly increased the basal [Ca2+]i of 0.1 mumol/L. Platelet aggregation (less than 5 seconds), assessed by single- particle counting, was not altered in platelets loaded with indo-1/AM (2.5 mumol/L).     

Surface markers of human eosinophils

Peripheral blood eosinophils from patients with eosinophilia and from healthy subjects were studied for surface immunoglobulins, receptors for the Fc region of IgG, complement receptors, and spontaneous rosette formation with sheep and mouse erythrocytes. Eosinophils were found to have receptors for complement and for aggregated IgG, and to have the same two types of complement receptors as do lymphocytes and monocytes. Immune adherence type receptors were specific for C4 or C3b, while C3d receptors were specific for C3d but unreactive with C4. Eosinophils differed from fully mature neutrophils in that the former had C3d receptors and relatively weak immune adherence (C4 or C3b) receptors, while the later did not have the C3d receptors and had strong immune adherence receptors. Eosinophil phagocytosis of complement-receptor bound erythrocytes was dependent on the presence of IgG in the antibody coating the red blood cells; this requirement for IgG resembled that found in neutrophil phagocytosis. No surface Ig or spontaneous erythrocyte rosette formation was observed with eosinophils.     

Human cytomegalovirus alters interleukin-6 production by endothelial cells

In an effort to study whether human cytomegalovirus (HCMV) can disrupt the balanced cytokine network that controls human hematopoiesis, we investigated the ability of a laboratory strain HCMV (AD169) to alter the production of interleukin-6 (IL-6) by cultured endothelial cells (HUVECs). ECs are important components of human bone marrow stroma and produce factors that stimulate the proliferation and differentiation of human hematopoietic progenitors. HCMV was able to greatly increase production of both mRNA and protein for IL-6 in unprimed HUVECs. When we discriminated between viral pellet and cleared viral supernatants, the supernatants induced an increase in mRNA at 30 minutes and protein by 2 hours, whereas an increase in IL-6 caused by virus itself did not become evident until 12 hours. The possibility that IL-6 induction was simply caused by the presence in the viral stock of endotoxin, IL-1 alpha, IL-1 beta, tumor necrosis factor alpha, or IL-4, all known inducers of IL-6 in HUVECs, was ruled out by the addition of polymyxin B and appropriate neutralizing antibodies. These findings show that HCMV is capable of directly and indirectly modulating the production by HUVECs of IL-6, one of the cytokines involved in the process of hematopoiesis.     

Immunologic characterization of a helper T-cell lymphoma

The lymphocytes of a patient with a T-cell non-Hodgkin's lymphoma with peripheral blood involvement and polyclonal hypergammaglobulinemia were characterized in terms of surface markers and immunologic functions. Using the fluorescence-activated cell sorter and employing various monoclonal antibodies against T-cell surface antigens, it was shown that almost all of the patient's peripheral blood lymphocytes were positive for OKT4 and 9.3, antibodies that recognize helper T-cell subset. The circulating lymphoma cells had typical characteristics for T cells; they formed spontaneous rosettes with sheep erythrocytes and stained with the pan-T-cell antibodies 9.6 and 10.2, but did not react with other anti-T-cell monoclonal reagents such as OKT3, UCHT-1, and 3A1. The cells appeared to be mature by the fact that they did not stain with OKT6, and terminal deoxynucleotidyl transferase was undetectable. Functionally, they were able to provide "help" for antibody production, and they could be stimulated to produce moderate amounts of interleukin-2, while unable to proliferate in response to mitogens. Morphologically, some of the lymphocytes showed a deeply cleaved nucleus.     

Massage therapy after heart surgery: possibly pleasant,but not effective

Albert NM, Gillinov AM, Lytle BW, Feng J, Cwynar R, Blackstone EH. A randomized trial of massage therapy after heart surgery. Heart Lung 2009; 38: 480–90.     

Therapeutic effect of topical ozonated oil on the epithelial healing of palatal wound sites: a planimetrical and cytological study

Aim: To evaluate the effect of ozonated oil on palatal wounds. Methods: Eighteen patients were randomized and allocated to either the ozone group (n = 8) or control (n = 10) group. Free gingival graft surgery was performed, and post‐harvested palatal wounds were treated with either 2 mL ozonated oil or control oil daily for 1 week. A planimetrical analysis analyzed the digital image for the wound sizes and shape factor at baseline, at 24 h, and days 5, 7, 14, 21, and 28, postoperatively. A cytological analysis used the keratinization and superficial cell indices at baseline, 24 h, and days 3, 7, 14, and 21 and the second and third months, postoperatively. Results: Planimetrical results showed a significant (P ≤ 0.05) improvement in wound size on days 5, 7, 14, 21, and 28, postoperatively, in the ozone group compared to the control group. Cytological results showed a significant (P ≤ 0.001) improvement in epithelial healing on days 7, 14, and 21, and the second and third months, postoperatively, after the application of ozonated oil compared to control oil. Conclusion: Our results showed significant improvement in wound size and epithelial healing after topical ozonated oil application compared to control oil on palatal wounds.     

Effect of mildronate on life quality of patients with chronic heart failure]

AIM: To study quality of life (QL) of patients with chronic heart failure (CHF) and QL changes resultant from mildronate therapy. MATERIALS AND METHODS: QL was studied in 30 IHD patients with CHF of NYHA class II-IV, ejection fraction > 45%; 40 IHD patients without CHF and 30 healthy subjects. CHF patients were treated for 30 days with oral mildronate (250 mg 4 times a day). QL was assessed according to the method SF-36 Health Status Survey. RESULTS: QL in CHF patients is much lower than that of the controls. Objective severity of the disease and subjective satisfaction with life do not always coincide. Mildronate in a dose 1 g/day per os may be beneficial for LQ of CHF patients. CONCLUSION: It is thought desirable to include QL in analysis of efficiency of managing patients with CHF. SF-36 is a tool able to follow up changes in QL of CHF patients within a short-term treatment period.