Terminal differentiation of human promyelocytic leukemic cells in primary culture in response to retinoic acid

The recent finding that retinoic acid induces terminal granulocytic differentiation of the human promyelocytic leukemia cell line, HL-60, prompted an investigation of the sensitivity to this inducer of human myelocytic leukemia cells in primary suspension culture. Of the 21 leukemic specimens, only cells from the two patients with acute promyelocytic leukemia differentiated in response to retinoic acid. After an incubation period of 5--7 days in 1 microM retinoic acid, the cells from these two patients showed extensive morphological and functional maturation. Thus, because it appears that retinoic acid specifically induces granulocytic differentiation of leukemic promyelocytes, this compound may have therapeutic utility in the treatment of acute promyelocytic leukemia.     

Biochemical and functional consequences of dissociation of the platelet membrane glycoprotein IIb-IIIa complex

The platelet membrane glycoproteins, IIb and IIIa, form a Ca2+- dependent heterodimer complex that functions as the fibrinogen receptor in activated platelets to mediate platelet aggregation. Little is known about factors that affect the IIb-IIIa complex within the platelet membrane. It has been observed that platelets incubated with ethylene glycol tetra-acetic acid (EGTA) at 37 degrees C are unable to aggregate or to bind monoclonal antibodies specific for the IIb-IIIa complex. To determine whether this is due to a dissociation of IIb from IIIa, we developed a method for quantitating the complex on nondenaturing, polyacrylamide gradient gels. Platelets were surface-labeled with 125I and then solubilized and electrophoresed in 0.2% Triton and 10 mmol/L CHAPS. Under these conditions and in the presence of 1 mmol/L Ca2+, glycoproteins IIb and IIIa migrated on the gels as a discrete band at Rf = 0.33. Protein that was eluted from this band bound to an immunoaffinity column specific for the IIb-IIIa complex. In contrast, when the IIb-IIIa complex was solubilized and then dissociated with EGTA, the discrete band at Rf = 0.33 was no longer present, and IIb and IIIa were now found in a broad band at Rf = 0.45 to 0.50. To study IIb and IIIa within the surface membrane, the 125I-labeled platelets were first incubated with 0.5 mmol/L EGTA (1 nmol/L free Ca2+) at 22 degrees C and then solubilized in the absence of EGTA. The IIb and IIIa from these platelets migrated at Rf = 0.33, indicating the presence of the intact IIb-IIIa complex. In contrast, when the platelets were incubated at 37 degrees C for one hour with the EGTA, the discrete band at Rf = 0.33 representing the IIb-IIIa complex gradually disappeared. This phenomenon could not be reversed by adding Ca2+ back to the platelets before solubilization and electrophoresis. This loss of the IIb-IIIa complex from intact platelets was accompanied by (a) a progressive and irreversible decrease in adenosine diphosphate (ADP)-induced platelet aggregation and (b) decreased binding of a complex-dependent monoclonal antibody to the platelets. These studies demonstrate that when platelets are exposed to low Ca2+ at 37 degrees C, the IIb-IIIa heterodimer complexes in their surface membranes are irreversibly disrupted. Because intact IIb-IIIa complexes are required for platelet aggregation, the loss of these complexes may account for the failure of these platelets to aggregate in response to ADP.     

噻托溴铵升级治疗成人未控制支气管哮喘

背景和目的:对于单用吸入糖皮质激素控制不佳的成人支气管哮喘(简称哮喘)患者,加用长效β-受体激动剂(LABA)可有效改善患者的症状及肺功能.但是最近对LABA长期治疗的安全性受到了美国食品药品管理局(FDA)及部分哮喘专家的质疑.另外,鉴于每个哮喘患者对治疗的反应不尽相同,因此寻求未控制哮喘患者的其他治疗方法是必要的.     

SB265610 is an allosteric,inverse agonist at the human CXCR2 receptor

Background and purpose:

In several previous studies, the C-X-C chemokine receptor (CXCR)2 antagonist 1-(2-bromo-phenyl)-3-(7-cyano-3H-benzotriazol-4-yl)-urea (SB265610) has been described as binding competitively with the endogenous agonist. This is in contrast to many other chemokine receptor antagonists, where the mechanism of antagonism has been described as allosteric.

