Quantitation of CD62, soluble CD62, and lysosome-associated membrane proteins 1 and 2 for evaluation of the quality of stored platelet concentrates

BACKGROUND: Platelets become activated during storage, which results in secretion of granules, vesiculation of microparticles, secretion of protein, and a number of other biochemical and morphologic processes that decrease the utility of platelet concentrates stored for transfusion. STUDY DESIGN AND METHODS: To evaluate the quality of stored platelet concentrates, the cell surface expression of specific activation-dependent antigens (CD62 and lysosome-associated membrane proteins 1 and 2 [LAMP-1, LAMP-2]) on platelets stored in a hospital blood bank over a 7-day period was examined. Relative microparticle counts and the expression of CD62 by microparticles, as well as platelet concentrate supernatant levels of soluble CD62, were determined. RESULTS: The percentage of platelets expressing CD62 increased significantly from Day 1 to Day 5 (p < 0.05) of storage; the mean fluorescence values for CD62 did not. In contrast, the mean fluorescence values of LAMP-1 and LAMP-2 rose significantly (p < 0.01 and p < 0.05, respectively) between Days 1 and 5. Significant declines in CD62, LAMP-1, and LAMP-2 percent expression and mean fluorescence were seen on Day 6 of storage (p < 0.001). Microparticle numbers increased significantly during storage and correlated with levels of CD62 protein (free and membrane-bound) (r = 0.95 vs. Day 2, p < 0.05; r = 0.88 vs. Day 5, p < 0.05). CONCLUSION: Flow cytometric evaluations of the expression of cell surface CD62, LAMP-1, and LAMP-2 are complementary tests that, especially when used in conjunction with the quantitation of CD62 protein, provided a simple and effective means of evaluating the quality of platelet concentrates stored for transfusion.     

Individual differences in self-regulatory failure and menstrual dysfunction predict upper respiratory infection symptoms and antibody response to flu immunization

Prior research indicates that cognitive priming manipulations that activate personal goals acutely increase or decrease natural killer cell cytotoxicity depending on whether individuals see themselves as making or failing to make progress toward their goals. Those findings in a laboratory setting revealed a psychobiological pathway whereby experiences of failure can influence health, but did not assess the impact of chronic perceived success/failure in goal pursuit on actual health outcomes. Three new studies investigated whether individual differences in perceived failure to attain personal goals influenced the self-reported symptoms of upper respiratory infections (URIs) as well as antibody response to flu immunization. Based on pilot data in young women, it also was hypothesized that the occurrence of menstrual dysfunction might interact with goal pursuit failure to more specifically predict cold and flu symptoms and optimal responses to vaccination. Perceived failure to attain goals did predict the reporting of URI symptoms as well as antibody levels post-immunization, both alone and in combination with menstrual dysfunction.     

Psychiatric morbidity in stroke patients attending a neurology clinic in Nigeria

Back ground

Stroke produces a wide range of mental and emotional disorders. Neuropsychiatric complications associated with stroke may have negative effects on the social functioning, overall quality of life and the recovery of motor functioning of stroke survivors.

Objective

To determine the prevalence and nature of psychiatric morbidity among stroke patients attending neurology outpatient clinic of the University of Ilorin Teaching Hospital (UITH), Ilorin-Nigeria.

Methods

All patients with stroke aged 18 years and above at an outpatient neurology clinic in Ilorin, Nigeria were assessed for mental and emotional disorders using the Schedule for Clinical Assessment in Neuropsychiatry (SCAN) over one year (March 2009 to February 2010).

Results

Overall prevalence of psychiatric morbidity was 36.0% (30/83) among 83 patients who constituted the study population. Specific diagnoses recorded were depression (19.2%), generalised anxiety disorder (9.6%), harmful alcohol use (2.4%); dementia, somatoform disorder, phobia and delusional disorder each had a prevalence of 1.2%. Clinical and sociodemographic variables were not significantly associated with psychiatric morbidity.

Conclusion

Psychiatric disorders are often associated with stroke. Identifying and treating stroke patients with these psychiatric co-morbidities could thus help to improve the overall quality of life of these patients.     

The functional and structural changes in the basilar artery due to overpressure blast injury

Overpressure blast-wave induced brain injury (OBI) leads to progressive pathophysiologic changes resulting in a reduction in brain blood flow, blood brain barrier breakdown, edema, and cerebral ischemia. The aim of this study was to evaluate cerebral vascular function after single and repeated OBI. Male Sprague-Dawley rats were divided into three groups: Control (Naive), single OBI (30 psi peak pressure, 1 to 2 msec duration), and repeated (days 1, 4, and 7) OBI (r-OBI). Rats were killed 24 hours after injury and the basilar artery was isolated, cannulated, and pressurized (90 cm H₂O). Vascular responses to potassium chloride (KCl) (30 to 100 mmol/L), endothelin-1 (10⁻¹² to 10⁻⁷ mol/L), acetylcholine (ACh) (10⁻¹⁰ to 10⁻⁴ mol/L) and diethylamine-NONO-ate (DEA-NONO-ate) (10⁻¹⁰ to 10⁻⁴ mol/L) were evaluated. The OBI resulted in an increase in the contractile responses to endothelin and a decrease in the relaxant responses to ACh in both single and r-OBI groups. However, impaired DEA-NONO-ate-induced vasodilation and increased wall thickness to lumen ratio were observed only in the r-OBI group. The endothelin-1 type A (ET_A) receptor and endothelial nitric oxide synthase (eNOS) immunoreactivity were significantly enhanced by OBI. These findings indicate that both single and r-OBI impairs cerebral vascular endothelium-dependent dilation, potentially a consequence of endothelial dysfunction and/or vascular remodelling in basilar arteries after OBI.     

