Contrasting effects of recombinant human granulocyte-macrophage colony- stimulating factor (CSF) and granulocyte CSF treatment on the cycling of blood elements in childhood-onset cyclic neutropenia

Recombinant human granulocyte colony-stimulating factor (G-CSF) treatment has been shown to increase average neutrophil counts substantially in patients with childhood-onset cyclic neutropenia (or "cyclic hematopoiesis"), but not to eliminate the cyclic oscillations of neutrophil counts or those of other blood elements (monocytes, platelets, eosinophils, and reticulocytes) that are characteristic of this hematopoietic disorder. Indeed, oscillations of neutrophil counts are amplified during G-CSF treatment. We have compared the effects of recombinant granulocyte-macrophage-CSF (GM-CSF) with those of G-CSF in three patients with this disease (2 men and 1 woman, 17, 30, and 32 years of age). These patients were treated with GM-CSF (2.1 micrograms/kg/day, subcutaneously) for 6 weeks, preceded and followed by 6 to 13 weeks of detailed observation to document changes in the cyclic oscillations of blood neutrophils and other blood elements; two of the patients were subsequently treated with G-CSF (5.0 micrograms/kg/d, subcutaneously) and observed for comparable periods of time. Unlike G-CSF treatment, which increased average neutrophil counts more than 20-fold, GM-CSF increased neutrophil counts only modestly, from 1.6- to 3.9-fold, although eosinophilia of varying prominence was induced in each patient. However, at the same time, GM-CSF treatment dampened or eliminated the multilineage oscillations of circulating blood elements (neutrophils, monocytes, platelets, and/or reticulocytes) in each of the patients. In contrast, G-CSF treatment of the same patients markedly amplified the oscillations of neutrophil counts and caused the cycling of other blood elements (monocytes in particular) to become more distinct. These findings support the conclusion that the distinctive cycling of blood cell production in childhood-onset cyclic neutropenia results from abnormalities in the coordinate regulation of both GM-CSF-responsive, multipotential progenitor cells and G-CSF-responsive, lineage-restricted, neutrophil progenitors.     

Inhibition of experimental autoimmune encephalomyelitis in Lewis rats by nasal administration of encephalitogenic MBP peptides: synergistic effects of MBP 68-86 and 87-99

Induction of mucosal tolerance by inhalation of soluble peptides with defined T cell epitopes is receiving much attention as a means of specifically down-regulating pathogenic T cell reactivities in autoimmune and allergic disorders. Experimental autoimmune encephalomyelitis (EAE) induced in the Lewis rat by immunization with myelin basic protein (MBP) and Freund's adjuvant (CFA) is mediated by CD4+ T cells specific for the MBP amino acid sequences 68-86 and 87-99. To further define the principles of nasal tolerance induction, we generated three different MBP peptides (MBP 68-86, 87-99 and the non- encephalitogenic peptide 110-128), and evaluated whether their nasal administration on day -11, -10, -9, -8 and -7 prior to immunization with guinea pig MBP (gp-MBP) + CFA confers protection to Lewis rat EAE. Protection was achieved with the encephalitogenic peptides MBP 68-86 and 87-99, MBP 68-86 being more potent, but not with MBP 110-128. Neither MBP 68-86 nor 87-99 at doses used conferred complete protection to gp-MBP-induced EAE. In contrast, nasal administration of a mixture of MBP 68-86 and 87-99 completely blocked gp-MBP-induced EAE even at lower dosage compared to that being used for individual peptides. Rats tolerized with MBP 68-86 + 87-99 nasally showed decreased T cell responses to MBP reflected by lymphocyte proliferation and IFN-gamma ELISPOT assays. Rats tolerized with MBP 68-86 + 87-99 also had abrogated MBP-reactive IFN-gamma and tumor necrosis factor-alpha mRNA expression in lymph node cells compared to rats receiving MBP 110-128 nasally, while similar low levels of MBP-reactive transforming growth factor-beta and IL-4 mRNA expressing cells were observed in the two groups. Nasal administration of MBP 68-86 + 87-99 only slightly inhibited guinea pig spinal cord homogenate-induced EAE, and passive transfer of spleen mononuclear cells from MBP 68-86 + 87-99-tolerized rats did not protect naive rats from EAE. Finally, we show that nasal administration of MBP 68-86 + 87-99 can reverse ongoing EAE induced with gp-MBP, although higher doses are required compared to the dosage needed for prevention. In conclusion, nasal administration of encephalitogenic MBP peptides can induce antigen-specific T cell tolerance and confer incomplete protection to gp-MBP-induced EAE, and MBP 68-86 and 87-99 have synergistic effects. Non-regulatory mechanisms are proposed to be responsible for tolerance development after nasal peptide administration.      