Experimental approach:

To determine whether it displays a unique mechanism among the chemokine receptor antagonists, the mode of action of SB265610 was investigated at the CXCR2 receptor using radioligand and [³⁵S]-GTPγS binding approaches in addition to chemotaxis of human neutrophils.

Key results:

In equilibrium saturation binding studies, SB265610 depressed the maximal binding of [¹²⁵I]-interleukin-8 ([¹²⁵I]-IL-8) without affecting the K_d. In contrast, IL-8 was unable to prevent binding of [³H]-SB265610. Kinetic binding experiments demonstrated that this was not an artefact of irreversible or slowly reversible binding. In functional experiments, SB265610 caused a rightward shift of the concentration-response curves to IL-8 and growth-related oncogene α, but also a reduction in maximal response elicited by each agonist. Fitting these data to an operational allosteric ternary complex model suggested that, once bound, SB265610 completely blocks receptor activation. SB265610 also inhibited basal [³⁵S]-GTPγS binding in this preparation.

Conclusions and implications:

Taken together, these data suggest that SB265610 behaves as an allosteric inverse agonist at the CXCR2 receptor, binding at a region distinct from the agonist binding site to prevent receptor activation, possibly by interfering with G protein coupling.     

Exploring quantitative methods for evaluation of lip function

Summary The objective was to explore quantitative methods for the measurement of lip mobility and lip force and to relate these to qualitative assessments of lip function. Fifty healthy adults (mean age 45 years) and 23 adults with diagnoses affecting the facial muscles (mean age 37 years) participated in the study. Diagnoses were Möbius syndrome (n = 5), Facioscapulohumeral muscular dystrophy (n = 6) and Myotonic dystrophy type 1 (n = 12). A system for computerised 3D analysis of lip mobility and a lip force meter were tested, and the results were related to results from qualitative assessments of lip mobility, speech (articulation), eating ability and saliva control. Facial expressions studied were open mouth smile and lip pucker. Normative data and cut‐off values for adults on lip mobility and lip force were proposed, and the diagnostic value of these thresholds was tested. The proposed cut‐off values could identify all inviduals with moderate or severe impairment of lip mobility but not always the milder cases. There were significant correlations between the results from quantitative measurements and qualitative assessments. The examined instruments for measuring lip function were found to be reliable with an acceptable measuring error. The combination of quantitative and qualitative ways to evaluate lip function made it possible to show the strong relation between lip contraction, lip force, eating ability and saliva control. The same combination of assessments can be used in the future to study if oral motor exercises aimed at improving lip mobility and strength could have a positive effect on lip function.     

Pain and fear in connection to orthodontic extractions of deciduous canines

International Journal of Paediatric Dentistry 2010; 20: 193–200 Background. Interceptive extractions of deciduous canines are, from a patient perspective, poorly investigated. Aims. To describe pain, discomfort, and dental fear in connection to extractions of the deciduous canines, indicated as an orthodontic treatment procedure. Design. Thirty‐two Swedish children aged 7–9 years had all four deciduous canines extracted over three occasions. The children rated procedural and postoperative pain on visual analogue scales. Acceptance of injections and extractions was assessed by the treating dentists. Analgesic consumption and recovery time for drinking and eating was reported by parents. Dental fear was assessed using the Children’s Fear Survey Schedule questionnaire. Results. Procedural pain showed low median levels, although some individuals reported high values. Boys reported significantly more pain at appointments when two (as opposed to one) canines were extracted. Postoperative pain levels were low and use of analgesics sparse. Dental fear paralleled norm values and did not increase from pre‐ to post‐extraction. Conclusions. Pain management routines during extractions of this kind should be revised. Single tooth extractions seem to be preferable to extractions of two canines at the same appointment. Extraction of four deciduous canines should not cause major postoperative inconvenience; these extractions neither triggered nor increased dental fear.