Regression of established tumors and metastases by potent vascular endothelial growth factor blockade

Vascular endothelial growth factor (VEGF) is a critical promoter of blood vessel growth during embryonic development and tumorigenesis. To date, studies of VEGF antagonists have primarily focused on halting progression in models of minimal residual cancer. Consistent with this focus, recent clinical trials suggest that blockade of VEGF may impede cancer progression, presumably by preventing neoangiogenesis. However, VEGF is also a key mediator of endothelial-vascular mural cell interactions, a role that may contribute to the integrity of mature vessels in advanced tumors. Here, we report that high-affinity blockade of VEGF, using the recently described VEGF-Trap, abolishes mature, preexisting vasculature in established xenografts. Eradication of vasculature is followed by marked tumor regression, including regression of lung micrometastases. Thus, the contribution of relatively low levels of VEGF to vessel integrity may be critical to maintenance of even very small tumor masses. Potent blockade of VEGF may provide a new therapeutic option for patients with bulky, metastatic cancers.     

骺板软骨细胞的体外培养及鉴定

目的:建立体外培养及鉴定骺板软骨细胞的方法。方法:实验于2005-03/2006-05在苏州大学附属儿童医院骨科实验室完成。选用3只生后14～28d的新西兰幼兔,空气栓塞处死,暴露股骨下端和胫骨上端,分别取2处的骺板组织,将其剪切成1～3mm3的小块,经胰蛋白酶消化,接种于含150g/L牛血清的1640培养基中,饱和湿度培养,传代。①细胞接种后3h在倒置显微镜下观察细胞生长情况,见细胞贴壁后每天观察2次。②第2代细胞达到80%～90%左右汇合时采用常规苏木精-伊红染色,光镜观察细胞爬片情况。③第3代细胞爬片至细胞达到80～90%左右汇合时,采用苏木素染色3min,3%亮绿染色5min,蕃红花“O”染色5min,光镜观察细胞产生蛋白聚糖情况。④采用PCR检测细胞Ⅱ型胶原的表达。⑤采用四唑盐MTT比色法检测细胞活性。结果:①骺软骨细胞刚接种后呈大小不等之圆形悬浮于培养液中,3h后见大部分细胞贴壁,24h后贴壁细胞呈短梭形、圆形、三角形和不规则形,见细胞分裂相。48h后见细胞伸展明显,细胞分裂相每高倍镜视野可见多个。经隔日换液细胞生长至第5天,细胞呈聚集生长,达到汇合状态,将细胞传代。接种后第1代细胞2d后呈梭形,培养4d传至第2代,第2代细胞长满瓶底后,90%呈胞膜较厚的圆形,10%为梭形,第3代、第4代亦如此。传至第5代见肥大细胞增多,细胞松散,折光性减弱,呈凋亡状态。②细胞爬片后观察骺软骨细胞形态以梭形居多,同时也有圆形、三角形和不规则形。可见细胞分裂及细胞中的分泌小泡。③见细胞呈红色,无绿色,证明蕃红花“O”-亮绿染色阳性,显示所培养的细胞可以分泌蛋白聚糖。④PCR检测术所养细胞含有Ⅱ型胶原,电泳带在440bp上。⑤四唑盐MTT比色法检测显示,第3代骺软骨细胞的生长曲线近似倒“S”形,在第4,5,6天细胞呈对数生长,约在7,8,9,10d达平台期,至第12天细胞出现生长抑制。结论:建立了骺板软骨细胞的体外培养方法,并证实所培养出的细胞分泌蛋白聚糖和Ⅱ型胶原,具有软骨细胞的共同特点。     

Skeletal myoblast based delivery of angiogenic growth factors: a comparison between angiopoietin-1 and VEGF gene delivery for therapeutic angiogenesis in the heart

Introduction Extensive cell death and an associated myocardial dys- function are the common features of chronic heart disease. Given the inadequate ability of the human heart to regenerate, a more recent approach to counter the remodel- ing process is to compensate for the loss of functioning cardiomyocyte number through stem cell transplantation with angiomyogenic potential.1,2 The novel approach of heart cell therapy is to repopulate the scar tissue with myogenic cells that may be functional…     

Validation of a prediction rule to maximize curative (R0) resection of early-stage pancreatic adenocarcinoma

Background:

The surgeon''s contribution to patients with localized pancreatic adenocarcinoma (PAC) is a margin negative (R0) resection. We hypothesized that a prediction rule based on pre-operative imaging would maximize the R0 resection rate while reducing non-therapeutic intervention.

Methods:

The prediction rule was developed using computed tomography (CT) and endoscopic ultrasound (EUS) data from 65 patients with biopsy-proven PAC who underwent attempted resection. The rule classified patients as low or high risk for non-R0 outcome and was validated in 78 subsequent patients.

Results:

Model variables were: any evidence of vascular involvement on CT; EUS stage and EUS size dichotomized at 2.6 cm. In the validation cohort, 77% underwent resection and 58% achieved R0 status. If only patients in the low-risk group underwent surgery, the prediction rule would have increased the resection rate to 92% and the R0 rate to 73%. The R0 rate was 40% higher in low-risk compared with high-risk patients (P < 0.001). High risk was associated with a 67% rate of non-curative surgery (unresectable disease and metastases).

Conclusion:

The prediction rule identified patients most likely to benefit from resection for PAC using pre-operative CT and EUS findings. Model predictions would have increased the R0 rate and reduced non-therapeutic interventions.