Sensitivity and specificity of four assays to detect human T− lymphotropic virus type I or type I/II antibodies

BACKGROUND: Assays that detect human T-lymphotropic virus type I and type II antibody (HTLV-I/II) are widely used in the routine screening of blood donors. STUDY DESIGN AND METHODS: Four commercially available anti-HTLV-I (Fujirebio and Organon Teknika) or -HTLV-I/II assays (Murex and Ortho) were evaluated in various serum panels: A) HTLV-I-positive specimens (n = 41), confirmed by Western blot and polymerase chain reaction; B) a commercially available anti-HTLV-I/II panel; C) serial dilutions of sera from HTLV-I-positive individuals (n = 30), confirmed by immunofluorescence assay and Western blot: D) serial dilutions of HTLV-II-positive blood donors (n = 20), confirmed by Western blot and polymerase chain reaction, and E) sera from first-time blood donors (n = 1055). RESULTS: All four assays elicited reactions in all 82 HTLV-I- positive samples in Panels A, B, and C. Of 32 HTLV-II-positive specimens in Panels B and D, 31 (96.9%) reacted in the Organon Teknika assay and all 32 reacted in the remaining tests. Probit analysis of test results in Panels C and D indicated that the Fujirebio test was the most sensitive assay, followed by Organon Teknika, Ortho, and Murex. The specificities of Fujirebio, Murex, Organon Teknika, and Ortho tests in 1055 first-time blood donors were 99.9, 100, 99.6, and 99.9 percent, respectively. CONCLUSION: All four studied assays for detecting HTLV-I or HTLV-I/II antibodies are appropriate as screening tests.     

Human marrow erythropoiesis in culture. I. Characterization of methylcellulose colony assay

We examined the morphological and functional characteristics of human marrow erythrocytes cultured with a recently developed methylcellulose colony assay technique. Erythrocytic cells in various stages of development were observed, and a significant degree of maturational synchrony within individual colonies was noted. By light microscopy, colonies consisting of late normoblasts appeared compact, had an orange hue attributable to their hemoglobin, and demonstrated pseudoperoxidase activity, whereas colonies composed of early erythroblasts grew less compact or in clusters of smaller cell aggregates and showed no reddish tinge. Colonies possessing intermediate features were also observed. Maturational synchrony of individual colonies was confirmed using ransmission and scanning electron microscopy. The ultrastructure and cytochemistry of most immature cells were normal. The mature erythrocytes, however, were severely microcytic and hypochromic and contained one to several Heinz bodies. These defects in the cytoplasmic maturation of erythrocytes corresponded with impaired granulocytic maturation in culture, which we observed previously, and suggest environmental or nutritional defects in culture. Linearity of the method was confirmed using five normal bone marrows. Erythropoietin dose-responses observed in ten normal marrows were comparable to the previously reported results and revealed significant variation in individual plating efficiencies.     

三维重建射频损伤区域观察猪舌根射频损伤体积与射频能量及时间的关系

目的：目前临床进行隧道法舌根射频治疗时,其作用参数的设置仍缺乏统一的标准,故通过计算机三维重建射频损伤区域,分析猪舌根射频损伤体积与射频能量、时间的关系,从中得出应用舌根隧道法射频治疗的最佳作用能级和作用时间。 方法：实验于2006-06/2007-05在上海交通大学耳鼻喉科研究所完成。将36只实验用猪以射频作用能级1,2,3,4,5,6随机分成6组,每组6头猪,各个猪舌的作用时间分别设置为2,5,10,15,20,25s。用Coblation射频发生仪及Reflex55刀头进行猪舌根射频操作。射频作用后的舌根组织行连续冰冻切片,苏木精-伊红染色后,进行序列组织切片的全貌二维图像采集,对拟重建的结构进行边界提取和图像分割。将提取分割图像导入Image-Pro Solution图像处理软件,利用3D Constructor插件进行三维重建,并根据设定参数进行体积计算。用SPSS10.0统计学软件对所测数据进行统计学分析。 结果：①作用能级固定时,舌根组织射频损伤体积随时间延长而增大,符合Logarithmic回归曲线。②作用时间固定时,舌根组织射频损伤体积随能级增大而增大,符合直线回归。③射频损伤体积随能量增大而增加亦符合Logarithmic回归曲线。④Coblation射频治疗系统在能级6时,在作用10s之前,损伤体积随作用时间增加而迅速增加,其后变化趋势平缓,超过20s后损伤体积无显著增加。 结论：①舌根区域射频治疗时,舌根组织射频损伤体积与时间或能量呈Logarithmic曲线相关,与能级呈直线相关。②Coblation射频治疗系统在能级6时,最佳作用时间范围为10-20s。     

针刺足三里和悬钟治疗缺血性脑卒中

目的:观察针刺足三里、悬钟2穴对缺血性脑卒中脑血管功能的影响,分析其可能的作用机制,并对临床疗效做出评价。方法:选择2004-11/2006-05湖北中医药高等专科学校附属古城医院针灸科、荆州市第五人民医院中医康复科、荆州市第三人民医院中医科3单位缺血性脑卒中患者合适病例160例,采用查随机数字表的方法,将其随机分为对照组和针刺组,各80例。对照组采用现代医学常规干预方法进行治疗:卧床,保持呼吸道通畅,预防感染,控制颅内压、血压,维持水电解质平衡。针刺组在此基础上加针刺足三里、悬钟2穴,采用慢速捻转进针法针刺,留针20～30min,每隔5min行针1次。1次/d。两组患者治疗30d。并以经颅多普勒检测观察缺血性脑卒中患者治疗前后脑血管舒缩反应能力、脑血流自动调节功能、大脑半球侧枝循环代偿功能的变化,同时以治疗前后神经功能缺损程度为指标评价其临床疗效。结果:160例病例全部进入结果分析。①针刺组与治疗前相比,脑血管舒缩反应能力明显加强,差异有显著性意义(t=2.97,P<0.05),且优于对照组(t=2.45,P<0.05)。②针刺组与治疗前相比,脑血流自动调节能力明显改善,差异有非常显著性意义(t=8.01,P<0.01),且优于对照组(t=7.67,P<0.05)。③针刺组与治疗前相比,大脑半球侧枝循环代偿功能得到加强,差异有显著性意义(t=3.15,P<0.05),且优于对照组(t=5.16,P<0.05)。④针刺组与治疗前相比,神经功能缺损积分明显降低,差异有非常显著性意义(t=4.83,P<0.01),且优于对照组(t=5.43,P<0.05)。结论:针刺足三里、悬钟2穴对缺血性脑卒中患者脑血管舒缩反应能力、脑血流自动调节功能、大脑半球侧枝循环代偿功能有明显改善作用,并能促进神经功能的恢复。     

Reactivity to alcohol assessment measures: an experimental test

Aims   Previous research has suggested that alcohol screening and assessment may affect drinking.
Design   This study was a randomized test of reactivity to alcohol assessment questionnaires among a group of heavy drinking college students.
Setting and participants   A total of 147 university students completed a screening questionnaire and were randomized to either immediate assessment or delayed assessment. The immediate assessment group completed a set of drinking questionnaires at baseline, 3, 6 and 12 months, while the delayed assessment group completed questionnaires only at 12 months.
Measurements   Primary outcomes included overall volume of drinking, risky drinking and use of risk reduction behaviors.
Findings   We found a significant effect of assessment on measures of risky drinking and risk reduction behaviors, but not on overall volume of drinking. Specifically, at 12 months, participants who had previously completed drinking assessments had a lower peak blood alcohol concentration (BAC) ( d  = −0.373), were more likely to report a low score on the Alcohol Use Disorders Identification Test (AUDIT; odds ratio = 2.55) and tended to use more strategies to moderate their alcohol consumption ( d  = 0.352). Risk reduction behaviors that were affected tended to be those that limited alcohol consumption, rather than those that minimized consequences.
Conclusions   These results may have implications for the development of brief interventions.